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Topic:Semen Analysis
Semen analysis involves the evaluation of semen quality to assess reproductive potential and health in horses. This process typically includes the examination of several parameters such as sperm concentration, motility, morphology, and volume. Semen analysis is used in breeding programs to determine the fertility of stallions and to identify any reproductive issues. The collected data can assist in making informed decisions about breeding management and interventions. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, findings, and implications of semen analysis in equine reproduction.
Testicular growth, hormone concentrations, seminal characteristics and sexual behaviour in stallions. Puberty was studied using 15 colts of Quarter Horse phenotype. Total scrotal width was measured every 8 weeks from 48 to 96 weeks. Blood samples were taken from 8 colts at 8, 16 and 24 weeks and then every 4 weeks until 100 weeks to measure changes in LH, FSH and testosterone concentrations. Seminal collections were attempted monthly from 48 to 64 weeks and every 2 weeks thereafter until puberty resumed every 3rd day from 96 weeks for 15 ejaculates. For all collections, times to erection, mount and ejaculation and seminal characteristics were recorded. Age at puberty was defined as the first e...
[Safety precautions during semen collections from stallions]. In the Federal Republic of Germany a lethal accident occurred recently during the semen collection from a stallion and it would be advisable to observe the safety rules of which there are three sections: hobbeling and if necessary twitching of the mare, security in the manner in which the stallion is led, precautions to be observed by the operator.
Diagnostic aids for the detection of urine in the equine ejaculate. An experiment was conducted to evaluate three commercially available test kits, the Azostix, Multistix and Uric-acid test, for the detection of urine in the equine ejaculate. Azostix, which tests for urea nitrogen, consistently detected urine in the equine ejaculate. Urine contamination was evident when a color change occurred in the reagent pad, going from yellow to green after 10 sec of exposure. The sensitivity of Azostix to urea nitrogen in contaminated samples was 39 mg/dl. The Multistix test kit also successfully detected urine in semen. In the Multistix nitrite pad the color changed fro...
A spectrophotometric procedure for the determination of objective measurements of equine spermatozoan motility. A spectrophotometric procedure was developed and evaluated for the objective measurement of equine spermatozoan motility. A 100 mul sample of a sperm suspension, prepared by the removal of seminal plasma, was layered under a column of optically clear medium in a specially designed spectrophotometric cuvette maintained at 37 degrees C. Changes in light transmittance above the interface of the sperm suspension and medium were recorded on chart paper. As sperm cells swam into the medium, a decrease in light transmittance was recorded as a deflection on the chart paper. Chart recordings were analy...
Semen selenium content and sperm mitochondrial volume in human and some animal species. Selenium (Se) and glutathione peroxidase (GSH-Px) were determined from the seminal plasma samples and spermatozoa of human and four different animal species. The human sperm Se concentration was 1.8 +/- 0.8 micrograms/g dry weight, which was about half of that in the bull. Abnormal sperm morphology and motility correlated with low sperm Se content. The volume of sperm mitochondrial sheath in human, bull and stallion was measured using transmission electron microscopy. In these species the sperm Se content was highly correlated with the volume of mitochondria. Among the five species studied, th...
Sperm granuloma in a stallion. A 7-year-old stallion with a history of abdominal pain after it fell was examined and found to have a swelling of the right testis and epididymis. Semen evaluation revealed an increase in secondary sperm abnormalities. The stallion was unilaterally castrated. The histologic diagnosis was sperm granuloma, with no evidence of infection. Periductal fibrosis was observed and appeared to have developed before the trauma occurred. The changes seen could be compatible with chronic blockade of efferent ductules, resulting in extravasation of spermatozoa.
Use of a monoclonal antibody to evaluate integrity of the plasma membrane of stallion sperm. Transmission electron microscopy was used to confirm that a monoclonal antibody (F79.3E2; class IgG1 kappa) was specifically localized to an antigen in the acrosomal ground substance of stallion sperm. This antibody was used to develop and validate an indirect immunofluorescent procedure to evaluate integrity of the plasma-acrosomal membranes of stallion sperm. The concept was that primary monoclonal antibody would be "shielded" from its acrosomal antigen by an intact plasma membrane. Conversely, sperm with damaged plasma-acrosomal membranes would exhibit green acrosomal fluorescence when view...
