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Topic:Semen Preservation

Semen preservation involves the collection, processing, and storage of stallion semen for future use in artificial insemination. This practice enables the extension of genetic material across geographical boundaries and temporal constraints. The preservation process typically includes semen evaluation, dilution with extenders, cooling, and sometimes cryopreservation. Factors such as semen quality, extender composition, and storage conditions influence the success of preservation. This page compiles peer-reviewed research studies and scholarly articles that explore techniques, challenges, and advancements in the field of equine semen preservation, focusing on optimizing fertility outcomes and extending the reproductive lifespan of stallions.
[Successful use of deep-frozen stallion sperm after 23 years of storage at -196 degrees C].
DTW. Deutsche tierarztliche Wochenschrift    December 1, 1990   Volume 97, Issue 12 544-545 
Krause HD, Sieme H, Merkt H, Bader H, Wöckener A.A reduction in the motility of the spermatozoa in stallion semen stored in pellet form for 23 years in liquid nitrogen at -196 degrees C could not be seen after thawing. The insemination of a mare with this semen resulted in a normal pregnancy. A normally developed, healthy male foal was born after a gestational period of 321 days.
[Successful use in horses of deep-frozen semen specimens stored for 18 years].
Berliner und Munchener tierarztliche Wochenschrift    June 1, 1990   Volume 103, Issue 6 211-212 
Braun J, Schefels W, Stolla R.In 1970 semen from a Haflinger-stallion was frozen by the pellet method. 18 years later semen samples were used to inseminate 4 mares. Inseminations were performed shortly after ovulation with a total number of motile spermatozoa between 150 and 636 x 10(6), the percentage of motile spermatozoa being 20% to 40%. Three mares conceived after a single insemination, one mare got pregnant after 4 inseminations during 3 oestrous periods. Meanwhile, 3 foals were born and one of the mares is still pregnant. The results demonstrate that long-term storage of frozen semen in liquid nitrogen does not impa...
Fertilizing capacity of equine spermatozoa stored for 24 hours at 5 or 20 degrees C.
Theriogenology    October 1, 1989   Volume 32, Issue 4 515-525 doi: 10.1016/0093-691x(89)90273-2
Varner DD, Blanchard TL, Meyers PJ, Meyers SA.A breeding trial was conducted to evaluate the effect of in vitro storage time and temperature on fertilizing capacity of equine spermatozoa. Semen obtained from one stallion and diluted with skim milk-glucose extender was used to artificially inseminate 45 estrussynchronized mares. The mares were assigned to one of three treatment groups (15 mares per group): 1) insemination with fresh semen (collected within 0.5 h of use), 2) insemination with semen stored for 24 h at 20 degrees C or 3) insemination with semen stored for 24 h at 5 degrees C. The mares were inseminated daily during estrus, fr...
[Preservation capability of horse semen by the use of two diluents and preservation temperatures].
DTW. Deutsche tierarztliche Wochenschrift    May 1, 1989   Volume 96, Issue 5 258-265 
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...
Changes in quality of stallion spermatozoa during cryopreservation: Plasma membrane integrity and motion characteristics.
Theriogenology    February 1, 1989   Volume 31, Issue 2 283-298 doi: 10.1016/0093-691x(89)90533-5
Blach EL, Amann RP, Bowen RA, Frantz D.Better procedures for freezing and thawing equine sperm are needed since variable fertility is obtained when cryopreserved sperm are used. To evaluate current methods of freezing equine sperm, we examined spermatozoal quality by means of two new techniques. These measured the integrity of plasma-acrosomal membranes by immunofluorescent analyses of binding of an antibody specific to the acrosome and evaluated eight parameters of spermatozoal motion using a fully automated computerized system. Five ejaculates from each of eight stallions were processed for freezing in egg yolk-lactose extender w...
Use of a monoclonal antibody to evaluate integrity of the plasma membrane of stallion sperm.
Gamete research    November 1, 1988   Volume 21, Issue 3 233-241 doi: 10.1002/mrd.1120210305
Blach EL, Amann RP, Bowen RA, Sawyer HR, Hermenet MJ.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...
[The effect of diluents, dose size and freezing speed on the survival rate of deep frozen stallion sperm].
DTW. Deutsche tierarztliche Wochenschrift    October 1, 1988   Volume 95, Issue 9 369-371 
Hellemann C, Hernandez C.No abstract available
Reproduction.
