Equine sperm refers to the male reproductive cells produced by stallions, essential for the process of fertilization and successful breeding in horses. The study of equine sperm encompasses various aspects, including morphology, motility, viability, and genetic integrity. These parameters are critical for assessing stallion fertility and improving breeding outcomes. Research in this field often focuses on understanding the factors that influence sperm quality, such as age, nutrition, and environmental conditions. Additionally, advancements in assisted reproductive technologies, such as artificial insemination and cryopreservation, rely heavily on the detailed study of sperm characteristics. This page compiles peer-reviewed research studies and scholarly articles that explore the biology, evaluation, and technological applications related to equine sperm.
Gross MK, Toscano DG, Toscano WA.Calmodulin (CaM), the calcium binding protein that modulates the activity of a number of key regulatory enzymes, is present at high levels in sperm. To determine whether CaM regulates adenylate cyclase in mammalian sperm, the actions of EGTA and selected CaM antagonists on a solubilized adenylate cyclase from mature equine sperm were examined. The activity of equine sperm adenylate cyclase was inhibited by EGTA in a concentration-dependent manner with a half-maximal inhibitory concentration (IC50) of 2 mM. Equine sperm adenylate cyclase was also inhibited in a concentration-dependent manner by...
Bélaïche D, Loir M, Kruggle W, Sautière P.Two protamines, St1 and St2, were isolated from stallion sperm nuclei, where they represent about 75 and 25%, respectively, of the total basic protein complement. The primary structure of protamine St1 (49 residues; Mr approximately equal to 6600) has been determined. The structure of this protamine is compared to the amino-acid sequence of other mammalian protamines already known.
Francl AT, Amann RP, Squires EL, Pickett BW.The effects of extender and storage at 20 degrees C on equine spermatozoa were evaluated in two experiments using embryo recovery as the end point. In both experiments, inseminations were every other day, starting on Day 2 or 3 of estrus or after a 35-mm follicle was detected, with 250 x 10(6) progressively motile cells (based on initial evaluation). In Experiment 1, semen from two stallions was used to compare the motility and fertility of spermatozoa maintained in a) heated skim milk extender at 37 degrees C with insemination in <1 h; b) E-Z Mixin extender at 37 degrees C with inseminatio...
Kosiniak K, Bittmar A.Physiological processes connected with sexual maturation of stallions were observed on 10 half-breed Anglo-Arab stallions beginning from 8 months of age, until 4.5 years of age. It was found that there is full somatic and sexual development in the stallion reached around the age of 3.5 years, and the sperm morphology stabilized in the range of the physiological norm around 3.0 years of age. On the other hand biochemical components of the semen plasma such as glycerylphosphorylcholine (GPC), ergothioneine (EGT), total protein (PRT), up to age 4.5 years, reach significantly lower value than in m...
Boyle MS, Cran DG, Allen WR, Hunter RH.The morphology of the uterotubal junction (UTJ) and caudal isthmus during the peri-ovulatory period, and the distribution of spermatozoa within the region, were studied in 10 Pony mares. The proximal tip of the uterine horn and caudal 1-2 cm of the isthmus were removed during oestrus or shortly after ovulation from animals mated or artificially inseminated within the previous 24 h. The tissues were incised longitudinally and fixed for scanning electron microscopy. Analysis of micrographs showed deep longitudinal and oedematous folds in the preovulatory samples. After ovulation, much of the fol...
Okólski A, Babusik P, Tischner M, Lietz W.Comparisons were made between 2 methods of oocyte recovery from the ovarian follicles of slaughtered mares: 500 oocytes (3 per mare) were obtained by aspiration of follicular fluid from ovaries of 162 mares, and 120 oocytes (8 per mare) by isolation and rupture of follicles from ovaries of 14 mares. In the oocytes recovered after rupture of follicles, 89.2% were morphologically unchanged, in comparison to 29.3% obtained by aspiration of follicular fluid. Stallion spermatozoa capacitated in vitro were tested on zona-free hamster oocytes. The stallion spermatozoa were washed in TCM-199 and prein...
