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Sample records for fresh ram sperm

  1. Cholesterol-loaded cyclodextrin improves ram sperm cryoresistance in skim milk-extender.

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    Salmon, Vianney M; Castonguay, François; Demers-Caron, Vincent; Leclerc, Pierre; Bailey, Janice L

    2017-02-01

    Cholesterol-loaded cyclodextrin (CLC) is known to improve ram sperm cryosurvival. This study expands on previous research to: (1) determine the mechanism by which CLC improves ram sperm cryosurvival and (2) compare the efficiency of a novel, skim milk-based extender containing CLC to a traditional egg yolk-based extender. Hypothesis #1 was that CLC enhances membrane cholesterol content to increase the resistance of ram sperm to cold and osmotic stress, thereby improving cryosurvival. We first assessed the ability of fresh sperm treated with CLC to withstand cold shock. Second, fresh sperm were treated with CLC to evaluate their tolerance to osmotic stress. Third, to confirm that cholesterol is incorporated into the sperm using CLC, we quantified sperm cholesterol. To test Hypothesis #2 that CLC is most effective in a medium without competing cholesterol, we compared sperm cryosurvival and fertility in skim milk-based extender containing CLC versus in a traditional egg yolk-based freezing extender without CLC. Our data confirmed that CLC treatment improves ram sperm cold shock and osmotic stress resistance, and augments sperm cholesterol content. Semen in skim milk-based extender containing CLC prior to freezing, had more motile sperm with intact acrosomes after thawing compared to semen in egg yolk-based extender. In contrast, sperm plasma membrane integrity and in vivo fertility of the semen cryopreserved in the skim milk-based extender with CLC did not differ from semen that was cryopreserved in egg yolk-based extender. Further research is warranted to combine CLC with other cryoprotection strategies or to modify the insemination protocol.

  2. Testicular Damage following Testicular Sperm Retrieval: A Ram Model Study

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    Jens Fedder

    2017-01-01

    Full Text Available The aim of this study was to evaluate the possible development of histological abnormalities such as fibrosis and microcalcifications after sperm retrieval in a ram model. Fourteen testicles in nine rams were exposed to open biopsy, multiple TESAs, or TESE, and the remaining four testicles were left unoperated on as controls. Three months after sperm retrieval, the testicles were removed, fixed, and cut into 1/2 cm thick slices and systematically put onto a glass plate exposing macroscopic abnormalities. Tissue from abnormal areas was cut into 3 μm sections and stained for histological evaluation. Pathological abnormalities were observed in testicles exposed to sperm retrieval (≥11 of 14 compared to 0 of 4 control testicles. Testicular damage was found independently of the kind of intervention used. Therefore, cryopreservation of excess sperm should be considered while retrieving sperm.

  3. Penicillamine prevents ram sperm agglutination in media that support capacitation.

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    Leahy, T; Rickard, J P; Aitken, R J; de Graaf, S P

    2016-02-01

    Ram spermatozoa are difficult to capacitate in vitro. Here we describe a further complication, the unreported phenomenon of head-to-head agglutination of ram spermatozoa following dilution in the capacitation medium Tyrodes plus albumin, lactate and pyruvate (TALP). Sperm agglutination is immediate, specific and persistent and is not associated with a loss of motility. Agglutination impedes in vitro sperm handling and analysis. So the objectives of this study were to investigate the cause of sperm agglutination and potential agents which may reduce agglutination. The percentage of non-agglutinated, motile spermatozoa increased when bicarbonate was omitted from complete TALP suggesting that bicarbonate ions stimulate the agglutination process. d-penicillamine (PEN), a nucleophilic thiol, was highly effective at reducing agglutination. The inclusion of 250 μM PEN in TALP reduced the incidence of motile, agglutinated spermatozoa from 76.7 ± 2.7% to 2.8 ± 1.4%. It was then assessed if PEN (1 mM) could be included in existing ram sperm capacitation protocols (TALP +1 mM dibutyryl cAMP, caffeine and theophylline) to produce spermatozoa that were simultaneously capacitated and non-agglutinated. This protocol resulted in a sperm population which displayed high levels of tyrosine phosphorylated proteins and lipid disordered membranes (merocyanine-540) while remaining motile, viable, acrosome-intact and non-agglutinated. In summary, PEN (1 mM) can be included in ram sperm capacitation protocols to reduce sperm agglutination and allow for the in vitro assessment of ram sperm capacitation.

  4. Differential proteome association study of freeze-thaw damage in ram sperm.

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    He, Yuxuan; Wang, Ke; Zhao, Xingxu; Zhang, Yong; Ma, Youji; Hu, Junjie

    2016-02-01

    In this study proteomics analysis was performed to investigate damage caused to ram sperm by the freeze-thaw process. Sperm motility, viability, reactive oxygen species (ROS) and adenosine triphosphate (ATP) content were measured to evaluate sperm quality. Compared with fresh groups, motility, viability and ATP content were all lower in freeze-thawed sperm (P < 0.001), and ROS content was higher (P < 0.001). Moreover, 25 differential protein spots were detected in two-dimensional gels using PDQuest 8.0 software and the corresponding proteins were identified using matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF-TOF MS) coupled with searching of the NCBI protein sequence database. Among these proteins, hexokinase1 (HXK1), the enzyme that catalyzes the first step of glycolysis in the sperm glycolytic pathway, is known to be associated with sperm motility. Casein kinase II subunit alpha (CSNK2A2), a serine/threonine-selective protein kinase, is associated with sperm apoptosis. We used immunoblotting and immunofluorescence to analyze the expression and localization of these two proteins. HXK1 and CSNK2A2 expression levels in fresh sperm were significantly higher than that in freeze-thawed sperm (P < 0.001). HXK1 and CSNK2A2 were detected in the main part of the sperm flagellum, and the immunofluorescence signal from these proteins was weakened in the freeze-thawed group. Decreased expression of HXK1 and CSNK2A2 may be associated with decreased sperm motility and viability following freeze-thawing.

  5. Ram seminal plasma proteins contribute to sperm capacitation and modulate sperm-zona pellucida interaction.

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    Luna, C; Colás, C; Casao, A; Serrano, E; Domingo, J; Pérez-Pé, R; Cebrián-Pérez, J A; Muiño-Blanco, T

    2015-03-01

    Incubation of ram spermatozoa in capacitating conditions with cAMP-elevating agents promotes a progressive time-dependent increase in the capacitated sperm subpopulation. In this study, the fertilizing capacity of ram spermatozoa (ability to bind to the zona pellucida, ZBA rate) capacitated in these conditions was determined. The results showed an increase (P ram spermatozoa. Likewise, the presence of two seminal plasma (SP) proteins able to protect sperm against cold shock (RSVP14 and RSVP20) was evidenced in both SP and the ram sperm surface, and their influence in the fertilizing ability of spermatozoa capacitated in basal medium or with cAMP-elevating agents was determined. The results verified that RSVP14 and RSVP20 act as decapacitating factors given that their addition to SP-free sperm samples previously to capacitation maintained high proportions of the noncapacitated sperm pattern with no increase in protein tyrosine phosphorylation. However, the obtained ZBA rate in the high-cAMP-containing samples was increased in the presence of RSVP20 (P < 0.05). These findings would indicate that the stimulating effect exerted by this protein on the sperm-oocyte binding occurs downstream from the cAMP generation and that the mechanisms by which RSVP20 promotes the zona pellucida binding might be independent of protein tyrosine phosphorylation.

  6. Heterologous recombinant protein with decapacitating activity prevents and reverts cryodamage in ram sperm: An emerging biotechnological tool for cryobiology.

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    Zalazar, L; Ledesma, A; Hozbor, F; Cesari, A

    2016-01-01

    During the last decades fundamental and applied aspects of mammalian ram sperm cryopreservation have been increasingly explored by scientists and biotechnologists. Many works report modifications in the composition of the freezing extenders and explore the beneficial and detrimental effects of seminal plasma or seminal plasma components in cryopreservation. Seminal plasma is known to contain stabilizing proteins, thereby this is a good start point to study the maintenance of membrane stability based on the basic knowledge of sperm physiology. However, seminal plasma composition is variable among rams and also the introduction of exogenous seminal plasma or its fractions to commercial semen can be associated with the transmission of viral diseases. Our work shows that a mouse protein, called SPINK3 (Serine Protease Inhibitor Kazal type 3) with decapacitating activity interacts with heterologous ram sperm when it is produced as a recombinant molecule. By immunocytochemistry assays we demonstrate that this protein (naturally expressed by mouse seminal vesicle under androgenic control) binds to the apical portion of both fresh and frozen ram sperm, the same localization described in mouse homologous sperm. Furthermore, it significantly improves sperm progressive motility compared to non-treated samples when it is added to freezing extenders and to dilution media after thawing. On the contrary, addition of SPINK3 does not modify sperm viability. The percentage of sperm with intact acrosome after ionophore induction was also significantly higher in sperm frozen in the presence of SPINK3 compared to control samples and the addition of SPINK3 after thawing significantly reduced both induced and non induced acrosomal loss, indicating that heterologous SPINK3 might act as a calcium inhibitor transport as described in mouse. Based on our results SPINK3 may find a place as a desirable biotechnological tool to achieve a higher proportion of competent sperm to fertilize.

  7. Architecture of the sperm whale forehead facilitates ramming combat

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    Olga Panagiotopoulou

    2016-04-01

    Full Text Available Herman Melville’s novel Moby Dick was inspired by historical instances in which large sperm whales (Physeter macrocephalus L. sank 19th century whaling ships by ramming them with their foreheads. The immense forehead of sperm whales is possibly the largest, and one of the strangest, anatomical structures in the animal kingdom. It contains two large oil-filled compartments, known as the “spermaceti organ” and “junk,” that constitute up to one-quarter of body mass and extend one-third of the total length of the whale. Recognized as playing an important role in echolocation, previous studies have also attributed the complex structural configuration of the spermaceti organ and junk to acoustic sexual selection, acoustic prey debilitation, buoyancy control, and aggressive ramming. Of these additional suggested functions, ramming remains the most controversial, and the potential mechanical roles of the structural components of the spermaceti organ and junk in ramming remain untested. Here we explore the aggressive ramming hypothesis using a novel combination of structural engineering principles and probabilistic simulation to determine if the unique structure of the junk significantly reduces stress in the skull during quasi-static impact. Our analyses indicate that the connective tissue partitions in the junk reduce von Mises stresses across the skull and that the load-redistribution functionality of the former is insensitive to moderate variation in tissue material parameters, the thickness of the partitions, and variations in the location and angle of the applied load. Absence of the connective tissue partitions increases skull stresses, particularly in the rostral aspect of the upper jaw, further hinting of the important role the architecture of the junk may play in ramming events. Our study also found that impact loads on the spermaceti organ generate lower skull stresses than an impact on the junk. Nevertheless, whilst an impact on the

  8. Effect of two ram sperm capacitating media on acrosome reaction and zona-free hamster oocyte penetration test

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    Silvia Ferrari

    2000-01-01

    Full Text Available An in vitro zona-free hamster oocyte penetration test was utilized in 24 trials in order to evaluate the capacitation media used for ram sperm. A pool of fresh semen was collected from three crossed breed rams. Two semen drops were washed by centrifugation and incubated in high ionic strength treatment (HIS or in a defined medium with HEPES, on heat ewe serum and heparin. After the incubation to promote capacitation, simplified triple-stain technique was used to evaluate the spontaneous acrosome reaction of the capacitated sperm. Superovulation in 96 golden hamsters was induced by PMSG and hCG. The oocytes were treated with hyaluronidase and trypsin to remove, respectively, the cumulus cells and the zona pellucida. Oocytes and capacitated sperm were incubated during 3 hours for further penetration. Then, oocytes were fixed and stained, being evaluated under phase contrast microscope. No significant statistical difference (p >; 0.05 was found between media, concerning the penetration rate of the capacitated sperm and between number of sperm viable with acrosome reaction after the capacitation treatment using two different media. It was concluded that both media utilized were effective in capacitating ram sperm.

  9. Seminal plasma proteins interacting with sperm surface revert capacitation indicators in frozen-thawed ram sperm.

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    Ledesma, Alba; Fernández-Alegre, Estela; Cano, Adriana; Hozbor, Federico; Martínez-Pastor, Felipe; Cesari, Andreína

    2016-10-01

    This study was conducted to evaluate the effects of interacting seminal plasma proteins (iSPP) obtained by AV or EE on frozen-thawed ram sperm in order to test the hypothesis whether this fraction could be sufficient to emulate the effect of complete seminal plasma (SP). Additionally, we evaluated whether these proteins have a differential effect between spermatozoa from high and low fertility rams and between breeding and non-breeding seasons. We assessed sperm motility, quality parameters (intracellular reactive oxygen species, membrane fluidity, plasma membrane permeability and mitochondrial activity) and capacitation status. The main findings from this work were: i) iSPP had no effect on sperm motility, whereas SP (AV or EE) addition produced the highest values of total motility (74.13±2.99 and 72.27±2.99 for AV and EE, respectively) and progressive motility (64.97±2.64 and 63.73±2.64 for AV and EE, respectively); ii) iSPP had no effect on sperm quality parameters (p>0.05), but whole SP improved all parameters evaluated. Moreover, SP collected by AV yielded significantly higher viability (44.60±2.87) and sperm with stable plasma membrane (44.56±2.49) comparing with the addition of SP collected by EE (35.80±2.47 and 36.67±1.71, respectively); iii) iSPP and SP collected by EE, but not by AV, reverted molecular signals of capacitation as protein tyrosine phosphorylation caused by freezing temperatures; iv) there were no effects of fertility or season in sperm quality parameters evaluated. This study demonstrated that, although the iSPP have a clear decapacitating effect, including the ability to revert cryo-capacitation indicators, they are not sufficient to emulate the effects of complete SP regarding sperm functional parameters.

  10. Protective effect of cholesterol-loaded cyclodextrin pretreatment against hydrogen peroxide induced oxidative damage in ram sperm.

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    Naseer, Zahid; Ahmad, Ejaz; Aksoy, Melih; Küçük, Niyazi; Serin, İlker; Ceylan, Ahmet; Boyacıoğlu, Murat; Kum, Cavit

    2015-08-01

    Three experiments were conducted to determine the protective effect of cholesterol-loaded cyclodextrin (CLC) against hydrogen peroxide (H2O2) or cryo-induced damage in ram sperm. In Experiment 1, the fresh ejaculates were either treated with CLC or remained untreated. Both CLC treated and untreated samples were then incubated with 0, 250 or 500 μM H2O2 at 35°C for 12 h. After incubation period of 12 h, the motility, viability and membrane integrity remained higher in CLC treated sperm even in the presence of 250 or 500 μM H2O2. The H2O2 treatment affected all the sperm parameters adversely (P0.05) in MDA level was observed among the groups at any stage of incubation. In conclusion, the CLC incorporation in ram sperm membrane may protects it against H2O2 or cryo-induced oxidative damage. The cryoprotective influence of CLC on ram sperm might be resulted from, at least partly, its antioxidative property.

  11. Induced lipid peroxidation in ram sperm: semen profile, DNA fragmentation and antioxidant status.

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    Hamilton, Thais Rose dos Santos; de Castro, Letícia Signori; Delgado, Juliana de Carvalho; de Assis, Patrícia Monken; Siqueira, Adriano Felipe Perez; Mendes, Camilla Mota; Goissis, Marcelo Demarchi; Muiño-Blanco, Teresa; Cebrián-Pérez, José Álvaro; Nichi, Marcílio; Visintin, José Antonio; D'Ávila Assumpção, Mayra Elena Ortiz

    2016-04-01

    Action of reactive oxygen species, protamination failures and apoptosis are considered the most important etiologies of sperm DNA fragmentation. This study evaluated the effects of induced lipid peroxidation susceptibility on native semen profile and identified the mechanisms involved in sperm DNA fragmentation and testicular antioxidant defense on Santa Ines ram sperm samples. Semen was collected from 12 adult rams (Ovis aries) performed weekly over a 9-week period. Sperm analysis (motility, mass motility, abnormalities, membrane and acrosome status, mitochondrial potential, DNA fragmentation, lipid peroxidation and intracellular free radicals production); protamine deficiency; PRM1, TNP1 and TNP2 gene expression; and determination of glutathione peroxidase (GPx), glutathione reductase, catalase (CAT) and superoxide dismutase activity and immunodetection in seminal plasma were performed. Samples were distributed into four groups according to the sperm susceptibility to lipid peroxidation after induction with ascorbate and ferrous sulfate (low, medium, high and very high). The results were analyzed by GLM test and post hoc least significant difference. We observed an increase in native GPx activity and CAT immunodetection in groups with high susceptibility to induced lipid peroxidation. We also found an increase in total sperm defects, acrosome and membrane damages in the group with the highest susceptibility to induced lipid peroxidation. Additionally, the low mitochondrial membrane potential, susceptible to chromatin fragmentation and the PRM1 mRNA were increased in the group showing higher susceptibility to lipid peroxidation. Ram sperm susceptibility to lipid peroxidation may compromise sperm quality and interfere with the oxidative homeostasis by oxidative stress, which may be the main cause of chromatin damage in ram sperm. © 2016 Society for Reproduction and Fertility.

  12. Use of antioxidants reduce lipid peroxidation and improve quality of crossbred ram sperm during its cryopreservation.

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    Banday, Mohamad Naiem; Lone, Farooz Ahmad; Rasool, Fabiha; Rashid, Muzamil; Shikari, Arif

    2017-02-01

    Ram sperm are subjected to extreme oxidative stress during their preservation at -196 °C resulting in reduced quality at post thaw. Therefore, the main objective of this study was to evaluate the effect of antioxidants taurine, quercetin and reduced glutathione on the post thaw quality of crossbred ram sperm. A total of twenty four ejaculates from six crossbred rams were collected and extended with tris-based extender with no antioxidant (Control), with taurine (40 mM), quercetin (5 μg/ml) and reduced glutathione (5 mM). The post thaw sperm quality was determined by percent sperm motility, live sperm count, intact acrosome and hypo-osmotic swelling test (HOST) reacted spermatozoa and lipid peroxidation was measured in terms of malondialdehyde (MDA) level both in seminal plasma and sperm cell. At post thaw, percent sperm motility and live sperm count were significantly (p  0.05) from quercetin. The percent HOST reacted spermatozoa were significantly higher for taurine than control, quercetin and reduced glutathione. Seminal plasma MDA level was significantly (p  0.05) among the control and antioxidants. In conclusion, taurine at 40 mM reduced lipid peroxidation and improved post thaw sperm quality of cryopreserved crossbred ram semen. Further, transportation time of semen samples in an ice chest at 4-5 °C may be included as a part of equilibration period, when collection shed and frozen semen unit are located at a distance. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Seminal plasma proteins modify the distribution of sperm subpopulations in cryopreserved semen of rams with lesser fertility.

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    Ledesma, Alba; Zalazar, Lucía; Fernández-Alegre, Estela; Hozbor, Federico; Cesari, Andreina; Martínez-Pastor, Felipe

    2017-09-01

    Any physiological mechanism involved in sperm selection and semen improvement has effects on heterogeneous sperm populations. This is mainly due to the fact that sperm populations within a single ejaculate have considerable heterogeneity for many variables, such as motility which is meaningful in terms of understanding how some sperm cells possess fertility advantages as compared with other cells. In the present research, initially there was a multivariate and clustering analysis used to assess sperm motility data from cryopreserved ram semen to identify subpopulations and compare the distribution of these clusters between rams with lesser and greater fertility. There were four classifications made of sperm subpopulations (clusters): CL1 fast/linear/progressive sperm; CL2 fast/non-linear sperm; CL3 very fast/linear sperm with vigorous beating and CL4 slow/non-linear sperm. Rams with greater fertility had a lesser proportion of sperm considered as "hyperactivated" (CL2) and a greater proportion of slow and non-linear sperm (CL4) than sperm of rams with lesser fertility. In addition, the effects were assessed for the capacity of seminal plasma (SP) and interacting SP proteins (iSPP) that were present during different seasons of the year to improve the distribution of sperm within subpopulations of semen from rams with lesser fertility. The iSPP and SP were obtained by artificial vagina (AV) and electroejaculation (EE) during breeding and non-breeding seasons and added to thawed semen. All the aggregates had a significant effect on the distribution of sperm subpopulations and effects differed among seasons of the year and depending on collection method used. Even though, future studies are needed to assess the contribution of each subpopulation on ram sperm fertility, it is important that a multivariate analysis be used to evaluate the effect of a treatment on sperm quality variables. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. ANALYSIS OF PLASMA MEMBRANE INEGRITY BY USING OF FLUORESCEIN LABELED ANNEXIN V AND ITS RELATIONSHIP TO MOTILITY AND VIABILITY OF RAM SPERM AFTER CRYOPRESERVATION

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    Maria IVANOVA-KICHEVA

    2006-05-01

    Full Text Available The present study was conducted to detect sperm apoptosis in fresh, capacitated and frozen ram semen and to determine its relationship with other seminal parameters as motility and viability. The cryopreservation process induces the plasma membrane disruptions, attended with increasing of phoshpatidylserine molecules expression on the extracellular surface, wherefore some of the live spermatozoa loss their biological potential to fertilize the egg.

  15. Intracytoplasmic Sperm Injection Outcomes with Freshly Ejaculated Sperms and Testicular or Epididymal Sperm Extraction in Patients with Idiopathic Cryptozoospermia

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    Ketabchi

    2016-01-01

    Background Cryptozoospermia (CO) is a situation in which spermatozoa cannot be observed in a fresh semen sample unless an extended centrifugation and microscopic search are performed. CO patients are suggested to use only intracytoplasmic sperm injection (ICSI) as infertility treatment. But still there is debate about the choice of sperm source in cryptozoospermic men candidate for ICSI. Objectives This study was conducted to eval...

  16. Effect of different extenders on ram sperm traits during storage

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    Zdeňka Hegedűšová

    2012-01-01

    Full Text Available The aim of the study was to test commercial extenders used for short-term and long-term sperm preservation. Semen was collected in the reproduction season, i.e. from June to December. The ejaculates were obtained from single services and the routine analysis of the semen was performed immediately after the collection. The examination included semen volume, colour and texture, sperm concentration and motility, ejaculate turbulence and percentage of sperm with abnormal morphology. The semen was diluted with an extender in the ratio of 1:4. The processed semen was transported in an insulated container at 16–18 °C to the laboratory and stored in a stationary thermostat under the same temperature. Sperm motility tests were performed 24, 48, 72 and 96 hours after the placement in to thermostat. Ejaculates diluted with Ovipro, Optidyl, Triladyl and Andromed CSS gave very good results of viability (81.23 %–83.41 % after 24 hours of storage. After 48 hours, Ovipro, Andromed, Optidyl and Triladyl gave values above 75 %. The Triladyl extender proved to be a good stabilizing agent, showing consistent results during a long-term storage. It was chosen as a control one for overall assessment. Other preservation media did not show any improving or worsening effects. The extender Ovipro showed a high motility effect in the first 48 hours only, and hence it appears to be the best solution for the short-term preservation.

