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.
Teuscher C, Kenney RM, Cummings MR, Catten M.In this study, 2 stallions were immunised with their own spermatozoa to ascertain whether an antisperm autoantibody response could be mounted. The results demonstrated that the stallion can recognise and respond to sperm autoantigens by producing circulating antisperm antibodies, primarily of the IgG class. Such autoantibodies appeared 2-4 weeks after inoculation and persisted for 6-20 weeks. Immunochemical characterisation by western blot identified two major sperm autoantigens, with molecular weights of 70 kD and 62 kD. Control pony stallions immunised with adjuvants alone failed to exhibit ...
Kotilainen T, Huhtinen M, Katila T.The objective of this study was to investigate the inflammatory reaction induced in the equine uterus by insemination with fresh and frozen semen. Eleven groups (6 to 8 mares per group) were studied during 2 breeding seasons. The mares were inseminated using raw semen, frozen semen, extended fresh and frozen semen, concentrated fresh semen, seminal plasma and seminal extenders only. One group was bred naturally. Six hours after insemination, the uteri were flushed with 50 ml of phosphate-buffered saline (PBS). Seventeen out of 104 samples (16%) exhibited slight bacterial growth. Neutrophil con...
Heiskanen ML, Huhtinen M, Pirhonen A, Mäenpää PH.Semen from 3 stallions was extended using 2 methods (Kenney extender and a modified Kenney extender), slowly cooled, and stored for 41 +/- 6 (s.d.) h before insemination. An insemination dose (40 ml) contained 1.5-2 billion spermatozoa. In the experiment, 26 mares were inseminated in 30 cycles. The pregnancy rate per cycle obtained with sperm stored in the Kenney extender was 87% (n = 15). When the semen was extended with the modified extender, centrifuged and stored, the pregnancy rate was 60% (n = 15). Inseminations were done every other day until ovulation was detected. If a mare ovulated m...
Hochi S, Fujimoto T, Choi YH, Braun J, Oguri N.Immature equine oocytes were frozen-thawed with ethylene glycol (EG), 1,2-propanediol (PD) or glycerol (GL) in PBS and cultured to assess the rate of in vitro maturation (Experiment 1). Compact-cumulus oocyte complexes were collected from slaughterhouse ovaries and equilibrated for 10 min in the freezing medium containing 10% (V/V) cryoprotectant and 0.1 M sucrose. The 0.25-ml straws, loaded with 10 to 30 oocytes, were seeded at -6 degrees C and cooled to -35 degrees C at 0.3 degrees C/min before being plunged into liquid nitrogen. The straws were thawed rapidly in a 37 degrees C waterbath for...
Davis RO, Gravance CG, Casey PJ.Tissue variation in microscope slides made for spermatozoon analysis and variation introduced by the subjective techniques used to analyze these slides reduce the statistical power of studies that seek to use spermatozoon morphology to predict fertility. A simple specimen preparation method was developed to standardize stallion spermatozoon morphologic smears, and a new, automated spermatozoa morphometry instrument was used to objectively analyze the efficacy of the specimen preparation technique. The method achieved a standard spermatozoon concentration and reduced field-to-field variation in...
Varner DD, Bowen JA, Johnson L.The onset of sperm capacitation/acrosome reaction was evaluated using heparin. Equine semen was incubated at 38 degrees C for 4.5 h in culture medium with and without 10 micrograms/mL heparin and with and without 0.1 microM of Ca2+ ionophore. Sperm acrosome reaction was detected using chlortetracycline fluorescence (CTC) and transmission electron microscopy (TEM). The CTC assay provided staining patterns that corresponded with the capacitation/acrosome reaction in other mammalian species (man, mouse, guinea pig). The percentages of incapacitated sperm (PUC), capacitated acrosome-intact sperm (...