Amann RP, Cristanelli MJ, Squires EL.Motility and fertility of frozen-thawed semen differs greatly amongst stallions. Differences in seminal plasma might be one cause of this variation. For 8 ejaculates from each of 17 stallions, seminal plasma was saved at -20 degrees C and spermatozoa were cryopreserved. Based on post-thaw sperm motility, seminal plasma samples from 7 stallions (2 good, 3 variable, 2 poor sperm motility) were selected for measurement of electrolytes, protein content and analysis by sodium dodecylsulphate gel electrophoresis (10% gel, Coomassie blue stain). Variation in seminal plasma was significant (P less tha...
Heiskanen ML, Pirhonen A, Koskinen E, Mäenpää PH.The role of various environmental conditions on sperm motility and ATP content was investigated by incubating raw and washed spermatozoa collected with an open-ended artificial vagina from 10 stallions in various biological and artificial media under different atmospheric conditions. Spermatozoa did not survive for more than 12 h when kept unextended in the original seminal fluid in any circumstances. The most favourable media tested for long-term sperm survival were Kenney's medium or Kenney's medium supplemented with 10 mM-theophylline and 10 mM-Hepes, pH 7.2. Centrifugation and slow cooling...
Arns MJ, Webb GW, Kreider JL, Potter GD, Evans JW.Bovine serum albumin (BSA) diluents containing lactose, raffinose or sucrose were not different (P greater than 0.05) in their ability to maintain stallion sperm viability, as determined by percentage motile spermatozoa (PMS) and their rate of forward movement (RFM), when stored at 37 or 5 degrees C for 24 h. These diluents did promote a higher (P greater than 0.05) PMS and RFM, when compared with BSA diluents containing arabinose or galactose. The BSA-arabinose and BSA-galactose diluents did not differ (P less than 0.05) in their ability to support sperm viability and were detrimental to sper...
Strzemienski PJ, Sertich PL, Varner DD, Kenney RM.Stallion semen was diluted in a Hepes-supplemented buffer (CM) (10(6) spermatozoa/ml) and placed in the upper well of a Sykes-Moore chemotaxis chamber. Chambers were incubated in a humidified atmosphere (5% CO2 in air) at 37 degrees C for 1 and 2 h and spermatozoa were allowed to swim through filters with a mean pore size of 3,5 or 8 micron. Spermatozoa entered filters of all three pore sizes. Distance travelled was greater for each increase in pore size (P less than 0.01) but did not differ (P greater than 0.05) between 1 and 2h of incubation. Extended semen from stallions of different fertil...
Klem ME, Kreider JL, Pruitt JB, Potter GD.Inclusion of either 1 or 3% (w/v) bovine serum albumin (BSA) in 8.6, 10, or 12% sucrose enhanced the maintenance of equine sperm motility in vitro at 38 degrees C for 8 h. There was a trend toward higher percent motile spermatozoa (PMS) at 16 and 24 h of incubation in semen samples containing BSA than in those that did not. The highest concentration of sucrose (12%) was slightly less effective in supporting PMS than either of the lower concentrations. However, sucrose concentrations had no apparent effect on rate of forward movement (RFM) of spermatozoa. Pregnancy and foaling rates were simila...
Garner DL, Pinkel D, Johnson LA, Pace MM.Spermatozoa from bulls, boars, dogs, horses, mice, and men were examined using a fluorogenic stain consisting of the membrane-permeant substrate carboxyfluorescin diacetate (CFDA) and the relatively membrane-impermeant nuclear stain propidium iodide (PI). Three distinct populations of spermatozoa were discernible in samples from each species upon microscopic examination. Individual spermatozoa, presumed to be viable because of their motility, retained products of the fluorescein chromophore throughout the cell. A second population of spermatozoa in which the nuclei stained red with PI retained...
Van Huffel XM, Varner DD, Hinrichs K, Garcia MC, Strzemienski PJ, Kenney RM.A photomicrographic method for evaluation of stallion spermatozoal motility was developed, and spermatozoal image and velocity characteristics were defined. The photomicrographic method was compared with visual estimation of motility in the same semen sample over time. Using photomicrography, velocities and percentages of individual spermatozoal image characteristics were determined. Although there was a high correlation between results of the 2 methods, results of the photomicrographic method were more repeatable than were those of the visual method.
