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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.
Biochemical components of seminal plasma and their correlation to the fresh seminal characteristics in Marwari stallions and Poitou jacks.
Veterinary world    February 18, 2017   Volume 10, Issue 2 214-220 doi: 10.14202/vetworld.2017.214-220
Talluri TR, Mal G, Ravi SK.To investigate various biochemical components of seminal plasma in Marwari stallions and Poitou Jacks and to find out their correlation with that of the seminal characteristics. Methods: In this study, semen was collected from six Marwari stallions and six Poitou jacks aged from 4 to 6 years and with known fertility status. The semen collection from the stallions were collected during the breeding season, i.e., between the months of April and June. From the collected semen ejaculates, we estimated the values of some biochemical components, viz., total protein content, total lipid content, and ...
Testosterone serum profile, semen characteristics and testicular biometry of Mangalarga Marchador stallions in a tropical environment.
Reproduction in domestic animals = Zuchthygiene    January 23, 2017   Volume 52, Issue 2 335-343 doi: 10.1111/rda.12918
Waddington B, Penitente-Filho JM, Neves J, Pinho RO, Chaya AY, Maitan PP, Silveira CO, Neves MG, Guimarães S, de Carvalho GR, Guimarães JD.This study was conducted to characterize the daily profile of testosterone secretion and its mean concentrations in the four seasons as well as to evaluate the semen characteristics and testicular biometry of Mangalarga Marchador stallions throughout the year in a tropical region. Three stallions were submitted to semen collections and evaluation of testicular biometry every 14 days along a year. Blood samples were collected once at the middle of each season, in a 20-min interval during 24 hr in order to evaluate the testosterone secretion profiles among seasons. Testosterone concentrations ...
Epigallocatechin-3-gallate (EGCG) and green tea polyphenols do not improve stallion semen parameters during cooling at 4°C.
Reproduction in domestic animals = Zuchthygiene    January 6, 2017   Volume 52, Issue 2 270-277 doi: 10.1111/rda.12891
Bucci D, Spinaci M, Mislei B, Gadani B, Rizzato G, Love CC, Tamanini C, Galeati G, Mari G.Stallion semen storage for artificial insemination is mainly based on liquid cooled storage. In many stallions this technique maintains sperm quality for an extended period of time (24-72 hr) at 7°C. While this technique is commonly used in the horse industry, there can be a decline in fertility in some stallions, due to an inability of their sperm to tolerate the cool storage process. The aim of the present work was to evaluate the effect of two natural antioxidants (epigallocatechin-3-gallate (EGCG) at 20, 60 and 120 μm and green tea polyphenols, and p at .001, .01 and .1 mg/ml) on some...
Modern Techniques for Semen Evaluation.
The Veterinary clinics of North America. Equine practice    November 5, 2016   Volume 32, Issue 3 531-546 doi: 10.1016/j.cveq.2016.07.006
Love CC.Stallion semen evaluation is an important part of the breeding soundness evaluation. The results of the semen evaluation cannot be interpreted without a thorough knowledge of the mare and management effects that may have played a role or may affect the potential fertility of the stallion evaluated. There are considerations and limitations that the clinician should understand about each test. Any sperm quality test must be interpreted with a clear understanding of how it relates to fertility.
The presence of seminal plasma, especially derived from stallion semen, helps preserve chilled Asian elephant (Elephas maximus) sperm motility.
Andrologia    October 26, 2016   Volume 49, Issue 6 doi: 10.1111/and.12690
Pinyopummin A, Mahasawangkul S, Kornkaewrat K, Rattanapirom S, Leartsang W, Kitkha S.The effects of seminal plasma (SP), derived from autologous, homologous and heterologous species (stallion, boar and dog) on chilled Asian elephant sperm quality, were determined. Semen was collected from eight males and samples with ≥30% motile spermatozoa were used in the study. Semen was diluted with Tris-glucose-egg yolk extender, supplemented with different SP types and preserved at 4°C for 48 hr. Experiment 1 (n = 31), showed that the presence of SP (autologous) helped to preserve sperm quality in terms of sperm motility and acrosome integrity (p < .05). Homologous SP did not ...