Vaillancourt D, Guay P, Higgins R.Semen from three stallions was used to evaluate the effectiveness of two antibiotics added to semen extender for samples stored at 20 degrees C or 5 degrees C for up to 48 hours. Each ejaculate was divided into six different treatments: semen+extender (SE); SE+gentamicin (100 micrograms/mL); SE+polymyxin B (1000 units/mL); and each of the above treatments inoculated with Pseudomonas aeruginosa ATCC 27853. Sampling of diluted semen for bacteriological analysis was performed after 2, 8, 24 and 48 hours of preservation at either temperatures. The presence of nonspecific bacteria was noted after t...
Ellington JE, Ignotz GG, Varner DD, Marcucio RS, Mathison P, Ball BA.Coculture of stallion sperm with monolayers of equine oviductal epithelial cells (OEC) was evaluated. Monolayers were obtained from frozen-thawed OEC. Live sperm attached to the OEC in vitro, whereas sperm killed by heat treatment or glutaraldehyde fixation did not. Sperm attached to OEC showed flagellar motion for 4 d in vitro, during which time they gradually became released. Scanning electron-micrographs showed an intimate association between the sperm and OEC. Incubation of sperm for 4 h with either control, heparinized or OEC-conditioned medium (Tyrode's albumin lactate phosphate) resulte...
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...
Casey PJ, Hillman RB, Robertson KR, Yudin AI, Liu IK, Drobnis EZ.An acrosomal staining technique that can differentiate between living and dead sperm was developed for equine sperm. The fluoresceinated lectin Pisum sativum agglutinin (FITC-PSA) was used to identify the presence or absence of acrosomal contents, while the supravital nuclear dye Hoechst 33258 (H258) was used to assess viability. The accuracy of the FITC-PSA acrosomal stain was tested by comparing the percentage of sperm that had lost their acrosomal contents, detected by the staining method, with that detected by transmission electron microscopy (TEM). Following capacitation in vitro, the acr...
Ellington JE, Ball BA, Yang X.The objective of this study was to determine whether coculture of stallion spermatozoa and mare oviductal (uterine tubal) epithelial cells induced sperm cell capacitation in vitro. Capacitation as determined by zona binding and chlortetracycline staining of the sperm cells was compared for stallion spermatozoa: (1) incubated with medium alone (negative control), (2) treated with calcium ionophore A23187 (positive control) or (3) cultured with mare oviductal epithelial cells (OEC) for 4 h. Chlortetracycline staining patterns of sperm cells bound to the zonae were used to group spermatozoa as un...
Massanyi L, Janisch R.According to the distribution of IMP, three different regions can be recognized on PF of the post-acrosomal plasma membrane of bull, ram, and boar spermatozoa. They are: (1) a region with linear aggregation of IMP, (2) a region with fewer and scattered IMP, and (3) a region with more numerous IMP. In the last two regions IMPs are randomly distributed or a clustering of certain particles is visible. In stallion spermatozoa the last two areas are undistinguishable. There are evident interspecies differences in the arrangement of linear aggregations of IMP which are characteristic for each specie...
Wöckener A, Colenbrander B.Two experiments were conducted to examine the effects of liquid storage extender and of a modified freezing protocol on motility and morphology parameters of 3-year-old pony stallions. In experiment 1 ejaculates were diluted 1 + 1 (v+v) with glycine-egg-yolk extender (D11) or skim milk extender (SME), centrifuged, resuspended in the corresponding extender and kept at +5 degrees C. Concerning motion characteristics, both progressive motility and average path velocity of semen stored in SME was significantly superior to semen stored in D11 after 6, 18 and 42 hrs. However, over time of storage th...
Casey PJ, Robertson KR, Liu IK, Espinoza SB, Drobnis EZ.Subfertility in stallions is common, and methodologies are needed to increase the fertility in these animals. In other species, removal of the dead sperm from semen increases the quality and fertility of semen. With horse semen we evaluated 48 combinations of column separation techniques using micro-spin chromatography columns. The greatest improvement in motility was observed with glass wool, whereas glass beads exhibited the greatest recovery of motile sperm. Although centrifugation time did not influence recovery rate or percent motility, a column length of 2 cm was superior for recovery of...