Zgórniak-Nowosielska I, Kosiniak K, Slagowska A.Eleven mycoplasma strains were isolated from the semen of 24 stallions. Eight of these strains were identified as Mycoplasma equigenitalium. Three strains which hydrolized arginine could not be identified. The growth inhibition test with immune sera against M. arginini and M. equirhinis was negative. Antibiotic sensitivity test showed that all strains were sensitive to four antibiotic of tetracycline group (oxytetracyclin, minocycline, transcycline and vibramycin). Lincomycin and gentamycin appeared to be the most active against all the strains. Comparative analysis of routine semen examinatio...
Province CA, Amann RP, Pickett BW, Squires EL.Two experiments were conducted to evaluate the effects of six extenders and three glycerol levels on the motility of sperm stored at 5 degrees C. Using a split-ejaculated design, semen from 10 dogs and 12 stallions was extended with egg-yolk-tris (EYT), egg-yolk-bicarbonate (EGB), Beltsville F-3 (BF-3), Cornell University (CUE), caprogen (CAP) and heated skim milk (SM) extenders. After cooling to 5 degrees C, additional extender containing 0% to 12% glycerol was added to provide a final concentration of 0%, 3% or 6% glycerol. Regardless of glycerol level, a higher (P<0.05) percentage of can...
Cochran JD, Amann RP, Froman DP, Pickett BW.Five experiments evaluated the effects of processing, freezing and thawing techniques on post-thaw motility of equine sperm. Post-thaw motility was similar for sperm frozen using two cooling rates. Inclusion of 4% glycerol extender was superior to 2 or 6%. Thawing in 75 degrees C water for 7 sec was superior to thawing in 37 degrees C water for 30 sec. The best procedure for concentrating sperm, based on sperm motility, was diluting semen to 50 x 10(6) sperm/ml with a citrate-based centrifugation medium at 20 degrees C and centrifuging at 400 x g for 15 min. There was no difference in sperm mo...
Livolant F.The fine structure of chromatin in sperm heads was investigated by different microscopic techniques: in vivo examinations in the polarizing microscope, thin sections and freeze-fracture replicas observed by transmission electron microscopy. The freeze-fractured chromatin appears to be formed of superimposed lamellae, each one 330 A thick. These lamellae are parallel to the flattening plane of the sperm head. This situation was already described in other mammal spermatozoa and in particular in the bull and the rabbit. This work presents a new interpretation of this lamellated aspect. The chroma...
Froman DP, Amann RP.The effects of four vaginal lubricants on progressive spermatozoal motility were evaluated. Neat semen was exposed to 0, 5, or 10% (w/v) of H-R, sterile K-Y, nonsterile K-Y or Maxilube lubricating jellies for 10 min at 37 degrees C and then extended to 10x10(6) spermatozoa/ml. Spermatozoal motility was evaluated after 0, 1, 2, 4 and 6 or 8 h of incubation at 37 degrees C. For bovine spermatozoa, sterile K-Y jelly at 10% suppressed motility (P<0.05), but nonsterile K-Y, H-R and Maxilube jellies had no effect. Maxilube was toxic (P<0.01) to canine spermatozoa and is not recommended for use...
Arriola J, Foote RH.Because microfloral content of stallion semen tends to be high, and strains may be resistant to commonly used antibiotics, amikacin was tested with stallion semen and compared with bull semen. Nine ejaculates to stallion semen were incubated at 37 C in egg yolk-tris extender for 0, 2, 4, 6, 8 and 10 h in the presence of amikacin concentrations of 0, 50, 100, 250, 500, 1,000 and 10,000 microgram/ml, with penicillin and penicillin-streptomycin as controls. Averaged over all incubations, spermatozoal motility was 44, 48, 49, 46, 45, 45 and 19%, for increasing concentrations of amikacin, compared ...
Bader H.A total of 23 mares were inseminated once within 0-6 h after clinical detection of ovulation, 14 with fresh and 9 with deep-frozen semen containing 0.1 x 10(9) to 4.7 x 10(9) motile spermatozoa. Within these two groups, the mares were slaughtered 2, 4 or 6 h after insemination and their genital tracts removed. The utero-tubal junction, isthmus and ampulla ipsilateral to the ovary in which ovulation occurred were flushed separately for sperm recovery. In 1 or 2 mares of each group, the uterine horn and corpus uteri, the cervix and vagina were also flushed. Tissue samples were collected from the...