Strategies for Processing Semen from Subfertile Stallions for Cooled Transport.
The Veterinary clinics of North America. Equine practice    October 7, 2016   Volume 32, Issue 3 547-560 doi: 10.1016/j.cveq.2016.07.007
Varner DD.Subfertility can be a confusing term because some semen of good quality can have reduced fertility following cooled transport if the semen is processed in an improper manner. General procedures aimed at processing stallion semen for cooled transport are well described. An array of factors could exist in reduced fertility of cool-transported semen. This article focuses on centrifugation techniques that can be used to maximize sperm quality of stallions whose semen is intended for cooled transport. Clinical cases are also provided for practical application of techniques.
Rosiglitazone Improves Stallion Sperm Motility, ATP Content, and Mitochondrial Function.
Biology of reproduction    September 28, 2016   Volume 95, Issue 5 107 doi: 10.1095/biolreprod.116.142687
Swegen A, Lambourne SR, Aitken RJ, Gibb Z.Media used for equine sperm storage often contain relatively high concentrations of glucose, even though stallion spermatozoa preferentially utilize oxidative phosphorylation (OXPHOS) over glycolysis to generate ATP and support motility. Rosiglitazone is an antidiabetic compound that enhances metabolic flexibility and glucose utilization in various cell types, but its effects on sperm metabolism are unknown. This study investigated the effects of rosiglitazone on stallion sperm function in vitro, along with the possible role of AMP-activated protein kinase (AMPK) in mediating these effects. Sp...
Equivalent seminal characteristics in human and stallion at first and second ejaculated fractions.
Andrologia    September 23, 2016   Volume 49, Issue 8 doi: 10.1111/and.12708
de la Torre J, Sánchez-Martín P, Gosálvez J, Crespo F.Sperm quality was assessed in normozoospermic human (n = 10) and Spanish breed stallion (n = 10) after sperm fractionation during ejaculation. The first ejaculated fraction was separated from the second. A third sample was reconstituted using equivalent proportion of both fractions (RAW). Fraction 1, Fraction 2 and RAW semen were incubated for 30 min at 37°C to homogenise the impact of iatrogenic damage between both species. Sperm concentration, motility and sperm DNA damage were assessed in each fraction and RAW semen. The results showed two important facts: (i) spermatozoa confined at ...
Reversible downregulation of the hypothalamic-pituitary-gonadal axis in stallions with a novel GnRH antagonist.
Theriogenology    July 26, 2016   Volume 86, Issue 9 2272-2280 doi: 10.1016/j.theriogenology.2016.07.021
Davolli GM, Ball BA, Esteller-Vico A, Claes AN, Canisso IF, Fedorka CE, Woodward EM, Troedsson MH, Squires EL.The GnRH antagonist, acyline, has not yet been investigated in the stallion. Our study aimed to: (1) evaluate the downregulation of the stallion hypothalamic-pituitary-gonadal axis by acyline through assessment of seminal parameters, testicular volume, and sexual behavior; (2) assess hormonal response of acyline-treated stallions to GnRH stimulation; and (3) verify reversibility after treatment. Stallions were assessed pretreatment and subsequently treated (every five days) for 50 days: acyline (n = 4; 330 μg/kg acyline) or control (n = 4, vehicle). The stallions were then monitored for 62Â...
Evaluation of the breeding behavior of Brazilian Pony stallions.
Animal reproduction science    July 21, 2016   Volume 172 137-142 doi: 10.1016/j.anireprosci.2016.07.011
Rua MA, Quirino CR, Bartholazzi Junior A, Santoro PN, Bastos R, Ribeiro Mda S, Matos LF, Vega WH.The aim of this study was to evaluate the breeding behavior of Brazilian Pony stallions during semen collection to examine the effect of age category (AC) and month of evaluation on the characteristics of breeding behavior. Nine stallions were evaluated during the breeding and non-breeding season, 5-8 (AC1) and 9-13 years (AC2) of age. The variables evaluated were reaction time (RT), mount time one (MT1) and two (MT2), mount and ejaculation time (MEjT), mounting without erection (MWEr), mounting without ejaculation (MWEj), flehmen, kicks, bites, smells, vocalizations and serum testosterone con...