Broussard JR, Goodeaux SD, Goodeaux LL, Thibodeaux JK, Moreau JD, Godke RA, Roussel JD.Control extender was incubated at 4 degrees C for 24 hours. Rubber or plastic syringe plungers were separately incubated in semen extender for 24 hours at 4 degrees C. Following incubation, the extender was stored at -20 degrees C until the time of semen collection. The treatments consisted of the following: Group A = equine semen plus control extender; Group B=equine semen plus extender incubated with rubber plungers and Group C=equine semen plus extender incubated in plastic plungers; Group D=equine semen plus control extended in rubber plunger syringes and Group E=equine semen plus control ...
White IG.When sperm of the ram, bull, boar and stallion are cold-shocked by rapid cooling to near freezing point, motility and metabolic activity are irreversibly depressed and the acrosome and plasma membrane disrupted. Ram sperm become susceptible to cold shock in the proximal corpus region of the epididymis when the cytoplasmic droplet has moved backwards to the distal portion of the sperm midpiece. The membrane constituents phospholipids and cholesterol are important in cold shock which causes loss of lipid from sperm. The susceptibility of sperm to cold shock is linked with a high ratio of unsatur...
Held JP, McCracken MD, Toal R, Latimer F.Aspermia was diagnosed in a 12-year-old Thoroughbred stallion with generalized lymphosarcoma. Invasion of the epididymus by neoplastic cells caused thickening and enlargement of both epididymes. The testes were not affected. The nodular ultrasonographic architecture was similar to that in previously reported cases of infectious epididymitis.
Farlin ME, Jasko DJ, Graham JK, Squires EL.The use of fluorescein-conjugated Pisum sativum agglutinin (FITC-PSA) was evaluated for its ability to distinguish acrosome-intact from acrosome-damaged stallion spermatozoa. Incubation of fresh (acrosome-intact) and frozen-thawed (acrosome-damaged) spermatozoa with FITC-PSA resulted in acrosome-intact spermatozoa that exhibited no fluorescence, while acrosome-damaged spermatozoa exhibited fluorescent staining over the rostral portion of the head and equatorial segment. Experiments using mixtures of various ratios of acrosome-intact and acrosome-damaged spermatozoa determined the precision (in...
Love CC.Ultrasonographic examination of the testis, epididymis, and spermatic cord of the stallion can be used to enhance the routine breeding soundness evaluation of the stallion. Normal ultrasonographic anatomy of the testes and associated structures are presented to aid the clinician in differentiating abnormalities of these structures.
Jasko DJ, Little TV, Lein DH, Foote RH.Information pertaining to evaluation of single ejaculates of semen and records for 2 consecutive breeding seasons were obtained. In all, data for 99 individual breeding seasons (n = 43 Standardbreds and 56 Thoroughbreds) were evaluated. Included in each semen evaluation was examination of semen characteristics and computer-aided analysis of spermatozoal movement characteristics. On the basis of the analysis of breeding records for 4,175 mares (7,017 estrous cycles), a per-estrous cycle fertility rate was calculated from data for 96 of the breeding seasons. Stallions with lower fertility than t...
Parks JE, Lynch DV.Composition and thermotropic phase behavior of sperm membrane lipids from species ranging in sensitivity to cold shock were determined. Lipids from whole sperm and sperm plasma membrane were fractionated into neutral lipid, glycolipid, and phospholipid fractions. Compositional analyses were completed for free sterols, phospholipids and phospholipid-bound fatty acids. Phase transition temperatures were determined for phospholipid and glycolipid fractions using differential scanning calorimetry. Cholesterol was the major sterol in sperm lipids of all species. Cholesterol to phospholipid molar ra...