  17. Effects of diluting medium and holding time on sperm motility analysis by CASA in ram.

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    Mostafapor, Somayeh; Farrokhi Ardebili, Farhad

    2014-01-01

    The aim of this study was to evaluate the effects of dilution rate and holding time on various motility parameters using computer-assisted sperm analysis (CASA). The semen samples were collected from three Ghezel rams. Samples were diluted in seminal plasma (SP), phosphate-buffered saline (PBS) containing 1% bovine serum albumin (BSA) and Bioexcell. The motility parameters that computed and recorded by CASA include curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), straightness (STR), linearity (LIN), amplitude of lateral head displacement (ALH), and beat cross frequency (BCF). In all diluters, there was a decrease in the average of all three parameters of sperms movement velocity as the time passed, but density of this decrease was more intensive in SP. The average of ALH between diluters indicated a significant difference, as it was more in Bioexcell in comparison with the similar amount in SP and PBS. The average of LIN in the diluted sperms in Bioexcell was less than two other diluters in all three times. The motility parameters of the diluted sperms in Bioexcell and PBS indicated an important and considerable difference with the diluted sperms in SP. According to the gained results, the Bioexcell has greater ability in preserving motility of sperm in comparison with the other diluters but as SP is considered as physiological environment for sperm. It seems that the evaluation of the motility parameters in Bioexcell and PBS cannot be an accurate and comparable evaluation with SP.

  18. D-penicillamine prevents ram sperm agglutination by reducing the disulphide bonds of a copper-binding sperm protein.

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    Leahy, T; Rickard, J P; Aitken, R J; de Graaf, S P

    2016-05-01

    Head-to-head agglutination of ram spermatozoa is induced by dilution in the Tyrode's capacitation medium with albumin, lactate and pyruvate (TALP) and ameliorated by the addition of the thiol d-penicillamine (PEN). To better understand the association and disassociation of ram spermatozoa, we investigated the mechanism of action of PEN in perturbing sperm agglutination. PEN acts as a chelator of heavy metals, an antioxidant and a reducing agent. Chelation is not the main mechanism of action, as the broad-spectrum chelator ethylenediaminetetraacetic acid and the copper-specific chelator bathocuproinedisulfonic acid were inferior anti-agglutination agents compared with PEN. Oxidative stress is also an unlikely mechanism of sperm association, as PEN was significantly more effective in ameliorating agglutination than the antioxidants superoxide dismutase, ascorbic acid, α-tocopherol and catalase. Only the reducing agents cysteine and DL-dithiothreitol displayed similar levels of non-agglutinated spermatozoa at 0 h compared with PEN but were less effective after 3 h of incubation (37 °C). The addition of 10 µM Cu(2+) to 250 µM PEN + TALP caused a rapid reversion of the motile sperm population from a non-agglutinated state to an agglutinated state. Other heavy metals (cobalt, iron, manganese and zinc) did not provoke such a strong response. Together, these results indicate that PEN prevents sperm association by the reduction of disulphide bonds on a sperm membrane protein that binds copper. ADAM proteins are possible candidates, as targeted inhibition of the metalloproteinase domain significantly increased the percentage of motile, non-agglutinated spermatozoa (52.0% ± 7.8) compared with TALP alone (10.6% ± 6.1).

  19. Virtual azoospermia and cryptozoospermia--fresh/frozen testicular or ejaculate sperm for better IVF outcome?

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    Hauser, Ron; Bibi, Guy; Yogev, Leah; Carmon, Ariella; Azem, Foad; Botchan, Amnon; Yavetz, Haim; Klieman, Sandra E; Lehavi, Ofer; Amit, Ami; Ben-Yosef, Dalit

    2011-01-01

    Men diagnosed as having azoospermia occasionally have a few mature sperm cells in other ejaculates. Other men may have constant, yet very low quality and quantity of sperm cells in their ejaculates, resulting in poor intracytoplasmic sperm injection (ICSI) outcome. It has not been conclusively established which source of sperm cells is preferable for ICSI when both ejaculate and testicular (fresh or frozen) sperm cells are available. It is also unclear whether there is any advantage of fresh over frozen sperm if testicular sperm is to be used. We used ejaculate, testicular (fresh or frozen) sperm cells, or both for ICSI in 13 couples. Five of these couples initially underwent ICSI by testicular sperm extraction, because the males had total azoospermia, and in later cycles with ejaculate sperm cells. Ejaculate sperm cells were initially used for ICSI in the other 8 patients, and later with testicular sperm cells. The fertilization rate was significantly higher when fresh or frozen-thawed testicular sperm cells were used than when ejaculated sperm cells were used. Likewise, the quality of the embryos from testicular (fresh and frozen) sperm was higher than from ejaculated sperm (65.3% vs 53.2%, respectively, P cryptozoospermia because of their superior fertility.

  20. Assessment of chromatin status (SCSA) in epididymal and ejaculated sperm in Iberian red deer, ram and domestic dog.

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    Garcia-Macias, Vanesa; Martinez-Pastor, Felipe; Alvarez, Mercedes; Garde, Jose Julian; Anel, Enrique; Anel, Luis; de Paz, Paulino

    2006-11-01

    Abnormal chromatin condensation is not detected using classical techniques for sperm analysis. SCSA has demonstrated its usefulness in sperm chromatin analysis in several species (human, bull, stallion and boar). In this work, we studied sperm samples from red deer, ram and dog to analyze the differentiation of chromatin structure applying SCSA in epididymal and ejaculated spermatozoa. Epididymal samples were obtained from the caput, corpus and cauda by means of cuts, and ejaculated ones were obtained by electroejaculation (deer), artificial vagina (ram) and digital manipulation (dog). SCSA results suggested different critical points in sperm maturation (spermatozoa with loose chromatin to more condensed chromatin) among species: from corpus to cauda in ram and from caput to corpus in deer and dog. Moreover, we also detected differences in ruminants and dog, reflected in the appearance of SCSA plots. Indeed, ram and deer samples rendered two peaks within the sperm main population (sperm with condensed chromatin), whereas only one was detected in dog. Although some differences were observed between cauda and ejaculated samples, SCSA parameters indicated good chromatin condensation, making these samples suitable for germplasm banking. Some species-dependent modifications in the analysis of the results may be necessary to take full advantage of its analytical power.

  1. Effects of Hoechst33342 staining on the viability and flow cytometric sex-sorting of frozen-thawed ram sperm.

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    Quan, Guo Bo; Ma, Yuan; Li, Jian; Wu, Guo Quan; Li, Dong Jiang; Ni, Yi Na; Lv, Chun Rong; Zhu, Lan; Hong, Qiong Hua

    2015-02-01

    Cytometric sorting of frozen-thawed sperm can overcome difficulties caused by the unavailability of sorting facilities on farms where semen is collected from male livestock. In order to optimize the cytometric sex-sorting procedure, effects of Hoechst33342 staining on the viability and cytometric sorting efficiency of frozen-thawed ram sperm were evaluated. The frozen-thawed sperm were stained with Hoechst33342 at various dye concentrations (80 μM, 120 μM, 160 μM, 200 μM, 240 μM, or 320 μM) for 45 min to evaluate effects of dye dose. The frozen-thawed sperm were stained with 160 μM Hoechst33342 for various durations (0 min, 15 min, 30 min, 45 min, 60 min, 75 min, or 90 min) to evaluate effects of staining duration. Sperm motility and moving velocity were analyzed using a computer-assisted sperm analysis system (CASAS). Acrosome status, membrane integrity, and distribution of phosphatidylserine (PS) in Hoechst33342-stained sperm were analyzed using flow cytometry after staining with fluorescein isothiocyanate-labeled lectin from pisum sativum (FITC-PSA), Annexin V, or propidium iodide (PI). The fertility of Hoechst33342-stained sperm was analyzed by in vitro fertilization (IVF). A high-speed cell sorter was used to evaluate effects of Hoechst33342 staining on cytometric sex-sorting of frozen-thawed sperm. The motility, moving velocity, membrane integrity, and PS distribution of Hoechst33342-stained sperm were significantly different from that of immediately thawed sperm (Pram sperm. Results of cytometric sorting indicated that frozen-thawed sperm can be efficiently sorted into two sperm populations with X and Y chromosome when the Hoechst33342 concentration was 160 μM. Moreover, when the staining duration was equal to or longer than 45 min, the frozen-thawed sperm can be successfully sorted in the presence of 160μM Hoechst33342. In conclusion, Hoechst33342 staining can detrimentally influence viability of frozen-thawed ram sperm except acrosome and in vitro

  2. Protective effects of Opuntia ficus-indica extract on ram sperm quality, lipid peroxidation and DNA fragmentation during liquid storage.

    Science.gov (United States)

    Allai, Larbi; Druart, Xavier; Öztürk, Mehmet; BenMoula, Anass; Nasser, Boubker; El Amiri, Bouchra

    2016-12-01

    The present study aimed to assess the phenolic composition of the acetone extract from Opuntia ficus indica cladodes (ACTEX) and its effects on ram semen variables, lipid peroxidation and DNA fragmentation during liquid storage at 5°C for up to 72h in skim milk and Tris egg yolk extenders. Semen samples from five rams were pooled extended with Tris-egg yolk (TEY) or skim milk (SM) extenders containing ACTEX (0%, 1%, 2%, 4% and 8%) at a final concentration of 0.8×10(9) sperm/ml and stored for up to 72h at 5°C. The sperm variables were evaluated at different time periods (8, 24, 48 and 72h). Sperm total motility and viability were superior in TEY than in SM whereas the progressive motility, membrane integrity, abnormality and spontaneous lipid peroxidation were greater in SM compared to TEY (PDNA fragmentation (P<0.05). In conclusion, SM and TEY supplemented with 1% of ACTEX can improve the quality of ram semen. Further studies are required to identify the active components in ACTEX involved in its effect on ram sperm preservation. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Fatty Acid Prof iles of Ram's Sperm after Removing Some Fatty Acid Sources from the Diets and Persistency of Fatty Acids in Sperm

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    Morteza Zarrabi

    2012-01-01

    Full Text Available Background: Mammalian spermatozoa are characterized by a high proportion of polyunsaturatedfatty acids (PUFAs, but reliable data concerning dietary effects on fatty acid (FA profile in ram'ssperm and the persistency of FA in the ration to the FA in sperm has not been reported. Therefore,the aim of this study was to determine the stability of saturated and unsaturated FAs in ram's spermdespite removing FA sources from their diet.Materials and Methods: Nine Kalkoohi rams were used in a completely randomized design and theywere assigned to 3 groups. The treatments were diet supplemented (35 g/d/ram by C16:0 (RP-10®,C18: 2 (Sunflower oil; SO and n-3 (Fish oil; FO with Vitamin E. Fifteen weeks after the start of thesupplemented diet, rams were offered a basal diet without any supplementary FA source for 35 dayswhen the sperm’s FA ratio was determined. The data were analyzed by ANOVA (Analysis of varianceusing the General Linear Model (GLM procedure of SAS Institute.Results: Thirty five days after removing the fat supplement from the diet, major FA in sperm consisted of:C14:0, C16:0, C18:0, C18:1 cis, C18:2 cis and C22:6 n-3 docosahexaenoic acid (DHA. The percentageof C14:0 (p=0.8 and C18:1 cis (P =0.4 were similar among all the treatments. Interestingly, 35 daysafter the removal of fatty acid source, the percentage of C22:6 was highest in the FO treated group.Conclusion: The different sperm FA profile among various groups suggests that dietary FA hadsignificant direct or indirect impacts on sperm FA profile after 35 days which might lead to physicaland chemical changes in sperm characteristics.

  4. Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm.

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    Pelufo, V; López Armengol, M F; Malcotti, V; Venturino, A; Aisen, E G

    2015-01-01

    Sperm deep freezing procedures for ram semen have considerable variations regarding the steps being employed for cooling, freezing, and addition of cryoprotectants. In this work, we evaluated the effects of the addition of glycerol and/or the disaccharides sucrose and trehalose to hypertonic diluents either before or after cooling from 30 °C to 5 °C in Merino Australian ram semen cryopreservation. Using optical and transmission electron microscopy techniques, we assessed that glycerol was beneficial to the cooling process independently of its addition at 30 °C or 5 °C in terms of sperm membrane integrity in different regions of the plasma membrane (acrosomal region, 14.5% higher integrity; postacrosomal region, 8.0% higher integrity [P ram semen cryopreservation in hypertonic disaccharide-containing diluents is the addition of glycerol and trehalose after the cooling process, at 5 °C.

  5. Helium-neon laser irradiation of cryopreserved ram sperm enhances cytochrome c oxidase activity and ATP levels improving semen quality.

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    Iaffaldano, N; Paventi, G; Pizzuto, R; Di Iorio, M; Bailey, J L; Manchisi, A; Passarella, S

    2016-08-01

    This study examines whether and how helium-neon laser irradiation (at fluences of 3.96-9 J/cm(2)) of cryopreserved ram sperm helps improve semen quality. Pools (n = 7) of cryopreserved ram sperm were divided into four aliquots and subjected to the treatments: no irradiation (control) or irradiation with three different energy doses. After treatment, the thawed sperm samples were compared in terms of viability, mass and progressive sperm motility, osmotic resistance, as well as DNA and acrosome integrity. In response to irradiation at 6.12 J/cm(2), mass sperm motility, progressive motility and viability increased (P < 0.05), with no significant changes observed in the other investigated properties. In parallel, an increase (P < 0.05) in ATP content was detected in the 6.12 J/cm(2)-irradiated semen samples. Because mitochondria are the main cell photoreceptors with a major role played by cytochrome c oxidase (COX), the COX reaction was monitored using cytochrome c as a substrate in both control and irradiated samples. Laser treatment resulted in a general increase in COX affinity for its substrate as well as an increase in COX activity (Vmax values), the highest activity obtained for sperm samples irradiated at 6.12 J/cm(2) (P < 0.05). Interestingly, in these irradiated sperm samples, COX activity and ATP contents were positively correlated, and, more importantly, they also showed positive correlation with motility, suggesting that the improved sperm quality observed was related to mitochondria-laser light interactions.

  6. Short communication. Evaluation of a commercial kit based on acridine orange/propidium iodide to assess the plasma membrane integrity of ram sperm

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    J. L. Yániz

    2013-03-01

    Full Text Available This study was designed to develop a semiautomatic computer assisted methodology to evaluate the membrane integrity of ram spermatozoa using a commercial kit based on acridine orange/propidium iodide (AO/PI labelling and ImageJ software. The study was divided into two experiments. In the first trial, the new computer-assisted method was validated by mixing fresh semen samples with different volumes of freeze killed spermatozoa to determine proportions of damaged spermatozoa in the final samples. The proportion of damaged spermatozoa in each sample determined by the automated procedure where highly correlated (R2=0.97, p<0.001 with the predicted theoretical values. In the second trial, the new method was compared with a previously validated method of membrane integrity assessment based on phase-contrast/propidium iodide (PH/PI methodology. Measurements by AO/PI were, on average, 4.0% larger than measurements by PH/PI (SD=7.02% and 1.79% smaller than measurements of sperm motility determined by CASA (SD=4.83. The AO/PI method was also more repeatable than the PH/PI. The double staining methodology coupled with the routine for image analysis allowing automatic determination of sperm membrane integrity means a reduction in processing time of 75% compared to the previously developed method using a single fluorochrome (3 vs 12 min on average if the incubation period was included. This facilitates its use when a large number of samples are analysed. Our results validate the new computer assisted method for assessing sperm membrane integrity in sheep. The new method developed, in addition to being a free tool, allows quick automatic determination of sperm viability, which facilitates its use in routine semen analysis.

  7. Evaluation of Lasting Effects of Heat Stress on Sperm Profile and Oxidative Status of Ram Semen and Epididymal Sperm

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    Thais Rose dos Santos Hamilton

    2016-01-01

    Full Text Available Higher temperatures lead to an increase of testicular metabolism that results in spermatic damage. Oxidative stress is the main factor responsible for testicular damage caused by heat stress. The aim of this study was to evaluate lasting effects of heat stress on ejaculated sperm and immediate or long-term effects of heat stress on epididymal sperm. We observed decrease in motility and mass motility of ejaculated sperm, as well as an increase in the percentages of sperm showing major and minor defects, damaged plasma and acrosome membranes, and a decrease in the percentage of sperm with high mitochondrial membrane potential in the treated group until one spermatic cycle. An increased enzymatic activity of glutathione peroxidase and an increase of stressed cells were observed in ejaculated sperm of the treated group. A decrease in the percentage of epididymal sperm with high mitochondrial membrane potential was observed in the treated group. However, when comparing immediate and long-term effects, we observed an increase in the percentage of sperm with low mitochondrial membrane potential. In conclusion, testicular heat stress induced oxidative stress that led to rescuable alterations after one spermatic cycle in ejaculated sperm and also after 30 days in epididymal sperm.

  8. Ethylene glycol, but not DMSO, could replace glycerol inclusion in soybean lecithin-based extenders in ram sperm cryopreservation.

    Science.gov (United States)

    Najafi, Abouzar; Daghigh-Kia, Hossein; Dodaran, Hossein Vaseghi; Mehdipour, Mahdieh; Alvarez-Rodriguez, Manuel

    2017-02-01

    The aim of this study was to evaluate the effects of glycerol, ethylene glycol or DMSO in a soybean lecithin extender for freezing ram semen. In this study, 20 ejaculates were collected from four Ghezel rams and diluted with soybean lecithin extender with glycerol (7%), ethylene glycol (3%, 5% and 7%) or DMSO (3%, 5% and 7%). Sperm motility (CASA), membrane integrity (HOS test), viability, total abnormality, mitochondrial activity (Rhodamine 123) and apoptotic features (Annexin V/Propidium iodide) were assessed after thawing. There was no significant difference between glycerol and ethylene glycol at different concentrations (3% and 5%) regarding sperm total and progressive motility, viability, and membrane integrity. The least percentages of mitochondrial functionality were observed in samples frozen with all different DMSO concentrations tested (Pethylene glycol could be a desirable substitute of glycerol in the freezing extender, in view of similar results obtained in post-thaw quality of ram semen cryopreserved in a soybean lecithin extender. We propose that glycerol in a soybean lecithin based extender could be replaced by ethylene glycol at 3% or 5% concentrations. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. THE PROPORTION OF X AND Y SPERM, VIABILITY AND MOTILITY OF RAM SPERMATOZOA AFTER SEPARATED WITH WHITE EGG ALBUMIN

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    Moh Takdir

    2017-02-01

    The aim of this research was to determine the proportion, viability and motility of X and Y ram spermatozoa separated with egg white albumin. Sperm samples derived from Garut ram, which was collected by using an artificial vagina. Observations were made on spermatozoa fraction above and below each medium fraction treatment. There are treatment egg white albumin as separation medium, each medium consisting of fractions top and bottom fraction with different concentration: 1 P0 = sperma before separation (control; 2 P1 = 10% above fraction + 30% lower fraction; P2 = 25% + 45%; P3 = 25% + 75%. Data proportion of X and Y, viability and motility were analyzed statistically by Completely Randomized Design patern in the direction followed by Duncan’s Multiple Range Test for data with a real difference. Separation with egg white albumin affect significantly increased the proportion of spermatozoa X and Y (P≤0.05, but tends to decrease the viability and motility of spermatozoa.The proportion of spermatozoa X and Y was highest in treatment P3,76.76% of spermatozoa X (fraction above 25% and 79.81% spermatozoa Y (75% lower fraction, with an average viability obtained respectively 68,9% (fraction above and 59,7% (bottom fraction, motility 77,5% (fraction above dan 84,0% (bottom fraction. It was concluded that the egg white albumin is very effective in changing the proportions of X and Y ram sperm with the quality of spermatozoa after separation feasible for applications insemination or processed into frozen semen.   (Keywords: Garut ram, White egg albumin, Spermatozoa X and Y

  10. Effect of semen extender and storage temperature on ram sperm motility over time

    Science.gov (United States)

    Storage of ram semen for long period of time depends on a number of factors, including type of extender and storage temperature. A study compared the effect of semen extender and storage temperature on motility of ram semen stored for 72 h. Semen collected via electroejaculator from 5 mature Katahd...

  11. Antioxidant effect of rosemary (Rosmarinus officinalis L.) extract in soybean lecithin-based semen extender following freeze-thawing process of ram sperm.

    Science.gov (United States)

    Motlagh, Mahdi Khodaei; Sharafi, Mohsen; Zhandi, Mahdi; Mohammadi-Sangcheshmeh, Abdollah; Shakeri, Malak; Soleimani, Masoud; Zeinoaldini, Saeed

    2014-10-01

    The aim of current study was to evaluate effect of rosemary aqueous extract on post-thawed ram sperm quality in a soybean lecithin-based (SL) extender. Ram semen samples were obtained, extended with SL extender and supplemented with 0% (SL-R0), 2% (SL-R2), 4% (SL-R4), 6% (SL-R6), and 8% (SL-R8) rosemary aqueous extract. Following equilibration, the straws were frozen, and then plunged into the liquid nitrogen. After thawing, sperm motility and velocity parameters, plasma membrane functionality, viability, acrosomal and capacitation status were evaluated. Membrane lipid peroxidation was also analyzed through the malondialdehyde (MDA) concentration. Our results showed that SL-R4 and SL-R6 groups resulted in higher (p 0.05) by different levels of rosemary aqueous extract. Lower (p ram sperm quality in a dose dependent manner.

  12. Refrigerated storage of ram sperm in presence of Trolox and GSH antioxidants: effect of temperature, extender and storage time.

    Science.gov (United States)

    Mata-Campuzano, María; Álvarez-Rodríguez, Manuel; Tamayo-Canul, Julio; López-Urueña, Elena; de Paz, Paulino; Anel, Luis; Martínez-Pastor, Felipe; Álvarez, Mercedes

    2014-12-30

    Antioxidants have a potential to improve the quality and fertility of refrigerated-stored ram semen. Reduced glutathione (GSH) and Trolox (0.2, 1 and 5mM) were evaluated in ram semen preserved at 15 and 5°C up to 48 and 96h, respectively. Extenders were also evaluated (15°C: Tris-citrate-fructose, TCF, without lipids, and TES-Tris-fructose 10% egg yolk, TTF-EY; 5°C: TTF-EY and 3.5% soybean lecithin, TTF-SL; INRA96 at both temperatures). Storage at 5°C resulted in poorer quality than 15°C up to 48h, while allowing acceptable quality at 96h. Antioxidants had few effects on sperm quality, with use of Trolox resulting in reduced motility and viability in TCF. Storage at 15°C in the TCF extender resulted in decreased motility, viability and mitochondrial activity compared with use of TTF-EY. Sperm quality when storage was at 5°C was similar, but storage in TTF-SL resulted in decreased motility and mitochondrial activity. Acrosomal status was only slightly affected by extender and antioxidant. Mitochondrial activity was improved by antioxidants in TTF-SL, and GSH at 5mM when semen was stored at 5°C in TTF-EY. A preliminary artificial insemination trial indicated that supplementation with GSH has the potential for improving lambing (Pram semen. Use of Trolox negatively impacted sperm quality and GSH had some positive impacts. The use of soybean lecithin requires further research to assess its impact on mitochondria.