Mitochondrial oxygen consumption is a unique indicator of stallion spermatozoal health and varies with cryopreservation media.
Theriogenology    May 6, 2016   Volume 86, Issue 5 1382-1392 doi: 10.1016/j.theriogenology.2016.04.082
Darr CR, Cortopassi GA, Datta S, Varner DD, Meyers SA.Mitochondrial oxygen consumption is a sensitive indicator of spermatozoal health in the context of cryopreservation. We investigated oxygen consumption of equine sperm mitochondria during incubation in four commercially available sperm cryopreservation extenders: modified INRA 96, BotuCrio, EZ Freezin-"LE" and "MFR5", in addition to several other parameters including motility, reactive oxygen species (ROS) production and viability. All experimental endpoints, with the exception of average path velocity, were affected significantly by freezing extender type after freezing and thawing. Sperm in ...
The effects of urine concentration, and cushion centrifugation to remove urine, on the quality of cool-stored stallion sperm.
Theriogenology    April 29, 2016   Volume 86, Issue 5 1294-1298 doi: 10.1016/j.theriogenology.2016.04.070
Voge J, Varner DD, Blanchard TL, Meschini M, Turner C, Teague SR, Brinsko SP, Love CC.Urine-contaminated stallion semen is a clinical problem due to a variety of causes. The effect of the level of urine contamination on the longevity of sperm quality has not been evaluated. The aim of this study was to determine the effects of urine concentration level (0%, 10%, 20%, 30%, and 40%) and cushioned centrifugation and resuspension of the sperm pellet in fresh extender, on measures of sperm quality, immediately after semen collection (T0), after 1 hour of storage at room temperature (T1), and after 24 hours of cooled storage (T24). In general, most sperm quality measures declined w...
New flow cytometry approaches in equine andrology.
Theriogenology    April 21, 2016   Volume 86, Issue 1 366-372 doi: 10.1016/j.theriogenology.2016.04.050
Peña FJ, Ortega Ferrusola C, Martín Muñoz P.Flow cytometry is currently recognized as a robust tool for the evaluation of sperm quality and function. However, within equine reproduction, this technique has not reached the sophistication of other areas of biology and medicine. In recent years, more sophisticated flow cytometers have been introduced in andrology laboratories, and the number of tests that can be potentially used in the evaluation of sperm physiology has increased accordingly. In this review, recent advances in the evaluation of stallion spermatozoa will be discussed. These new techniques in flow cytometry are able to simul...
Aldehyde Dehydrogenase Plays a Pivotal Role in the Maintenance of Stallion Sperm Motility.
Biology of reproduction    April 20, 2016   Volume 94, Issue 6 133 doi: 10.1095/biolreprod.116.140509
Gibb Z, Lambourne SR, Curry BJ, Hall SE, Aitken RJ.Although stallion spermatozoa produce significant quantities of reactive oxygen species, a lag between 4-hydroxynonenal (4HNE) adduction and the loss of motility in stallion spermatozoa suggests the presence of a robust aldehyde detoxification mechanism. Because there is a paucity of studies characterizing the role of aldehyde dehydrogenase (ALDH) in sperm functionality, the aim of this study was to ascertain the relationship between 4HNE production and motility and ALDH expression by stallion spermatozoa. PCR analysis revealed the presence of the ALDH1A3, ALDH1B1, and ALDH2 isoforms in these ...
Effect of various concentrations of butylated hydroxyanisole and butylated hydroxytoluene on freezing capacity of Turkman stallion sperm.
Animal reproduction science    April 19, 2016   Volume 170 108-113 doi: 10.1016/j.anireprosci.2016.04.010
Seifi-Jamadi A, Kohram H, Zareh-Shahne A, Dehghanizadeh P, Ahmad E.The present study aimed to determine the effect of different concentrations of butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) on post-thaw stallion sperm quality. The ejaculates collected from four healthy mature Turkmen stallions were pooled and divided into eight aliquots. The samples were diluted with extenders containing different concentrations (0.5, 1 or 2mM/mL) of BHA or BHT. The positive control (PC) samples were diluted with extender containing 0.5% ethanol (v/v) whereas; the negative control (NC) samples were diluted with basic extender only. Semen samples were fro...
