Semen

Distribution by Scientific Domains
Distribution within Medical Sciences

Kinds of Semen

  • fresh semen
  • human semen

  • Terms modified by Semen

  • semen analysis
  • semen characteristic
  • semen collection
  • semen cryopreservation
  • semen donor
  • semen extender
  • semen parameter
  • semen profile
  • semen quality
  • semen sample
  • semen specimen
  • semen volume

  • Selected Abstracts


    Relation between Holstein bulls' proofs for milk in USA and the survival and body weights up to 18 months of their F1 zebu progeny in Venezuela

    JOURNAL OF ANIMAL BREEDING AND GENETICS, Issue 3 2003
    J. R. Pacheco
    Summary The purpose of the study was to determine the relation between Holstein bulls' proofs for milk in USA, and the survival and body weights of their F1 progeny under lowland grazing conditions in Venezuela. Semen was selected from sires with the highest proofs available locally (class H, n = 23) or with proofs close to the breed average (class A, n = 18). Mean proofs were 934 kg (H) and 237 kg (A), respectively, when semen was purchased (1989,93), falling to 204 and ,386 kg when updated in 2001. Dams were Brahman-type zebus on seven farms, raising the F1 progeny (n = 866) on native or cultivated pasture with only mineral supplements. Records of survival to 8 and 18 months and weights at birth, 8 and 18 months of age were analysed using linear models. Sire class had no effect on survival or weight at birth, but H progeny were slightly heavier than A progeny at 8 (4.0%, p = 0.06) and 18 months (2.6%, p < 0.05). The interaction class × farm was only significant for survival, with H progeny equal or superior to A groups on six farms. The effect of sire within class was highly significant for survival to 8 months and weight at 18 months. Rank correlations using sires' milk proofs and F1 progeny weights and survival were not significant. It was concluded that the use of Holstein bulls with high milk proofs may lead to slightly higher 18-month weights in their F1 progeny in tropical grazing systems, covering the additional cost of H semen, but that sire evaluation for weight gain and progeny survival rates may be justified under these conditions. Zusammenfassung Beziehungen zwischen den Zuchtwerten für Milch von Holstein-Bullen in den USA und dem Überleben und dem Körpergewicht bis zu 18 Monaten ihrer F1-Zebu-Nachkommen in Venezuela Der Zweck dieser Studie war die Ermittlung der Beziehungen zwischen den Milch-Zuchtwerten von Holstein-Bullen in den USA und dem Überleben und dem Körpergewicht von F1-Nachkommen unter den Weidebedingungen des Flachlandes in Venezuela. Ausgewählt wurde das Sperma von lokal verfügbaren Bullen mit den höchsten Zuchtwerten (Klasse H, n = 23) oder mit Zuchtwerten, die eng am Populationsdurchschnitt (Klasse A, n = 18) lagen. Beim Erwerb des Spermas (1989,1993) betrugen die mittleren Zuchtwerte 934 kg (H) beziehungsweise 237 kg (A), die bei der Neubewertung 2001 auf 204 kg und ,386 kg abfielen. Die Kühe waren Zebus im Brahman-Typ von sieben Farmen, die die F1-Nachkommen (n = 866) auf natürlichen oder kultivierten Weiden nur mit zusätzlichem Mineralfutter aufzogen. Das Überleben mit acht und 18 Monaten und das Gewicht bei der Geburt, mit acht und mit 18 Monaten wurde mittels linearer Modelle analysiert. Die Bullenklasse zeigte keinen Effekt auf das Überleben und das Geburtsgewicht, aber die H-Nachkommen waren mit acht (4%, p = 0,06) und 18 Monaten (2,6%, p < 0,05) etwas schwerer als die A-Nachkommen. Die Interaktion "Klasse × Farm" war nur für das Überleben signifikant, mit H-Nachkommen gleich oder besser als die A-Gruppen auf sechs Farmen. Der Effekt des Vaters innerhalb der Klasse war hochsignifikant für das Überleben der ersten acht Monate und das Gewicht mit 18 Monaten. Rang-Korrelationen mit den Milch-Zuchtwerten der Bullen und den Gewichten und dem Überleben der F1-Nachkommen waren nicht signifikant. Die Verwendung von Holstein-Bullen mit hohen Zuchtwerten für Milch kann zu einer leichten Erhöhung des Gewicht der F1-Nachkommen mit 18 Monaten in tropischen Weidesystemen führen, welche die zusätzlichen Kosten des H-Spermas abdeckt, aber die Bewertung der Bullen hinsichtlich Gewichtszunahme und Überlebensrate der Nachkommen müsste unter diesen Bedingungen berichtigt werden. [source]


    Relationship between sperm density, spermatocrit, sperm motility and spawning date in wild and cultured haddock

    JOURNAL OF FISH BIOLOGY, Issue 2 2004
    R. M. Rideout
    Semen was collected repeatedly from captive haddock Melanogrammus aeglefinus and the effect of seasonality on various sperm parameters was investigated. No differences in sperm traits were observed for wild and cultured haddock. A highly significant positive relationship existed between spermatocrit and spermatozoa density. A significant increase in mean spermatocrit occurred throughout the spawning season but the amount of variability explained by collection date was low (35·1%) due to variability between males. Each of 10 males sampled repeatedly throughout the spawning season demonstrated an increase in spermatocrit. No relationship existed between spermatocrit and proportion of motile spermatozoa when spermatocrit was ,70%. Motility was reduced in semen samples with spermatocrits >70%. The proportion of spermatozoa that were motile decreased with time since activation. Some motility was still observed after 60 min in sea water (0·1,15·2%) for sperm collected at all times within the spawning season. Of those spermatozoa that were motile, the proportion that exhibited forward swimming motion decreased and the proportion that had only vibratory movement increased with time post-activation. The speed of forward swimming spermatozoa showed no significant relationship with spermatocrit at any time between 0 and 60 min after activation. Swimming speed was negatively related to time since activation, decreasing from 174,240 ,m s,1 at 0 min to 80,128 ,m s,1 at 60 min after activation. [source]


    Nelumbinis Semen reverses a decrease in hippocampal 5-HT release induced by chronic mild stress in rats

    JOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 5 2005
    Moonkyu Kang
    Depression is associated with a dysfunctional serotonin system. Recently, several lines of evidence have suggested that a very important evoking factor in depression may be a serotonin deficit in the hippocampus. This study assessed the antidepression effects of Nelumbinis Semen (NS) through increasing serotonin concentrations under normal conditions and reversing a decrease in serotonin concentrations in rat hippocampus with depression-like symptoms induced by chronic mild stress (CMS). Using an in-vivo microdialysis technique, the serotonin-enhancing effect of NS on rat hippocampus was investigated and its effects compared with those of two well-known antidepressants, Hypericum perforatum (St John's wort) and fluoxetine (Prozac). Rats were divided into five groups: saline-treated normal, without CMS; saline-treated stress control; NS-, St John's wort- and fluoxetine-treated rats under CMS for 8 weeks or no stress treatment. NS and fluoxetine significantly increased serotonin in normal conditions and reversed a CMS-induced decrease in serotonin release in the hippocampus (P< 0.05 compared with normal group or control group under CMS). These results suggest that NS increases the serotonin levels normally decreased in depression, resulting in an enhancement of central serotonergic transmission and possible therapeutic action in depression. It is suggested that NS may present an antidepressant effect through enhancement of serotonin. [source]


    Female and Male Condoms Offer Similar Protection Against Exposure to Semen

    PERSPECTIVES ON SEXUAL AND REPRODUCTIVE HEALTH, Issue 4 2007
    D. Hollander
    No abstract is available for this article. [source]


    Quality Parameters for Alpaca (Vicugna pacos) Semen are Affected by Semen Collection Procedure

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 4 2010
    KM Morton
    Contents Artificial insemination (AI) is poorly developed in camelids owing to the difficulty in collecting high quality semen and the highly viscous nature of the semen. Semen collected by artificial vagina (AV) is often of low quality and must be improved before any further development of AI technology can occur. The present study investigated the effects of adding a cervix-like stricture to the AV, presence of females, collecting semen into Androhep®, skim-milk or Tris diluents, and catalase supplementation (0, 100, 200 or 600 units/ml) of Tris diluent on alpaca semen quality parameters. The addition of a cervix-like stricture increased mating length (p < 0.05), whilst the presence of females during semen collection did not improve semen quality parameters (p > 0.05). Collection of semen into Tris diluent improved sperm motility (58.0 ± 11.9%) compared with the control (34.0 ± 10.8%; p < 0.05), Androhep® (33.5 ± 10.7%) and skim-milk diluents (28.2 ± 10.4%). Semen viscosity was reduced by collection into Androhep® (4.6 ± 1.7 mm) and skim-milk diluents (3.6 ± 1.3 mm) compared with Tris diluent (5.7 ± 2.1 mm) and no collection medium (9.3 ± 3.5 mm; p < 0.05). Tris diluent supplemented with 100, 200 or 600 units/ml catalase increased semen viscosity (5.0 ± 3.2 and 4.9 ± 3.2 mm). Collection of alpaca semen by AV into Tris diluent increased semen quality facilitating further development of AI technology in alpacas. [source]


    Effect of Docosahexaenoic Acid on Quality of Cryopreserved Boar Semen in Different Breeds

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2010
    K Kaeoket
    Contents During the cryopreservation process, the level of polyunsaturated fatty acids, especially docosahexaenoic acid (DHA), in the sperm plasma membrane decreases significantly because of lipid peroxidation, which may contribute to sperm loss quality (i.e. fertility) of frozen,thawed semen. The aim of this study was to investigate the effect of supplementation of DHA (fish oil) in freezing extender II on frozen,thawed semen quality. Semen from 20 boars of proven motility and morphology, were used in this study. Boar semen was split into four groups, in which the lactose,egg yolk (LEY) extender used to resuspend the centrifuged sperm pellet was supplemented with various levels of fish oil to reach DHA level of 1X (group I, control, no added fish oil), 6X (group II), 12X (group III) and 18X (group IV). Semen solutions were frozen by using a controlled rate freezer. After cryopreservation, frozen semen was thawed and evaluated for progressive motility, viability by using SYBR-14/Ethidiumhomodimer-1 (EthD-1) staining and acrosome integrity by using FITC-PNA/EthD-1 staining. There was a significantly higher (p < 0.001) percentage of progressive motility, viability and acrosome integrity in DHA (fish oil) supplemented groups than control group. Generally, there seemed to be a dose-dependent effect of DHA, with the highest percentage of progressive motility, viability and acrosome integrity in group-III. In conclusion, supplementation of the LEY extender with DHA by adding fish oil was effective for freezing boar semen as it resulted in higher post-thaw plasma membrane integrity and progressive motility. [source]


    Protocol Optimization for Long-Term Liquid Storage of Goat Semen in a Chemically Defined Extender

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 6 2009
    B-T Zhao
    Contents A specific problem in the preservation of goat semen has been the detrimental effect of seminal plasma on the viability of spermatozoa in extenders containing egg yolk or milk. The use of chemically defined extenders will have obvious advantages in liquid storage of buck semen. Our previous study showed that the self-made mZAP extender performed better than commercial extenders, and maintained a sperm motility of 34% for 9 days and a fertilizing potential for successful pregnancies for 7 days. The aim of this study was to extend the viability and fertilizing potential of liquid-stored goat spermatozoa by optimizing procedures for semen processing and storage in the mZAP extender. Semen samples collected from five goat bucks of the Lubei White and Boer breeds were diluted with the extender, cooled and stored at 5°C. Stored semen was evaluated for sperm viability parameters, every 48 h of storage. Data from three ejaculates of different bucks were analysed for each treatment. The percentage data were arcsine-transformed before being analysed with anova and Duncan's multiple comparison test. While cooling at the rate of 0.1,0.25°C/min did not affect sperm viability parameters, doing so at the rate of 0.6°C/min from 30 to 15°C reduced goat sperm motility and membrane integrity. Sperm motility and membrane integrity were significantly higher in semen coated with the extender containing 20% egg yolk than in non-coated semen. Sperm motility, membrane integrity and acrosomal intactness were significantly higher when coated semen was 21-fold diluted than when it was 11- or 51-fold diluted and when extender was renewed at 48-h intervals than when it was not renewed during storage. When goat semen coated with the egg yolk-containing extender was 21-fold diluted, cooled at the rate of 0.07,0.25°C/min, stored at 5°C and the extender renewed every 48 h, a sperm motility of 48% was maintained for 13 days, and an in vitro -fertilizing potential similar to that of fresh semen was maintained for 11 days. [source]