The Veterinary clinics of North America. Equine practice    August 1, 1988   Volume 4, Issue 2 161-336 
No abstract available
Studies of stallion sperm survival: preservation of progressive motility of stallion spermatozoa by low ionic strength media.
Gamete research    June 1, 1988   Volume 20, Issue 2 215-222 doi: 10.1002/mrd.1120200211
Bize I, Driscoll DM.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...
Effect of insemination timing on the fertilizing capacity of frozen/thawed equine spermatozoa.
Theriogenology    February 1, 1988   Volume 29, Issue 2 429-439 doi: 10.1016/0093-691x(88)90245-2
Kloppe LH, Varner DD, Elmore RG, Bretzlaff KN, Shull JW.A breeding trial was conducted to evaluate the effect of insemination timing on the fertility of mares bred with frozen/thawed equine semen. One stallion and 60 reproductively sound, estrous-synchronized mares were included in the study. Mares were assigned to one of three groups (n = 20): 1) insemination with fresh semen every other day during estrus from detection of a 35-mm follicle until ovulation, 2) insemination with frozen/thawed semen every day during estrus from detection of a 35-mm follicle until ovulation or 3) insemination with frozen/thawed semen once, within 6 h after ovulation. ...
Reproductive efficiency in domestic animals.
Annals of the New York Academy of Sciences    January 1, 1988   Volume 541 697-705 doi: 10.1111/j.1749-6632.1988.tb22307.x
First NL, Eyestone WH.No abstract available
Effects of cooling rate and storage temperature on equine spermatozoal motility parameters.
Theriogenology    January 1, 1988   Volume 29, Issue 5 1043-1054 doi: 10.1016/s0093-691x(88)80028-1
Varner DD, Blanchard TL, Love CL, Garcia MC, Kenney RM.Two experiments were conducted to examine the effects of cooling rate and storage temperature on motility parameters of stallion spermatozoa. In Experiment 1, specific cooling rates to be used in Experiment 2 were established. In Experiment 2, three ejaculates from each of two stallions were diluted to 25 x 10(6) sperm/ml with 37 degrees C nonfat dry skim milk-glucose-penicillin-streptomycin seminal extender, then assigned to one of five treatments: 1) storage at 37 degrees C, 2) storage at 25 degrees C, 3) slow cooling rate to and storage at 4 degrees C, 4) moderate cooling rate to and storag...
[Sperm received in shipment versus fresh sperm in relation to fertilization results].
Tijdschrift voor diergeneeskunde    December 15, 1987   Volume 112, Issue 24 1410-1412 
Berghuis GA.The conception rates of semen intended for shipment and those of recently obtained semen are compared in the present paper. Conception rates using recently obtained semen were significantly superior to those obtained with semen intended for shipment. A number of factors to which this difference could be due are briefly discussed.
Effects of semen fractionation and dilution ratio on equine spermatozoal motility parameters.
Theriogenology    November 1, 1987   Volume 28, Issue 5 709-723 doi: 10.1016/0093-691x(87)90288-3
Varner DD, Blanchard TL, Love CL, Garcia MC, Kenney RM.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 fract...
[Establishment of modern equine insemination centers].
DTW. Deutsche tierarztliche Wochenschrift    September 1, 1987   Volume 94, Issue 8 478-480 
Klug E.No abstract available
[The effect of nitrofurazone on the state of sperm in the stallion].
DTW. Deutsche tierarztliche Wochenschrift    September 1, 1987   Volume 94, Issue 8 483-484 
Bader H, Wöckener A, Heilkenbrinker T.No abstract available
A modified phantom (dummy mount) for stallion semen collection.
Equine veterinary journal    July 1, 1987   Volume 19, Issue 4 339-341 doi: 10.1111/j.2042-3306.1987.tb01427.x
Volkmann DH.No abstract available
Motility and fertility of equine spermatozoa in a milk extender after 12 or 24 hours at 20 degrees C.
Theriogenology    March 1, 1987   Volume 27, Issue 3 517-525 doi: 10.1016/0093-691x(87)90239-1
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...
Effects of stallion seminal plasma on hydrogen peroxide release by leukocytes exposed to spermatozoa and bacteria.
Journal of reproductive immunology    February 1, 1987   Volume 10, Issue 2 157-166 doi: 10.1016/0165-0378(87)90073-8
Hansen PJ, Hoggard MP, Rathwell AC.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.