Rideout MI, Burns SJ, Simpson RB.Sterile filtrates were prepared from equine isolates of Klebsiella pneumoniae (genitalium), Proteus mirabilis, Pseudomonas aeruginosa, Escherichia coli, Streptococcus zooepidemicus, Streptococcus equisimilis, Actinobacillus equuli, and Corynebacterium equi and mixed individually with extended stallion semen. When diluted in the extended semen the filtrates represented bacterial populations of 0.5 x 10(6), 1 x 10(6), 2 x 10(6) and 4 x 10(6) cells/ml. pH values were recorded for each filtrate. Specimens were monitored for percentage motile spermatozoa at 30-min intervals until they reached 10% o...
Guay P, Rondeau M, Boucher S.The effect of different glycerol concentrations (0 to 5.3 per cent) on motility, viability and aspartate aminotransferase (AST) release of stallion spermatozoa was studied before and after deep-freezing. Addition of glycerol to a TRIS-fructose-egg yolk diluent used to extend stallion semen had no effect on motility and viability of spermatozoa and it did not increase AST release. Inclusion of glycerol in the extender only partially preserved the motility and viability of stallion semen during deep-freezing. A fertility trial revealed that concentrating stallion semen by centrifugation, followe...
Dixon KE, Kreider JL.Fifty ejaculates, ten from each of 5 mature stallions, were utilized to study the effects of calcium and fatty acids on equine spermatozoa which were isolated in 3% Bovine Serum Albumin (BSA). The ejaculates were evaluated for percent motile spermatozoa, rate of forward movement, debris, primary abnormalities and secondary abnormalities. The isolation procedure consisted of layering 2 ml of diluted semen (100 x 10(6) spermatozoa/ml) over 6 ml of 3% BSA in 13 x 125 mm columns in a water bath (37 degrees C). After 30 min., the top semen layer and upper half of the BSA layer were withdrawn from a...
Chacon-Arellano JT, Woolley DM.Smooth muscle cells are present in the tunica albuginea testis of the horse, pig and sheep. typical fusiform muscle cells constitute a distinct layer up to 0.3 micrometer thick in the horse; there are fewer muscle cells, mainly of the branched form, in the pig; whereas in the sheep the muscle component is least well developed, with some cells intermediate in form between smooth muscle cells and fibroblasts (myofibroblasts). Attention is drawn to the continuity of this capsular muscle with the smooth muscle associated with the vasculature of the spermatic cord in the horse. This association sug...
Johnson L, Amann RP, Pickett BW.Spermatozoa from four regions of the epididymis and from ejaculated semen were evaluated for their resistance to cold shock, progressive motility, and structural stability. Spermatozoa were incubated at 38 C and their percentage of eosinophilia was compared with that of spermatozoa cooled to 0 C in 2 minutes, 10 C in 12 minutes, or 4 C in 22 minutes. Spermatozoa motility was estimated visually under phase-contrast microscopy and was recorded by cinematography. Structural stability of spermatozoa incubated in 0.05 M sodium borate buffer, 0.035 M sodium dodecyl sulfate (SDS), 0.002 M dithiothrei...
Shideler RK, Squires EL, Pickett BW, Anderson EW.Twenty-three geldings were used to determine changes in seminal characteristics following castration and the effect of frequency of ejaculation on these seminal characteristics. In Exp. 1, semen was collected from 8 geldings every other day after castration until the number of spermatozoa per ejaculate was below 1% of the precastration value. An average of 3 ejaculates was required to reduce the number of spermatozoa below this level. In Exp. 2, 15 stallions were castrated and each stallion was assigned to 1 of 3 groups for seminal collection at 7, 14 or 21 days post-castration. The ejaculates...
Martins HS, Souza MR, Penna CF, da Silva GC, Côrtes SF, Stahlberg R, Lagares MA.Cooled semen has been used routinely to prolong sperm viability until artificial insemination time. However, spermatozoa are subjected to oxidative stress. The aim of the present work was to investigate the protective and antioxidant effect of the milk proteins lactoferrin (Lf) and caseinate added to equine semen cooling extenders. Semen from six stallions was cooled at 5 °C after resuspension with C1) milk- and glucose-based, C2) 0.6% caseinate, C3) C2 + Lf 200 μg ml-1 , C4) C2 + Lf 500 μg ml-1 and C5) C2 + Lf 1000 μg ml-1 extenders, and kept at 5 °C for 24 h. Sperm motili...