    Effect of Oxytocin Treatment on Artificial Insemination with Frozen,Thawed Semen in Murciano,Granadina Goats

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 4 2009
    MP Viudes-de-Castro
    Contents The site where the semen is deposited appears to be one of the most important factors affecting pregnancy of inseminated goats. In Murciano,Granadina (MG) goats, post-cervical insemination is achieved in a limited number of females. An effective way to increase fertility rate could be by increasing post-cervical inseminations. Effect of exogenous oxytocin application to facilitate the cervical penetration and its effect on kidding rate and prolificacy in MG goats were investigated. Oestrus was synchronized using progesterone-impregnated sponges for 11 days. Females were randomly divided into three groups (n = 190) and received either an i.v. injection of 100 or 200 IU of oxytocin or saline solution 15 min before being inseminated. Data on semen deposition depth were recorded for each animal using a catheter scaled in centimetres (up to 4 cm). Depth of semen deposition was affected by the oxytocin treatment (p < 0.05). Oxytocin enhanced cervical passage only with the dose of 200 IU compared with the control group, increasing the deposition depth (2.9 cm vs 1.9 cm). No significant effect of oxytocin treatment on kidding rate and prolificacy was detected. Depth of semen deposition affected kidding rate (p < 0.01). In conclusion, oxytocin treatment improved the depth of semen deposition in AI of MG goats, but kidding rate and prolificacy was not affected. More studies must be conducted to assess the minimal effective dose required for sufficient cervical dilation, and to determine the effects of such doses of oxytocin on uterine motility, sperm transport and fertility in goats. [source]


    Comparison of the Effect of the Aromatase Inhibitor, Anastrazole, to the Antioestrogen, Tamoxifen Citrate, on Canine Prostate and Semen

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 2009
    G Gonzalez
    Contents This study compared the efficiency of the aromatase inhibitor, anastrazole, with the antioestrogenic receptor blocker, tamoxifen, on normal (NRL) and hyperplastic prostate glands. Forty healthy dogs were classified as NRL (n = 18) or abnormal (ABN) with benign prostate hyperplasia (n = 22). The dogs were randomly assigned to one of the following six groups, treated for 60 days; oral placebo for normal (NRL-PLC; n = 6) and abnormal (ABN-PLC; n = 6), oral anastrazole 0.25,1 mg/day, for normal (NRL-ANZ, n = 6) and abnormal (ABN-ANZ, n = 8) and oral tamoxifen citrate 2.5,10 mg/day for normal (NRL-TMX; n = 6) and abnormal (ABN-TMX; n = 8) dogs. The dogs were evaluated before treatment and then monthly for 4 months. At the end of the treatment, the prostatic volume decreased by 28.5 ± 4.3%, 21.6 ± 6.3% and 0.7 ± 1.0% in the ABN-TMX, ABN-ANZ and ABN-PLC (p < 0.01), respectively. From then on, prostatic volume began to increase without reaching pre-treatment values at the end of the study. In the ABN animals, there were no differences for this parameter between ANZ and TMX treatment (p > 0.1), whereas in the NRL animals ANZ produced a less pronounced decrease (p < 0.05), libido, testicular consistency and scrotal diameter decreased during treatment in the TMX group (p > 0.05). These parameters and sperm volume, count, motility and morphological abnormalities remained unaltered throughout the study in the ANZ and PLC groups (p > 0.05). There were no haematological nor biochemical side effects. Anastrazole might offer a safe and effective alternative for the medical management of dogs with benign prostatic hyperplasia. [source]


    Influence of Ascorbic Acid and Glutathione Antioxidants on Frozen-Thawed Canine Semen

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 2009
    JC Monteiro
    Contents Poor sperm viability post-thaw has resulted in constant research into methods of cryopreservation of canine semen. One factor that may be involved in poor viability is sperm oxidative stress caused by excessive formation of reactive oxygen species. The present study was performed in order to evaluate the effect of different concentrations of ascorbic acid (AA) and glutathione (Glu) added to an extender for the freeze-thawing of dog sperm. Semen from five mature dogs was collected and frozen in two studies. Prior to and after freezing, sperm motility, morphology and membrane status were examined. In addition, sperm motility was examined up to 120 min after thawing to evaluate thermo-resistance. In study I, semen was collected twice from each dog. On both occasions, semen was divided into three aliquots: control, Glu 1 mm and Glu 5 mm. In study II, semen was collected twice and divided into three aliquots; control, AA 50 ,m and AA 250 ,m. Initial sperm motility was significantly higher in sperm diluted with AA 50 ,m; sperm longevity, however, measured by a thermal-resistance test (TRT), was higher for Glu treatments. Higher concentration of Glu produced significant improvement in TRT and membrane status, whereas higher concentration of AA had a negative impact in sperm longevity. Antioxidant supplementation to semen freezing extenders improved semen quality post-thaw. Moreover, Glu had the most beneficial effect when supplemented at 5 mm. [source]


    DNA Status on Thawed Semen from Fighting Bull: A Comparison Between the SCD and the SCSA Tests

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2009
    F Martínez-Pastor
    Contents The assessment of sperm chromatin status is compulsory in a complete spermiogram. Here we applied the sperm chromatin structure assay (SCSA) and the sperm chromatin dispersion (SCD) test to assess the chromatin status of three fighting bulls. Cryopreserved semen (two straws/bull) were analysed by duplicate after thawing and after 6 h at 37°C with and without oxidative stress (1 mm FE2+). Results (SCD: percentage of spermatozoa with halo; SCSA: SD-DFI, %DFI and HDS) were analysed for differences between bulls and treatments, sensitivity and specificity (receiver operating characteristic curves) and repeatability (repeatability coefficients as 2SD of duplicate differences).%DFI for the three bulls was below 2% at 0 h, indicating no risk for fertility according to previous reports. It increased slightly for two of the bulls after FE2+ treatment (%DFI < 5%) and more pronouncedly for the other bull (C, %DFI,10%), which merits further investigation. SCD rendered higher percentage of halos for bull C, but could not discriminate between samples with and without oxidizing treatment (AUC: 0.52). SCSA (%DFI) showed a high discriminating ability between treatments (AUC: 0.96). The repeatability coefficient was also higher for SCD (5.9) than for %DFI (1.8), indicating lower repeatability for SCD. Overall, %DFI might be the most useful parameter for assessing sperm chromatin on fighting bull. SCD might yield different information than SCSA, hence further research is warranted. [source]