Polskie archiwum weterynaryjne    January 1, 1987   Volume 27, Issue 1 5-21 
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...
Practicalities of insemination of mares with deep-frozen semen.
Journal of reproduction and fertility. Supplement    January 1, 1987   Volume 35 121-125 
Müller Z.From 341 stallions examined for sperm quality, 61% of warm-blooded stallions and 47% of cold-blooded stallions fulfilled the pre-existing criteria for their occasional use in insemination. From these stallions 51-71% of acceptable ejaculates were obtained. Altogether 959 mares were inseminated in an average of 1.36 oestrous cycles. For the insemination of one mare in one oestrous cycle on the average 2.2 insemination doses were used. These inseminations were carried out by 41 cattle insemination technicians trained in mare insemination. A pregnancy rate of 56% and a foaling rate of 48% were ac...
Proteins in stallion seminal plasma.
Journal of reproduction and fertility. Supplement    January 1, 1987   Volume 35 113-120 
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...
Influence of season and frequency of ejaculation on production of stallion semen for freezing.
Journal of reproduction and fertility. Supplement    January 1, 1987   Volume 35 127-133 
Magistrini M, Chanteloube P, Palmer E.In an attempt to define optimal season and ejaculation frequency for frozen semen, semen was collected from 6 stallions (3 horses and 3 ponies) 3 times per week or every day, alternating every week, for 1 year. The semen was evaluated and frozen. All the samples were thawed at the end of the experiment. At collection, fresh semen evaluations showed that winter (as opposed to spring and summer) was associated with low sexual behaviour, small volumes of spermatozoa and gel, high sperm concentration and lower motility. The high ejaculation frequency yielded a decreased volume, concentration of sp...
Influence of chlorhexidine on seminal patterns in stallions.
Journal of reproduction and fertility. Supplement    January 1, 1987   Volume 35 109-112 
Rath D, Leiding C, Klug E, Krebs HC.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...
Fertility of stallion semen frozen in 0.5-ml straws.
Journal of reproduction and fertility. Supplement    January 1, 1987   Volume 35 143-148 
Volkmann DH, van Zyl D.Semen of 2 pony stallions was frozen by 2 methods in 0.5 ml PVC straws. The fertility of the frozen-thawed semen was evaluated by inseminating 60 mares during 69 oestrous cycles. An overall single cycle pregnancy rate of 55% was achieved. Freezing method, stallion, insemination during steroid-synchronized oestrus or insemination only every 2nd day during oestrus did not significantly influence pregnancy rates. Pregnancy rates were significantly improved from a mean 44% to a mean 73% when the mean number of progressively motile spermatozoa per insemination was increased from 175 x 10(6) to 249 ...
Motility and ATP content of extended equine spermatozoa in different storage conditions.
Journal of reproduction and fertility. Supplement    January 1, 1987   Volume 35 103-107 
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...
Use of different nonglycolysable sugars to maintain stallion sperm viability when frozen or stored at 37 degrees C and 5 degrees C in a bovine serum albumin medium.
Journal of reproduction and fertility. Supplement    January 1, 1987   Volume 35 135-141 
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...
Demonstration of the carrier state in naturally acquired equine arteritis virus infection in the stallion.
Research in veterinary science    September 1, 1986   Volume 41, Issue 2 279-280 
Timoney PJ, McCollum WH, Roberts AW, Murphy TW.The chronic carrier state was virologically confirmed in 15 thoroughbred stallions naturally infected with equine arteritis virus based on the results of test matings and, or, isolations of the virus from semen. Carrier stallions were shown to shed equine arteritis virus in the semen for at least one to two years. Existence of a short-term or convalescent carrier state was also demonstrated in five additional stallions. The frequency of the long-term carrier state in stallions naturally infected with equine arteritis virus was 35 per cent; it varied considerably between groups of stallions on ...
[Comparative studies on stallion sperm after repeated sperm collection and subsequent deep-freeze conservation].
Zentralblatt fur Veterinarmedizin. Reihe A    February 1, 1986   Volume 33, Issue 2 155-159 
von Frey W, Bernal A, de los Reyes M.No abstract available
Assessment of spermatozoal function using dual fluorescent staining and flow cytometric analyses.
Biology of reproduction    February 1, 1986   Volume 34, Issue 1 127-138 doi: 10.1095/biolreprod34.1.127
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...