  13. Use of Relief Contrast(®) objective to improve sperm morphometric analysis by Isas(®) casa system in the ram.

    Science.gov (United States)

    Yániz, J L; Capistrós, S; Vicente-Fiel, S; Soler, C; Núñez de Murga, M; Santolaria, P

    2013-12-01

    The aim of this study was to develop a new method for morphometric assessment of the sperm head and acrosome in the ram. Ejaculates from 10 adult males were collected using an artificial vagina. For each ejaculate, 10 semen smears were prepared, air-dried and divided (in pairs) into the following five treatment groups: (i) washed in distilled water and allowed to dry without further processing (DRY); (ii) fixed in 50% methanol (MET); (iii) fixed in 2% glutaraldehyde (GLUT); (iv) fixed and stained with Hemacolor(®) (HEM) and (v) fixed and stained with SpermBlue(®) (SB). The prepared slides were examined with a 40 × Relief Contrast(®) objective (RC) and processed with ISAS(®) commercial software. The use of RC optics increased the contrast between acrosome and sperm head, allowing capture and morphometric analysis by ISAS of sperm heads and the acrosome, even in non-stained samples. MET and GLUT groups resulted in a lower number of acceptable, that is, correctly delineated, sperm heads than those in the SB, and SB and HEM groups, respectively (p  HEM≥ MET≥ SB> DRY. When studying the proportion of the sperm head covered by the acrosome, the relation between treatments was: MET> DRY = GLUT = SB> HEM. It was concluded that the new method for sperm morphometric assessment allows the simultaneous assessment of sperm head and acrosome in the ram by the first time, even in unprocessed semen smears.

  14. Sperm cell population dynamics in ram semen during the cryopreservation process.

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    Manuel Ramón

    Full Text Available BACKGROUND: Sperm cryopreservation has become an indispensable tool in biology. Initially, studies were aimed towards the development of efficient freezing protocols in different species that would allow for an efficient storage of semen samples for long periods of time, ensuring its viability. Nowadays, it is widely known that an important individual component exists in the cryoresistance of semen, and efforts are aimed at identifying those sperm characteristics that may allow us to predict this cryoresistance. This knowledge would lead, ultimately, to the design of optimized freezing protocols for the sperm characteristics of each male. METHODOLOGY/PRINCIPAL FINDINGS: We have evaluated the changes that occur in the sperm head dimensions throughout the cryopreservation process. We have found three different patterns of response, each of one related to a different sperm quality at thawing. We have been able to characterize males based on these patterns. For each male, its pattern remained constant among different ejaculates. This latter would imply that males always respond in the same way to freezing, giving even more importance to this sperm feature. CONCLUSIONS/SIGNIFICANCE: Changes in the sperm head during cryopreservation process have resulted useful to identify the ability of semen of males for freezing. We suggest that analyses of these response patterns would represent an important tool to characterize the cryoresistance of males when implemented within breeding programs. We also propose follow-up experiments to examine the outcomes of the use of different freezing protocols depending on the pattern of response of males.

  15. CRYOPRESERVATION OF RAM SPERM FROM AUTOCHTHONOUS BREEDS DURING A NON-MATING SEASON

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    Milko SABEV

    2007-07-01

    Full Text Available It is possible to collect and successfully cryopreserve ejaculates in a non-mating season from rams of the autochthonous breeds Karakachan, Cooper-red Shumen and Karnobat-local, raised in Bulgaria. Studies are in progress aiming the elaboration of optimal cryoprotective extenders and freezing technology.

  16. The identification of proteomic markers of sperm freezing resilience in ram seminal plasma.

    Science.gov (United States)

    Rickard, J P; Leahy, T; Soleilhavoup, C; Tsikis, G; Labas, V; Harichaux, G; Lynch, G W; Druart, X; de Graaf, S P

    2015-08-03

    The source and composition of seminal plasma has previously been shown to alter the ability of spermatozoa to survive cryopreservation. In the present study, the ionic and proteomic composition of seminal plasma from rams with high (HSP; n = 3) or low (LSP; n = 3) freezing resilient spermatozoa was assessed. 75 proteins were identified to be more abundant in HSP and 48 proteins were identified to be more abundant in LSP. Individual seminal plasma proteomes were established for each of the six rams examined. For each ram, correlations were conducted between previously recorded freezing resilience [1] and individual spectral counts in order to identify markers of freezing resilience. 26S proteasome complex, acylamino acid releasing enzyme, alpha mannosidase class 2C, heat shock protein 90, tripeptidyl-peptidase 2, TCP-1 complex, sorbitol dehydrogenase and transitional endoplasmic reticulum ATPase were found to be positively correlated (r(2) > 0.7) with freezing resilience. Cystatin, zinc-2-alpha glycoprotein, angiogenin-2-like protein, cartilage acidic protein-1, cathepsin B and ribonuclease 4 isoform 1 were found to be negatively correlated (r(2) > 0.7) with freezing resilience. Several negative markers were found to originate from the accessory sex glands, whereas many positive markers originated from spermatozoa and were part of or associated with the 26S proteasome or CCT complex.

  17. Seminal and spermatic characteristics of fresh semen and the effects of sperm cooling in Steindachneridion melanodermatum (Garavello, 2005

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    Ronan Maciel Marcos

    2015-12-01

    Full Text Available This study describes the seminal and spermatic characteristics of fresh semen of Steindachneridion melanodermatum and investigates the effects of dilution, temperature, and storage period on its spermatic parameters. Sperm samples were collected from nine hormonally-induced males. The following parameters in fresh sperm were analyzed: seminal plasma osmolality (OSM, seminal pH, sperm motility (MOT, sperm velocity (SV (including sperm curvilinear velocity (CVV, sperm straight-line velocity (SLV, and sperm average path velocity (APV, total time of sperm motility (TEMP, sperm concentration (CONC, and index of sperm normality (NORM. Sperm samples from each male were diluted in a solution containing 5% fructose and 5% powdered milk, and stored at 10°C and 25°C. The same was carried out for sperm samples not subjected to dilution. From these samples, MOT, CVV, SLV, APV, SV, and TEMP were measured after 0 h, 5 h, 9 h, 18 h, 27 h, 36 h, 45 h, and 54 h. Males released 11.74 ± 5.38 mL of sperm, with an osmolality of 258.78 ± 29.36 mOsm.kg-1 and pH of 7.11 ± 0.31. The sperm presented a MOT of 99.86 ± 0.31% at a concentration of 1.03 × 1010 ± 3.65 × 109 spermatozoa.mL-1 with CVV of 185.58 ± 14.11 ?m.s-1, SLV of 49.15 ± 4.66 ?m.s-1, APV of 87.02 ± 4.13 ?m.s-1, SV of 106.52 ± 4.45 ?m.s-1, TEMP of 79.31 ± 5.62 s, NORM of 75.81 ± 5.71%. The results indicate that sperm motility, sperm velocity, and total time of sperm activation were affected by dilution, storage temperature, and storage period (p < 0.05. Procedures for semen storage should be performed with undiluted sperm cooled at 10°C, or kept undiluted at 25°C for up to 27 h.

  18. Peroxisome proliferator-activated receptors (PPARα, PPARγ and PPARβ/δ) gene expression profile on ram spermatozoa and their relation to the sperm motility.

    Science.gov (United States)

    Kadivar, Ali; Heidari Khoei, Heidar; Hassanpour, Hossein; Ghanaei, Hamid; Golestanfar, Arefeh; Mehraban, Hossein; Davoodian, Najmeh; Dehghani Tafti, Roohollah

    2016-01-01

    Peroxisome proliferator-activated receptors (PPARs) are a member of nuclear receptors superfamily, which mainly regulate the expression of target genes involved in lipid and energy metabolism. These receptors are divided to three isotypes: PPARα, PPARγ and PPARβ/δ. Each isotype has a distinct tissue distribution relating to the distinct functions. In this study, the mRNA abundance for PPARα, PPARγ and PPARβ/δ was evaluated and compared with high and low motile ram spermatozoa. Semen samples from 6 adult rams were fractionated on a two layer discontinuous Percoll gradient to high and low motile sperm and quantitative parameters of sperm motility were determined by CASA. Total RNA was extracted and the mRNA abundance for each gene was measured by relative quantification technique with Real time PCR. The levels of three isotypes of PPAR transcripts were significantly higher in high motile semen samples using quantitative RT-PCR. Some of sperm motility indices were also significantly correlated with PPARα and PPARγ relative expression. This study revealed the novel association of PPAR gene isotypes with sperm motility. Data from our study suggested PPARs are one of the possible factors that can be studied in male infertility.

  19. Characteristics of reproductive performance of Garut rams

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    Muhammad Rizal

    2003-06-01

    Full Text Available Basic information on reproductive potency of Garut rams is necessary in order to identify the capacity of rams in producing chilled or frozen semen. Eight Garut rams (three to five years old were used in this study. The male sexual behaviors were observed and semen was collected once a week using artificial vagina. Semen quality was evaluated and its potency to produce frozen semen was calculated. Results of this study indicated that first, second, and third ejaculations were at the 29, 87 and 176th seconds, respectively. Fresh semen volume, sperm concentration, motility, intact acrosomal cap, and intact plasma membrane were 0.99 ml, 3224 million/ml; 76.67; 86.13 and 87.73%, respectively. Protein value, fructose, vitamin C, vitamin E, sodiu, potassium, calcium, magnesium, phosphor, chloride, and mangan in seminal plasma of fresh semen were 4140, 180, 3.2, 24, 180, 117, 9, 6.12, 60, 104, and 5 mg/ml, respectively. Measurement of head length, width, and length of sperm tail were 6.59, 3.99, and 42.65 μm, respectively. Length and width measurement of right and left testes, and scrotal circumference were 12.71, 6.5, and 32.36 cm, respectively. Capacity of each Garut rams to produce frozen semen from three consecutive ejaculations are 35.88 mini straw with the cencentration of 200 million motile sperm per 0.25 ml.

  20. Effects of supplemental conjugated linoleic acids (CLA) on fresh and post-thaw sperm quality of Holstein bulls.

    Science.gov (United States)

    Karimi, R; Towhidi, A; Zeinoaldini, S; Rezayazdi, K; Mousavi, M; Safari, H; Martinez-Pastor, F

    2017-02-07

    This study was designed to investigate the effects of feeding-protected conjugated linoleic acid (CLA) on the semen production and sperm freezability in Holstein bulls. Twelve bulls were randomly assigned to two groups (n = 6 per group). Bulls received the normal diet (control group) or the normal diet top-dressed with 50 g of CLA (treated group) for 10 weeks. The control group received 40 g/day calcium soap of fatty acid. Fresh and post-thaw semen quality was assessed on ejaculates collected at the 0, 4, 6, 8 and 10 week of supplementation. Semen evaluations including sperm concentration, motion characteristics (subjective and computer-assisted), viability (Eosin-Nigrosin), membrane integrity (hypo-osmotic swelling test) and abnormality were conducted. Semen volume, sperm concentration and total sperm output were not affected by dietary treatment (p > .05). The proportion of spermatozoa with abnormal morphology in fresh semen significantly increased (p CLA-fed group compared to control group. Also, in CLA-fed group, the proportion of post-thaw spermatozoa with abnormal morphology at week 10 of trial was significantly higher in CLA than control group (p CLA-fed group, although dietary supplementation did not affect other CASA parameters or viability in fresh and frozen-thawed sperm. In this study, CLA supplementation had little positive effect on fresh or post-thaw sperm quality of Holstein bulls.

  1. Role of amino acids as additives on sperm motility, plasma membrane integrity and lipid peroxidation levels at pre-freeze and post-thawed ram semen.

    Science.gov (United States)

    Sangeeta, Sharon; Arangasamy, A; Kulkarni, S; Selvaraju, S

    2015-10-01

    The possibility of including amino acids for cryopreservation of ram semen to improve the quality of frozen semen was explored in this study in sheep model. 24 samples were collected in triplicate from 8 rams of 2-3 year old Bannur cross bred rams maintained at the Institute Experimental Livestock Unit. Semen was diluted in tris-egg yolk glycerol diluent and made into 7 aliquots as follows: aliquot 1 served as control, "l-alanine" was added at 100 and 135mM in the aliquots 2 and 3, "l-glutamine" was added at 20 and 25mM in the aliquots 4 and 5 and "l-proline" was added at 25 and 50mM in the aliquots 6 and 7, respectively. Diluted semen was filled in 0.25ml French straws and frozen in LN2. Inclusion of "l-proline" and "l-glutamine" in the diluent increased the percent live sperm (Pram semen as they prevented cryoinjuries to sperm and improved the pre-freeze and post-thaw semen characteristics.

  2. Evaluation of maize grain and polyunsaturated fatty acid (PUFA) as energy sources for breeding rams based on hormonal, sperm functional parameters and fertility.

    Science.gov (United States)

    Selvaraju, Sellappan; Raju, Priyadarshini; Rao, Somu Bala Nageswara; Raghavendra, Subbarao; Nandi, Sumantha; Dineshkumar, Dhanasekaran; Thayakumar, Allen; Parthipan, Shivashanmugam; Ravindra, Janivara Parameswaraiah

    2012-01-01

    The objective of the present study was to elucidate the effect of different sources of dietary energy (maize vs polyunsaturated fatty acid (PUFA) on semen functional parameters and fertility of adult rams. Eighteen adult rams were divided into two groups (maize and PUFA, n=9). The main energy source for the rams in the maize group was coarsely ground maize grain, whereas in the PUFA group it was sunflower oil (rich in 18:2 linoleic acid, an omega-6 acid). The ration was fed for a minimum period of 60 days and thereafter semen was collected for evaluation. The proportion of progressive forward motility was significantly (P<0.05) higher in the PUFA group compared with the maize group. Sperm lipid peroxidation as measured by malondialdehyde formation (µM per 1×10(9) spermatozoa) was significantly (P<0.05) higher in the PUFA group compared with the maize group. When the semen was diluted with Tris-egg yolk-citrate buffer and incubated for 24h at 4°C, the proportions of plasmalemma integrity, the sperm subpopulation positive for functional membrane and acrosomal integrities, and mitochondrial membrane potential were significantly (P<0.05) higher in PUFA-fed than in maize-fed animals. The different sources of energy did not influence the serum and seminal plasma IGF-I levels. The cleavage rate (percentage) did not differ significantly between PUFA- (45.4±4.91) and maize- (44.63±6.8) fed animals. In conclusion, PUFA feeding influenced sperm quality by altering or stabilising membrane integrity. The present study indicates that PUFA may improve semen quality but did not improve in vitro fertilisation.

  3. Method of euthanasia influences the oocyte fertilization rate with fresh mouse sperm.

    Science.gov (United States)

    Hazzard, Karen C; Watkins-Chow, Dawn E; Garrett, Lisa J

    2014-11-01

    In vitro fertilization (IVF) is used to produce mouse embryos for a variety of reasons. We evaluated the effect of the method of euthanasia on the fertilization rate in 2 different IVF protocols. Oocytes collected from C57BL/6J female mice euthanized by CO2 inhalation or cervical dislocation were used in IVF with fresh sperm from either wild-type or genetically engineered C57BL/6J. Compared with CO2 inhalation, cervical dislocation improved the resulting rate of fertilization by 18% in an IVF method using Cook media and by 13% in an IVF method using methyl-B cyclodextrin and reduced glutathione. The lower fertilization rate due to euthanasia by CO2 inhalation was accompanied by changes in blood pH and body temperature despite efforts to minimize temperature drops. In our hands, euthanasia by cervical dislocation improved fertilization rates and consequently reduced the number of egg-donor mice required.

  4. Method of Euthanasia Influences the Oocyte Fertilization Rate with Fresh Mouse Sperm

    Science.gov (United States)

    Hazzard, Karen C; Watkins-Chow, Dawn E; Garrett, Lisa J

    2014-01-01

    In vitro fertilization (IVF) is used to produce mouse embryos for a variety of reasons. We evaluated the effect of the method of euthanasia on the fertilization rate in 2 different IVF protocols. Oocytes collected from C57BL/6J female mice euthanized by CO2 inhalation or cervical dislocation were used in IVF with fresh sperm from either wild-type or genetically engineered C57BL/6J. Compared with CO2 inhalation, cervical dislocation improved the resulting rate of fertilization by 18% in an IVF method using Cook media and by 13% in an IVF method using methyl-B cyclodextrin and reduced glutathione. The lower fertilization rate due to euthanasia by CO2 inhalation was accompanied by changes in blood pH and body temperature despite efforts to minimize temperature drops. In our hands, euthanasia by cervical dislocation improved fertilization rates and consequently reduced the number of egg-donor mice required. PMID:25650969

  5. Protective effect of esterified glucomannan on aflatoxin-induced changes in testicular function, sperm quality, and seminal plasma biochemistry in rams.

    Science.gov (United States)

    Ataman, Mehmet Bozkurt; Dönmez, Hasan Hüseyin; Dönmez, Nurcan; Sur, Emrah; Bucak, Mustafa Numan; Coyan, Kenan

    2014-02-01

    The aim of this study was to determine the effect of aflatoxin (AF) on spermatologic, biochemical, and testis parameters in rams, and the protective efficiency of esterified glucomannan (EG) co-administered with AF. Thirty-two Merino rams (12-14 months old) were used. The experimental design consisted of four dietary treatments. The control group was fed commercial feed. The AF group was fed with commercial feed plus 250 μg/d of total AF. The EG group received commercial feed plus 2 g/d of EG. The AF + EG group was given commercial feed plus 250 μg/d of total AF and 2 g/d of EG. There were treatment, time, and treatment-by-time interaction effects on sperm motility, abnormal spermatozoa, damaged acrosome, and dead spermatozoa (P glutathione (GSH) levels were determined to have significantly decreased in the AF group in comparison to the control, it was observed that, in the group co-administered with AF and EG, particularly after week 7, the GSH levels, which had decreased owing to AF, were largely ameliorated (P < 0.05). In conclusion, AF adversely affected spermatologic, biochemical, and testis parameters, and the combined administration of EG with AF reversibly eliminated these adverse effects in rams.

  6. Integridade das membranas plasmática, nuclear e mitocondrial de espermatozóides ovinos criopreservados com etileno glicol Integrity of sperm plasm membrane, nucleus and mitochondria after freezing ram semen with ethylene glycol

    Directory of Open Access Journals (Sweden)

    Luciana Bignardi de Soares Brisola

    1999-09-01

    to 5, sperm cell concentration (3x10(6 cells/mm³ and normal sperm morphology (80%. Each pool of semen from ejaculates of two rams was divided into two equal subsamples. One subsample was frozen with ethylene glycol and the other with glycerol. The parameters used to evaluate the semen were sperm motility, vigor, acrossome status and membrane integrity. Sperm motility was evaluated in fresh semen, cooled semen, frozen semen, after 5 hours at 38°C or after 30 minutes at 45°C. No difference was observed between ethylene glycol and glycerol for acrossome status and sperm motility. The sperm cells that were preserved with ethylene glycol showed more integrity of the plasmatic, nuclear and mitochondrial membranes. From the viewpoint of cell membrane integrity, it can be concluded that ethylene glycol gives higher protection to the sperm cell than glycerol.

  7. Expression, cellular localization, and involvement of the pentose phosphate pathway enzymes in the regulation of ram sperm capacitation.

    Science.gov (United States)

    Luna, C; Serrano, E; Domingo, J; Casao, A; Pérez-Pé, R; Cebrián-Pérez, J A; Muiño-Blanco, T

    2016-08-01

    Spermatozoa require substantially more ATP than other cells, not only for sustaining sperm motility but also for regulating protein phosphorylation during capacitation. In this study, we have reported for the first time the presence of the two key enzymes of the pentose phosphate pathway (PPP), glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in ovine spermatozoa by indirect immunofluorescence, Western blotting, in-gel activity, and reverse transcription polymerase chain reaction analysis. We found that the activity of both enzymes significantly increased after in vitro capacitation in the presence of high-cAMP levels, with a concomitant increase in protein tyrosine phosphorylation and in the proportion of sperm-capacitated pattern assessed by the chlortetracycline staining. These results suggest that PPP is related with the progress of capacitation and that a relationship between calcium compartmentalization, protein tyrosine phosphorylation and PPP seems to exist. This is the first report that shows a connection between the PPP, cAMP/PKA signaling pathways and sperm capacitation. These findings can be of high-biological importance to improve our knowledge of the biochemical mechanisms involved in the acquisition of mammalian sperm functional competence and, ultimately, fertility.

  8. Early Morphokinetic Monitoring of Embryos after Intracytoplasmic Sperm Injection with Fresh Ejaculate Sperm in Nonmosaic Klinefelter Syndrome: A Different Presentation

    Directory of Open Access Journals (Sweden)

    Ali Sami Gurbuz

    2015-01-01

    Full Text Available The patient was diagnosed with nonmosaic 47, XXY Klinefelter Syndrome with the AZF deletion absent and SRY+. The nonmosaic 47, XXY karyotype was confirmed on a skin biopsy chromosomal analysis. Using only ejaculate motile sperms, 11 oocytes underwent ICSI and were placed rapidly in a time lapse (Embryoscope © with a specific culture dish. Biopsies were performed on six embryos on the 3rd day, and numerical chromosomal abnormalities were observed using the FISH test before transfer. PGS results were normal in only two embryos with normal morphokinetics in the Embryoscope. For clinical confirmation of pregnancy, ultrasonographic examination was performed during the 7th week of pregnancy, and two gestational sacs and fetal heart beat were observed.

  9. In Vitro Comparison of Soybean Lecithin Based-Extender with Commercially Available Extender for Ram Semen Cryopreservation

    Directory of Open Access Journals (Sweden)

    Ahmad reza Javaheri

    2009-01-01

    Full Text Available Background: Egg yolk is the main cryoprotectant agent in semen freezing extenders which is usedin order to protect spermatozoa against cold shock. However, elimination of animal bioproductsfrom the cryopreservation protocol is becoming mandatory. Therefore, the aim of this study is tocompare a previously studied, homemade soya bean lecithin based extender with a commerciallyavailable extender for ram sperm cryopreservation.Materials and Methods: Samples from three rams were pooled and split into two equal aliquots anddiluted (1:20 with i%lecithin - 7%glycerol (L1G7 and Bioxcell®. The effects of L1G7 and Bioxcell® onsperm parameters and the in vitro fertilization ability of frozen-thawed ram spermatozoa were assessed.Results: The results of this study revealed no difference between the two extenders in terms of motility,viability, and capacitation status. The results of in vitro fertilization in terms of rate of blastocyst formationwere similar for both extenders, but significantly lower than that of freshly processed ram sperm.Conclusion: We conclude that both extenders are suitable for ram sperm cryopreservation.