    Prognostic Value of Various Spermatological Attributes as Predictors of Zona Binding and Zona Penetration of Buffalo (Bubalus bubalis) Semen

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2009
    S Selvaraju
    Contents Twenty-four ejaculates from six (four ejaculates each) Surti buffalo bulls aged 4,8 years were used to assess various attributes of spermatozoa influencing the zona-binding and zona-penetration tests. Ejaculates from each bulls were subjected to in vitro sperm--zona binding and sperm--zona penetration tests (four replicates per bull) using immature buffalo oocytes. The average number of spermatozoa bound per oocyte was 27.79 ± 5.90. The average number of spermatozoa penetrated per oocyte was 3.35 ± 0.64. The average number of zona-bound and -penetrated spermatozoa differed significantly between animals. Significant difference (p < 0.05) was observed between the plasmalemma integrity as assessed by eosin--nigrosin stain and hypo-osmotic swelling (HOS) test. Furthermore, the percentage of cells positive for the HOS test, i.e. functional membrane integrity (51.25 ± 2.32) was significantly (p < 0.05) higher than hypo-osmotic swelling-Giemsa (HOS-G) test, i.e. the subpopulation of spermatozoa positive for functional membrane and acrosomal integrities (42.87 ± 4.56). The HOS test had significant correlations with plasmalemma integrity as measured by the vital stain, eosin--nigrosin (r = 0.85, p < 0.05). The HOS-G test also had significant correlation with plasmalemma integrity measured by vital stains such as eosin--nigrosin (r = 0.90, p < 0.05) and fluorogenic stains [carboxyfluorescein diacetate (CFDA) and propidium iodide (PI); r = 0.92, p < 0.01] and HOS test (r = 0.93), acrosomal integrity (r = 0.86, p < 0.05) and mitochondrial membrane potential (r = 0.99, p < 0.01). The plasmalemma integrity (fluorogenic stain), functional membrane integrity (HOS test), subpopulation of spermatozoa positive for functional membrane and acrosomal integrities (HOS-G test) and mitochondrial membrane potential had significant (p < 0.05) correlation with sperm zona binding and penetration. The present study indicates that these parameters could represent important determinants of sperm quality influencing zona binding and penetration. [source]


    Testing Usability of Butylated Hydroxytoluene in Conservation of Goat Semen

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2008
    TAA Khalifa
    Contents The objective of this study was to investigate whether butylated hydroxytoluene (BHT) could be used as a suitable supporter or alternative of egg yolk during preservation of goat spermatozoa. Three in vitro experiments and a fertility test were conducted to evaluate the effect of BHT on viability of chilled-stored semen as well as motility and kidding rate of frozen-thawed spermatozoa. In the first two experiments, ejaculates (n = 30/experiment) were collected from 10 bucks, split, diluted with egg yolk-based and egg yolk-free extenders supplemented with or without 0.3, 0.6, 2, 5 and 8 mm BHT and stored at 5°C for 168 h. In the third experiment, 30 ejaculates were collected from the above-mentioned bucks, split and diluted with egg yolk-free extenders supplemented with or without 0.3, 0.6 and 0.9 mm BHT and egg yolk-based extenders supplemented with or without 5 mm BHT. Diluted semen was cooled to 5°C over a period of 4 h, frozen and thawed in the form of 0.3-ml pellets. In the fertility test, 75 ejaculates were collected from two proven fertile bucks, split, diluted with egg yolk-free extenders containing 0.6 mm BHT and egg yolk-based extenders supplemented with or without 5 mm BHT, frozen and thawed as described above. An insemination volume of 0.6 ml containing 120,140 × 106 progressively motile spermatozoa was used for a single cervical insemination of cloprostenol-synchronized does (n = 230). The results showed that addition of 5 mm BHT to egg yolk-deficient (2.5%) extenders significantly improved viability of chilled-stored semen together with motility (48.5%) and fertility (62.5%) of frozen-thawed spermatozoa. Replacement of egg yolk in semen extenders by 0.6 mm BHT could sustain not only viability of chilled-stored semen but also post-thaw motility (47.5%) and fertility (53.75%) of frozen-thawed spermatozoa. In conclusion, supplementation of semen diluents with BHT can ameliorate preservability of goat sperm. [source]


    CASA Assessment of Kinematic Parameters of Ram Spermatozoa and their Relationship to Migration Efficiency in Ruminant Cervical Mucus

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 4 2008
    I Robayo
    Contents Sperm motility is an indicator of male fertility because of its importance for sperm migration through the female genital tract and for gamete interaction at fertilization. This study analyses the relationship between computer assisted semen analysis (CASA) motility patterns and sperm migration of rams in ruminant cervical mucus. In experiment 1, spermatozoa extended with sperm analysis medium (SAM) and seminal plasma were compared in terms of motility. In experiment 2, 56 semen samples were collected either with artificial vagina (AV) or electroejaculator to be compared in terms of motility performance. In experiment 3, 104 ejaculates collected by AV from 26 males were analysed via the CASA system to characterize their motility patterns. In experiment 4, ejaculates from pairs of rams (20 rams in total) were simultaneously assessed for mucus migration (ovine, caprine, bovine) and motility patterns to evaluate the correlations between both parameters. Semen collected by AV and extended in SAM allows the most reliable assessment for sperm motility. Ram spermatozoa move fast and follow a linear trajectory compared with other ruminants. Continuous line velocity (VCL) and average path velocity (VAP) are the only sperm kinematic parameters that presented significant positive correlations with the ability to migrate in sheep cervical mucus (p < 0.05). Continuous line velocity, VAP, straight line velocity and linearity are highly significantly related with migration efficiency in goat cervical mucus (p < 0.01) and only lateral head displacement is negatively related to sperm migration in bovine cervical mucus (p < 0.05). These results suggest that specific kinematic parameters confer the ability of spermatozoa to colonize and migrate through epithelial mucus with different rheological properties. [source]