Lipidomics of equine sperm and seminal plasma: Identification of amphiphilic (O-acyl)-ω-hydroxy-fatty acids.
Theriogenology    April 16, 2016   Volume 86, Issue 5 1212-1221 doi: 10.1016/j.theriogenology.2016.04.012
Wood PL, Scoggin K, Ball BA, Troedsson MH, Squires EL.Using a nontargeted lipidomics analysis of equine sperm and seminal plasma, we were able to characterize a diverse array of individual lipids including ethanolamine and choline ether lipids and seminolipids essential to membrane raft function. We also detected, for the first time in sperm, the presence of (O-acyl)-ω-hydroxy-fatty acids (OAHFA) with up to 52 carbon chain lengths, which were localized to the head and not the tail of sperm. The only previous identification of OAHFAs has been in meibomian glands and their sebaceous secretions. The identities of these lipid amphiphiles were valida...
Development of a new fertility prediction model for stallion semen, including flow cytometry.
Theriogenology    April 11, 2016   Volume 86, Issue 4 1111-1131 doi: 10.1016/j.theriogenology.2016.04.001
Barrier Battut I, Kempfer A, Becker J, Lebailly L, Camugli S, Chevrier L.Several laboratories routinely use flow cytometry to evaluate stallion semen quality. However, objective and practical tools for the on-field interpretation of data concerning fertilizing potential are scarce. A panel of nine tests, evaluating a large number of compartments or functions of the spermatozoa: motility, morphology, viability, mitochondrial activity, oxidation level, acrosome integrity, DNA integrity, "organization" of the plasma membrane, and hypoosmotic resistance, was applied to a population of 43 stallions, 33 of which showing widely differing fertilities (19%-84% pregnancy rat...
Effects of α-tocopherol and freezing rates on the quality and heterologous in vitro fertilization capacity of stallion sperm after cryopreservation.
Theriogenology    March 19, 2016   Volume 86, Issue 4 957-962 doi: 10.1016/j.theriogenology.2016.03.019
de Vasconcelos Franco JS, Faheem M, Chaveiro A, Moreira da Silva F.The effects of supplementation of α-tocopherol and different freezing rates (FRs) on the ability of stallion sperm to fertilize bovine oocytes with intact zona pellucida were investigated, in an attempt to develop a model to assess cryopreserved sperm function. Semen was obtained from four purebred Lusitano stallions (n = 4). Each ejaculate was subjected to cryopreservation with a commercial extender (Ghent, Minitub Iberia, Spain), without any supplementation (control) or supplemented with 2-mM α-tocopherol. The semen was exposed to two different FRs between 5 °C and -15 °C: slow (5 °C...
The autophagy-related protein LC3 is processed in stallion spermatozoa during short-and long-term storage and the related stressful conditions.
Animal : an international journal of animal bioscience    March 2, 2016   Volume 10, Issue 7 1182-1191 doi: 10.1017/S1751731116000240
Aparicio IM, Martin Muñoz P, Salido GM, Peña FJ, Tapia JA.Use of cooled and frozen semen is becoming increasingly prevalent in the equine industry. However, these procedures cause harmful effects in the sperm cell resulting in reduced cell lifespan and fertility rates. Apoptosis and necrosis-related events are increased during semen cryopreservation. However, a third type of cell death, named autophagy, has not been studied during equine semen storage. Light chain (LC)3 protein is a key component of the autophagy pathway. Under autophagy activation, LC3-I is lipidated and converted to LC3-II. The ratio of LC3-II/LC3-I is widely used as a marker of au...
Comparison of cushioned centrifugation and SpermFilter filtration on longevity and morphology of cooled-stored equine semen.