  10. Saturated, omega-6 and omega-3 dietary fatty acid effects on the characteristics of fresh, frozen-thawed semen and blood parameters in rams.

    Science.gov (United States)

    Esmaeili, V; Shahverdi, A H; Alizadeh, A R; Alipour, H; Chehrazi, M

    2014-02-01

    The aim of this study was to investigate the effects of several dietary fatty acids (FAs) on semen quality and blood parameters in rams. We gave diet-supplemented treatments (35 g day(-1) ram(-1)) by C16:0 (palm oil), C18:2 [sunflower oil (SO)] and an n-3 source [fish oil (FO)] to 12 rams, who were fed for 15 weeks during their breeding season. Semen was collected once per week. Semen samples were extended with Tris-based cryoprotective diluents, then cooled to 5 °C and stored in liquid nitrogen. Positive responses were seen with FO after 4 weeks. The mean prefreezing semen characteristics improved with the intake of FO (P oil 5.3 × 10(9). Rams that received FO had the highest total testosterone concentrations (11.3 ng ml(-1) for FO, 10.8 ng ml(-1) for SO and 10.2 ng ml(-1) for palm oil) during the experiment (P characteristics after thawing (P oils.

  11. Seasonal variation in semen quality of Dorper rams using different ...

    African Journals Online (AJOL)

    Botswana

    2014-02-08

    Feb 8, 2014 ... 2 Department of Animal, Wildlife and Grassland Sciences, ... semen wave motion, sperm motility, sperm cell concentration, sperm ... sheep numbers in South Africa, but also realises superior prices at sales and auctions (Milne, 2000; ..... Possibly the rams had become accustomed to the stress inflicted by ...

  12. Memoria RAM

    DEFF Research Database (Denmark)

    Cruz Suárez, Juan Carlos

    2013-01-01

    The present study intends to open a new theoretical means of analysis within the field of comparative memories. Through the metaphorical designation of RAM memory we will try to observe the continuous circulation of social discourses related to the collective memory of a particular community. Thi...

  13. Memoria RAM

    DEFF Research Database (Denmark)

    Cruz Suárez, Juan Carlos

    2013-01-01

    The present study intends to open a new theoretical means of analysis within the field of comparative memories. Through the metaphorical designation of RAM memory we will try to observe the continuous circulation of social discourses related to the collective memory of a particular community. Thi...

  14. Role of cAMP in the reactivation of demembranated ram spermatozoa.

    Science.gov (United States)

    San Agustin, J T; Witman, G B

    1994-01-01

    Ejaculated ram sperm were demembranated with Triton X-100, separated from the detergent-soluble matrix, and reactivated [San Agustin and Witman (1993): Cell Motil. Cytoskeleton 24:264-273]. The percent motility of models prepared from freshly washed sperm was comparable to that of the washed sample before demembranation, regardless of whether cAMP was included in the reactivation medium. However, demembranated models derived from aging or metabolically inhibited sperm exhibited a lower percent reactivation and required cAMP to attain the level of motility of freshly washed sperm. Cyclic AMP was approximately 100 times more effective than cGMP. The requirement for cAMP could be bypassed by addition of porcine heart cAMP-dependent protein kinase (PKA) catalytic subunit to the reactivation medium, demonstrating that cAMP was acting via PKA. The cAMP stimulation of reactivation was not affected by inclusion of the PKA inhibitor PKI(5-24) in the reactivation medium, but was decreased when the models were preincubated with PKI(5-24) prior to reactivation. The cytosol-free models retained > 90% of the sperm PKA activity; therefore, the PKA appears to be anchored to internal sperm structures. This PKA could not be extracted by cAMP or Triton X-100 alone, but only by cAMP and Triton X-100 in combination. We conclude that cAMP-dependent protein phosphorylation is critical for sperm motility, but that the essential protein phosphate sites turn over slowly under our reactivation conditions, so that the cAMP requirement is apparent only in models prepared from sperm having a low internal ATP or cAMP content. Interestingly, reactivation was rapidly blocked by the peptide arg-lys-arg-ala-arg-lys-glu, which has been reported to be a selective inhibitor of cGMP-dependent protein kinase.

  15. Validation of merocyanine 540 staining as a technique for assessing capacitation-related membrane destabilization of fresh dog sperm

    NARCIS (Netherlands)

    Steckler, D; Stout, T A E; Durandt, C; Nöthling, J O

    2015-01-01

    The aim of this study was to determine whether flow cytometric evaluation of combined merocyanine 540 and Yo-Pro 1 (M540-YP) staining would identify viable dog sperm that had undergone membrane stabilization known to be associated with capacitation in other species, and whether such destabilization

  16. Differences in the ovine HSP90AA1 gene expression rates caused by two linked polymorphisms at its promoter affect rams sperm DNA fragmentation under environmental heat stress conditions.

    Science.gov (United States)

    Salces-Ortiz, Judit; Ramón, Manuel; González, Carmen; Pérez-Guzmán, M Dolores; Garde, J Julián; García-Álvarez, Olga; Maroto-Morales, Alejandro; Calvo, Jorge H; Serrano, M Magdalena

    2015-01-01

    Heat shock (HS) is one of the best-studied exogenous cellular stresses. Almost all tissues, cell types, metabolic pathways and biochemical reactions are affected in greater or lesser extent by HS. However, there are some especially thermo sensible cellular types such as the mammalian male germ cells. The present study examined the role of three INDELs in conjunction with the -660G/C polymorphism located at the HSP90AA1 promoter region over the gene expression rate under HS. Specially, the -668insC INDEL, which is very close to the -660G/C transversion, is a good candidate to be implied in the transcriptional regulation of the gene by itself or in a cooperative way with this SNP. Animals carrying the genotype II-668 showed higher transcription rates than those with ID-668 (FC = 3.07) and DD-668 (FC = 3.40) genotypes for samples collected under HS. A linkage between gene expression and sperm DNA fragmentation was also found. When HS conditions were present along or in some stages of the spermatogenesis, alternative genotypes of the -668insC and -660G/C mutations are involved in the effect of HS over sperm DNA fragmentation. Thus, unfavorable genotypes in terms of gene expression induction (ID-668GC-660 and DD-668GG-660) do not produce enough mRNA (stored as messenger ribonucleoprotein particles) and Hsp90α protein to cope with future thermal stress which might occur in posterior stages when transcriptional activity is reduced and cell types and molecular processes are more sensible to heat (spermatocytes in pachytene and spermatids protamination). This would result in the impairment of DNA packaging and the consequent commitment of the events occurring shortly after fertilization and during embryonic development. In the short-term, the assessment of the relationship between sperm DNA fragmentation sensitivity and ram's fertility will be of interest to a better understanding of the mechanisms of response to HS and its consequences on animal production and

  17. 大黄鱼(Pseudosciaena crocea)精子冷冻前后的活力及超微结构变化%VITALITY AND ULTRASTRUCTURE OBSERVATION OF FRESH AND CRYOPRESERVATED SPERM IN PSEUDOSCIAENA CROCEA

    Institute of Scientific and Technical Information of China (English)

    程顺; 闫家强; 竺俊全; 姜建湖; 吴雄飞; 史会来

    2013-01-01

    采用两步降温法超低温冷冻保存大黄鱼精子,并用透射电镜技术研究了精子的超微结构损伤.结果表明,大黄鱼冻精的激活率、运动时间及寿命与鲜精相比无显著差异.鲜精中28.5%的精子形态结构异常,冻精中37%的精子形态结构异常.形态结构正常的精子表现为质膜与核膜结构完整、无膨胀现象,袖套、轴丝及中心粒结构正常,线粒体形态完整、嵴较发达;形态结构异常的精子表现为质膜破裂、脱落,质膜膨胀,核膜破裂、脱落,核局部受损伤,线粒体膨胀、嵴退化或消失,线粒体移位或脱落.结果显示,以Cortland溶液为稀释液,10% DMSO为抗冻剂,对大黄鱼精子具有较好的抗冻保护作用.%A two-step cooling procedure was employed to cryopreserve Pseudosciaena crocea sperm, and the sperm ultrastructure after which was observed under transmission electron microscopy. The results show that there were no significant differences between frozen-thawed sperm and fresh sperm by comparing the activation rate, moving time and life-span. Both the fresh sperm and cryopreserved sperm had ultrastructural damages in various degree. The deformation rate of the fresh and cryopreservated sperms were 28.5% and 37%, respectively. The following aspects of cryopreserved sperm with normal morphology was observed: the plasma and nuclear membrane; the sleeve, axoneme and centriole; the mitochondrion obtained integrity and with well-developed cristae. On the contrary, the sperm cryodamages were observed as follows: swelled or disrupted plasma and nuclear membrane; partially damaged nucleus; the swelled, dislocated or disarticulated mitochondrion with degenerated or vanished cristae. The results show that Cortland solutions and 10% DMSO are the best choice for extender and cryoprotecant, which are helpful for improving the frozen-thawed P. crocea sperm quality.

  18. Successful ram semen cryopreservation with lyophilized egg yolk-based extender.

    Science.gov (United States)

    Alcay, Selim; Berk Toker, M; Gokce, Elif; Ustuner, Burcu; Tekin Onder, N; Sagirkaya, Hakan; Nur, Zekariya; Kemal Soylu, M

    2015-10-01

    The aim of this study was to evaluate the effects of lyophilized egg yolk extender on ram semen cryopreservation. Ejaculates with a thick consistency, rapid wave motion (3-5 on a 0-5 scale) and >75% initial motility were pooled. Sperm were diluted to final concentration of 1/5 (semen/extender) in lyophilized egg yolk or fresh egg yolk extenders using two-step dilution method. The equilibrated semen was frozen in 0.25 mL straws. Semen samples were assessed for sperm motility, plasma membrane functional integrity using hypoosmotic swelling test (HOST), damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) and DNA integrity using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) at three time points: after dilution with extender A, equilibration and post-thaw. The results showed that freezing and thawing procedures (dilution, equilibration and thawing) had negative effects on motility (Pram semen.

  19. Blastocyst transfer ameliorates live birth rate compared with cleavage-stage embryos transfer in fresh in vitro fertilization or intracytoplasmic sperm injection cycles: reviews and meta-analysis.

    Science.gov (United States)

    Wang, Shan-Shan; Sun, Hai-Xiang

    2014-05-01

    Blastocyst transfer has been recommended to raise the implantation rate without affecting the pregnancy rate. The objective of this meta-analysis is to systematically evaluate whether the live birth rate and other pregnancy outcomes can be improved by blastocyst transfer compared with cleavage-stage embryos transfer. EMBASE and MEDLINE databases were searched for papers published between March 2004 and March 2013. An extensive range of the electronic databases yielded initially 317 studies from which seven trials met the inclusion criteria for further analysis. Our outcome measures were the live birth rate, clinical pregnancy rate, implantation rate, ongoing pregnancy rate, multiple pregnancy rate, first trimester miscarriage rate and ectopic pregnancy rate. Fixed effects models were chosen to calculate the odds ratio (OR). Seven trials (n=1446 cases) were finally analyzed. Compared with cleavage-stage embryos transfer, the blastocyst transfer was statistically significantly associated with an increase in clinical pregnancy rate [OR 1.43; 95% confidence interval (CI), 1.15-1.78], implantation rate (OR 1.38; 95% CI, 1.09-1.74) and ongoing pregnancy rate (OR 2.15; 95% CI, 1.57-2.94), and also a reduction in the probability of first trimester miscarriage rate (OR 0.51; 95% CI, 0.30-0.87). The improvement in the live birth rate was also observed (OR 1.77; 95% CI, 1.32-2.37). Moreover, there was no evidence of difference in multiple pregnancy and ectopic pregnancy rates. The available evidences suggest that live birth and other pregnancy outcomes after fresh in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) are significantly improved following blastocyst transfer as compared to cleavage-stage embryo transfer.

  20. Effects of pre-incubation of eggs in fresh water and varying sperm concentration on fertilization rate in sterlet sturgeon, Acipenser ruthenus

    DEFF Research Database (Denmark)

    Siddique, Mohammad Abdul Momin; Butts, Ian; Psěnička, Martin

    2015-01-01

    Standardization of fertilization protocols for sterlet Acipenser ruthenus is crucial for improving reproductive techniques and for conservation purposes. Our objectives were to determine the number of sperm (tested 430,000:1, 43,000:1, 4300:1, 430:1 sperm to egg) required to fertilize eggs...

  1. Soy lecithin interferes with mitochondrial function in frozen-thawed ram spermatozoa.

    Science.gov (United States)

    Del Valle, I; Gómez-Durán, A; Holt, W V; Muiño-Blanco, T; Cebrián-Pérez, J A

    2012-01-01

    Egg yolk and milk are the 2 major membrane cryoprotectants commonly used in freezing media for the long-term preservation of semen (alone or in combination with others). However, in recent years, there have been increasing arguments against the use of egg yolk or milk because of the risk of introducing diseases through the use of cryopreserved semen. In this study, we analyzed the protective effect of lecithin as an alternative to egg yolk for the cryopreservation of ram semen, using a range of functional markers for sperm viability, motility, apoptosis, and mitochondrial functionality analyses (mitochondrial inner membrane surface [MIMS], mitochondrial inner membrane potential [MIMP], and cell membrane potential) as methods of assessment in samples diluted in 3 different media: Tris-citrate-glucose as control and 2 media supplemented with soy lecithin or egg yolk. The results showed that lecithin was able to effectively protect certain sperm quality characteristics against freezing-induced damage. However, lecithin induced loss of mitochondrial membrane potential or mitochondrial loss that was not reflected by modifications in sperm motility in fresh semen. MIMS and MIMP values decreased in thawed lecithin-treated samples, concomitant with a lower (P lecithin may have affected the inner mitochondrial membrane in frozenthawed spermatozoa and confirmed that sublethal damages that seriously affect sperm functionality, not detected by classic sperm quality analyses, can be evidenced by changes in the inner mitochondrial membrane surface. These findings strengthen the relationship between mitochondrial membrane potential and motility and show that the mitochondrial alterations induced by the cryopreservation process could be specific targets for the improvement of semen cryopreservation protocols.

  2. RAM Technology Study.

    Science.gov (United States)

    1980-01-03

    resistor load cell can be mitigated by replacing the load resistors by pnp transistors as shown in Figure 9. 22 I *- . . ’’ - - - - 1...1 LIST OF ILLUSTRATIONS Figure Page 1 Three- transistor dynamic RAM cell, dual input output lines. 14 2 Single transistor (1-T) dynamic RAM cell. 14 3a...resistor load static n-MOS memory cell. 20 6 Six transistor CMOS RAM cell. 21 7 Five transistor CMOS RAM cell. 23 8 Bipolar ECL resistor coupled cell with

  3. Implications of the pH and temperature of diluted, cooled boar semen on fresh and frozen-thawed sperm motility characteristics

    Science.gov (United States)

    Boar semen is typically collected, diluted and cooled for AI use over numerous days, or frozen immediately after shipping to capable laboratories. The storage temperature and pH of the diluted, cooled boar semen could potentially influence the fertility of boar sperm. Therefore, the purpose of thi...

  4. Effects of pre-incubation of eggs in fresh water and varying sperm concentration on fertilization rate in sterlet sturgeon, Acipenser ruthenus.

    Science.gov (United States)

    Siddique, Mohammad Abdul Momin; Butts, Ian Anthony Ernest; Psenicka, Martin; Linhart, Otomar

    2015-08-01

    Standardization of fertilization protocols for sterlet Acipenser ruthenus is crucial for improving reproductive techniques and for conservation purposes. Our objectives were to determine the number of sperm (tested 430,000:1, 43,000:1, 4300:1, 430:1 sperm to egg) required to fertilize eggs and explore how pre-incubation of eggs in freshwater for 0min, 0.5min, 1min, and 10min interacts with different sperm ratios. Fertilization success ranged from 29.7% at 430:1 to 84.2% at 430,000:1. Pre-incubation time had no effect on fertilization success at 430,000:1 and 43,000:1 sperm to egg ratios, while it was significant at the 4300:1 and 430:1 ratios. The use of adequate experimental suboptimal sperm to egg ratio revealed a positive effect of pre-incubation time, such that at the 430:1 ratio, 0.5min pre-incubation increased the fertilization rate than 10min. At 0min pre-incubation the proportion of fertilized eggs increased at the 430,000:1 ratio, while at 1min fertilization increased at the 4300:1 ratio. At the 10min pre-incubation time, fertilization increased at the 43,000:1 ratio. Moreover, at the 0.5min pre-incubation time, the 43,000:1 ratio increased the fertilization rate than the 430:1 ratio. Generally, for 430:1 ratio, the fertilization rate is lower than in control. Transmission electron microscopy showed that pre-incubation of eggs in water for fertilization can enhance fertilization rate of sterlet. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Sperm-Egg Interaction: Evidence for Boar Sperm Plasma Membrane Receptors for Porcine Zona Pellucida

    Science.gov (United States)

    Peterson, Rudolph N.; Russell, Lonnie; Bundman, Donna; Freund, Matthew

    1980-01-01

    Freshly ejaculated, noncapacitated boar sperm bind rapidly and in large numbers to pig egg zona pellucida in vitro. In the present study, the number of sperm bound decreased sharply when sperm motility was lowered by energy poisons or by reducing the temperature. Highly motile sperm from humans, guinea pigs, and rats, added at concentrations ten times higher than control sperm, did not bind to the porcine zona. At the same high concentration, a small number of hamster and bull sperm bound to the zona. Binding of boar sperm to the zona pellucida was blocked almost completely by diluted whole antiserum to sperm plasma membranes and by univalent (Fab) antibody to these membranes. When antibody to sperm plasma membrane was first absorbed with plasma membrane vesicles, sperm binding was not inhibited. These results provide direct evidence for the existence of sperm plasma membrane receptors for the zona pellucida of the pig.

  6. Effect of Three Days Storage of Coated Spermatozoa at Cooling and Adding Seminal Plasma on Ram Fertility

    Directory of Open Access Journals (Sweden)

    Alireza Vaferi

    2016-08-01

    Full Text Available Introduction Artificial insemination (AI has only been used as a supplement to natural mating. AI, when used in conjunction with accurate progeny testing schemes, can substantially increase the rate of genetic progress compared with that of natural service. Moreover, the use of AI causes the limitation of the transmitted diseases. Cervical insemination with frozen-thawed ram semen has not been widely adopted, probably because of the relative poor fertility obtained. Thus using fresh and diluted semen is only approach for performing AI. AI is currently limited by the poor fertility achieved after cervical insemination with the storage of liquid semen at sub-ambient temperature. The success of this procedure in sheep is restricted by the short length of time that ram sperm can be stored in a liquid state. Moreover, the effect of cooling on sperm differs depending on species. It is also well known that ram spermatozoa are more sensitive to cold-shock stress than those of other species. Seminal plasma, as physiological secretion, is a complex mixture of secretions originating from testis, epididymis and accessory sex glands which is mixed with epididymal sperm at ejaculation; it serves as the carrier of sperm to the female genital tract. This mixture contains numerous factors such as organic and nonorganic material which play an important role in the final maturation of the spermatozoa through hormonal, enzymatic and surface-modifying events. During natural mating, a mechanism may be activated to separate spermatozoa from seminal plasma. After being ejaculated into the vagina, sperm swim through cervical mucus and enter the uterus within minutes (>30 min; cervical mucus acts as a barrier for seminal plasma. In the artificial insemination industry, seminal plasma with all the useful and harmful components is not removed from semen and is in contact with sperm throughout cooling, freezing and storage. On the other hand, it was demonstrated that the

  7. Feline spermatozoa from fresh and cryopreserved testicular tissues have comparable ability to fertilize matured oocytes and sustain the embryo development after intracytoplasmic sperm injection.

    Science.gov (United States)

    Buarpung, S; Tharasanit, T; Comizzoli, P; Techakumphu, M

    2013-01-01

    Cryopreservation of testicular tissue associated with intracytoplasmic sperm injection (ICSI) is a critical tool that still needs to be explored for preserving the fertility of endangered species. Using the domestic cat as a model for wild felids, the study aimed at determining the effect of different cryoprotectants and freezing techniques (two-step freezing vs. controlled slow freezing) on the sperm quality (membrane and DNA integrity). Then, spermatozoa were extracted from frozen-thawed testicular tissues and used for ICSI to assess early gamete activation or developmental competence in vitro. The percentage of spermatozoa with intact plasma membrane was not different (P > 0.05) among nonfrozen control, glycerol-, and ethylene glycol-frozen tissues (63.2 ± 2%, 58.2 ± 2.6%, 53.3 ± 2.3%, respectively). However, these percentages were significantly lower (P sperm membrane integrity (55.0 ± 2.7%) when compared with other freezing techniques. The incidence of DNA fragmentation was found to be low (0.2%-1.1%) in all freezing protocols. After ICSI with frozen testicular spermatozoa, male and female gametes were asynchronously activated and the percentages of normal fertilization at 6, 12, and 18 hours were 11.2%, 20.6%, and 22.1%, respectively. Metaphase II-arrested oocytes containing or not a decondensed sperm head were predominantly found after ICSI with frozen testicular spermatozoa. Although two-pronucleus formation could be observed as soon as 6 hours post ICSI (10%), the rate increased dramatically after 12 and 18 hours post ICSI (17.2% and 19.5%, respectively). ICSI using frozen-thawed testicular spermatozoa yielded cleavage (32.7%), morula (6.5%), and blastocyst (4.4%) percentages similar to nonfrozen control (P > 0.05). It is concluded that conventional freezing technique with glycerol as a principle cryoprotectant is simplified and applicable for cat testicular tissue cryopreservation. We also demonstrate for the first time that feline spermatozoa

  8. Chemical forms of selenium present in rat and ram spermatozoa.

    Science.gov (United States)

    Alabi, N S; Beilstein, M A; Whanger, P D

    2000-08-01

    In vivo and in vitro studies were conducted to investigate the chemical forms by ion-exchange chromatography of selenium (Se) present in rat and ovine spermatozoa. After injection with 75Se-selenite, the form of 75Se in rat sperm was selenocysteine, but selenocysteine and selenomethionine (SeMet) were present in ovine sperm. Presumably, synthesis of SeMet by rumen microbes are responsible for its presence in ovine sperm. In vitro incubation of ram sperm with selenocysteine or SeMet produced no changes, but incubation with selenite produced a compound that eluted one fraction before SeMet from the ion-exchange column. After treatment of this fraction with mercaptoethanol, it eluted in a later fraction upon rechromatography, suggesting it to be selenodicysteine. This compound is apparently formed because of high levels of cysteine in semen. Cysteine, reduced glutathione, and oxidized glutathione were also found in semen. The significance of the results is discussed.