    Effects of Oxygen Exposure and Gentamicin on Stallion Semen Stored at 5 and 15°C

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2008
    S Price
    Contents This study was undertaken to investigate the effects of storage of stallion semen in a defined milk protein extender at 5 and 15°C under either anaerobic or aerobic conditions, with or without addition of the antibiotic gentamicin. Semen samples were collected from eight fertile stallions and stored for 96 h (day 0,4) and assessed daily for motility, velocity and membrane integrity (viability) using a CASA system. Samples for bacteriology assessment were taken on day 2 of storage. No significant (p > 0.05) differences in motility, velocity or viability were observed between treatments on days 0,2. On days 3 and 4, semen stored without gentamicin at 5°C had a significantly (p < 0.05) better semen quality compared with storage at 15°C without gentamicin, irrespective of oxygen exposure. On days 3 and 4, motility and velocity were greater in samples stored at 15°C with gentamicin, compared with the corresponding treatments without antibiotic (p < 0.05). This effect was also evident for viability on day 4. The decline in semen quality observed at 15°C most likely resulted from the effect of bacterial growth. Bacterial growth was the greatest in samples stored at 15°C without gentamicin, under both anaerobic and aerobic conditions (p < 0.05). Bacterial growth was inhibited by adding of gentamicin at 15°C, which accordingly reduced the decline in semen quality. Addition of antibiotic to samples stored at 5°C had no significant effect on any parameter analysed. In conclusion, storage at 15°C can be achieved by using an extender containing the antibiotic gentamicin. Storage at 5°C tended to maintain better semen quality irrespective of oxygen exposure, and did not necessitate an antibiotic treatment. [source]


    Multivariate Cluster Analysis Regression Procedures as Tools to Identify Motile Sperm Subpopulations in Rabbit Semen and to Predict Semen Fertility and Litter Size

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2007
    A Quintero-Moreno
    Contents Computerized motility analysis (CASA) shows that four separate subpopulations of spermatozoa with different motility characteristics co-exist in rabbit ejaculates. There were significant (p < 0.01) differences in the distribution of these subpopulations among separate genetic lines, total sperm abnormalities and the percentage of altered acrosomes. Furthermore, logistic and linear multivariate regressions among several parameters of rabbit semen quality analysis were tested for use as predictive tools for the fertilizing ability of a specific artificial insemination semen sample. Logistic regression analysis rendered two mathematical, significant (p < 0.01) models: one between sperm viability and conception rate and the other between total sperm abnormalities and conception rate. Multiple linear regression analyses also yielded some significant relationships between both fertility (p < 0.001) and litter size (p < 0.05), with respect to some semen characteristics. Our results support the hypothesis that the predictive in vivo fertility use of the standard rabbit semen quality analysis coupled with a CASA determination could be reasonably achieved by applying linear and logistic regression analyses among several parameters of rabbit semen quality analysis. [source]


    Effects of Constant, 9 and 16-h Light Cycles on Sperm Quality, Semen Storage Ability and Motile Sperm Subpopulations Structure of Boar Semen

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2006
    MM Rivera
    Contents This study was performed to test the effect that two separate, daily, constant-light regimes of both 9 and 16 h could have on the main parameters of boar-semen quality analysis, as well as on the motile sperm subpopulations structure and the ability of its conservation at 16°C. Results show that both luminous regimes have slight, specific effects on the main parameters of boar-semen quality analysis, as well as on the motile sperm subpopulations structure. Furthermore, the conservation ability at 16°C of boar semen was not significantly different between both photoperiods. When a temporal study was performed, results showed that semen quality and motility parameter changes were stabilized at nearly constant values from the second month of the study to the last month in both luminous regimes, indicating a rapid light-related effect on testicular function. Our results indicate that light regimes oscillating from 9 h daily to 16 h daily are of little importance in the control of boar-semen quality in a farming environment. [source]


    Comparative Study on Five Different Commercial Extenders for Boar Semen

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2004
    P Vyt
    Contents Increasing interest in a longer preservation of diluted boar sperm raises questions in the field concerning the choice of the extender. The aim of this study was to evaluate the longevity of boar sperm extended in currently used commercial semen extenders. Three long-term extenders and two short-term extenders were compared for different semen quality parameters that can be assessed under routine laboratory conditions. Sperm morphology, motility, pH and bacteriological contamination were investigated during a 7-day period. The number of dead spermatozoa did not differ significantly among the extenders (p > 0.05). Sperm motility was not only related with storage period but most of all with pH, especially in long-term extenders. Differences between the different extenders were prominent (p < 0.05); the sperm preserved in only one long-term extender showed good motility during the whole test period. In all cases, the pH of the extended semen increased by 0.3,0.5 in the first days of storage and was significantly correlated with a decrease in motility. Bacteriological quality had no significant influence on motility or pH of the semen. In conclusion, we can state that in both short-term extenders and in only one long-term extender, sperm longevity, as evaluated by the parameters used in this study, was sufficient during the preservation period. To preserve the quality of diluted boar semen during long-term storage, the choice of the long-term extender is important. In addition, the monitoring of the pH of extended boar semen in our study emphasizes the importance of the buffering capacity of semen extenders. [source]


    Assessing in vivo Fertilizing Capacity of Liquid-Preserved Boar Semen According to the ,Hanover Gilt Model'

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 2 2003
    F Ardón
    Contents The goal of this study was to determine the ability of the Hanover gilt model to assess in vivo fertilizing capacity of preserved sperm and to consider whether any modifications to this model were needed. This model evaluates the fertilizing capacity of semen based on the fertilization rate, the rate of normal embryos and the accessory sperm count of 3,5-day embryos. Its distinguishing characteristics are the use of one-time insemination of sperm in reduced numbers, of spontaneously ovulating gilts and of ovulation detection through ultrasound examination of ovaries. Reduced sperm numbers allow for an accurate evaluation of the fertilizing potential of different semen treatments, thereby avoiding the compensatory effect of doses calibrated to maximize fertility. The model's usefulness was assessed in a trial run designed to compare the fertilizing capacity of liquid boar semen diluted into two different extenders. The diluent, the boar and the backflow, had no significant effect on any of the parameters studied. Gilts inseminated less than 24 h before ovulation had a significantly higher (p < 0.01) fertilization rate and accessory sperm cell count (p < 0.05) than those inseminated more than 24 h before ovulation. Very good/good embryos from homogeneous litters (only very good/good embryos were present) had a significantly higher (p < 0.01) accessory sperm count than those from heterogeneous litters (at least one embryo was of a different quality and/or oocytes were present). Both very good/good and degenerated/retarded embryos from heterogeneous litters had low accessory sperm numbers. This suggests that accessory sperm count is significantly related to the quality of the litter, but not to the quality of the embryo within gilts. It can be concluded that the Hanover gilt model is sensitive enough to show fertility differences (in this study, those associated with in vivo ageing of semen), while using relatively few gilts and little time. [source]