Ellington JE, Ball BA, Blue BJ, Wilker CE.Reliable capacitation of equine spermatozoa has been a major obstacle in the development of equine in vitro fertilization. Experiments were done to compare in vitro capacitation of equine spermatozoa by use of heparin/caffeine, calcium ionophore, uterine tube epithelial cell (UTEC)-conditioned medium, and direct culturing of spermatozoa with UTEC (coculturing). Capacitation-like changes, as determined by chlortetracycline membrane staining patterns, developed with UTEC-conditioned medium and coculturing, equivalent to that with calcium ionophore. Both of these treatments induced more (P < 0.05...
Gil L, Olaciregui M, Luño V, Malo C, González N, Martínez F.In recent years, there has been an increased interest in new preservation techniques that facilitate sperm storage and distribution, with freeze-drying (FD) having been proposed as an alternative method for sperm preservation and maintenance of genetic resources in different animal species. FD is a method in which frozen material is dried by sublimation of ice, thereby involving a direct transition from a solid (ice) to a vapour (gas) phase. One of the main advantages of FD is that nitrogen and dry ice are no longer required for the storage and shipment of frozen sperm, which can be stored at ...
Lagares MA, Martins HS, Carvalho IA, Oliveira CA, Souza MR, Penna CF, Cruz BC, Stahlberg R, Henry MR.Extenders with a defined composition containing only components with clearly protective effects on sperm during storage would be an advantage. The aims of the present work were to assess whether caseinate, improves cooled and frozen equine semen quality. Semen from six stallions were suspended with four different cooling extenders C1) Kenney extender; C2) 0.6 % caseinate; C3) 2.7 % caseinate ; and C4) C1 + 2.1 % caseinate, and frozen extenders: F1) INRA 82 extender; F2) 1.35 % caseinate; and F3) 2.7 % caseinate. Although there was no significant difference between the motility rate among the c...
Merkies K, Buhr MM.Spermatozoal function is affected by the ability to regulate intracellular calcium concentrations ([Ca2+]i), and may be influenced by epididymal maturation as well as environmental components. Regulation of [Ca2+]i in ejaculated and epididymal stallion spermatozoa was monitored over time in various media. Spermatozoa from each of 5 pony stallions (3 ejaculate samples and 1 caput and cauda sample) were labeled with the fluorescent calcium indicator probe Indo-1 in a calcium-free modified Tyrode's buffer. Fluorescent emissions were monitored by a dual wavelength spectrofluorometer over 5 h. Calc...
Bresnahan DR, Lupole RE, Stilz CR, Carnevale EM.Zona pellucida (ZP) proteins are important for fertilization and sperm binding and are closely associated with cumulus cells. Communication between cumulus and oocytes is facilitated by intracellular membrane channels composed of connexins. The extent aging impacts potential differences in fertilization and reductions in fertility is not well understood. This study characterized age-related differences in transcript abundance of ZP proteins and connexins in cells from ovarian follicles. Additionally, differences in sperm binding to oocytes from old and young mares was evaluated. For experiment...
Ortiz I, Quiñones-Pérez C, Hidalgo M, Consuegra C, Diaz-Jimenez M, Dorado J, Vega-Pla JL, Crespo F.Dynamic assessment of sperm DNA fragmentation (SDF) has shown to give fuller understanding of stallion semen quality; however, there have been limited attempts to use this parameter to investigate seasonal changes in productive functions. The aims of this study were to: (a) establish a reliable mathematical model to describe the longevity of cooled-stored sperm DNA integrity; (b) to examine the effect of seasonal variations on SDF. Ejaculates were cooled to 5°C, and SDF was analysed after 0, 6 and 24 hr of storage. The coefficient of determination (R ) was calculated after fine-tuning linear...
Azuma T, Choi YH, Hochi S, Oguri N.The development of in-vitro matured and microfertilized horse oocytes was examined in vitro. Fertilized oocytes were produced by 20-h insemination of in-vitro matured and partially zona-removed oocytes with frozen spermatozoa that had been treated with caffeine/calcium ionophore A23187 (fertilization rate 34.2%, monospermy rate 76.9%). Embryonic development was assessed by the number of nuclei stained with Giemsa solution. In Experiment 1, a continuous 8-day culture of the microfertilized oocytes in TCM199 or modified synthetic oviduct fluid (m-SOF) supplemented with 10% fetal bovine serum or ...