Bittmar A, Kosiniak K.Experiments conducted on the freezability of 400 ejaculates collected from 64 stallions demonstrate the possibility of predicting the semen's ability to withstand the freezing/thawing process. If the sperm concentration, AspAT activity and total protein content in the seminal plasma of raw ejaculates are determined before freezing, the effects of freezing may be forecast in about 80% of the ejaculates.
Power MM, Gustavsson I, Switoński M, Plöen L.Synaptonemal complex analysis by electron microscopy of a trisomy 28 in a male horse demonstrated a trivalent or a bivalent plus a univalent in primary spermatocytes. Two of the chromosomes making up the trivalent were, most often, completely paired with each other and only partially paired or associated with the third one. Half of the spermatocytes analysed demonstrated heterologous pairing or association between the free axis of the trivalent and the sex bivalent. The pairings remained, to a large extent, into diakinesis-metaphase I. In most pachytene cells one autosomal bivalent showed prox...
Liu DY, Lopata A, Pantke P, Baker HW.The present study demonstrates that horse and marmoset monkey sperm can bind to the human zona of salt-stored oocytes that failed to fertilize in vitro. Marmoset monkey sperm are also able to penetrate the salt-stored human zona. In contrast, human sperm do not bind to the zona of either horse or marmoset monkey oocytes. These results suggest that human sperm binding to the zona pellucida is more strictly species-specific than it is for horse and marmoset monkey sperm. In contrast, horse and marmoset monkey sperm contain receptors recognized by the human zona.
Padilla AW, Foote RH.Slow-cooled stallion spermatozoa, with and without seminal plasma removed by centrifugation, were diluted in Kenney's extender (KE) containing nonfat dry skim milk with glucose and antibiotics or in KE supplemented by adding a modified high-potassium Tyrode's medium (KMT). Four ejaculates from each of four stallions were collected and divided factorially across these four treatments. Percentage of motile sperm, path velocity, and linearity immediately after treatment (0 h) and after storage at 4 degrees C for 24, 48, and 72 h were evaluated objectively by use of a HTM-2030 sperm motility analy...
Janett F, Aebi L, Burger D, Imboden I, Hässig M, Kindahl H, Thun R.The objective of this study was to evaluate the effect of the non-steroidal anti-inflammatory drug (NSAID) vedaprofen (Quadrisol) on quality and freezability of stallion semen. Experiments were performed using 22 Franches Montagnes stallions from the National Stud in Avenches (Switzerland) randomly divided into a control and test group. Vedaprofen was given orally to all stallions of the test group at the recommended therapeutic dose (initial dose of 2mg/kg followed by 1mg/kg body weight every 12h) for 14 days. Control animals received the same amount of carrier substance. During treatment, bl...
Scarlet D, Budik S, Aurich C.A new device for storage and shipping of cell cultures--the Petaka G3 cell management device--was tested for its applicability for cooled-storage of equine semen. Semen from three stallions was processed with EquiPro extender either without antibiotics (three ejaculates per stallion) or with gentamicin (250 mg/l; three ejaculates per stallion). Semen was either stored at five (anaerobic conditions) or 15 °C (aerobic conditions) in syringes or cell culture devices. Total and progressive motility, as well as membrane integrity of spermatozoa, were evaluated from days 1 to 7 after collection wit...
Najjar A, Ben Said S, Benaoun B, Chetoui C, Ezzaouia M, Ben Mrad M.The study was undertaken in order to evaluate sperm morphology features of post-thawed semen of Tunisian Arab stallions. Forty two ejaculates was collected and frozen, during years 2009 and 2010, from 9 stallions aged between 9 to 24 years. After thawing, sperm morphology was studied after eosin-nigrosin stain. The percentages of abnormal head, mid piece, flagella, sperm with droplets and the total abnormal sperm were determined. Analysis of variance was carried out using SAS software. The results showed that all sperm morphology features varied among ejaculates within stallion and among stall...