    Validation of the FACSCount AF System for Determination of Sperm Concentration in Boar Semen

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 6 2002
    C Hansen
    Contents A flow cytometric method has been developed for rapid determination of sperm concentration in semen from various mammalian species., All cells containing DNA are stained with SYBR-14 or propidium iodide (PI) and sperm concentration is determined in relation to an internal standard of fluorescent microspheres (beads). Satisfactory staining can be achieved within 2,3 min and the following flow cytometric analysis on the FACSCount AF System rapidly provides the user with a precise and accurate assessment of the sperm concentration. In this study, the FACSCount AF System and Sperm Counting Reagent (BD Biosciences) was compared with microscopic counting using a Bürker,Türk haemocytometer. In addition, sperm concentration was determined using the Corning 254 spectrophotometer which is used routinely by Danish artificial insemination stations for boars. The results show that the agreement between flow cytometry and microscopic counting is very high. The slope for the regression line was 1.12 (SE = 0.03) with an estimated intercept with the Y-axis of 22 × 106sperm/ml (SE = 10 × 106 sperm/ml) and an estimated error of the model of 10 × 106 sperm/ml. For the spectrophotometer, the slope of the regression line was 1.09 (SE = 0.07) with an estimated intercept of 137 × 106 sperm/ml (SE = 25 × 106 sperm/ml). The average error made by the spectrophotometer was 55 × 106 sperm/ml. In addition, the results obtained using flow cytometry was highly repeatable (CV = 2.7%) in comparison with the spectrophotometric method (CV = 6.3%). These results indicate that the FACSCount AF System is a valuable tool for precise and accurate assessment of sperm concentration in boar semen and that use of this system may lead to production of more uniform insemination doses containing a specific number of sperm per dose. [source]


    Characteristics of Buck Semen with Regard to Ejaculate Numbers, Collection Intervals, Diluents and Preservation Periods

    REPRODUCTION IN DOMESTIC ANIMALS, Issue 2 2000
    M Shamsuddin
    Contents To determine the number of ejaculates which can be collected within a 20-min period after the smallest number of days of sexual rest, and a good diluent to preserve semen for routine AI, five mature Black Bengal bucks were used in three experiments. In experiment 1, semen from the bucks were collected by using artificial vagina at homosexual mounts as many times as possible during 20 min. The ejaculate numbers 1, 3 and 4 (or 5 when the buck could produce it) were examined for important semen characteristics. The mean ejaculate volume, density, mass activity, sperm motility, sperm concentrations, total spermatozoa/ejaculate, proportion of spermatozoa with normal acrosome, midpiece and tail, and the proportion with normal head morphology varied between 267 and 342 µl, 4.1,4.5 (1,5 scale), 4.1,4.2 (1,5 scale), 77,79%, 4187 × 106,5064 × 106/ml, 1140 × 106,1746 × 106, 91,94% and 99%, respectively, depending on the collection number of the ejaculate. The difference between the ejaculates was significant only with respect to volume (p < 0.05). In experiment 2, semen was collected from the bucks successively during 20 min after 1, 2, 3 and 4 day intervals, and the first ejaculates were evaluated for the above-mentioned semen characteristics. Semen collected after 2 or more day intervals had significantly higher volume, sperm concentration and total spermatozoa/ejaculate (p < 0.05). In experiment 3, pools of two to three ejaculates were diluted (1 : 5; semen : diluent) in splits with glucose-citrate-egg yolk (GCEY), Tris-fructose-egg yolk (TFEY) or skim milk (SM) and preserved at +4 to +7°C. Before chilling or after 0 (15 min chilling), 1, 2, 3 and 4 days of preservation, semen was evaluated for motility and proportion of normal spermatozoa with respect to acrosome, midpiece and tail. In data pooled across the bucks, the sperm motility was better in GCEY and TFEY than in SM, and the proportion of normal spermatozoa was higher in SM than in the others (p < 0.05). However, the differences in proportion of normal spermatozoa between diluents were not significant when the data were analysed separately within preservation periods. The sperm motility consistently dropped after 1 day of preservation (p < 0.01); the motility remained 50% or more up to 4 days in TFEY, 3 days in GCEY and only 2 days in SM. The proportion of spermatozoa with normal acrosome, midpiece and tail, which was generally quite high ( 90%), decreased after 3 days of preservation (p < 0.01). We conclude that Black Bengal bucks can be collected three times during 20 min, every 3 days, and that buck semen holds good motility and proportion of normal spermatozoa up to 3 days in GCEY or TFEY at 4 to 7°C. [source]


    ORIGINAL ARTICLE: Expression of IL-6, IL-8, TNF-,, IL-10, HSP-60, Anti-HSP-60 Antibodies, and Anti-sperm Antibodies, in Semen of Men with Leukocytes and/or Bacteria

    AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 3 2010
    Elizabeth Martínez-Prado
    Citation Martínez-Prado E, Bermúdez MIC. Expression of IL-6, IL-8, TNF-,, IL-10, HSP-60, anti-HSP-60 antibodies, and anti-sperm antibodies, in semen of men with leukocytes and/or bacteria. Am J Reprod Immunol 2010; 63: 233,243 Problem, Different cellular and biochemical markers have been proposed as indicators of infection-inflammation of male genital tract. Method of study, Semen samples from 80 men attending an andrologic clinic were evaluated to determine the presence of leukocyte, bacteria, antibodies against Chlamydia trachomatis, levels of IL-6, IL-8, IL-10, and TNF-,, HSP-60, anti-HSP-60 antibodies, and anti-sperm antibodies. Results, Leukocytes in semen significantly correlated with an increase in IL-6, IL-8, and TNF-,. The simultaneous presence of pathogens and leukocytes was associated with high levels of IL-8 and TNF-,, whereas IL-6 was more associated with the presence of leukocytes. Anti-HSP-60 antibodies positively correlated with IL-6 and IL-8. The presence of anti-sperm antibodies highly associated with an increase in anti-HSP-60 antibodies. Conclusions, The type of cytokines present in the semen will depend on the single or simultaneous presence of leukocytes and/or pathogens. Chronic male genital tract infections could be associated with the development of anti-HSP-60 antibodies and anti-sperm antibodies. [source]