Studies of stallion sperm survival: preservation of progressive motility of stallion spermatozoa by low ionic strength media. Preservation of stallion sperm forward motility was studied using a video recording system in semen diluted with media of different ionic strength and sodium content. After 8 hr of incubation at room temperature, semen diluted in a low ionic strength media containing sucrose displayed 65 +/- 9% motility with 68 +/- 3% of the motile sperm showing forward motility (diameter of head trajectory greater than or equal to 80 microns). In contrast, sperm populations diluted and incubated with a normal ionic strength media containing sodium had 56 +/- 7% motile sperm of which only 36 +/- 7% displayed f...
Bilateral seminal vesiculitis and ampullitis in a stallion. A Thoroughbred stallion suspected of having venereal disease was found to have an infection of the accessory sex glands. Purulent debris, blood, and Pseudomonas aeruginosa were recovered from all ejaculates. Treatment with gentamicin sulfate, tobramycin, and amikacin sulfate was unsuccessful in eliminating the infection. The stallion's seminal plasma, collected during treatment with gentamicin sulfate, did not contain any appreciable antibacterial activity. Apparently, negligible amounts of gentamicin diffused across the mucosal cell borders of the accessory sex glands into the seminal plasma....
Effects of semen fractionation and dilution ratio on equine spermatozoal motility parameters. Two experiments were conducted to examine the effects of semen fractionation and dilution ratio on motility parameters of stallion spermatozoa. In Experiment 1, three ejaculates from each of three stallions were divided into sperm-rich (SR) and sperm-poor (SP) fractions to determine the difference in sperm concentration. Mean sperm concentration in SR fractions (349.5 x 10(6)/ml) was greater (P < 0.001) than that of SP fractions (96.9 x 10(6)/ml). In Experiment 2, three ejaculates from each of two stallions were divided into SR and SP fractions. Fifty percent of the original volume of SR fr...
Motility and fertility of equine spermatozoa in a milk extender after 12 or 24 hours at 20 degrees C. 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...
Influences of season and artificial photoperiod on stallions: testicular size, seminal characteristics and sexual behavior. To investigate the influence of daylength on the seasonal reproductive cycle of stallions, 21 stallions were assigned to one of three treatments: control, ambient (natural) photoperiod; S-L, 8 h light and 16 h dark (8:16) for 20 wk beginning July 16, 1982 then 16:8 from December 2, 1982 until March 5, 1984; S-S, 8:16 from July 16, 1982 until March 1984. Temperature was not controlled and was similar for all groups. Total scrotal width (TSW) was measured every 4 wk throughout the experiment. During 10 periods, semen was collected and evaluated every other day for 3 wk and sexual behavior was as...
Effects of stallion seminal plasma on hydrogen peroxide release by leukocytes exposed to spermatozoa and bacteria. The ability of stallion seminal plasma to modify phagocytosis of spermatozoa and Streptococcus zooepidemicus was examined. Phagocytosis was monitored indirectly as the H2O2 produced by peripheral blood leukocytes after addition of spermatozoa or bacteria. Hydrogen peroxide production after addition of ejaculated spermatozoa was greater (P less than 0.01) than after addition of epididymal sperm. Furthermore, pre-incubation of epididymal sperm with 6.25-50% seminal plasma caused a dose-dependent increase in subsequent H2O2 production by leukocytes (P less than 0.05). In addition, equine serum wa...
Analysis of the physiological processes connected with sexual maturation of stallions. 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...
The carrier state in equine arteritis virus infection in the stallion with specific emphasis on the venereal mode of virus transmission. The carrier state has been confirmed virologically in Thoroughbred and non-Thoroughbred stallions naturally infected with equine arteritis virus (EAV). Short-term or convalescent and long-term carriers occur. The frequency rate of the long-term carrier state in Thoroughbreds was high, averaging 33.9% among the three groups of stallions under study. While the convalescent carrier state only lasted a few weeks after clinical recovery, the long-term carrier state could persist for years. There was evidence, however, that not all such carriers might remain persistently infected for life. Carrier s...