The Veterinary record    February 23, 2016   Volume 178, Issue 10 241 doi: 10.1136/vr.103607
Roach J, Schnobrich M, Ellerbrock R, Feijo L, Bradecamp E, Alvarenga MA, Kline K, Canisso I.This study compares two methods for seminal plasma removal by evaluating sperm recovery rates, and motility and morphology of cooled-stored semen. Ejaculates were divided into three groups: control, filtration and cushioned centrifugation. Semen was extended to 25 million sperm/ml using a skim-milk-based extender and stored at 5°C for all groups. Sperm motility (total motility (%TM) and progressive motility (%PM)) was determined at 0, 24, 48 and 72 hours by a computer-assisted sperm analyser. Sperm morphology was assessed using differential interference microscopy. Overall, %TM of the cen...
Effects of a long-day light programme on the motility and membrane integrity of cooled-stored and cyropreserved semen in Shetland pony stallions.
Animal reproduction science    February 8, 2016   Volume 167 68-73 doi: 10.1016/j.anireprosci.2016.02.008
Deichsel K, Schrammel N, Aurich J, Aurich C.Increasing day length in spring stimulates reproductive functions in horses. In this study, we have analysed the effect of artificial long days on the quality of cooled-stored and cryopreserved semen in Shetland stallions. Stallions of the treatment group (AL, n = 8) were exposed to 16 h light and 8h darkness from 15th December to 20th March while control stallions (CON, n = 7) were kept under natural photoperiod. Semen was collected once weekly and processed for cooled-storage and cryopreservation once per month. Total and progressive motility and percentage of membrane intact spermatozoa wer...
Effect of clinically-related factors on in vitro blastocyst development after equine ICSI.
Theriogenology    December 28, 2015   Volume 85, Issue 7 1289-1296 doi: 10.1016/j.theriogenology.2015.12.015
Choi YH, Velez IC, Macías-García B, Riera FL, Ballard CS, Hinrichs K.Equine intracytoplasmic sperm injection (ICSI) is being used clinically for foal production, but little information is available on factors affecting the efficiency of this procedure. We examined factors that may influence blastocyst development when ICSI is performed clinically, i.e., on oocytes recovered from live mares by transvaginal ultrasound-guided follicle aspiration (TVA), using sperm from the stallion of the client's choice. In a clinical setting, there may be a delay from the time of TVA to isolation of oocytes from the aspirated fluid. In a preliminary study, oocytes from fluid hel...
Analysis of breed effects on semen traits in light horse, warmblood, and draught horse breeds.
Theriogenology    December 10, 2015   Volume 85, Issue 8 1375-1381 doi: 10.1016/j.theriogenology.2015.11.030
Gottschalk M, Sieme H, Martinsson G, Distl O.In the present study, systematic effects on semen quality traits were investigated in 381 stallions representing 22 breeds. All stallions were used for AI either at the Lower Saxon National Stud Celle or the North Rhine-Westphalian National Stud Warendorf. A total of 71,078 fresh semen reports of the years 2001 to 2014 were edited for analysis of gel-free volume, sperm concentration, total number of sperm, progressive motility, and total number of progressively motile sperm. Breed differences were studied for warmblood and light horse breeds of both national studs (model I) and for warmblood b...
The cholesterol transporter ABCA1 is expressed in stallion spermatozoa and reproductive tract tissues.
Theriogenology    November 30, 2015   Volume 85, Issue 6 1080-1089 doi: 10.1016/j.theriogenology.2015.11.019
Merkl M, Ertl R, Handschuh S, Aurich C, Schäfer-Somi S.In the present study, we assessed the presence of the ATP-binding-cassette (ABC) transporter molecules ABCA1 in spermatozoa of adult stallions and in testicular and epididymal tissue of prepubertal and adult stallions. For this purpose, semen samples from six fertile Shetland pony stallions aged 4 to 19 years were collected. Semen was collected from each stallion on three consecutive days. Ejaculates were analyzed immediately after collection, and only ejaculates meeting minimal requirements for fertile stallions were further evaluated. ABCA1 immunosignal was localized after staining of semen ...
Ejaculate Characteristics Depend on Social Environment in the Horse (Equus caballus).