    ORIGINAL ARTICLE: Sperm Antibodies, Intra-Acrosomal Sperm Proteins, and Cytokines in Semen in Men from Infertile Couples

    AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 3 2009
    Zdenka Ulcova-Gallova
    Problem, The aim of this study was to investigate seminal sperm-agglutinating antibodies, intra-acrosomal proteins, sperm head abnormalities, and cytokines (IL-1,, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70 TNF-,, and IFN-,) in men from infertile couples. Method of study, The direct mixed anti-immunoglobulin reaction test for IgG, IgA, and IgE in semen, and immunocytochemical method using monoclonal antibodies and indirect immunofluorescence for the examination of intra-acrosomal proteins in the spermatozoa were used. Cytokines in seminal plasma were determined by multiplex immunoanalytic xMAP (LUMINEX) technology. Results, Sperm-agglutinating antibodies, IgG and IgA, in seminal plasma were found to be more in asthenospermatic and oligoasthenospermatic men than in normospermatic men. Sperm head pathology and very low amounts of acrosomal proteins were frequently detected in pathologic semen samples. Cytokine levels defined as ,high' (based on the 75 percentile for each cytokine in all groups) were obtained especially for IL-8, IL-5, IL-6, and IL-10. The high cellularity in semen was correlated with higher IL-5. Conclusion, Immunologic cause of male infertility is a very important risk factor in the pathogenesis of sperm cells. Sperm autoantibodies and the presence of intra-acrosomal factors must be studied together, cytokines according to accessory cellularity in the semen. [source]


    Litter Characteristics of Gilts Artificially Inseminated with Transforming Growth Factor-,

    AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 3 2006
    Michelle Rhodes
    Problem, Semen is a rich source of transforming growth factor- , (TGF- ,) and it has been proposed that this molecule promotes embryonic survival by modifying immune responses to promote tolerance toward paternal antigens and by inducing release of cytokines that promote embryonic development. The role of TGF- , was tested using pigs by evaluating whether its addition to washed sperm improves conceptus survival and fetal growth. Methods of study, At estrus, gilts were artificially inseminated twice at 12-hr intervals with 100 mL of either washed semen resuspended in a commercial semen extender supplemented with 2 mg/mL of gelatin or washed semen in the same extender containing 65 ng/mL of TGF- ,1. Three boars were used as semen donors. At day 80 (±4 days) of gestation, gilts were sacrificed and reproductive tracts harvested. Results, Treatment had no effect (P > 0.10) on total or live fetuses per litter, implantation rate, fetal survival or percentage of corpora lutea resulting in live fetuses at day 80. Insemination with TGF- ,1 also did not affect total or average fetal weight or total placental weight. There was a tendency (P = 0.09) for average placental weight of live fetuses to be lower for pregnancies established in gilts treated with TGF- ,1. Also, placental efficiency (mass of fetus/mass of placenta) was greater (P < 0.05) for pregnancies established in gilts treated with TGF- ,1. The high fertility in control gilts (80% implantation rate and 11.5 live fetuses per litter) is indicative that soluble seminal factors are not necessary for the establishment of pregnancy. Conclusions, Within the ranges tested, concentration of TGF- , in the fluid phase of the inseminate is not an important determinant of conceptus survival or fetal and placental growth to day 80 of gestation in the pig. [source]


    Distinct Expression Pattern of Cytokines in Semen of Men with Genital Infection and Oligo-Terato-Asthenozoospermia

    AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 3 2002
    Ioannis Matalliotakis
    PROBLEM: The objective of this study was to evaluate the possible relevance of cytokines in seminal plasma (SP) of patients with accessory gland infection and oligo-terato-asthenozoospermia. METHOD OF STUDY: Semen samples were obtained by masturbation from 90 men and were examined for the presence of interleukin (IL)-2, IL-6, IL-8, IL-11 and soluble CD23 (sCD23) by enzyme-linked immunosorbent assay. Five groups were included: (1) fertile men (n=20), (2) infertile men with varicocele and oligo-terato-asthenozoospermia (V-OTA, n=20), (3) infertile men with genital infection and OTA (INF-OTA, n=20), (4) infertile men with idiopathic testicular lesion and OTA (ITL-OTA, n=20) and (5) infertile men with azoospermia (AZOO, n=10). RESULTS: We found that the mean level of IL-2 was higher in SP from infertile men compared with SP from fertile men (P < 0.05). Mean levels of IL-6, IL-8, IL-11 in SP of INF-OTA were higher than that of all other groups (P < 0.05, P < 0.05, P < 0.001, respectively). However, no significant differences could be detected between other groups. A significant increase was noted in sCD23 levels in SP from men with ITL-OTA compared with all other groups (P < 0.01). We have not observed any correlations between IL-2, IL-6, IL-8, IL-11 and sCD23 levels in SP and semen parameters. Spearman's correlation coefficient revealed that there was a significant association between IL-6, IL-8, IL-11 levels in men with INF-OTA. CONCLUSION: The measurement of each cytokine separately in the SP of men with INF-OTA, in spite of the existing significant differences, does not have a diagnostic value in male infertility. However, a combined determination of IL-6, IL-8, IL-11 in the SP of men with genital infection and oligo-terato-asthenozoospermia may provide clinically useful information for the diagnosis of male accessory gland infection. [source]


    Cryopreservation of Black Bengal buck semen: Effects of diluents and freezing on sperm motility and morphology