Ertmer F, Oldenhof H, Schütze S, Rohn K, Wolkers WF, Sieme H.If the physiological balance between production and scavenging of reactive oxygen species (ROS) is shifted towards production of ROS this may result in accumulation of cell damage over time. In this study stallion spermatozoa were incubated with xanthine and xanthine oxidase (X-XO) to artificially generate defined levels of superoxide and hydrogen peroxide resulting in sub-lethal oxidative damage. The effects of X-XO treatment on various sperm characteristics were studied. Special emphasis was placed on sperm osmotic tolerance pre-freeze and its correlation with cryosurvival, given that cryopr...
Yánez-Ortiz I, Catalán J, Rodríguez-Gil JE, Miró J, Yeste M.Sperm cryopreservation is one of the most important procedures in the development of biotechnologies for assisted reproduction. In some farm animals, the use of cryopreserved sperm has so many benefits for which relevance has become more evident in recent decades. Values for post-thaw sperm quality, however, are variable among species and within individuals of the same species. There is no standardized methodology for each of the stages of the cryopreservation procedure (andrological examination, semen collection, dilution, centrifugation, resuspension of the pellet with the freezing medium, p...
Vanderwall DK, Woods GL.To determine the effect on fertility of large-volume uterine lavage with lactated Ringer's solution (LRS) performed immediately prior to insemination in mares. Methods: Prospective randomized controlled study. Methods: 20 mares. Methods: Control mares (n = 10) were inseminated with 1 billion (estimated before cooling) progressively motile spermatozoa that had been cooled in a passive cooling unit for 24 hours. Mares (n = 10) in the treatment group were inseminated with 1 billion progressively motile spermatozoa (cooled as described for control mares) immediately after uterine lavage with 4 L o...
Folin M, Gennari G, Jori G.The irradiation of horse and sperm-whale Fe” or Fez’ myoglobins with visible light
showed that axial ligands that render the heme diamagnetic (e.g. 02, CO or CN-) endow the
hemoproteins with a marked photosensitivity. In contrast, high-spin myoglobins are unaffected by
visible light. These findings appear to be of general validity for all hemo-proteins and are in agreement
with the involvment of the triplet state of the heme as the reactive intermediate. In all cases, the overall
photoprocess occurs within a very narrow spatial range, leading to specific modification of these
photoox...
Retamal CA, Dias AJ, Brasil FC, Lanzana FR, López ML.The expression of α-D-mannosidase activity was fluorometrically and electrophoretically assessed in spermatozoa, epididymal fluid and homogenates of stallion epididymal tissue. Enzyme activity had regional differences; it was higher (P<0.05) in samples from the cauda epididymal region than in samples from the proximal caput region (largely composed of efferent ducts). Based on enzyme activity, as a function of pH of the assay substrate, electrophoretic analysis in native and native/SDS-PAGE conditions, and the effect of inhibitors or activators, we inferred the presence of at least two cat...
McDonnell SM.This study is part of ongoing work toward developing pharmacological methods for enhancing and inducing ejaculation in stallions with ejaculatory dysfunction or disabilities that interfere with normal breeding behavior. The objective was to evaluate a treatment regimen involving oral imipramine followed by intravenous xylazine that, in uncontrolled field clinical trials, had shown promise for a higher rate of ejaculation and fewer side effects using a more easily obtained and administered form of imipramine. Eight stallions each underwent eight trials in which treatment consisted of imipramine...
Love CC, Thompson JA, Brinsko SP, Rigby SL, Blanchard TL, Lowry VK, Varner DD.Relationships between sperm motility parameters and viability were evaluated using two fluorescent staining techniques in fresh extended semen (fresh and after 24 h storage at 5 degrees C) that had various concentrations of dead sperm added to simulate different levels of viable and nonviable sperm. Both protocols incorporated SYBR-14 and propidium iodide (PI) while the second protocol added the mitochondrial probe JC-1. The relationship between total sperm motility and percent viable sperm was high between staining protocols (r = 0.98). Time (0 h versus 24 h, P<0.0001) and treatment (0, 10, 2...
Tekin N, Wöckener A, Klug E.The effect of a skim milk extender and a glycine-containing extender on sperm motility and acrosome morphology of stallion semen was examined. There was no difference concerning acrosome morphology. After 24 hours of preservation motility of the ejaculates diluted with glycine extender was significantly superior to those handled with skim milk extender. Storage at 5 degrees C in all cases gave better results than storage at room temperature. Skim milk extender is an appropriate diluent when the semen is used for al on the day of its collection, whereas the glycine-containing extender offers th...