PloS one    November 24, 2015   Volume 10, Issue 11 e0143185 doi: 10.1371/journal.pone.0143185
Burger D, Dolivo G, Wedekind C.Sperm competition theory predicts semen characteristics to be affected by the social environment. We used the polygamous horse (Equus caballus) to experimentally study within-subject plasticity in response to different social environments. Stallions were sequentially exposed, over a period of 8 weeks each, to other stallions and then singly to mares, or vice versa (in adjacent boxes separated by grills). Ejaculates were collected to determine semen characteristics. Highest sperm numbers were found in stallions that were first exposed to other stallions and then to mares, while lowest sperm num...
A long-day light program accelerates seasonal coat changes but is without effect on semen and metabolic parameters in Shetland pony stallions.
Theriogenology    November 12, 2015   Volume 85, Issue 5 946-953 doi: 10.1016/j.theriogenology.2015.11.003
Schrammel N, Deichsel K, Aurich J, Aurich C.Horses are seasonal breeders, and robust breeds may exhibit a winter hypometabolism when kept under semiferal conditions. In this study, we analyzed the effects of artificial long days on rectal temperature, heart rate, heart rate variability, hematology, coat changes, semen parameters, and plasma testosterone concentrations in Shetland stallions stabled overnight and assigned to a control group (CON, n = 9) kept under natural photoperiod, and a treatment group exposed to a long-day light program from 15 December to 20 March (AL, n = 9). During the 8-month study, rectal temperature, heart rate...
Coenzyme Q10 and α-Tocopherol Prevent the Lipid Peroxidation of Cooled Equine Semen.
Reproduction in domestic animals = Zuchthygiene    October 22, 2015   Volume 50, Issue 6 1003-1010 doi: 10.1111/rda.12627
Nogueira BG, Sampaio BF, Souza MI, Costa E Silva EV, Zúccari CE.Biotechnology applied for equine semen increases the levels of reactive oxygen species and reduces the natural antioxidant defence, by both dilution and removal of seminal plasma. Therefore, the aims of this study were to evaluate the effect of adding coenzyme Q10 (CoQ10) and α-tocopherol (α-TOH) to the cooling extender, singly or in combination, on sperm parameters, and their effectiveness in preventing lipid peroxidation (LPO) of equine semen during cooling at 5°C for 72 h. Ten adult stallions of proven fertility were used, using two ejaculates each, subjecting them to the treatments with...
Draft Genome Sequence of Taylorella equigenitalis Strain ERC_G2224 Isolated from the Semen of a Lipizzaner Stallion in South Africa.
Genome announcements    October 15, 2015   Volume 3, Issue 5 doi: 10.1128/genomeA.01205-15
May CE, Schulman ML, Howell PG, Lourens CW, Gouws J, Joone C, Monyai MS, le Grange M, Bezuidt OK, Harper CK, Guthrie AJ.Taylorella equigenitalis is the causative agent of contagious equine metritis (CEM), a sexually transmitted infection of horses. We report here the genome sequence of T. equigenitalis strain ERC_G2224, isolated in 2015 from a semen sample collected in 1996 from a Lipizzaner stallion in South Africa.
Cryopreservation of stallion spermatozoa using different cryoprotectants and combinations of cryoprotectants.
Animal reproduction science    October 3, 2015   Volume 163 75-81 doi: 10.1016/j.anireprosci.2015.09.020
Wu Z, Zheng X, Luo Y, Huo F, Dong H, Zhang G, Yu W, Tian F, He L, Chen J.The present study investigates the effects of five cryoprotectants (CPAs) and cryoprotectant combinations on the post-thaw total motility, progressive motility, viability, mitochondrial membrane potential and acrosome integrity in stallion spermatozoa. In Experiment 1, the objective was to compare the impact of different concentrations (2.5%, 3.5% and 5%) of a single CPA, including glycerol (Gly), ethylene glycol (EG), dimethyl sulphoxide (DMSO), methyl formamide (MF), and dimethylformamide (DMF) for stallion spermatozoa cryopreservation. In Experiment 2, two or more CPAs were used to assess w...
Characteristics of frozen epididymal spermatozoa from stallions that died 12 to 36 hours after colic surgery.
Theriogenology    September 11, 2015   Volume 85, Issue 2 345-350 doi: 10.1016/j.theriogenology.2015.09.006
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...
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