  9. Differences in the Ovine HSP90AA1 Gene Expression Rates Caused by Two Linked Polymorphisms at Its Promoter Affect Rams Sperm DNA Fragmentation under Environmental Heat Stress Conditions

    Science.gov (United States)

    González, Carmen; Pérez-Guzmán, M. Dolores; Garde, J. Julián; García-Álvarez, Olga; Maroto-Morales, Alejandro; Calvo, Jorge H.; Serrano, M. Magdalena

    2015-01-01

    Heat shock (HS) is one of the best-studied exogenous cellular stresses. Almost all tissues, cell types, metabolic pathways and biochemical reactions are affected in greater or lesser extent by HS. However, there are some especially thermo sensible cellular types such as the mammalian male germ cells. The present study examined the role of three INDELs in conjunction with the -660G/C polymorphism located at the HSP90AA1 promoter region over the gene expression rate under HS. Specially, the -668insC INDEL, which is very close to the -660G/C transversion, is a good candidate to be implied in the transcriptional regulation of the gene by itself or in a cooperative way with this SNP. Animals carrying the genotype II-668 showed higher transcription rates than those with ID-668 (FC = 3.07) and DD-668 (FC = 3.40) genotypes for samples collected under HS. A linkage between gene expression and sperm DNA fragmentation was also found. When HS conditions were present along or in some stages of the spermatogenesis, alternative genotypes of the -668insC and -660G/C mutations are involved in the effect of HS over sperm DNA fragmentation. Thus, unfavorable genotypes in terms of gene expression induction (ID-668GC-660 and DD-668GG-660) do not produce enough mRNA (stored as messenger ribonucleoprotein particles) and Hsp90α protein to cope with future thermal stress which might occur in posterior stages when transcriptional activity is reduced and cell types and molecular processes are more sensible to heat (spermatocytes in pachytene and spermatids protamination). This would result in the impairment of DNA packaging and the consequent commitment of the events occurring shortly after fertilization and during embryonic development. In the short-term, the assessment of the relationship between sperm DNA fragmentation sensitivity and ram’s fertility will be of interest to a better understanding of the mechanisms of response to HS and its consequences on animal production and

  10. Effects of seasons on some semen parameters and bacterial contamination of Awassi ram semen.

    Science.gov (United States)

    Azawi, O I; Ismaeel, M A

    2012-06-01

    The objectives of the present study were to determine the effects of season on some semen parameters and bacterial contamination of Awassi ram semen. Semen samples from six mature Awassi rams were used in this study. Semen collection was performed with artificial vagina every week, from September 2009 to October 2010. Volume, sperm concentration, mass motility, individual motility, percentage live sperm and sperm abnormalities were evaluated. Moreover, determination of viable bacterial count of the rams was also recorded weekly. Higher (p highest (p highest values and differed significantly (p highest value of the mean sperm acrosomal defects (13.33 ± 0.63%) was recorded in December. The highest value of the mean viable bacterial count (138.3 ± 21.6) was recorded in July (summer). A significant decrease (p bacterial count was observed from the middle of winter towards the end of spring. The lowest bacterial count was noted in January (60.5 ± 2.98). It could be concluded from the results of the present study that there is an effect of season on ram semen quality, and summer high temperature in northern Iraq has no effect on Awassi ram semen. There is a significant effect of season on bacterial count on Awassi ram semen.

  11. Variations in semen characteristics rams of Ouled Djellal breed have received an important dietary supplement after regular and intensive collection

    Institute of Scientific and Technical Information of China (English)

    Litim Miloud; Bereksi Reguig Karima

    2015-01-01

    Objective: To evaluate the effect of an important dietary supplement on sperm quantity and quality breeding rams of artificial insemination center Belhandjir in region of Naama in Algeria. Methods:Ten breeding rams of the white arab breed, called Ouled Djellal are divided into two groups CR and SR, and subjected to two different diets, the first group CR followed a regime R based of barley and fodder and the second SR group to follow a regime R +a vitamin and mineral dietary supplement for a 15 weeks of regular collections followed by 12 weeks of intensive collections. Analysis of sperm quality and quantity were performed to normal and intensive collection. Results:The quantitative effect is more significant in rams supplemented SR, volume and concentration were higher in rams SR (respectively+40%and±13%), therefore the doses produced differ significantly compared to the group of control rams CR (+70%). Results of qualitative parameters were more mixed, motility showed a significant difference in rams supplemented with against the rate of dead semen and the rate of anomalies did not show significant differences in sperm. Conclusion:supplementation could significantly improve the quantitative semen characteristics in rams breeding Ouled Djellal. Collections of semen on the rams may be daily without sperm quantity is affected.

  12. Organizational Impact of RAMS.

    Science.gov (United States)

    Staebler, Mel

    A series of observations is made regarding the potential impact of the Remotely Accessible Management System (RAMS) upon the administrative structure of the Pontiac, Michigan City School System. Nine major results of the district wide needs assessment are reported, and evidence is advanced to support the conclusion that modern educational…

  13. The Ram's Horn.

    Science.gov (United States)

    Rassias, John A., Ed.; And Others

    1983-01-01

    The summer-fall and winter-spring numbers of the journal, "The Ram's Horn," contain these articles: "The Text as Dramatic Departure"; "The Dartmouth Language Outreach Approach to Spanish for Police Action"; "The Dartmouth Intensive Language Model (DILM) in Florida: John Rassias with High School Teachers"; "The Flexibility of Using Drama Techniques…

  14. Analysis of the Differential Proteomics Between Fresh and Frozen-Thawed Sheep Sperm%绵羊精子冻融前后差异蛋白质组学研究

    Institute of Scientific and Technical Information of China (English)

    李鸿岩; 赵兴绪; 张勇; 贺钰垣; 杨永新

    2011-01-01

    [目的]检测和分析绵羊精子冻融前后的差异蛋白质.[方法]用二维凝胶电泳分离精子蛋白,凝胶用考马斯亮蓝染色,用PDQues t8.0软件检测分析差异蛋白质斑点并经液相色谱串联离子阱质谱鉴定.[结果]鲜冻精凝胶图谱分析后得到14个差异蛋白斑点,在国际生物信息学中心数据库搜索到相对应的5种蛋白质:推定的热休克蛋白70.1、转录因子AP-2 α蛋白、烯醇酶、烯醇酶1和血清白蛋白前体.在解冻精子中,推定的热休克蛋白70.1、烯醇酶和烯醇酶1含量减少,转录因子AP-2α蛋白含量增加,血清白蛋白前体来源于精浆.[结论]绵羊精子经冷冻、解冻后,蛋白质存在差异,对差异蛋白质的研究有利于探索绵羊精子在冷冻过程中的生理状态,为进一步研究绵羊精子蛋白质组学及揭示精子冻伤机理提供依据.%[Objective] To investigate the differential proteins between fresh and frozen-thawed sheep sperm. [Method] The current research presents the protein changes using two-dimensional gel electrophoresis (2-DE) after stained with commassie blue, differential proteins were detected by PDQuest 8.0 software and subjected to ion trap mass spectrometer equipped with a Surveyor HPLC system, and differential spots of protein were identified. [Result] A search against protein sequences in the National Center for Biotechnology Information databases indicated that the differential fourteen proteins correspond to five proteins including putative heat shock protein 70.1, transcription factor AP-2-alpha, enolase, enolasel and serum albumin precursor that protein content of putative heat shock protein 70.1, enolase, enolasel were decreased and protein content of transcription factor AP-2-alpha was increased in frozen-thawed spermatozoa, serum albumin precursor was seminal plasma protein. [Conclusion] It was concluded there was a significant difference at protein level in sheep sperm during frozen and frozen

  15. Hydraulic rams; a comparative investigation

    NARCIS (Netherlands)

    Tacke, J.H.P.M.

    1988-01-01

    A mathematical model describing the essential features of hydraulic ram operation is developed in order to clarify the possibilities and limitations of the ram relative to its site and its adjustments. The model distinguishes three different periods in the pumping cycle of the hydraulic ram: acceler

  16. The utility of nanowater for ram semen cryopreservation.

    Science.gov (United States)

    Murawski, Maciej; Schwarz, Tomasz; Grygier, Joanna; Patkowski, Krzysztof; Oszczęda, Zdzisław; Jelkin, Igor; Kosiek, Anna; Gruszecki, Tomasz M; Szymanowska, Anna; Skrzypek, Tomasz; Zieba, Dorota A; Bartlewski, Pawel M

    2015-05-01

    Nanowater (NW; water declusterized in the low-temperature plasma reactor) has specific physicochemical properties that could increase semen viability after freezing and hence fertility after artificial insemination (AI) procedures. The main goal of this study was to evaluate ram semen quality after freezing in the media containing NW. Ejaculates from 10 rams were divided into two equal parts, diluted in a commercially available semen extender (Triladyl®; MiniTüb GmbH, Tiefenbach, Germany) prepared with deionized water (DW) or NW, and then frozen in liquid nitrogen. Semen samples were examined for sperm motility and morphology using the sperm class analyzer system and light microscopy. Cryo-scanning electron microscopy (cryo-SEM) was employed to determine the size of extracellular water crystals in frozen semen samples. Survival time at room temperature, aspartate aminotransferase (AspAT) and alkaline phosphatase (ALP) concentrations post-thawing as well as conception/lambing rates after laparoscopic intrauterine AI of 120 ewes were also determined. There were no significant differences between DW and NW groups in sperm progressive motility (26.4 ± 12.2 and 30.8 ± 12.4%) or survival time (266.6 ± 61.3 and 270.9 ± 76.7 min) after thawing and no differences in the percentages of spermatozoa with various morphological defects before or after freezing. There were, however, differences (P ram semen and lamb productivity of inseminated ewes.

  17. Single-embryo transfer reduces clinical pregnancy rates and live births in fresh IVF and Intracytoplasmic Sperm Injection (ICSI cycles: a meta-analysis

    Directory of Open Access Journals (Sweden)

    Oliveira João

    2009-04-01

    Full Text Available Abstract Background It has become an accepted procedure to transfer more than one embryo to the patient to achieve acceptable ongoing pregnancy rates. However, transfers of more than a single embryo increase the probability of establishing a multiple gestation. Single-embryo transfer can minimize twin pregnancies but may also lower live birth rates. This meta-analysis aimed to compare current data on single-embryo versus double-embryo transfer in fresh IVF/ICSI cycles with respect to implantation, ongoing pregnancy and live birth rates. Methods Search strategies included on-line surveys of databases from 1995 to 2008. Data management and analysis were conducted using the Stats Direct statistical software. The fixed-effect model was used for odds ratio (OR. Fixed-effect effectiveness was evaluated by the Mantel Haenszel method. Seven trials fulfilled the inclusion criteria. Results When pooling results under the fixed-effect model, the implantation rate was not significantly different between double-embryo transfer (34.5% and single-embryo transfer group (34.7% (P = 0.96; OR = 0.99, 95% CI 0.78, 1.25. On the other hand, double-embryo transfer produced a statistically significantly higher ongoing clinical pregnancy rate (44.5% than single-embryo transfer (28.3% (P Conclusion Meta-analysis with 95% confidence showed that, despite similar implantation rates, fresh double-embryo transfer had a 1.64 to 2.60 times greater ongoing pregnancy rate and 1.44 to 2.42 times greater live birth rate than single-embryo transfer in a population suitable for ART treatment.

  18. Effect of increased testicular temperature on seminal plasma proteome of the ram.

    Science.gov (United States)

    Rocha, David R; Martins, Jorge André M; van Tilburg, Mauricio F; Oliveira, Rodrigo V; Moreno, Frederico B; Monteiro-Moreira, Ana Cristina O; Moreira, Renato A; Araújo, Airton A; Moura, Arlindo A

    2015-11-01

    The present study evaluated the effects of heat stress on the ram seminal plasma proteome. Six Morada Nova rams were scrotal insulated for 8 days. Scrotal circumference, sperm parameters, and seminal fluid proteins were evaluated before (Day 0) and twice during scrotal insulation (Days 4 and 8), and weekly until semen parameters returned to preinsulation values (normal). Seminal proteins were analyzed by two-dimensional SDS-PAGE and mass spectrometry. Scrotal circumference decreased from 30 ± 0.4 cm on Day 0 to 22.6 ± 0.6 cm on Day 36 (P Rams were azoospermic between Days 29 and 64, and sperm concentration came back to normal on Day 92. The number of spots/two-dimensional gel reduced from 256 ± 31 on Day 0 to 104 ± 14 on Day 29 (when rams were azoospermic) and then increased to 183 ± 9 on Day 113 (P rams, indicating alterations in both spermatogenesis and sperm maturation. Changes of seminal plasma proteome were coincidental with variations in semen parameters. Proteins affected by heat challenge are potentially involved in sperm protection, maturation, and fertilization.

  19. Sperm motility of externally fertilizing fish and amphibians.

    Science.gov (United States)

    Browne, R K; Kaurova, S A; Uteshev, V K; Shishova, N V; McGinnity, D; Figiel, C R; Mansour, N; Agney, D; Wu, M; Gakhova, E N; Dzyuba, B; Cosson, J

    2015-01-01

    We review the phylogeny, sperm competition, morphology, physiology, and fertilization environments of the sperm of externally fertilizing fish and amphibians. Increased sperm competition in both fish and anurans generally increases sperm numbers, sperm length, and energy reserves. The difference between the internal osmolarity and iconicity of sperm cells and those of the aquatic medium control the activation, longevity, and velocity of sperm motility. Hypo-osmolarity of the aquatic medium activates the motility of freshwater fish and amphibian sperm and hyperosmolarity activates the motility of marine fish sperm. The average longevity of the motility of marine fish sperm (~550 seconds) was significantly (P fish sperm (~150 seconds), with the longevities of both marine and freshwater fish being significantly (P fish (140 μm/s) or freshwater fish (135 μm/s) sperm. The longevity of the sperm of giant salamanders (Cryptobranchoidea) of approximately 600 seconds was greater than that of freshwater fish sperm but much lower than anuran sperm. Our research and information from the literature showed that higher osmolarities promote greater longevity in anuran sperm, and some freshwater fish sperm, and that anuran and cryptobranchid sperm maintained membrane integrity long after the cessation of motility, demonstrating a preferential sharing of energy reserves toward the maintenance of membrane integrity. The maintenance of the membrane integrity of anuran sperm in fresh water for up to 6 hours showed an extremely high osmotic tolerance relative to fish sperm. The very high longevity and osmotic tolerance of anuran sperm and high longevity of cryptobranchid sperm, relative to those of freshwater fish, may reflect the complex fertilization history of amphibian sperm in general and anurans reversion from internal to external fertilization. Our findings provide a greater understanding of the reproductive biology of externally fertilizing fish and amphibians, and a

  20. Use of spin labels to evaluate effects of cold shock and osmolality on sperm

    Energy Technology Data Exchange (ETDEWEB)

    Hammerstedt, R.H.; Keith, A.D.; Snipes, W.; Amann, R.P.; Arruda, D.; Griel, L.C. Jr.

    1978-05-01

    Spin labels were used to evaluate the effects of butylated hydroxytoluene (BHT), rapid cooling to 0/sup 0/C and osmolality on the integrity of sperm membranes. In vitro incubation of rabbit sperm with 0.5 mM BHT prior to artificial insemination did not alter the fertilizing ability of the sperm. Sperm from 6 species were ranked in terms of susceptibility to membrane damage caused by rapid cooling to 0/sup 0/C. The integrity of bull and ram sperm membranes was destroyed by the rapid cooling; BHT protected membranes of these spermatozoa from cold-induced lysis. Boar sperm membranes were porous after rapid cooling and BHT did not prevent this membrane damage. Membranes of rabbit and rooster sperm were not damaged by rapid cooling to 0/sup 0/C. Stallion sperm could not be analyzed because their membranes were altered by addition of reagents necessary to use the technique. The responses of bull, ram and rabbit sperm membranes to hyper- and hypo-osmotic conditions were determined. Hypotonic treatment (less than 200 mOsm) resulted in a 50 percent expansion of the volume of the aqueous compartment of sperm while hypertonic (700 mOsm) conditions compressed the volume of the aqueous compartment to 25 to 30 percent of the volume measured at 300 mOsm. Bull sperm, but not rabbit or ram sperm, responded as ''perfect osmometers'' between 300 and 700 mOsm.

  1. A transmission electron microscopic study of impala (Aepyceros melampus sperm from the Kruger National Park

    Directory of Open Access Journals (Sweden)

    D.J. Ackerman

    1996-02-01

    Full Text Available Since knowledge of sperm morphology can play an important role in semen evaluation and fertilisation, baseline data are required. Live spermatozoa were collected from the cauda epididymis of 64 impala rams in the Kruger National Park and studied by trans- mission electron microscopy. The morphology of normal sperm was documented. The impala sperm shares characteristics with other members of the Bovidae. The occurrence of appendages on the cytoplasmic droplet of the flagellum of impala sperm is described for the first time. A total of 31 micrographs, showing typical features of impala sperm, in sections through various planes of the sperm, are presented.

  2. On RAM priority queues

    Energy Technology Data Exchange (ETDEWEB)

    Thorup, M. [Univ. of Coepnhagen (Denmark)

    1996-12-31

    Priority queues are some of the most fundamental data structures. They are used directly for, say, task scheduling in operating systems. Moreover, they are essential to greedy algorithms. We study the complexity of priority queue operations on a RAM with arbitrary word size. We present exponential improvements over previous bounds, and we show tight relations to sorting. Our first result is a RAM priority queue supporting insert and extract-min operations in worst case time O(log log n) where n is the current number of keys in the queue. This is an exponential improvement over the O({radical}log n) bound of Redman and Willard from STOC`90. Our algorithm is simple, and it only uses AC{sup 0} operations, meaning that there is no hidden time dependency on the word size. Plugging this priority queue into Dijkstra`s algorithm gives an 0(mloglogm) algorithm for the single source shortest path problem on a graph with m edges, as compared with the previous O(m {radical} log m) bound based on Redman and Willard`s priority queue.

  3. Sperm DNA fragmentation is related to sperm morphological staining patterns.

    Science.gov (United States)

    Sá, Rosália; Cunha, Mariana; Rocha, Eduardo; Barros, Alberto; Sousa, Mário

    2015-10-01

    In this prospective comparative study, sperm DNA fragmentation (sDNAfrag) was compared at each step of a sequential semen preparation, with semen parameters according to their degree of severity. At each step (fractions) of the sequential procedure, sDNAfrag was determined: fresh (Raw), after gradient centrifugation, washing, and swim-up (SU) for 70 infertile men enrolled in intracytoplasmic sperm injection cycles. sDNAfrag significantly (P = 0.04; P < 0.0001) decreased throughout the steps of semen preparation, with centrifugation and washing not increasing it. A negative correlation to sperm motility was observed in Raw and SU fractions, and a higher sDNAfrag was observed in samples with lower semen quality. Our results confirm that the steps of the sequential procedure do not compromise sperm DNA integrity and progressively decreased sDNAfrag regardless of the sperm abnormality and that semen parameters with lower quality present higher sDNAfrag. Four distinct patterns were observed, of which the entire sperm head staining was the pattern most expressed in all studied fractions. Additionally, the sperm head gene-rich region staining pattern was reduced by the procedure. This suggests that pattern quantification might be a useful adjunct when performing sDNAfrag testing for male infertility. Copyright © 2015 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  4. Do increased levels of progesterone and progesterone/estradiol ratio on the day of human chorionic gonadotropin affects pregnancy outcome in long agonist protocol in fresh in vitro fertilization/intracytoplasmic sperm injection cycles?

    Directory of Open Access Journals (Sweden)

    Neeta Singh

    2015-01-01

    Full Text Available Background: The effect of elevated levels of serum progesterone (P 4 and estradiol (E 2 on the day of human chorionic gonadotropin and their cut-off value on in vitro fertilization (IVF outcomes is still not clear. Aims: The aim was to evaluate the association between serum P 4 , E 2 and progesterone/estradiol ratio (P 4 /E 2 on pregnancy outcome in IVF/intracytoplasmic sperm injection (ICSI cycles with long agonist protocol. Setting and Design: Retrospective, single center, cohort study. Materials and Methods: A review of complete data of 544 women undergoing fresh IVF/ICSI cycles (539 cycles with long agonist protocol from January 2012 to February 2014 was done. Data were stratified into Three groups according to the number of oocytes retrieved: low (≤4 oocytes obtained, intermediate (5-19 oocytes obtained, and high ovarian response (≥20 oocytes obtained. Statistical Analysis: Fishers exact test/Chi-square was carried for comparing categorical data. Receiver operating characteristics analysis was performed to determine the cut-off value for P 4 and P 4 /E 2 detrimental for pregnancy. Results: A negative association was observed between pregnancy rate (PR and serum P 4 and P 4 /E 2 levels with no effect on fertilization and cleavage rate. The overall cut-off value of serum P 4 and P 4 /E 2 ratio detrimental for pregnancy was found to be 1.075 and ≥0.35, respectively. Different P 4 threshold according to the ovarian responders were calculated, 1.075 for intermediate and 1.275 for high responders. Serum E 2 levels were not found to be significantly associated with PR. Conclusion: Serum P 4 levels and P 4 /E 2 ratio are a significant predictor for pregnancy outcome without affecting cleavage and fertilization rate while serum estradiol levels do not seem to affect PR.

  5. Remotely Accessible Management System (RAMS).

    Science.gov (United States)

    Wood, Rex

    Oakland Schools, an Intermediate School District for Administration, operates a Remotely Accessible Management System (RAMS). RAMS is composed of over 100 computer programs, each of which performs procedures on the files of the 28 local school districts comprising the constituency of Oakland Schools. This regional service agency covers 900 square…

  6. Externally Verifiable Oblivious RAM

    Directory of Open Access Journals (Sweden)

    Gancher Joshua

    2017-04-01

    Full Text Available We present the idea of externally verifiable oblivious RAM (ORAM. Our goal is to allow a client and server carrying out an ORAM protocol to have disputes adjudicated by a third party, allowing for the enforcement of penalties against an unreliable or malicious server. We give a security definition that guarantees protection not only against a malicious server but also against a client making false accusations. We then give modifications of the Path ORAM [15] and Ring ORAM [9] protocols that meet this security definition. These protocols both have the same asymptotic runtimes as the semi-honest original versions and require the external verifier to be involved only when the client or server deviates from the protocol. Finally, we implement externally verified ORAM, along with an automated cryptocurrency contract to use as the external verifier.

  7. Evaluation of semen extenders for short-term storage of ram semen at 4° C

    Science.gov (United States)

    Preliminary studies found that progressive motility of ram sperm declined ~75% when stored at 4° C for 24 h, and continued to decline over time when using extenders supplemented with 5% egg yolk. The current study evaluated the effects of different combinations of extenders, ethylene glycol (EG), eg...

  8. Intrinsic Hydrophobicity of Rammed Earth

    Science.gov (United States)

    Holub, M.; Stone, C.; Balintova, M.; Grul, R.