Serafini R, Ghosh S, Love CC, Medrano JMR, Teague SR, LaCaze KA, Varner DD.Commercially available vaginal lubricants, typically labeled as non-spermicidal, are used to lubricate equine artificial vaginas prior to semen collection. Improper type or amount of lubricant might affect stallion sperm quality, either after short-time exposure or following cooled storage of extended semen previously exposed to lubricant. The aim of this study was to evaluate stallion sperm quality following exposure to lubricant-containing extender for 1 h (T1h) or 24 h (T24h). Three ejaculates were collected from each of four stallions using a small volume of petrolatum to lubricate a...
Minelli A, Moroni M, Trinari D, Mezzasoma I.Ectoenzymic activities capable of hydrolyzing ATP sequentially to adenosine are present on equine epidydimal spermatozoa membranes. Kinetic parameters for ATPase, ADPase and 5'-nucleotidase were obtained by analysis of progress reactions curve when ATP, ADP and AMP were supplied as initial substrates. These values are not different from those found when the substrates were supplied from the preceding reactions. Feed-forward inhibition on 5'-nucleotidase by ATP/ADP was taken into account to fit simulated data to the experimental results. None of the substrates supplied by the preceding reaction...
Mogielnicka-Brzozowska M, Woźniak MJ, Cichowska AW, Fraser L, Kraziński B, Strzezek R, Zielińska D.Seminal plasma (SP) proteins are responsible for sperm functional quality. Developing a reliable method to determine the degree of oxidative damage of these proteins is important for establishing semen fertilizing ability. The main aim of the study was to verify the applicability of protein carbonyl derivatives measurement in the SP of canine and stallion, using a method with 2,4-dinitrophenylhydrazine (DNPH). The research material consisted of ejaculates obtained from eight English Springer Spaniels, and from seven half-blood stallions during the breeding and non-breeding season. The content ...
Teuscher C, Kenney RM, Cummings MR, Catten M.In this study, 2 stallions were immunised with their own spermatozoa to ascertain whether an antisperm autoantibody response could be mounted. The results demonstrated that the stallion can recognise and respond to sperm autoantigens by producing circulating antisperm antibodies, primarily of the IgG class. Such autoantibodies appeared 2-4 weeks after inoculation and persisted for 6-20 weeks. Immunochemical characterisation by western blot identified two major sperm autoantigens, with molecular weights of 70 kD and 62 kD. Control pony stallions immunised with adjuvants alone failed to exhibit ...
Vĕzník Z, Svecová D, Pospísil L, Diblíková I.Frequency of elementary and reticular chlamydial bodies was investigated by direct immunofluorescence tests in ejaculates collected from 52 men, 60 stallions, 42 bulls, and 66 boars using the kits of Progen Biotechnic GmbH and the microscope Labophot-2 Nikon. At the same time, qualitative semen tests, including ejaculate volume, sperm motility, percentage of live and dead sperms and morphological' analyses (Vĕzník and Svecová, 1992) were done. Repeatability of the findings was checked in a group of nine bulls housed at the institute and sampled at weekly intervals for 3 to 4 months (Tab. 1)...
Landim-Alvarenga FC, Alvarenga MA, Seidel GE, Squires EL, Graham JK.Experiments evaluated the ability of follicular fluid (FF), dilauroylphosphatidylcholine (PC12) and the calcium ionophore A23187 (A23187) to induce capacitation in stallion and bull spermatozoa, determined by the ability of the spermatozoa to penetrate zona-free hamster, bovine and equine oocytes. Spermatozoa suspensions were incubated at 37 degrees C in one of the following treatments: 1) a modified Tyrode's medium (BGM3) alone; 2) BGM3 + FF; 3) BGM3 + PC12; 4) BGM3 + FF + PC12; 5) BGM3 + A23187; and 6) BGM3 + FF + A23187. Treated spermatozoa were incubated with zona-free hamster, bovine and ...