Influence of chlorhexidine on seminal patterns in stallions. Ejaculates were collected at 3-day intervals before, during and after a washing procedure with chlorhexidine (2%). Semen motility and pathology were determined before and after deep-freezing. Blood samples were taken before and within 1 h after washing procedures and then extracted in ether. This was followed by HPL chromatography. Chlorhexidine concentrations in blood and seminal plasma were distinctly higher in the treated stallions than in control groups. Concentrations in the control groups were below the detection limit of the column. Significant correlations between decreasing semen qual...
Evaluation of cellulose acetate/nitrate filters for the study of stallion sperm motility. 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...
Assessment of spermatozoal function using dual fluorescent staining and flow cytometric analyses. 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...
Proteinase inhibitors of horse seminal plasma. A high molecular mass, acid-soluble proteinase inhibitor. Horse seminal plasma does not possess a proteinase inhibitor corresponding to human HUSI-I (human seminal plasma inhibitor). Instead a protein complex of high relative molecular mass (Mr) containing proteinase inhibitory activity was detected, which was called horse seminal plasma protein complex or HSPC. The compound had a broad enzyme-inhibiting spectrum. Its Mr was estimated to be 800 000 and it was composed of 7 different polypeptides with Mr values ranging from 11 000 to 30 000. Its carbohydrate content was between 3.5% and 5%. Despite the high molecular mass, the complex was soluble in d...
Photomicrographic evaluation of stallion spermatozoal motility characteristics. 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.
Microtubular mass defect of spermatozoa in the stallion. A microtubular mass (MM) defect was found in the spermatozoa of 7 Standardbred stallions; 3 stallions were sons of the same sire. Two of these 3 stallions and 2 other stallions (for a total of 4 out of the 7 stallions) were considered subfertile when the defect was first observed. Fertility improved with time, either during the first breeding season or when a given stallion was used less frequently; however, the MM defect persisted, consisting of tortuous arrays of small abnormal microtubules visible only by transmission electron microscopy. The MM probably contained the protein tubulin as ind...
[Breeding ability of the stallion depending on bacteriological findings]. No abstract available
Seasonal and circadian changes of testosterone levels in the peripheral blood plasma of stallions and their relation to semen quality. Three stallions were bled each hour for 25 h at 28-day intervals throughout 1 year. Testosterone levels were pulsatile. Pulse frequency and pulse amplitude were higher in the summer months than at other times (P less than 0.01). The number of testosterone pulses also varied throughout the day, with the greatest frequency occurring in the afternoon (14.00-17.00 h) and at night (22.00-01.00 h). Mean testosterone levels were highest in the summer (P less than 0.01) but showed a secondary, smaller increase in the autumn. Semen characteristics were assessed from measurements of 222 stallions. Semen...
Inhibition of motility of bovine, canine and equine spermatozoa by artificial vagina lubricants. 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...
Effects of amikacin sulfate on the motility of stallion and bull spermatozoa at different temperatures and intervals of storage. 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 ...
Effect of anabolic steroids on reproductive function of young stallions. Thirty-two stallions were used to determine the effect of anabolic steroids on reproductive function. Stallions were assigned to one of the four treatments: 1) .23 ml sesame oil/kg of body weight (BW; control, C); 2) 4.4 mg boldenone undecylenate/kg BW (4E); 3) 1.1 mg boldenone undecylenate/kg BW (1E) and 4) 1.1 mg nandrolone decanoate/kg BW (D). Injections were given at 3-wk intervals for 15 wk. Semen was collected every other day for 3 wk before the first injection and at the same frequency during d 85 through 105 (d 0 = day of first injection). Libido was assessed on the basis of reaction t...
Prostaglandins in stallion semen. The purpose of the experiment was to obtain preparatory information about the presence of prostaglandins in semen collected from various types of horses after different periods of sexual rest. Semen was collected with an artificial vagina. Prostaglandin-like activity was estimated by the bioassay procedure described by Vane (1). Results are expressed in ng/ml PGE(2) of seminal plasma. The total concentration of prostaglandins in the full ejaculate averaged 43.73 +/- 4.93 ng/ml of plasma while the total amount of prostaglandins in the ejaculate was 1076 ng. Taking into consideration the period ...