    2015-11-01

    Rammed earth is well known for its vapour diffusion properties, its ability to regulate humidity within the built environment. Rammed earth is also an aesthetically iconic material such as marble or granite and therefore is preferably left exposed. However exposed rammed earth is often coated with silane/siloxane water repellents or the structure is modified architecturally (large roof overhangs) to accommodate for the hydrophilic nature of the material. This paper sets out to find out optimal hydrophobicity for rammed earth based on natural composite fibres and surface coating without adversely affecting the vapour diffusivity of the material. The material is not required to be waterproof, but should resist at least driving rain. In order to evaluate different approaches to increase hydrophobicity of rammed earth surface, peat fibres and four types of repellents were used.

  9. Effect of donkey seminal plasma on sperm movement and sperm-polymorphonuclear neutrophils attachment in vitro.

    Science.gov (United States)

    Miró, Jordi; Vilés, Karina; García, Wilber; Jordana, Jordi; Yeste, Marc

    2013-08-01

    To evaluate the effect of seminal plasma in endometrial inflammation in donkeys, samples from fresh pure, fresh diluted and frozen-thawed semen of three different jackasses were co-incubated in water bath at 37°C with uterine Jennie's secretions collected 6h after artificial insemination with frozen-thawed donkey semen. Individual sperm movement parameters using the computerised sperm analysis system (CASA) and sperm-polymorphonuclear neutrophils (sperm-PMN) attachment observed in Diff-Quick stained smears were evaluated at 0, 1, 2, 3 and 4h of co-incubation. Controls consisted of incubating diluted or frozen-thawed sperm in the absence of uterine secretions. For data analyses, a repeated measures ANOVA was performed with incubation time as intra-subject factor and with treatment and donkey as inter-subject factor, followed by a post-hoc Bonferroni's test. Greater values (Pdonkey semen. As a result, spermatozoa with the greater motility characteristics are selected.

  10. Cryopreservation of European catfish Silurus glanis sperm: sperm motility, viability, and hatching success of embryos.

    Science.gov (United States)

    Linhart, Otomar; Rodina, Marek; Flajshans, Martin; Gela, David; Kocour, Martin

    2005-12-01

    The aim of this study was to elaborate cryopreservation methods for ex situ conservation of European catfish. The success of sperm cryopreservation was evaluated by post-thaw sperm motility and velocity, percentage of live spermatozoa and fertility (hatching rates) using frozen/thawed sperm. The best hatching rates of 82-86% were obtained with sperm stored for 5 h before freezing in immobilizing solution and frozen with Me2SO in concentrations of 8, 10, and 12%, or with a mixture of 5% Me2SO and 5% propandiole. These results did not significantly differ from the fresh sperm control sample. The percentage of live spermatozoa in frozen/thawed sperm did not correlate with hatching rate or motility of spermatozoa, but was negatively correlated with velocity of spermatozoa (r=-0.47, P=0.05). The percentage motility in frozen/thawed sperm ranged from 8 to 62%, when sperm was stored in immobilizing solution 5h before freezing. The average value in the fresh sperm (control) was 96%. The frozen/thawed sperm motility rate significantly correlated with the hatching rate (r=0.76, P=0.0002), but not with the percentage of live spermatozoa (r=0.16, P=0.52) or the sperm velocity (r=0.07, P=0.79). The velocity of frozen/thawed spermatozoa ranged from 37 to 85 microm/s, whereby methanol concentrations of 7.5 and 10% resulted in highest velocities. Freezing sperm volumes of 1-4 ml did not affect the quality of frozen/thawed sperm.

  11. A scanning electron microscopic study of impala (Aepyceros melampus sperm from the Kruger National Park

    Directory of Open Access Journals (Sweden)

    D.J. Ackerman

    1996-02-01

    Full Text Available Since knowledge of sperm morphological characteristics can play an important role in semen evaluation and fertilisation, baseline data on sperm ultrastructure are required. Live spermatozoa were collected from the cauda epididymis from 64 impala rams in the Kruger National Park and 5082 spermatozoa from 40 of these impala were studied by scanning electron microscopy. The mean length of impala sperm was 59.23 @ 2.7 um. The morphology of normal sperm as well as the occurrence of abnormalities were documented. The morphology of impala sperm were compared with those of other mammals. New findings on appendages of the cytoplasmic droplet are described and interpreted.

  12. Seasonal changes in reproductive activity, sperm variables and sperm freezability in Blanca Andaluza bucks

    Directory of Open Access Journals (Sweden)

    Lourdes Gallego-Calvo

    2015-12-01

    Full Text Available Interest in the preservation of endangered breeds such as the Blanca Andaluza goat, has increased and some steps should be therefore taken to ensure it. The study was designed to determine the seasonal reproductive pattern of Blanca Andaluza bucks, and whether this affects the quality of their semen and its freezability over the year. Seven bucks were used and their body weight, testicular weight, plasma testosterone concentration and fresh sperm quality determined every week. The collected sperm was cryopreserved and stored; it was then thawed and the same sperm quality variables measured every fortnight. High plasma testosterone concentrations were recorded during the summer and autumn, and low concentrations were recorded during winter and spring (p<0.001. No differences were seen between seasons in terms of the percentage of bucks ejaculating, the percentage of active bucks, or ejaculate volume. However, the sperm concentration, the total number of sperm per ejaculate, and the values for most fresh sperm variables were lower during the winter period (at least p<0.05. After freezing-thawing, the quality of winter-collected sperm was better, in some respects, than that of summer-collected sperm (at least p<0.05. These results reveal that Blanca Andaluza bucks show seasonal reproductive activity in terms of their plasma testosterone concentration, but no clear change in their sexual behaviour between seasons was observed. The values of fresh sperm variables also vary over the year, reaching their lowest during winter. However, after freezing-thawing, winter-collected sperm is of overall better quality than sperm collected during the summer.

  13. Seasonal changes in reproductive activity, sperm variables and sperm freezability in Blanca Andaluza bucks

    Energy Technology Data Exchange (ETDEWEB)

    Gallego-Calvo, L.; Gatica, M.C.; Santiago-Moreno, J.; Guzmán, J.L.; Zarazaga, L.

    2015-07-01

    Interest in the preservation of endangered breeds such as the Blanca Andaluza goat, has increased and some steps should be therefore taken to ensure it. The study was designed to determine the seasonal reproductive pattern of Blanca Andaluza bucks, and whether this affects the quality of their semen and its freezability over the year. Seven bucks were used and their body weight, testicular weight, plasma testosterone concentration and fresh sperm quality determined every week. The collected sperm was cryopreserved and stored; it was then thawed and the same sperm quality variables measured every fortnight. High plasma testosterone concentrations were recorded during the summer and autumn, and low concentrations were recorded during winter and spring (p<0.001). No differences were seen between seasons in terms of the percentage of bucks ejaculating, the percentage of active bucks, or ejaculate volume. However, the sperm concentration, the total number of sperm per ejaculate, and the values for most fresh sperm variables were lower during the winter period (at least p<0.05). After freezing-thawing, the quality of winter-collected sperm was better, in some respects, than that of summer-collected sperm (at least p<0.05). These results reveal that Blanca Andaluza bucks show seasonal reproductive activity in terms of their plasma testosterone concentration, but no clear change in their sexual behaviour between seasons was observed. The values of fresh sperm variables also vary over the year, reaching their lowest during winter. However, after freezing-thawing, winter-collected sperm is of overall better quality than sperm collected during the summer. (Author)

  14. Effect of Egg Yolk and Cooling on Storage of Ram Coated Spermatozoa

    Directory of Open Access Journals (Sweden)

    A. Mohammadi-Nohdehy

    2013-08-01

    Full Text Available This study was conducted to evaluate the effect of egg yolk and cooling on ram coated spermatozoa. Semen was collected from three ram by artificial vagina contacted with a tube containing Tris- fructose-egg yolk 15%. Samples were pooled, centrifuged by 700 g for 10 min and removed supernatant. Then, samples were diluted by Tris-glucose and centrifuged again to remove seminal plasma and egg yolk. Aliquots split into two fractions and each one was split into 5 parts and added egg yolk 0, 5, 10, 15 and 20%. The half of the treatments were gradually cold and other ones were encountered with cold shock then samples were incubated at 5C for 72 h. Progressive sperm motility, plasma membrane integrity, viability (by Hoechst 33258 fluorescent staining and acrosome reaction (by PNA-Alexa flur-488 were investigated at 0, 24, 48 and 72 h. The results showed that there was no difference between 15% and 20% egg yolk in the progressive sperm motility but they were higher than 0% and 5% egg yolk. There was highest difference between 0% and 20 % egg yolk in the progressive sperm motility. There was no difference among the treatments containing egg yolk in plasma membrane integrity and acrosome reaction. In both cooling rate, there was no difference among the treatments containing egg yolk in the sperm viability. It was suggested that 20% egg yolk was superior to keep the function of ram coated spermatozoa for storage at 5C.

  15. Sperm competition, sperm numbers and sperm quality in muroid rodents.

    Directory of Open Access Journals (Sweden)

    Laura Gómez Montoto

    Full Text Available Sperm competition favors increases in relative testes mass and production efficiency, and changes in sperm phenotype that result in faster swimming speeds. However, little is known about its effects on traits that contribute to determine the quality of a whole ejaculate (i.e., proportion of motile, viable, morphologically normal and acrosome intact sperm and that are key determinants of fertilization success. Two competing hypotheses lead to alternative predictions: (a sperm quantity and quality traits co-evolve under sperm competition because they play complementary roles in determining ejaculate's competitive ability, or (b energetic constraints force trade-offs between traits depending on their relevance in providing a competitive advantage. We examined relationships between sperm competition levels, sperm quantity, and traits that determine ejaculate quality, in a comparative study of 18 rodent species using phylogenetically controlled analyses. Total sperm numbers were positively correlated to proportions of normal sperm, acrosome integrity and motile sperm; the latter three were also significantly related among themselves, suggesting no trade-offs between traits. In addition, testes mass corrected for body mass (i.e., relative testes mass, showed a strong association with sperm numbers, and positive significant associations with all sperm traits that determine ejaculate quality with the exception of live sperm. An "overall sperm quality" parameter obtained by principal component analysis (which explained 85% of the variance was more strongly associated with relative testes mass than any individual quality trait. Overall sperm quality was as strongly associated with relative testes mass as sperm numbers. Thus, sperm quality traits improve under sperm competition in an integrated manner suggesting that a combination of all traits is what makes ejaculates more competitive. In evolutionary terms this implies that a complex network of genetic

  16. Sperm flagellum volume determines freezability in red deer spermatozoa

    National Research Council Canada - National Science Library

    Ros-Santaella, José Luis; Domínguez-Rebolledo, Alvaro Efrén; Garde, José Julián

    2014-01-01

    ... provides sperm motility. Here, for the first time, we determined the volumes of the flagellum structures in fresh epididymal red deer spermatozoa using a stereological method under phase contrast microscopy...

  17. Laser-propelled ram accelerator

    Energy Technology Data Exchange (ETDEWEB)

    Sasoh, A. [Tohoku Univ., Sendai (Japan). Inst. of Fluid Science

    2000-11-01

    The concept of 'laser-propelled ram accelerator (L-RAMAC)' is proposed. Theoretically it is capable of achieving a higher launch speed than that by a chemical ram accelerator because a higher specific energy can be input to the propellant gas. The laser beam is supplied through the muzzle, focused as an annulus behind the base of the projectile. The performance of L-RAMAC is analized based on generalized Rankine-Hugoniot relations, suggesting that a superorbital muzzle speed is achievable out of this device. (orig.)

  18. Tris buffer improves fluorescence yield of ram spermatozoa when evaluating membrane integrity.

    Science.gov (United States)

    Yániz, Jesús Luis; Mateos, José Angel; Santolaria, Pilar

    2012-04-01

    This study was designed to evaluate the effect of various buffers on the fluorescence signal intensity of two fluorochromes (IP and CFDA) when used to assess the membrane integrity of ram sperm. Second ejaculates (18) from nine adult males were collected using an artificial vagina and diluted in either MOPS, TRIS, TES, HEPES, citrate, or phosphate-based extenders. Semen samples were stored at 15°C and the membrane integrity was assessed within the first 24 h of storage. Mean fluorescence intensity (FI) of PI- and CDFA-labeled sperm heads and fluorescence background noise (FBN) were determined quantitatively using Image J software. Fluorescence contrast (FC) was expressed as the difference between FI and FBN. Significantly, higher FI and FC were recorded when TRIS diluent was used, rather than the other diluents, both in the propidium- and fluorescein-labeled cells. The citrate and phosphate-based extenders showed intermediate results of FC between those of TRIS and zwitterionic (MOPS, TES and HEPES) groups for the PI-labeled sperm. However, in the CFDA-labeled sperm, the lower values of FC were obtained in the citrate and phosphate groups due to increased levels of FBN. For the membrane-damaged sperm, fluorescent labeling was limited to the sperm heads when TRIS-buffer was used, whereas in the other groups, the sperm tail was also frequently observed. It was concluded that TRIS buffer solution markedly increases the fluorescence yield of IP/CFDA-labeled sperm cells in the ram and that this should be considered when evaluating their membrane integrity.

  19. In vitro effects of nonylphenol on motility, mitochondrial, acrosomal and chromatin integrity of ram and boar spermatozoa.

    Science.gov (United States)

    Uguz, C; Varisli, O; Agca, C; Evans, T; Agca, Y

    2015-10-01

    The objective of this study was to determine the effects of nonylphenol (NP) on viability of ram and boar sperm in vitro. Ram or boar spermatozoa were exposed to 1, 10, 100, 250 and 500 μg NP ml(-1) for 1, 2, 3 or 4 h. Computer-assisted sperm motility analysis (CASA) system was used to evaluate sperm motility characteristics. Flow cytometry was used to determine mitochondrial membrane potential (MMP) and chromatin integrity, while epifluorescent microscopy was used to determine sperm acrosomal status. Exposure of both species spermatozoa to 250 and 500 μg NP ml(-1) was detrimental to progressive motility (P boar spermatozoa with high MMP declined drastically after exposures to ≥250 μg ml(-1) NP (P boar spermatozoa and 10 μg ml(-1) NP for ram spermatozoa. These data show adverse effects of NP on ram and boar spermatozoa and thus its potential harmful effects on male reproduction as NP is found in fruits, vegetables, human milk, fish and livestock products.

  20. Determination of fatty acid profile in ram spermatozoa and seminal plasma.

    Science.gov (United States)

    Díaz, R; Torres, M A; Bravo, S; Sanchez, R; Sepúlveda, N

    2016-08-01

    Fatty acids are important in male reproductive function because they are associated with membrane fluidity, acrosome reaction, sperm motility and viability, but limited information exists about the fatty acid profile of ram semen. Our aim was to determine the fatty acid composition in ram spermatozoa and seminal plasma. Sixty ejaculates were obtained from three ram (20 ejaculates/ram) using artificial vagina. Ram spermatozoa (RS) and seminal plasma (SP) were separated using centrifugation, and the fatty acids were analysed by gas chromatography. Total lipids obtained in ram spermatozoa were 1.8% and 1.6% in seminal plasma. Saturated fatty acid (SFA) was proportionally major in SP (66.6%) that RS (49.9%). The highest proportions of SFA corresponded to C4:0 (RS = 16.3% and SP = 28.8%) and C16:0 (RS = 16.3% and PS = 20%). The most important unsaturated fatty acid (UFA) was docosahexaenoic acid (DHA), 44.9% in RS and 31.5% in SP. The profile of fatty acid and their proportions showed differences between spermatozoa and seminal plasma.

  1. Lime-Crusted Rammed Earth: Materials Study

    OpenAIRE

    Mileto, Camilla; Vegas López-Manzanares, Fernando; Alejandre, Francisco Javier; Martín, Juan Jesús; Garcia Soriano, Lidia

    2013-01-01

    This study analyses the durability of rammed-earth wall construction techniques. The analysis focuses on three medieval masonry types from the Castle of Villavieja (Castellón, Spain) using two variations of lime-reinforced rammed earth in its walls: lime-crusted rammed earth and brick-reinforced rammed earth. Materials analysis reveals the good properties of the materials used in the outer wall facing despite its age. It also clearly shows how deterioration depends more on the construction t...

  2. Does Vitamin E or Vitamin E plus Selenium improve reproductive performance of rams during hot weather?

    Directory of Open Access Journals (Sweden)

    Basilio Floris

    2010-01-01

    Full Text Available The objective of this study was to determine the effect of Vitamin E (Vit E and Selenium (Se on semen quality, sexual activity, packed cell volume (PCV, and white blood cell counts (WBC of Awassi rams during the hot season. Twelve Awassi rams were subdivided into three groups and treated for 90 days. Rams in the 1st group (T1 were treated twice weekly with 175 mg/ram vitamin E at intervals of 12h; rams in the 2nd group (T2 were treated on the same schedule with 70 mg/ram vitamin E plus 2800 mg selenium; and rams in 3rd group (T3 served as controls. Sperm quality (percentage of motile cells and percentage of morphologically normal cells and quantity (sperm volume, and concentration were recorded weekly. Sexual activity was tested by using the pen libido test at monthly intervals. Blood samples were taken before treatment and after the 1st, 2nd, and 3rd months. Semen quality was significantly affected by treatments: the ejaculate volume increased in T2 vs T1, and T3; mass activity and individual motility were increased in T1 and T2 vs T3. Sperm concentration was increased in T2 and T1 vs T3, and the percentage of dead and abnormal spermatozoa was reduced in T1 and T2 vs T3, though the differences were not statistically significant. Pen libido test showed reduced reaction time for the first mount in the 1st month in T2 and T1 vs T3. The number of serves was increased in groups T1 and T2 vs T3. There were no significant differences in PCV among groups. After 3 months from the beginning of treatments, the percentage of lymphocytes increased in T1 and T2 vs. T3, while the percentage of neutrophils was reduced in T2 vs T3 in the 2nd and 3rd months. The results of this experiment indicate that treatments with vitamin E alone or in combination with selenium improved semen characteristics and reproductive performance of Awassi rams during the hot season.

  3. Effect of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem.

    Science.gov (United States)

    Kaabi, M; Paz, P; Alvarez, M; Anel, E; Boixo, J C; Rouissi, H; Herraez, P; Anel, L

    2003-10-15

    Post-mortem spermatozoa recovery is an important technique for obtaining germplasm reserves from genetically valuable animals or endangered species. However, there are many factors that influence the outcome of this technique. We have studied the effect of the interval between animal's death and sperm recovery (0, 24 or 48 h) on the quality and freezability of ram spermatozoa from cauda epididymidis. Storage temperature of epididymis (room temperature or 5 degrees C) was also analysed. Spermatozoa were diluted with Tes-Tris-Fructose solution supplemented with egg yolk (10%) and glycerol (4%), and frozen using a programmable biofreezer (-20 degrees C/min). Pre-freeze and post-thaw sperm samples showed viable spermatozoa up to 48 h after the animal's death, although their quality declined significantly as post-mortem storage time increased. Epididymis sperm stored at 5 degrees C showed better motility and a lower percentage of abnormal forms than epididymis stored at room temperature after 24 and 48 h. The fertilizing ability of cauda epididymis ram spermatozoa obtained at 0 and 24h after the animal's death is similar to that of ejaculated spermatozoa. Therefore, a good protocol for post-mortem semen collection in rams when epididymal spermatozoa cannot be collected immediately, is to preserve the epididymis at 5 degrees C and process the samples in the first 24h after the animal's death.

  4. Estrous sheep serum enables in vitro capacitation of ram spermatozoa while preventing caspase activation.

    Science.gov (United States)

    Del Olmo, E; García-Álvarez, O; Maroto-Morales, A; Ramón, M; Jiménez-Rabadán, P; Iniesta-Cuerda, M; Anel-Lopez, L; Martinez-Pastor, F; Soler, A J; Garde, J J; Fernández-Santos, M R

    2016-01-15

    Estrous sheep serum (ESS) is considered the most efficient agent for in vitro capacitation of ram spermatozoa. We have explored the relationship between caspase activation and capacitation in ram. Semen samples from 17 rams were cryopreserved. In vivo fertility was evaluated after intrauterine artificial insemination. Samples were submitted to four treatments: control, ESS (10%), caspase inhibitor (Z-VAD-FMK), and estrous ewe serum plus caspase inhibitor (I + E). Sperm samples were incubated for 30 minutes at 38.5 °C and 5% CO2 and analyzed with flow cytometry for mitochondrial membrane potential (MitoTracker deep red), sperm viability and apoptosis-like changes (YO-PRO-1/propidium iodide), acrosomal status (peanut agglutinin-fluorescein isothiocyanate), membrane fluidity (merocyanine 540), and caspase activity (Vybrant FAM kits for polycaspases, caspase-8, and caspases 3-7). Estrous sheep serum induced changes compatible with capacitation, doubling the proportion of viable spermatozoa with increased merocyanine 540 and increasing YO-PRO-1(+) and acrosome-reacted spermatozoa (P ram spermatozoa by using ESS suggests a downregulation in apoptotic pathways. However, males with the lowest fertility showed parameters similar to high-fertility males, suggesting that other factors were involved apart from capacitation and/or caspase activation.

  5. The quality of Garut ram liquid semen in Tris egg yolk extender to the sucrose supplementation

    Directory of Open Access Journals (Sweden)

    Yulnawati

    2016-01-01

    Full Text Available The successful program of artificial insemination in sheep is determined by the quality of sperm that are used. Therefore, it is important to maintain the quality of sperm during and after storage in low temperature. The research was conducted to study the influence of sucrose in the Tris-yolk extender in maintaining the quality of Garut ram sperm during preservation in reaction tube for four days at 5°C. Ejaculated sperm was collected once a week for five weeks using artificial vagina from same ram. Semen was divided into four groups of extenders, i.e. Tris egg yolk 20% (TKT, TKT + 0,1% sucrose, TKT + 0,3% sucrose and TKT + 0,5% w/v sucrose. The results showed that the percentage of motility on day four (D-4 of storage in TKT (41.00 ± 2.0% was lower (P0.05 from TKT + 0.1% (45.00 ± 3.16%. Whereas the percentage of live cells (%H on D-4 in TKT, TKT + 0.1%, TKT + 0.3% and TKT + 0.5% were 55.00 ± 2.19%; 57.20 ± 2.79%; 59.20 ± 3.25% and 61.20 ± 3.60%, respectively. Meanwhile, the percentage of membrane integrity (MPU in TKT (51.80 ± 1.94% was significantly different (P0.05 from TKT + 0.1% (54.80 ± 2.86%. In conclusion, the addition of sucrose 0.3% w/v into TKT extender could maintain the quality of Garut ram sperm more efficiently and better than TKT extender.

  6. Turbine ram jet engine. Turbinen-Staustrahltriebwerk

    Energy Technology Data Exchange (ETDEWEB)

    Jabs, A.

    1991-04-25

    A turbine ram jet engine is described with a ram compressed air duct, which is formed between an outer duct wall and a housing-like jacket of a basic turbine engine and is to be shut off on the incoming air side with means of shutting off, by which the ram compressed air duct is shut off with the basic engine switched on and the ram jet engine switched off with simultaneous release of suction air into the basic engine. The jacket should form an air shaft opening out polygonally in the direction of the end of the ram compressed air duct on the inlet side and should have penetrations, where the means of shutting off at the penetrations are hinged flaps, which open up the penetrations with the ram jet engine switched on and thus form a shut-off body of the air shaft symmetrically opposite the ram air flow.