Leopold S, Samper JC, Curtis E, Buhr MM.Movement of Ca2+ into spermatozoa is a critically important event for capacitation and the acrosome reaction. In the present study, the nature of Ca2+ movement in fresh equine spermatozoa was established and the effects of oviductal cell conditioned medium (OCM) and cryopreservation on Ca2+ flux were investigated. The ability of fresh and cryopreserved stallion spermatozoa to regulate Ca2+ concentration over time was evaluated in Ca2+ -free PBS. Intracellular Ca2+ concentrations were higher in cryopreserved spermatozoa than in fresh spermatozoa. However, extracellular Ca2+ concentrations were ...
Zhang JJ, Muzs LZ, Boyle MS.Three experiments were conducted to assess the structural and functional changes of stallion spermatozoa in response to the calcium ionophore A23187, and to determine individual variation between stallions. In Experiment 1, changes in the acrosome of spermatozoa exposed to 7.14 microM A23187 for fixed times between 0 and 120 min were examined. There was a steady increase with time in the number of spermatozoa undergoing the acrosome reaction although the rate of increase differed between stallions. Sperm motility decreased sharply when incubation was extended beyond 30 min. In Experiment 2, th...
McKinnon AO, Lacham-Kaplan O, Trounson AO.The use of intracytoplasmic sperm injection (ICSI) for in vitro fertilization of equine oocytes and the developmental potential of these oocytes after transfer to the Fallopian tubes of synchronized mares were examined. Oocytes were aspirated from mature follicles 39 h after injection of a GnRH analogue and transported 190 km at 39 degrees C. Semen from a fertile and an infertile stallion was frozen and prepared for injection. Successfully injected oocytes were transferred surgically into the ampulla of the Fallopian tube either: (i) 4-8 h after semen injection; or (ii) after 24-48 h culture b...
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...
Gil L, Galindo-Cardiel I, Malo C, González N, Alvarez C.Cholesterol and Equex-STM are frequently added to different commercial and experimental extenders improving postthawing sperm quality. Doses of 125-150 mM of cholesterol from pig liver and 0.5-0.7% of Equex-STM were evaluated in a standard eggyolk extender (Martin et al., 1979). Six ejaculates per stallion from six pure Spanish stallions (6-8 years old) were collected in Martin's extender (B) and different mixtures of 125 mM-0.5% (I), 125 mM-0.7% (II), 150 mM-0.5% (III), and 150 mM-0.7% (IV) were added to original Martin's extender. Samples were frozen in 0.5 mL straws (100 × 10(6...
Thomas PG, Ball BA.To facilitate the study of interactions between equine spermatozoa and homologous oviduct epithelial cells, we developed an assay to count labelled spermatozoa bound to oviduct epithelial cell (OEC) monolayers and used the assay to compare the binding ability of spermatozoa from different stallions. Washed spermatozoa from three stallions were incubated with the fluorochrome Hoechst 33342 (5 micrograms/ml) for 1 min. Spermatozoa were then layered over confluent monolayers of oviduct epithelial cells in 2 cm2 culture wells. Coculture treatments comprised five concentrations of spermatozoa (10(5...
Spirito S, Campi S, Boquet M, Fernández H, Ferrari M.The aims of this study were to estimate the ability of the Feulgen reaction to identify equine sperm nuclei with different ploidy levels, to determine the frequency of haploid, diploid and polyploid sperm nuclei in the semen of fertile equines and to evaluate the relationship between the nuclear DNA content and the nuclear area. Determination of the ploidy level of Feulgen-stained spermatozoa using a scanning microspectrophotometer was very similar to the subjective estimations made with a light microscope. This indicates that the Feulgen reaction is a simple, inexpensive and reliable techniqu...