Balao da Silva CM, Ortega Ferrusola C, Gallardo Bolaños JM, Plaza Dávila M, Martín-Muñoz P, Morrell JM, Rodriguez Martínez H, Peña FJ.Flow cytometry is considered the only reliable method for the separation of X and Y chromosome bearing spermatozoa in equines. The MoFlo SX DP sorter is highly efficient, allowing the production of foals of the desired sex. However, to achieve acceptable pregnancy rates the currently used protocol requires working with fresh semen obtained close to, or at, the sorting facility. An alternative protocol was tested during two consecutive breeding seasons. Fresh stallion semen was cooled for 20 h, during which staining with Hoechst 33342 took place. On the following day, this sample was flow sort...
Aurich J, Kuhl J, Tichy A, Aurich C.Differences in the cryotolerance of spermatozoa exist among stallions, but it remains to be determined to what extent such differences are affected by breed. In this study, post-thaw semen quality in stallions presented for semen cryopreservation was analysed retrospectively (1012 ejaculates from 134 stallions of 5 breeds). The percentage of frozen-thawed ejaculates acceptable for artificial insemination (AI) and the number of insemination doses per cryopreserved ejaculate was calculated. Logistic regression analysis revealed sperm motility in raw semen as the most important explanatory variab...
Tkachev AV, Tkacheva OL.The article presents the results of the studies of cytotoxic effect of zearalenone and T-2 toxin on sperm of horses and bulls during incubation and after thawing according to the technology of sperm obtaining and cryopreservation in Kharkov. We first have shown in vitro toxic effects of different concentrations of zearalenone and T-2 toxin (from 0.5 to 0.01 mM) on the membrane stability, as well as quantitative and qualitative indicators of semen in stallions and bulls before and after freezing and thawing. It has been found that the biological activity of the native sperm in 1 h after additio...
Hoshiba H, Sinowatz F.Spermadhesins are proteins with various functions in sperm capacitation and zona pellucida binding. In this study the cellular localization of the spermadhesin AWN-1 has been examined in the equine male genital tract. Results obtained by immunohistochemical methods reveal that in the horse AWN-1 is synthesized in spermatogonia, in the rete testis, the ductus epididymidis and the seminal vesicles. These findings indicate that the cellular origin of spermadhesins is species-specific.
Nagy S, Kovacs B, Johannisson A.The aim of the present study was to test a rapid, robust flow cytometric technique for the detection of sperm head abnormalities of domestic bulls and stallions. The so-called PulSA approach detects the pulse profiles of propidium-iodide labelled spermatozoa. In the first experiment, species-specific threshold values were established on sperm samples that were tested for sperm head abnormalities with a classic visual morphology analysis. In the second experiment, serial mixtures of bull and stallion spermatozoa mimicking different percentages of sperm head abnormalities were analysed. Non-metr...
Maclellan LJ, Albertini DF, Stokes JE, Carnevale EM.The impact of vitrification on oocyte developmental competence as a function of donor age remains an important issue in assisted reproductive technologies (ARTs). Methods: Equine germinal vesicle (GV) or metaphase II (M(II) oocytes were vitrified using the Cryotop® method. Spindle organization and chromosome alignment were evaluated from confocal imaging data sets of in vivo (IVO) or in vitro (IVM) matured oocytes subjected to vitrification or not. Intracytoplasmic sperm injection (ICSI) from the same groups was used to assess developmental potential. Results: An increase in chromosome misali...
Díaz Rojas E, Carrillo Moreno DI, Contreras Villarreal V, Arellano Rodríguez F, Alvarado Espino AS, Ángel García O.The use of reproductive biotechnologies in equine practice has shown that some stallions are subfertile, so ways to improve fertility have been sought. This study aimed to evaluate the effect of nutraceutical supplementation on improving semen quality in Quarter Horse stallions. Semen from six Quarter Horse stallions was assessed for 4 months every 20 days using the computer-assisted semen analysis system. They were evaluated for 60 days before supplementation; then, the same stallions were re-evaluated for 60 days with nutraceutical supplementation (30 g/day). Volume showed no significant...