  7. Effect of cholesterol-loaded-cyclodextrin on sperm viability and acrosome reaction in boar semen cryopreservation.

    Science.gov (United States)

    Lee, Yong-Seung; Lee, Seunghyung; Lee, Sang-Hee; Yang, Boo-Keun; Park, Choon-Keun

    2015-08-01

    This study was undertaken to examine the effect of cholesterol-loaded-cyclodextrin (CLC) on boar sperm viability and spermatozoa cryosurvival during boar semen cryopreservation, and methyl-β-cyclodextrin (MBCD) was treated for comparing with CLC. Boar semen treated with CLC and MBCD before freezing process to monitor the effect on survival and capacitation status by flow cytometry with appropriate fluorescent probes. Sperm viability was higher in 1.5mg CLC-treated sperm (76.9±1.01%, Psperm before cryopreservation (58.7±1.31% and 60.3±0.31%, respectively). For CTC patterns, F-pattern was higher in CLC treated sperm than MBCD-treated sperm, for B-pattern was higher in CLC-treated sperm than fresh sperm (Psperm) was lower in CLC-treated sperm than MBCD-treated sperm (Pboar semen.

  8. Cryopreservation of epididymal sperm in tree shrews (Tupaia belangeri).

    Science.gov (United States)

    Ping, S; Wang, F; Zhang, Y; Wu, C; Tang, W; Luo, Y; Yang, S

    2011-07-01

    Cryopreservation of sperm from tree shrews, which are considered primitive primates, would enhance genetic management and breeding programs. Epididymal sperm were surgically harvested from male tree shrews, cryopreserved in two Tes-Tris-based cryodiluents, and used in four experiments. In Experiment 1, there were no significant differences in motility and acrosome integrity among five concentrations of egg yolk in TTE after cooling to 4 °C. However, sperm frozen in TTE containing 20% egg yolk at -172 °C/min had better (P 0.05) among treatments. In Experiment 3, sperm frozen in TTE diluent had higher (P 0.05) in the fertilization rate of oocytes and the proportion of tree shrews yielding fertilized oocytes, following AI with fresh versus frozen sperm. In conclusion, tree shrew epididymal sperm were successfully cryopreserved, as assessed by post-thaw motility, acrosome integrity, and fertilizing ability.

  9. Effects of vitrification on ram spermatozoa using free-egg yolk extenders.

    Science.gov (United States)

    Jiménez-Rabadán, Pilar; García-Álvarez, Olga; Vidal, Ana; Maroto-Morales, Alejandro; Iniesta-Cuerda, María; Ramón, Manuel; del Olmo, Enrique; Fernández-Santos, Rocío; Garde, J Julián; Soler, Ana Josefa

    2015-08-01

    The present study aimed to examine the behavior of ram spermatozoa subjected to a vitrification process in free-egg yolk diluents in relation with conventional diluents and cryopreservation protocol used in this species. Previously it was investigated the toxicity of cryoprotectants, sucrose and glycerol, based on different concentrations (sucrose at 0.03 M, 0.05 M, 0.15 M and 0.25 M; and glycerol at 3%, 7%, 14% and 18%) compared to a commercial extender (Biladyl® with 20% egg yolk and 7% glyerol). Cryoprotectants which reported less toxicity were chosen to perform the vitrification and results were compared with the conventional cryopreservation. Semen from three rams was collected by electroejaculation. The sperm evaluation was carried out at 0, 2 and 4h through the incubation time at 37°C for the experiment of toxicity and, at thawing when cryopreservation was performed. The sperm quality throughout the incubation time always resulted lower (P⩽0.05) for the free-egg yolk diluents in relation to Biladyl® (control), obtaining the lowest values of sperm quality with the highest concentrations of sucrose and glycerol. The vitrification was carried out with combinations of sucrose and glycerol (sucrose at 0.03 and 0.05 M with 3% and 7% of glycerol, respectively) and with Biladyl® (at different sperm concentrations). The vitrification decreased drastically (P⩽0.05) the sperm quality when combinations of sucrose and glycerol were used. Nevertheless, the sperm samples vitrified with Biladyl® at the lowest sperm concentration showed acceptable values of viability, acrosome integrity and DFI, although the sperm motility was strongly decreased. In conclusion, the use of vitrification with diluents based on combinations of sucrose and glycerol did not work for semen cryopreservation of ram. Promising results were obtained when diluents with egg yolk were used in the vitrification procedure, although more studies are necessary to improve this technique and the use

  10. High resolution DNA content measurements of mammalian sperm

    Energy Technology Data Exchange (ETDEWEB)

    Pinkel, D.; Lake, S.; Gledhill, B.L.; Van Dilla, M.A.; Stephenson, D.; Watchmaker, G.

    1982-01-01

    The high condensation and flat shape of the mammalian sperm nucleus present unique difficulties to flow cytometric measurement of DNA content. Chromatin compactness makes quantitative fluorescent staining for DNA difficult and causes a high index of refraction. The refractive index makes optical measurements sensitive to sperm head orientation. We demonstrate that the optical problems can be overcome using the commercial ICP22 epiillumination flow cytometer (Ortho Instruments, Westwood, MA) or a specially built cell orientating flow cytometer (OFCM). The design and operation of the OFCM are described. Measurements of the angular dependence of fluorescence from acriflavine stained rabbit sperm show that it is capable of orienting flat sperm with a tolerance of +-7/sup 0/. Differences in the angular dependence for the similarly shaped bull and rabbit sperm allow discrimination of these cells. We show that DNA staining with 4-6 diamidino-2-phenylindole (DAPI) or an ethidium bromide mithramycin combination allows resolution of the X and Y populations in mouse sperm. They have also been successful with sperm from the bull, ram, rabbit, and boar. Reliable results with human sperm are not obtained. The accuracy of the staining and measurement techniques are verified by the correct determination of the relative content of these two populations in sperm from normal mice and those with the Cattanach (7 to X) translocation. Among the potential uses of these techniques are measurement of DNA content errors induced in sperm due to mutagen exposure, and assessment of the fractions of X and Y sperm in semen that may have one population artifically enriched.

  11. Studies on Freezing RAM Semen in Absence of Glycerol.

    Science.gov (United States)

    Abdelnaby, Abdelhady Abdelhakeam

    1988-12-01

    Glycerol is widely used as a major cryoprotective agent for freezing spermatozoa of almost all species. However, it reduces fertility of sheep inseminated cervically compared with intrauterine insemination. Studies were conducted to develop a method and procedure for freezing ram semen in the absence of glycerol. Post -thaw survival of ram spermatozoa frozen in the absence of glycerol was affected by time and temperature after collection and before dilution and time after dilution and before freezing. Increase in time at 5^ circC before or after dilution and before freezing increased both post-thaw motility and number of cells passing through Sephadex filter. A cold dilution method was developed. Slow cooling of fresh ram semen and diluting at 5^circ C 2-3 hr. after collection, then freezing 1 hr. after dilution improved both post-thaw motility and number of cells passing through Sephadex filter compared with immediate dilution at 30-37^circC after collection and freezing 3-4 hr. later (P synchronization during breeding season resulted in higher heat response and lambing rate than two injections given 10 days apart.

  12. Ability of Catalonian donkey sperm to penetrate zona pellucida-free bovine oocytes matured in vitro.

    Science.gov (United States)

    Taberner, E; Morató, R; Mogas, T; Miró, J

    2010-04-01

    An experiment was designed to study the interaction between fresh/frozen-thawed donkey spermatozoa and zona pellucida (ZP)-free bovine oocytes in an attempt to develop a model for assessing cryopreserved Catalonian donkey sperm function. Semen from five donkeys was collected using an artificial vagina. Sperm motility and viability were immediately assessed and the semen sample cryopreserved. Sperm viability and motility were then reassessed immediately after thawing. The motion characteristics of the fresh and frozen-thawed spermatozoa were determined using a computer-assisted sperm analysis system. In vitro-matured cow oocytes were inseminated with different percent live donkey sperm (high (>60%) or low (donkey sperm). After 18h of co-incubation, the oocytes were fixed, stained with 4',6-diamidino-2-phenylindole (DAPI) and examined for sperm penetration, the number of penetrated spermatozoa per oocyte, and male pronucleus formation. Frozen-thawed spermatozoa from high viability semen showed significantly lower VCL, VAP and mean ALH values than did high viability fresh spermatozoa. In contrast, frozen-thawed spermatozoa of low viability had significantly higher velocity values than fresh spermatozoa of low viability. A significant positive correlation (Pdonkey spermatozoa were able to fuse with the oolema and even to decondense and form the male pronucleus (85-94%). Larger numbers of penetrated spermatozoa per oocyte were recorded when high viability sperm samples were used, whether fresh (3.02 vs. 1.12 for low viability sperm) or frozen-thawed (3.41 vs. 1.47). Consequently, low viability sperm samples showed higher percentages of monospermic penetration (91.17% and 61.97% for fresh and frozen-thawed sperm samples respectively). These findings suggest that bovine oocytes provide a useful model for assessing the penetration potential of frozen-thawed donkey sperm.

  13. HyRAM Testing Script.

    Energy Technology Data Exchange (ETDEWEB)

    Parkins, Owen

    2014-12-01

    The testing script is to provide a method of inspections to HyRAM (v1.0.0.244 Alpha) application features. This document will lead participants through the use of the application to make sure the application performs as designed. If a feature of the application becomes non-working, this script will relay useful information back to the designers of the application so that the feature can be fixed. This is essential to keep the application updated and performing as designed so that the users of this program can be satisfied. There will be frequent updates of this document to ensure proper testing of future application versions.

  14. Effect of argan oil on liquid storage of ram semen in Tris or skim milk based extenders.

    Science.gov (United States)

    Allai, Larbi; Druart, Xavier; Contell, Jesus; Louanjli, Noureddine; Moula, Anass Ben; Badi, Abdelmoughit; Essamadi, Abdelkhalid; Nasser, Boubker; El Amiri, Bouchra

    2015-09-01

    Due to its high antioxidant content, the argan oil could play a beneficial role in liquid storage of ram semen. The aim of this study was to investigate effects of different concentration of argan oil (ARO) on spermatologic parameters, lipid peroxidation and DNA fragmentation during liquid storage of ram semen until 48 h. Also effects of extenders and temperature on same parameters were assessed. For these aims, semen samples were collected from Boujaâd rams, extended with Tris egg yolk or skim milk extenders without (control) or supplemented with different concentrations of ARO (1%, 2%, 5% and 10% v/v) at a final concentration of 0.8 × 10(9) sperm/mL and stored until 48 h at 5 °C or 15 °C. The sperm quality assessments were performed at different intervals during storage (0, 8, 24 and 48 h). Sperm progressive motility started to decrease after 8h of storage in all temperatures--extenders combinations and dropped steadily during the 8-48 h interval. However, sperm viability, progressive motility and membrane integrity were markedly higher in ARO groups (especially in 1% in Tris and 5% in skim milk) until 24h and 48 h storage at both temperatures compared to controls. The argan oil also decreased the level of spontaneous and induced malondialdehyde (MDA) and the sperm DNA fragmentation until 48 h storage. In conclusion, it was determined that addition of argan oil to conventional extenders may improve the quality of ram semen during liquid storage in different temperatures.

  15. Sperm immobilization activity of Allium sativum L. and other plant extracts

    Institute of Scientific and Technical Information of China (English)

    KausikiChakrabarti; SulagnaPal; AsokK.Bhattacharyya

    2003-01-01

    Aim: To identify possible spermicidal agents through screening a number of edible medicinal plants with antimicrobial activity. Methods: Initial screening was made on the basis of ram cauda epididymal sperm immobiliza-tion immediately after addition of extracts. The most potent extract was selected and was evaluated on both ram and human spermatozoa. To unravel its mode of action several sperm functional tests were carried out, namely viability of cells, hypo-osmotic swelling test for membrane integrity and assays of membrane-bound enzyme 5'-nucleotidase and acrosomal marker enzyme acrosin. Results: The crude aqueous extract of the bulb ofAllium sativum L. Showed the most promising results by instant immobilization of the ram epididymal sperm at 0.25 g/mL and human ejaculated sperm at 0.5 g/mL. Sperm immobilizing effects were irreversible and the factor of the extract responsible for immobilization was thermostable up to 90℃. On boiling at 100℃ for 10 minutes, this activity was markedly reduced. Moreover, this extract was able to cause aggregation of ram sperms into small clusters after 30 minutes of incubation at 37℃. However this property was not found in human spermatozoa. More than 50 % reduction in sperm viability and hypo-osmotic swelling occurred in treated sperm as compared with the controls, indicating the possibility of plasma membrane disintegration which was further supported by the significant reduction in the activity of membrane bound 5''-nucleotidase and acrosomal acrosin. Conclusion: The crude aqueous extract of A. Sativum bulb possesses spermicidal activity in vitro. (Asian J Androl 2003 Jun, 5:131-135 )

  16. Oestrous sheep serum balances ROS levels to supply in vitro capacitation of ram spermatozoa.

    Science.gov (United States)

    Del Olmo, E; García-Álvarez, O; Maroto-Morales, A; Ramón, M; Iniesta-Cuerda, M; Martinez-Pastor, F; Montoro, V; Soler, A J; Garde, J J; Fernández-Santos, M R

    2016-10-01

    Reactive oxygen species (ROS) are fundamental for intracellular signalling. In spermatozoa, they are involved both to apoptosis and to capacitation, and changes in ROS levels can alter the balance between these two processes. Oestrous sheep serum (OSS) is considered an efficient agent for in vitro capacitation of ram spermatozoa. We have explored the effects of OSS on ram sperm physiology, especially on ROS production, during in vitro capacitation. Semen samples from 15 rams were cryopreserved. After thawing, samples were submitted to four treatments: control (CTL), 10% OSS supplementation for in vitro sperm capacitation, caspase inhibitor (INH, Z-VAD-FMK 100 μM) and OSS (10%) plus caspase inhibitor (I + E). Sperm samples were incubated for 30 min at 38.5°C and 5% CO2 and evaluated motility and kinetic parameters by computer-assisted semen analysis (CASA) and viability (propidium iodide), apoptotic-like membrane changes (YO-PRO-1), acrosomal status (PNA-FITC), intracellular calcium (FLUO-3), membrane fluidity (M540) and ROS production (CM-H2 DCFDA) by flow cytometry. OSS induced changes in kinetic parameters compatible with capacitation, with a decrease in the percentage of progressive motility and linearity, and an increase in the amplitude of the lateral displacement of the sperm head (p < .05). Moreover, OSS increased the proportion of M540+ viable spermatozoa, YO-PRO-1+ and acrosome-reacted spermatozoa (p < .05). After incubation, OSS and I+E achieved lower ROS levels (p < .05). Ca(2+) levels did not change with the incubation, but were slightly higher (p < .05) when both OSS and the inhibitor were present. We suggest that OSS may modulate ROS levels, allowing intracellular signalling for capacitation to occur while preventing higher levels that could trigger apoptosis.

  17. Comparison of reproductive outcome in oligozoospermic men with high sperm DNA fragmentation undergoing intracytoplasmic sperm injection with ejaculated and testicular sperm.

    Science.gov (United States)

    Esteves, Sandro C; Sánchez-Martín, Fernando; Sánchez-Martín, Pascual; Schneider, Danielle T; Gosálvez, Jaime

    2015-12-01

    To investigate the effectiveness of intracytoplasmic sperm injection (ICSI) using testicular sperm as a strategy to overcome infertility in men with high sperm DNA fragmentation (SDF). Prospective, observational, cohort study. Private IVF centers. A total of 147 couples undergoing IVF-ICSI and day 3 fresh ETs whose male partner has oligozoospermia and high SDF. Sperm injections were carried out with ejaculated sperm (EJA-ICSI) or testicular sperm (TESTI-ICSI) retrieved by either testicular sperm extraction (TESE) or testicular sperm aspiration (TESA). SDF levels were reassessed on the day of oocyte retrieval in both ejaculated and testicular specimens. Percentage of testicular and ejaculated spermatozoa containing fragmented DNA (%DFI) and clinical pregnancy, miscarriage, and live-birth rates. The %DFI in testicular sperm was 8.3%, compared with 40.7% in ejaculated sperm. For the TESTI-ICSI group versus the EJA-ICSI group, respectively, the clinical pregnancy rate was 51.9% and 40.2%, the miscarriage rate was 10.0% and 34.3%, and the live-birth rate was 46.7% and 26.4%. ICSI outcomes were significantly better in the group of men who had testicular sperm used for ICSI compared with those with ejaculated sperm. SDF was significantly lower in testicular specimens compared with ejaculated counterparts. Our results suggest that TESTI-ICSI is an effective option to overcome infertility when applied to selected men with oligozoospermia and high ejaculated SDF levels. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  18. Sperm flagellum volume determines freezability in red deer spermatozoa.

    Science.gov (United States)

    Ros-Santaella, José Luis; Domínguez-Rebolledo, Alvaro Efrén; Garde, José Julián

    2014-01-01

    The factors affecting the inter-individual differences in sperm freezability is a major line of research in spermatology. Poor sperm freezability is mainly characterised by a low sperm velocity, which in turn is associated with low fertility rates in most animal species. Studies concerning the implications of sperm morphometry on freezability are quite limited, and most of them are based on sperm head size regardless of the structural parts of the flagellum, which provides sperm motility. Here, for the first time, we determined the volumes of the flagellum structures in fresh epididymal red deer spermatozoa using a stereological method under phase contrast microscopy. Sperm samples from thirty-three stags were frozen and classified as good freezers (GF) or bad freezers (BF) at two hours post-thawing using three sperm kinetic parameters which are strongly correlated with fertility in this species. Fourteen stags were clearly identified as GF, whereas nineteen were BF. No significant difference in sperm head size between the two groups was found. On the contrary, the GF exhibited a lower principal piece volume than the BF (6.13 µm3 vs 6.61 µm3, respectively, p = 0.006). The volume of the flagellum structures showed a strong negative relationship with post-thawing sperm velocity. For instance, the volume of the sperm principal piece was negatively correlated with sperm velocity at two hours post-thawing (r = -0.60; psperm principal piece results in poor freezability, and highlights the key role of flagellum size in sperm cryopreservation success.

  19. DNA fragmentation and sperm head morphometry in cat epididymal spermatozoa.

    Science.gov (United States)

    Vernocchi, Valentina; Morselli, Maria Giorgia; Lange Consiglio, Anna; Faustini, Massimo; Luvoni, Gaia Cecilia

    2014-10-15

    Sperm DNA fragmentation is an important parameter to assess sperm quality and can be a putative fertility predictor. Because the sperm head consists almost entirely of DNA, subtle differences in sperm head morphometry might be related to DNA status. Several techniques are available to analyze sperm DNA fragmentation, but they are labor-intensive and require expensive instrumentations. Recently, a kit (Sperm-Halomax) based on the sperm chromatin dispersion test and developed for spermatozoa of different species, but not for cat spermatozoa, became commercially available. The first aim of the present study was to verify the suitability of Sperm-Halomax assay, specifically developed for canine semen, for the evaluation of DNA fragmentation of epididymal cat spermatozoa. For this purpose, DNA fragmentation indexes (DFIs) obtained with Sperm-Halomax and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) were compared. The second aim was to investigate whether a correlation between DNA status, sperm head morphology, and morphometry assessed by computer-assisted semen analysis exists in cat epididymal spermatozoa. No differences were observed in DFIs obtained with Sperm-Halomax and TUNEL. This result indicates that Sperm-Halomax assay provides a reliable evaluation of DNA fragmentation of epididymal feline spermatozoa. The DFI seems to be independent from all the measured variables of sperm head morphology and morphometry. Thus, the evaluation of the DNA status of spermatozoa could effectively contribute to the completion of the standard analysis of fresh or frozen semen used in assisted reproductive technologies. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. Sperm release pathway

    Science.gov (United States)

    ... called seminiferous tubules, which are the sites of sperm production. The structure on top of the seminiferous tubules in the testes is the epididymis. The sperm migrate from of the seminiferous tubules to the ...

  1. Intracytoplasmic sperm injection

    Science.gov (United States)

    Intracytoplasmic sperm injection, or ICSI, is a form of in vitro fertilization in which fertilization occurs outside of the ... laboratory dish. Within a few hours, a single sperm is injected through a fine needle into the ...

  2. New Schemes for Self-Testing RAM

    CERN Document Server

    Bodean, Gh; Labunetz, A

    2011-01-01

    This paper gives an overview of a new technique, named pseudo-ring testing (PRT). PRT can be applied for testing wide type of random access memories (RAM): bit- or word-oriented and single- or dual-port RAM's. An essential particularity of the proposed methodology is the emulation of a linear automaton over Galois field by memory own components.

  3. Combined Effect of Trolox and EDTA on Frozen-Thawed Sperm Quality

    Directory of Open Access Journals (Sweden)

    Sara Keshtgar

    2016-05-01

    Full Text Available The freezing and thawing process not only is associated with serious damage to sperm such as damage to the plasma membrane and the acrosomal membrane but also changes the membrane permeability to some ions including calcium. Also, the generation of oxygen free radicals is increased during the freezing-thawing process. The purpose of this study was to evaluate of the effects of Trolox as an antioxidant and edetic acid (EDTA as a calcium chelator on frozen-thawed (FT sperm and compare these effects with those on fresh sperm. This study was done on these men of 25 healthy men, who referred to Shiraz Infertility Centerbetween2012 and2013. Normal samples were transferred to the ReproductivePhysiology Laboratory, Department of Physiology,Shiraz University of Medical Sciences, Shiraz. The samples were divided into two groups randomly: fresh and FT sperm groups. Each group was divided into five subgroups: control group, the solvent group (0.1%dimethyl sulfoxide [DMSO], Trolox group (200μM, EDTA group (1.1mM, and Trolox+EDTA group. The percentages of motility, viability, and acrosome-reacted sperm were tested. The percentages of motility and viability in the FT sperm were lower than those in the fresh sperm. The progressive motility of the FT sperm was improved nonsignificantly with Trolox+EDTA. However, the effect of Trolox+EDTA on the progressive motility of the FT sperm was much more than that on the fresh sperm. The fewest acrosome-reacted sperm were observed in the EDTA-containingFT sperm. Antioxidant supplementation or omission of extracellular calcium may partly improve motility and also reduce acrosomal damage in FT sperm.