Ecot P, Vidament M, de Mornac A, Perigault K, Clément F, Palmer E.In the present study, the interactions among stallions, semen extenders and cooling treatments before stallion semen samples were frozen were studied. In Expt 1, the effects of four cooling treatments and three semen extenders were investigated (11 stallions x four split ejaculates), whereas in Expt 2, the effects of two semen extenders, two egg yolk concentrations and two glycerol concentrations were investigated (six stallions x five split ejaculates). Sperm motility after thawing was evaluated. In Expt 1, the extender x cooling treatment interaction was significant. Centrifugation and addit...
Hatami M, Qasemi-Panahi B, Daghigh Kia H, Moghaddam G, Janmohammadi H.Equine semen cryopreservation is one of the major procedures for the genetic conservation of rare and endangered genotypes. The current study was conducted to evaluate the effect of egg yolk plasma (EYP) enriched with β-carotene as an antioxidant supplement on INRA-96 extender regarding freezing Arabic stallion sperm. For this purpose, β-carotene various concentrations were utilized as a supplementary ingredient in formulating the diets of laying hens. Birds were randomly divided into four groups, fed with 0 (control), 500, 1000 and 2000 mg/kg in a supplemented diet with β-carotene. Subse...
Jakop U, Hensel B, Czirják GÁ, Quirino M, Schröter F, Jung M, Schulze M.During insemination, bacterial contamination of the ejaculate can lead to reduced sperm quality and transmission of pathogens to the female, thus should be avoided. The semen of a variety of animal taxa possess antimicrobial properties against a wide range of bacterial species through antimicrobial molecules, such as lysozyme, but their variance and the factors influencing it are unknown for most species. In this study, the antibacterial defence (bacterial killing activity (BKA) against Escherichia (E.) coli and Staphylococcus (S.) aureus as well as lysozyme concentration) was studied in semin...
Lindsey AC, Bruemmer JE, Squires EL.Mares are generally inseminated with 500 million progressively motile fresh sperm and approximately 1 billion total sperms that have been cooled or frozen. Development of techniques for low dose insemination would allow one to increase the number of mares that could be bred, utilize stallions with poor semen quality, extend the use of frozen semen, breed mares with sexed semen and perhaps reduce the incidence of post-breeding endometritis. Three low dose insemination techniques that have been reported include: surgical oviductal insemination, deep uterine insemination and hysteroscopic insemin...
Torres-Boggino F, Sato K, Oka A, Kanno Y, Hochi S, Oguri N, Braun J.Seminal characteristics, fertility and the response to semen preservation (liquid storage and cryopreservation) were evaluated in 4 Draft stallions (Percheron 2, Breton 2). Seminal characteristics (gel-free volume, sperm concentration, sperm morphology, percentage of motile spermatozoa) were assessed in 5 ejaculates from each of the 4 stallions. The fertility of the stallions was calculated retrospectively as the accumulated pregnancy rate over 3 breeding seasons. Five ejaculates from each of the stallions were subjected to liquid storage at 5 degrees C. The percentage of motile spermatozoa (P...
Ferrer MS, Canisso IF, Podico G, Ellerbrock RE, Hurley DJ, Palomares R.Different stallions exhibit a high level of variation in the ability of their sperm to survive cryopreservation. A large fraction of stallions show poor post-thaw sperm motility, and their semen is not suitable for commercial freezing. In this study, we hypothesized that the presence of sperm-bound antisperm antibodies (ASAs) was associated with poor cryosurvival of stallion sperm. Our objective was to assess the level of ASA binding to stallion sperm, and determine if it was associated with good or poor sperm cryosurvival. In Experiment 1, cooled shipped semen from 27 stallions was frozen usi...
Braun J, Torres-Boggino F, Hochi S, Oguri N.The objective of this experiment was to examine the effect of seminal plasma on motion characteristics of epididymal and ejaculated equine spermatozoa during storage at 5 degrees C. Epididymal spermatozoa were flushed with either seminal plasma or a skim milk-glucose extender. Ejaculated spermatozoa were collected with extender added 10 minutes after semen collection and addition of extender during ejaculation by placing 50 ml extender in the collection bottle. Semen samples were centrifuged and resuspended with a skim milk-glucose extender containing seminal plasma (0, 5 and 25%; v/v), prepar...