    ANIMAL SCIENCE JOURNAL, Issue 5 2008
    Sadia AFROZ
    ABSTRACT Semen from Black Bengal bucks was collected to establish a cooling protocol (to ,196°C) for buck semen preservation, and to study the effect of freezing on sperm motility and morphology. Semen was diluted with diluents (Triladyl & Tris) and cryoprotectants, filled into straws, sealed, cooled (to 5°C) and equilibrated. After dilution, motility ranged from 75.00% to 76.67% and from 73.33% to 80.00% in Triladyl and Tris diluents, respectively. Motility of sperm after cooling to 5°C in Triladyl and Tris diluents ranged from 65.00% to 66.67% and from 63.33% to 70.00%, respectively. After equilibration in straws, the semen was subjected to a freezing protocol in a computer-controlled biofreezer CL-3000 (cooling at 10°C per minute, from 5°C to ,80°C) and plunged into liquid nitrogen. Sperm motility of re-thawed semen varied from 38.33% to 43.33% and from 6.00% to,6.67% in Triladyl and Tris diluents, respectively. Sperm morphology of re-thawed semen was studied and head damage or cryoinjury was found in 2,3% of sperm in Triladyl diluents and 3,6% in Tris diluents. Whether the differences of sperm motility and head damage reflect fertility after artificial insemination is yet unknown and needs to be studied further. [source]


    Cryopreservation of silver barb Puntius gonionotus (Bleeker) spermatozoa: effect of extender composition, cryoprotective agents and freezing rate on their postthawing fertilization ability

    AQUACULTURE RESEARCH, Issue 15 2008
    Padmanav Routray
    Abstract The effects of extender composition, cryoprotectant concentration and freezing and thawing on the fertilization efficiency of cryopreserved spermatozoa of Puntius gonionotus were evaluated. Computer-aided motility analysis of semen was conducted to check the suitability of spermatozoa for cryopreservation after mixing with different extenders and cryoprotective agents (CPAs). Extender-4 with an osmolality 260 mOsmol kg,1and pH 7.6 was used for the cryopreservation study. Among the CPAs, dimethyl sulphoxide (DMSO) was least toxic and more than 60% fertilization was achieved when used at 1.4 M at 0 °C for 10 and 30 min, whereas the toxicity of all CPAs to spermatozoa was evident when tested at 30 °C. Semen frozen at ,16 °C min,1 with 1.4 M DMSO showed 70% fertilization, which was significantly higher (P<0.05) than other freezing rates. Samples thawed at 35 °C water showed a fertilization rate comparable with that of fresh semen. Computer-assisted semen analysis of fresh and frozen semen after thawing showed variations in different types of motility in spermatozoa and in their class. There was no significant difference in motility before or after cryopreservation; however, significant differences could be observed in the average path velocity (VAP), straight line velocity (VSL) and curve linear velocity (VCL). Semen of silver barb could be cryopreserved with extender-4 by addition of 1.4 M DMSO to a final cryopreservation medium (MED 2) cooled at a rate of ,16 °C min,1, stored in liquid nitrogen (,196 °C) and utilized after thawing at 35±2 °C. [source]


    Effect of extender composition and freezing rate on post-thaw motility and fertility of Arctic char, Salvelinus alpinus (L.), spermatozoa

    AQUACULTURE RESEARCH, Issue 9 2006
    Nabil Mansour
    Abstract The effects of extender composition and freezing rate on motility and fertility of frozen-thawed Arctic char, Salvelinus alpinus, spermatozoa were investigated. Three freezing rates, two semen diluents and three cryoprotectants were tested. Semen frozen in 0.3 mol L,1 glucose diluent with 10% methanol as a cryoprotectant or in a diluent described by Lahnsteiner with 10%N,N- dimethylacetamide (DMA) resulted in the highest sperm motility. Fertility was the highest for semen frozen in a glucose,methanol extender but was not significantly different than that for semen frozen in Lahnsteiner's diluent with 10% DMA. Dimethyl sulphoxide (DMSO) at 10% was a relatively ineffective cryoprotectant with either semen diluent. Semen frozen at 6 cm above the surface of liquid nitrogen resulted in a higher post-thaw sperm motility and fertility than semen frozen at 5 cm. The addition of 7% fresh egg yolk to glucose diluent containing methanol or DMSO did not improve the fertility of frozen-thawed spermatozoa. However, the addition of 7% fresh egg yolk to glucose,DMA extender significantly improved the fertilization percentages of frozen-thawed spermatozoa. In conclusion, dilution of semen 1:3 in 0.3 mol L,1 glucose with 10% methanol and freezing 6 cm above the surface of liquid nitrogen (freezing rate of 40±8°C min,1, mean±SD from ,5 to ,55°C) is a promising protocol for cryopreservation of Arctic char semen. [source]


    Birth of twins using in vitro matured oocytes and cryopreserved semen from a male with carcinoma of the penis

    AUSTRALIAN AND NEW ZEALAND JOURNAL OF OBSTETRICS AND GYNAECOLOGY, Issue 4 2000
    Tony T Y Chiu
    Summary: Semen cryopreserved from a male prior to treatment for carcinoma of the penis was used for intra-cytoplasmic sperm injection (ICSI) of in vitro matured (IVM) oocytes, resulting in the birth of healthy, non-identical twin girls. [source]


    Freezing equine semen: the effect of combinations of semen extenders and glycerol on post-thaw motility

    AUSTRALIAN VETERINARY JOURNAL, Issue 7 2009
    J Scherzer
    Objective We evaluated combinations of two commercial semen extenders and three concentrations of glycerol to determine the combination that yielded the highest post-thaw sperm motility. Design A randomised 2 × 3 block design was used. Procedure Semen was collected from four stallions (6 collections per stallion). The sample was diluted with either a dried skim-milk glucose extender (EZ Mixin Original Formula) or a chemically defined, milk-free diluent (INRA 96), and each was used in combination with 2%, 3% or 4% glycerol in standard commercial freezing medium. Sperm motility was assessed by microscopy in fresh and post-thaw semen. Results There was a significant difference between the two extenders in the motility of spermatozoa after cryopreservation (48.9% for INRA 96; 38.6% for EZ Mixin OF; P < 0.0001). Glycerol at 4% in freezing medium yielded the highest post-thaw motility, significantly better than 2% (P < 0.05). Three of four stallions had significantly higher post-thaw motility using INRA 96 relative to EZ Mixin OF (P < 0.01), and two of four stallions had significantly higher post-thaw motility using 4% glycerol (P < 0.05). The combination of INRA 96 and 4% glycerol in freezing medium gave the highest average post-thaw motility of 51.5%. Conclusion In this study, INRA 96 combined with 4% glycerol yielded an average recovery of progressively motile sperm consistently above the 35% target. [source]