Hedia M, Angel-Velez D, Papas M, Peere S, Gerits I, De Coster T, Van den Branden E, Govaere J, Van Soom A, Leroy JLMR, Smits K.The in vitro production of equine embryos via ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) has increased rapidly. There is a marked effect of the individual mare on the outcome of OPU-ICSI, but little is known about the influence of the mare's health condition. This study aimed to investigate the potential associations between the concentrations of interleukin-6 (IL-6), reactive oxygen metabolites (d-ROMs), and biological antioxidant potential (BAP) in serum of oocytes' donor mares and the subsequent embryonic development. Just before OPU, a blood sample was collected from 28...
Ebel FA, Liaudat AC, Blois DA, Capella V, Broglia MF, Barbero CA, Rodríguez N, Bosch P, Rivarola CR.Novel soft materials based on hydrogel are proposed to enhance the selection of high-quality stallion sperm based on their adhesion capacity. The hydrogel surfaces are derived from polyacrylamide (PAAm), which is copolymerized with neutral and ionic co-monomers to modify the interfacial properties. The hydrogels undergo characterization through FTIR spectroscopy, assessment of swelling capacity, and wettability under various experimental conditions. Sperm adhesion capacity on the hydrogels is examined through several parameters including the percentage of bound sperm (%Sp) to hydrogels, tail o...
Egyptien S, Deleuze S, Ledeck J, Ponthier J.Stallion sperm analysis is indicated for infertility diagnosis, pre-sale expertise, production of fresh or frozen doses, and frozen straw quality control. Various collection methods are described, and numerous assays can be performed on semen. Determining an approach for each of these cases is challenging. This review aims to discuss how to obtain relevant clinical results, answering stallion owners' concerns. Semen can be collected with an artificial vagina on a phantom or a mare, by electro-ejaculation under anesthesia, or after pharmacological induction. The collection method influences the...
Fracaro PL, Corcini CD, Souza Gatti NL, Filho JS, Acosta IB, Filho JS, Hartwig FP, Rosa Curcio BD, Varela Junior AS.Glycerol is a cryoprotectant widely used in the freezing of mammalian semen, but no study has demonstrated its optimum concentration and the appropriate exposure time for equine species. Objective: To demonstrate that the exposure time (15, 30, 45, 60, 75 and 90 min) versus concentration (2, 3, 4 and 5%) of the cryoprotectant glycerol influences the freezing success of equine semen. Methods: The ejaculate of 12 stallions were frozen in different glycerol concentrations following different exposure times. The thawed sperm was evaluated for kinetic parameters using a Computer Assisted Semen Anal...
Talluri TR, Jhamb D, Paul N, Singh J, Dedar RK, Mehta SC, Legha RA, Pal Y.The recovery of spermatozoa from the cauda epididymis may be the only option to obtain genetic material from elite stallions that had undergone castration or sudden death due to colic or severe injury. Objective: To evaluate two different protocols for retrieval of stallion epididymal spermatozoa and to evaluate different cryoprotectants on the freezability of the epididymal spermatozoa. Methods: Six epididymides from three stallions were collected immediately after routine castration under general anesthesia. In the first experiment, each epididymis (of two testes) of the same stallion were p...
Catalán J, Yánez-Ortiz I, Martínez-Rodero I, Mateo-Otero Y, Nolis P, Yeste M, Miró J.Previous research revealed that several seminal plasma (SP) metabolites are related to sperm functionality, fertility, and preservation. While it is understood that variations between species exist, whether the SP metabolome differs between donkeys and horses has not been previously investigated. The aim of this work, therefore, was to characterize and compare donkey and horse SP metabolites using nuclear magnetic resonance (NMR) spectroscopy, and relate them to sperm viability and motility. For this purpose, ejaculates from 18 different donkeys and 18 different horses were collected and separ...
Shakeel M, Yoon M.The threat posed by increased surface temperatures worldwide has attracted the attention of researchers to the reaction of animals to heat stress. Spermatogenesis in animals such as stallions is a temperature-dependent process, ideally occurring at temperatures slightly below the core body temperature. Thus, proper thermoregulation is essential, especially because stallion spermatogenesis and the resulting spermatozoa are negatively affected by increased testicular temperature. Consequently, the failure of thermoregulation resulting in heat stress may diminish sperm quality and increase the li...