Frazer GS, Bucci DM.The aims of this project were to document the protein profile of equine seminal plasma and determine the variability between stallions in the relative composition of proteins in the ejaculate. A single ejaculate was obtained from 14 stallions of varying breed and age. The gel fraction was removed by an in-line filter. The semen was centrifuged and the supernatant seminal plasma aspirated without disturbing the sperm pellet. The seminal plasma was recentrifuged and stored in cryovials at -70 degrees C. Samples were thawed, recentrifuged, assayed for protein concentration (BCA protein assay), di...
Bedford SJ, McDonnell SM, Tulleners E, King D, Habecker P.A 14-year-old Arabian stallion was examined because of acute hemospermia. The stallion was used in an artificial breeding program and had a 6-year history of low-grade hemospermia and a 4-year history of self-mutilation behavior. During previous examinations, minor irritation of the urethral process was identified as the source of the bleeding. Physical examination revealed a mucosal ulceration in the distal portion of the urethra. Histologic examination of a biopsy specimen from this area revealed low-grade squamous cell carcinoma. The urethral process was excised, and the hemospermia resolve...
Gloria A, Carluccio A, Petrizzi L, Noto F, Contri A.Equine spermatozoa from the cauda epididymis were previously collected and frozen, and the fertility was assessed. Most studies were performed on healthy stallions that had undergone routine castration or on the epididymis collected at the abattoir, but there are no studies on the quality of epididymal semen in subjects which have died from colic or which underwent intensive care. The present study was designed to verify whether a severe illness could affect epididymal semen quality and freezability in the stallion. Therefore, epididymal semen characteristics during the freezing process in sta...
Evenson DP, Jost LK, Varner DD.Data from the sperm chromatin structure assay (SCSA), a flow cytometric measurement of susceptibility of sperm nuclear DNA to denaturation, show strong correlation with the fertility potential of bulls, boars, men and stallions. Previous studies showed a strong relationship between stallion spermatozoa with denatured DNA and the presence of DNA strand breaks. In the present study, the relationship between stallion sperm DNA denaturation and the redox status of -SH groups on the cysteine residues of sperm nuclear protamines that are thought to stabilize chromatin was investigated. Semen samples...
Wessel MT, Ball BA.Osmotic stress is an important component of the damage to spermatozoa during cryopreservation. Osmotic injury, due to hyperosmolar freezing extenders, changes in relative solute concentration in the extra cellular medium during freezing and differences in the relative permeabilities of penetrating cryoprotectants, such as glycerol, and water occur when cryopreserved spermatozoa are diluted into isosmotic media or when spermatozoa are placed in the female reproductive tract. The purpose of the study reported here was to evaluate the effect of step-wise dilution for the removal of the permeating...
Morris LH.The generally recommended minimum number of spermatozoa required for conventional artificial insemination in the mare is in excess of 200 x 10(6) progressively motile spermatozoa. Recent developments in different insemination techniques such as deep uterine, hysteroscopic and oviductal insemination, which have been designed to use significantly fewer spermatozoa, are reviewed in this paper. A number of studies have demonstrated that ultrasound guided deep uterine insemination of 5 x 10(6) fresh spermatozoa can produce satisfactory pregnancy rates. The use of hysteroscopic insemination enables ...
Clément F, Vincent P, Mahla R, Meriaux JC, Palmer E.The aim of the present study was to determine which artificial insemination results in fertilization when mares are inseminated several times before ovulation. Mares in oestrus were inseminated over 62 cycles with fresh semen at 48 h intervals from when a follicle > or =30 mm in diameter was detected until ovulation. The number of inseminations was limited to three. Three fertile stallions were used and a different stallion was used for each artificial insemination. The order of the three stallions was changed for each cycle. Embryos were collected between day 10 and day 12 after ovulation and...