  4. The RAM network in pathogenic fungi.

    Science.gov (United States)

    Saputo, Sarah; Chabrier-Rosello, Yeissa; Luca, Francis C; Kumar, Anuj; Krysan, Damian J

    2012-06-01

    The regulation of Ace2 and morphogenesis (RAM) network is a protein kinase signaling pathway conserved among eukaryotes from yeasts to humans. Among fungi, the RAM network has been most extensively studied in the model yeast Saccharomyces cerevisiae and has been shown to regulate a range of cellular processes, including daughter cell-specific gene expression, cell cycle regulation, cell separation, mating, polarized growth, maintenance of cell wall integrity, and stress signaling. Increasing numbers of recent studies on the role of the RAM network in pathogenic fungal species have revealed that this network also plays an important role in the biology and pathogenesis of these organisms. In addition to providing a brief overview of the RAM network in S. cerevisiae, we summarize recent developments in the understanding of RAM network function in the human fungal pathogens Candida albicans, Candida glabrata, Cryptococcus neoformans, Aspergillus fumigatus, and Pneumocystis spp.

  5. The effect of quercetin on fertility of frozen-thawed ram epididymal ...

    African Journals Online (AJOL)

    mz

    2017-03-13

    Mar 13, 2017 ... However, the blastocyst rate of fresh sperm was significantly higher ... unexpected death of endangered animals or animals with genetic .... ai is the fixed effect of different quercetin concentrations (i = 0, 5, 10, .... In agreement to the current results, in a study on red deer epididymal spermatozoa after freezing.

  6. Intracytoplasmic Sperm Injection (ICSI)

    Science.gov (United States)

    ... sperm must attach to the outside of the egg. Once attached, the sperm pushes through the outer layer to the inside ... in vitro fertilization (IVF) to help fertilize the egg. During ICSI, a single sperm is injected directly into the cytoplasm the egg. ...

  7. Sperm preparation for fertilization

    NARCIS (Netherlands)

    Gadella, B.M.

    2014-01-01

    Description This book contains 19 chapters that discuss theoretical and applied andrology for domestic, zoo and wild animals. Topics include semen and its constituents; sperm production and harvest; determinants of sperm morphology; sperm preparation for fertilization; practical aspects of semen cry

  8. Study on the effect of prostaglandin F2α treatment on semen characteristics and enzymatic activates of Awassi rams in breeding and non breeding seasons

    Directory of Open Access Journals (Sweden)

    Osama Ibrahim Azawi,

    2011-05-01

    Full Text Available The purpose of this research work was to determine the effects of PGF2α, given immediately before semen collection, on semen characteristics and libido in Awassi rams during breeding and non breeding season. The experiment was conducted in late summer to early autumn when major breeding activities commence and winter during the non breeding season at Mosul region in northern Iraq at the Animal Research and Practice Farm of the College of The Veterinary Medicine, University of Mosul. Twelve mature Awassi rams were used in this study. Animals were randomly allocated into two equal groups, the first group was administered 7.5 mg IM of PGF2αweekly and the second group as a control group received 1 ml of N-saline solution. Semen samples were collected from the Awassi rams 24 h after IM administration. Scrotal circumference (SC and testicular volume were measured weekly during the study period. Semen ejaculates were evaluated for semen volume, sperm concentration, sperm concentration/ejaculate, mass motility, individual motility, percentage live sperm, sperm abnormalities, and sperm acrosomal defects. Samples of seminal plasma were analyzed for the estimation of alanine amino transferase (ALT, aspartate amino transferase (AST, acid phosphatase (ACP, alkaline phosphatase (ALP and lactic dehydrogenase (LDH. Results of the present showed that PGF2α treatment to Awassi rams did not improve most semen characteristics in both breeding and non breeding seasons compared with the group. The only improvement of Awassi semen quality observed was in sperm concentration in the breeding season. The testicular volume showed a significant increase (P<0.05 in Awassi rams treated with PGF2α in breeding season compared to the control group and PGF2α treated group in the non breeding season. The mean activity of LDH enzyme estimated in the PGF2αtreated group and control group showed a significant difference (P<0.05 between the two groups in the breeding season and

  9. Effect of postthaw storage time and sperm-to-egg ratio on fertility of cryopreserved brook trout sperm.

    Science.gov (United States)

    Nynca, J; Dietrich, G J; Dobosz, S; Zalewski, T; Ciereszko, A

    2015-01-15

    The aim of this study was to test the influence of postthaw storage time on sperm motility parameters of brook trout (n = 9). Furthermore, we examined the effect of sperm-to-egg ratios of 300,000:1 and 600,000:1 on fertility of postthaw, cryopreserved, brook trout sperm. The application of a cryopreservation procedure produced very high postthaw sperm motility (56.8 ± 4.0%). The cryopreserved sperm of brook trout could be stored up to 60 minutes without loss of the percentage of sperm motility (52.0 ± 9.0%). The fertilization capacity of brook trout postthaw sperm was comparable with the fertilization rate of fresh semen at a sperm-to-egg ratio as low as 300,000:1 (42.4 ± 14.0% and 36.5 ± 11.0% for eyed and hatched stages, respectively). The possibility of postthaw semen storage for the prolonged time and the obtainment of high fertilization rate at low sperm-to-egg ratio can lead to the significant improvement of brook trout semen cryopreservation procedure.

  10. Changes in sexual behaviour and semen quality associated with age and type of enclosure of Saint Croix rams in different seasons of the year

    Directory of Open Access Journals (Sweden)

    Sandra Ivon Santos

    2015-12-01

    Full Text Available The objective of this study was to evaluate sexual behaviour and semen quality of St. Croix ram lambs bred in a semi-desert region. In Experiment 1, 24 yearling rams were assigned to two groups: T1 with 12 males confined for 11 months; T2 with 12 males confined individually for the same period of time. In Experiment 2, 20 four-month-old ram lambs were assigned to two groups of 10 animals each. In each experiment, sexual behaviour and semen quality were evaluated every 2 weeks over 11 months. In Experiment 1, rams that were individually confined showed better sexual activity than lambs confined together (P<0.05, whereas in Experiment 2 ram lambs confined individually only showed shorter time to first mount with ejaculation (P<0.05. In experiment 1, sperm with better progressive motility (P<0.05 was recorded per yearling ram confined individually (63.4±0.2% in comparison with rams confined together (51.60.1%. In Experiment 2, ram lambs confined together showed better semen quality (P˂0.05. In both trials, an effect of the season was observed on both sexual behaviour and semen quality (P<0.05. In conclusion, this study of St. Croix males showed different sexual behaviours according to age and type of enclosure.

  11. The development of cat testicular sperm cryopreservation protocols: Effects of tissue fragments or sperm cell suspension.

    Science.gov (United States)

    Chatdarong, Kaywalee; Thuwanut, Paweena; Morrell, Jane M

    2016-01-15

    In endangered animals that have been found dead or sterilized for medical reasons, testis is the ultimate source of haploid DNA or sperm. Thus, preservation of testicular sperm may be performed to rescue their genetics. The aim of this study was to evaluate protocols for testicular sperm freezing: as tissue fragments or cell suspension in domestic cats as a model. A pair of testes from each cat (n = 9) were cut into eight equal pieces. Four randomly selected pieces were cryopreserved as: (1) tissue pieces using two-step freezing; (2) tissue pieces using a slow passive cooling device (CoolCell); (3) sperm suspension after single-layer centrifugation (SLC) through colloids; and (4) sperm suspension without being processed through SLC. A testicular piece from each cat served as fresh control. Testicular sperm membrane and DNA integrity were evaluated before, and after, the cryopreservation process. In addition, spermatogenic cell types (testicular sperm, spermatogonia, spermatocyte, and spermatid) present in the suspension samples were counted before and after SLC. The results found that testicular sperm membrane integrity in the suspension after SLC process was higher than that in the fragment form neither using the two-step nor CoolCell freezing, both before and after freezing (before freezing: 92.3 ± 3.4 vs. 81 ± 4.5 and 80.0 ± 7.0; after freezing: 84.5 ± 4.6 vs. 71.2 ± 12 and 76.2 ± 4.6; P ≤ 0.05). Testicular sperm DNA integrity was, however, not different among groups. Furthermore, the samples processed through the SLC had higher ration of sperm cells: other spermatogenic cells than those were not processed through the SLC (88.9 ± 3.8 vs. 30 ± 7.9; P ≤ 0.05). In summary, testicular sperm cryopreserved as a minced suspension is considered suitable in terms of preventing sperm membrane integrity, and SLC is considered a selection tool for enriching haploid sperm cells from castrated or postmortem cats.

  12. Fertilization after intracytoplasmic sperm injection with cryopreserved testicular spermatozoa.

    Science.gov (United States)

    Romero, J; Remohí, J; Mínguez, Y; Rubio, C; Pellicer, A; Gil-Salom, M

    1996-04-01

    To assess the possibility of cryopreserving testicular tissue extracted sperm for intracytoplasmic sperm injection (ICSI). A report of two cases. Our study was approved by the Ethical Committee at the Instituto Valenciano de Infertilidad. In vitro fertilization program at the Instituto Valenciano de Infertilidad. Two azoospermic patients with severe spermatogenic failure but with focal spermatogenesis on testicular biopsies. In both cases, a first ICSI attempt with fresh testicular biopsy extracted sperm was unsuccessful. Cryopreservation of testicular spermatozoa in 100-micro L "pills." Intracytoplasmic sperm injection with thawed testicular spermatozoa. Fertilization rate, cleavage rate, embryo quality, clinical pregnancy. Fertilization rates were 36 percent and 100 percent after ICSI with fresh testicular spermatozoa, and 63 percent and 57 percent after ICSI with cryopreserved testicular sperm. In both cases, cleavage rates and embryo quality were similar when using fresh and cryopreserved testicular spermatozoa. No clinical pregnancies were achieved. High fertilization rates can be obtained after ICSI with frozen-thawed testicular tissue extracted spermatozoa. Cryopreservation of testicular sperm may avoid repetition of testicular biopsies in azoospermic patients in whom the only source of spermatozoa is the testicle.

  13. Changes in the structures of motile sperm subpopulations in dog spermatozoa after both cryopreservation and centrifugation on PureSperm(®) gradient.

    Science.gov (United States)

    Dorado, J; Alcaráz, L; Duarte, N; Portero, J M; Acha, D; Hidalgo, M

    2011-05-01

    The aims of the present study were to: (1) determine if discrete motile sperm subpopulations exist and their incidence in fresh dog ejaculates, (2) evaluate the effects of cryopreservation on the distribution of spermatozoa within the different subpopulations, and (3) determine the effect of the discontinuous PureSperm(®) gradient on the sperm subpopulation structure of frozen-thawed dog spermatozoa. Semen from 5 dogs were collected and cryopreserved following a standard protocol. After thawing, semen samples were selected by centrifugation on PureSperm(®). Sperm motility (assessed by computerized-assisted semen analysis, CASA) was assessed before freezing, just after thawing and after preparation on the PureSperm(®) gradients. Cryopreservation had a significant (Psperm suspensions with improved motility (Psperm subpopulation structure was perfectly maintained after freezing and thawing. The selected sperm samples was enrich in Subpopulation 4, reaching a proportion of 31.9% of the present spermatozoa, in contrast with the unselected sperm samples, where this sperm subpopulation accounted for 24.9% of the total. From these results, we concluded that four well-defined motile sperm subpopulations were present either in fresh semen, in unselected sperm samples or in selected preparations from dogs. The discontinuous PureSperm(®) gradient is a simple method to improve the quality of canine frozen-thawed semen samples, since Subpopulation 4 (high-speed and progressive spermatozoa) was more frequently observed after preparation on the gradient. Finally, this study also demonstrated that the general motile sperm structure present in dog remains constant despite the effect caused by either cryopreservation or separation on PureSperm(®) gradient.

  14. Improvement in in vitro fertilization rate, decrease in reactive oxygen species and spermatozoa death incidence in rams by dietary fish oil.

    Science.gov (United States)

    Matini Behzad, A; Ebrahimi, B; Alizadeh, A R; Esmaeili, V; Dalman, A; Rashki, L; Shahverdi, A H

    2014-08-01

    Our aim was to evaluate the effects of fish oil feeding on sperm classical parameters, level of reactive oxygen spices (ROS), spermatozoa death incidence and in vitro fertilization (IVF) rate in rams. We randomly assigned nine rams, into two experimental groups (isoenergetic and isonitrogenous rations with constant level of vitamin E supplement): control (CTR; n = 5) and fish oil (FO; n = 4, 35 g/day/ram). Diets were fed for 70 days during the physiological breeding season. After a 21-day dietary adaptation period, semen was collected weekly from each ram by an artificial vagina. Sperm classical parameters were determined by the computer-assisted sperm analyzer system (CASA), and it was prepared for IVF process by swim-up technique. These evaluations were performed during the first and last weeks of sampling. Intracellular ROS level and spermatozoa death incidence were detected by flow cytometry on a weekly basis after adaptation. Data were analysed with SPSS 15. The volume, concentration (3.6 and 2.7 × 10(9) /ml) and sperm progressive motility (60 and 48%) were significantly improved in the FO group compared with the CTR (p < 0.05). A comparison of two-cell stage embryos following IVF in the two groups showed a significantly higher fertilization rate in the FO group (56%) compared with the CTR (49%). Superoxide anion (O2 (-) ) rate was significantly lower (p < 0.05) at the third week of sampling in the FO. Although the H2 O2 rate was numerically lower in the FO group compared with the CTR, this difference was not significant. In addition, apoptosis showed a significant difference in the third week of sampling (15 and 30% for FO and CTR, respectively; p < 0.05). Overall, adding fish oil to the ram diet not only improved sperm quality and IVF results, it also could reduce oxygen-free radicals and the incidence of spermatozoa death.

  15. 日粮锌水平对辽宁绒山羊种公羊精液品质的影响%Effects of dietary zinc levels on semen quality of breeding rams of Liaoning Cashmere Goat

    Institute of Scientific and Technical Information of China (English)

    杨毅强; 刘海英; 刘国华; 宋先忱; 李丰田; 刘兴伟; 孙亚波

    2012-01-01

    为探讨日粮锌水平对辽宁绒山羊种公羊精液品质的影响。本试验选用成年健康、精液品质相近的辽宁绒山羊种公羊20只,随机分为4组,对照组饲喂基础饲粮,试验组分别在基础饲粮中添加20、40和80mg/kg的锌。结果表明:①日粮锌水平对种公羊的精液量、精子密度、精子活力和精子解冻后活力影响显著(P〈0.05);②日粮锌水平对种公羊的精液pH值、美蓝褪色时间没有显著影响(P〈0.05);③日粮锌水平对种公羊的降低鲜精和冻精畸形率有一定的促进作用(P〉0.05)。综合鲜精、冻精品质指标,本试验日粮条件下,日粮锌添加量为40mg/kg辽宁绒山羊种公羊的精液品质最优。%The study was conducted to investigate the effect of dietary zinc levels on semen quality of breeding rams of Liaoning Cashmere Goat. A total of 20 breeding rams with weight similar and age similar were randomly divided into 4 groups. The control group was fed a basic diet, and the test groups were 20, 40 and 80 mg/kg of zinc added in the basal diet. The result showed that: (1)There was significant effect by adding dietary zinc to breeding rams on semen volume, sperm concentration, sperm motility and post-thaw- ing motility (P〈0. 05). (2)The pH of semen and methylene blue fading time were not markedly affected by dietary zinc levels (P〈0. 05). (3)Dietary zinc levels on breeding rams has a certain role in promoting to reduce the fresh semen and frozen semen malformation rate (P〉0. 05). Based on the quality indicators of fresh semen and frozen semen under the conditions of the experimental diets, semen quality of breeding rams of Liaoning Cashmere Goat is highest with 40 mg/kg of dietary zinc added in the basal diet.

  16. Gamete evolution and sperm numbers: sperm competition versus sperm limitation.

    Science.gov (United States)

    Parker, Geoff A; Lehtonen, Jussi

    2014-09-22

    Both gamete competition and gamete limitation can generate anisogamy from ancestral isogamy, and both sperm competition (SC) and sperm limitation (SL) can increase sperm numbers. Here, we compare the marginal benefits due to these two components at any given population level of sperm production using the risk and intensity models in sperm economics. We show quite generally for the intensity model (where N males compete for each set of eggs) that however severe the degree of SL, if there is at least one competitor for fertilization (N - 1 ≥ 1), the marginal gains through SC exceed those for SL, provided that the relationship between the probability of fertilization (F) and increasing sperm numbers (x) is a concave function. In the risk model, as fertility F increases from 0 to 1.0, the threshold SC risk (the probability q that two males compete for fertilization) for SC to be the dominant force drops from 1.0 to 0. The gamete competition and gamete limitation theories for the evolution of anisogamy rely on very similar considerations: our results imply that gamete limitation could dominate only if ancestral reproduction took place in highly isolated, small spawning groups. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

  17. Daily sperm production

    DEFF Research Database (Denmark)

    Kyjovska, Zdenka Orabi; Boisen, Anne Mette Zenner; Jackson, Petra

    2013-01-01

    instillation with Printex90. Body and testicle weight, sperm content per g testicular parenchyma and daily sperm production (DSP) were assessed. The protocol for assessment of DSP was optimized for application in mice (C57BL/6J) and the influence of different parameters was studied. Maternal particulate...... exposure did not affect DSP statistically significantly in the F1 generation, although TiO2 tended to reduce sperm counts. Overall, time-to-first F2 litter increased with decreasing sperm production. There was no effect on sperm production in the F2 generation originating after TiO2 exposure. F2 offspring......, whose fathers were prenatally exposed to Printex90, showed lowered sperm production. Furthermore, we report statistically significant differences in sperm production between mouse strains....

  18. Evaluation of sperm DNA damage in bulls by TUNEL assay as a parameter of semen quality

    OpenAIRE

    TAKEDA, Kumiko; UCHIYAMA, Kyoko; KINUKAWA, Masashi; Tagami, Takahiro; Kaneda, Masahiro; Watanabe, Shinya

    2015-01-01

    Sperm DNA damage affects the conception rate resulting from human assisted reproduction technology. The objective of this study was to adapt the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay to provide a quality parameter for bull semen based on the detection of sperm DNA damage. Fresh semen was collected from two Japanese Black bulls (A, B) several times over the course of a year, and the percentage of TUNEL-positive spermatozoa (sperm TUNEL index) was d...

  19. Evaluation of cholesterol- treated dromedary camel sperm function by heterologous IVF and AI.

    Science.gov (United States)

    Crichton, Elizabeth G; Malo, Clara; Pukazhenthi, Budhan S; Nagy, Peter; Skidmore, Julian A

    2016-11-01

    Cholesterol (cholesterol-loaded cyclodextrins: CLC) treatment of dromedary camel sperm prior to freezing enhances cryosurvival. The present study first validated the efficacy of a heterologous zona-free goat oocyte assay (n=115 oocytes) to evaluate camel sperm function in vitro (Experiment 1: n=6 bulls), then examined the effects of CLC treatment (1.5mg/mL CLC; CLC+) versus no treatment (0 CLC) of fresh (Experiment 2: n=4 bulls) and frozen-thawed (Experiment 3: n=5 bulls) camel sperm to penetrate, de-condense and form pro-nuclei in in vitro-matured goat oocytes. Finally, the ability of fresh 0 CLC and CLC+ sperm to fertilize in vivo was studied by artificially inseminating super-ovulated females (n=7-9 per treatment) and examining embryo production (Experiment 4: n=4-5 bulls/treatment). Camel spermatozoa penetrated (60%) and formed pro-nuclei (33%) in goat oocytes demonstrating the utility of this heterologous system for assessing sperm function in vitro. For fresh spermatozoa, 0 CLC-treated sperm performed better than their CLC+ counterparts for all parameters measured (P<0.05). In contrast, cryopreservation resulted in a sharp decline in sperm-oocyte interaction in 0 CLC aliquots but remained unaltered in CLC+ aliquots demonstrating a protective effect of cholesterol treatment. There was no difference between treatments in the in vitro fertilizing ability of frozen-thawed sperm or in the numbers of embryos retrieved following AI with fresh 0 CLC or CLC+ sperm. We conclude that although CLC treatment of dromedary camel sperm improves sperm motility it fails to confer an advantage to them in terms of improved in vitro sperm-oocyte interaction or in vivo fertilization under the conditions tested. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Negative effect of combined cysteine and glutathione in soy lecithin-based extender on post-thawed ram spermatozoa.

    Science.gov (United States)

    Zhandi, Mahdi; Sharafi, Mohsen

    2015-09-01

    This study was conducted to evaluate the effect of combined cysteine and glutathione in soy lecithin-based semen extender on post-thawed ram sperm quality. A total of 28 ejaculates were collected twice a week (from four rams) during breeding season. In each replicate, semen samples (n = 4, one ejaculate for each ram) were pooled and divided into three equal parts, and each part was diluted with one of following extender: (1) soy lecithin-based extender containing no cysteine and no glutathione (C0-G0), (2) soy lecithin-based extender containing cysteine (5 mM) and glutathione (5 mM) (C5-G5), and (3) soy lecithin-based extender containing cysteine (10 mM) and glutathione (10 mM) (C10-G10). After freeze-thawing process, motility and velocity parameters, plasma membrane integrity and functionality, mitochondrial activity, and apoptosis features of spermatozoa were evaluated. The obtained results showed that total and progressive motility, plasma membrane integrity and functionality, and live post-thawed spermatozoa was lower in C10-G10 extender compared to C0-G0 and C5-G5 extenders (P 0.05). In conclusion, it seems that high concentration of combined cysteine and glutathione in soy lecithin-based semen extender has a detrimental effect of post-thawed ram sperm quality.

  1. Fertility prediction of frozen boar sperm using novel and conventional analyses

    Science.gov (United States)

    Frozen-thawed boar sperm is seldom used for artificial insemination (AI) because fertility is lower than fresh or cooled semen. Despite the many advantages of AI including reduced pathogen exposure and ease of semen transport, cryo-induced damage to sperm usually results in decreased litter sizes a...

  2. RAMS (Risk Analysis - Modular System) methodology

    Energy Technology Data Exchange (ETDEWEB)

    Stenner, R.D.; Strenge, D.L.; Buck, J.W. [and others

    1996-10-01

    The Risk Analysis - Modular System (RAMS) was developed to serve as a broad scope risk analysis tool for the Risk Assessment of the Hanford Mission (RAHM) studies. The RAHM element provides risk analysis support for Hanford Strategic Analysis and Mission Planning activities. The RAHM also provides risk analysis support for the Hanford 10-Year Plan development activities. The RAMS tool draws from a collection of specifically designed databases and modular risk analysis methodologies and models. RAMS is a flexible modular system that can be focused on targeted risk analysis needs. It is specifically designed to address risks associated with overall strategy, technical alternative, and `what if` questions regarding the Hanford cleanup mission. RAMS is set up to address both near-term and long-term risk issues. Consistency is very important for any comparative risk analysis, and RAMS is designed to efficiently and consistently compare risks and produce risk reduction estimates. There is a wide range of output information that can be generated by RAMS. These outputs can be detailed by individual contaminants, waste forms, transport pathways, exposure scenarios, individuals, populations, etc. However, they can also be in rolled-up form to support high-level strategy decisions.

  3. HyRAM Testing Strategy and Quality Design Elements.

    Energy Technology Data Exchange (ETDEWEB)

    Reynolds, John Thomas

    2014-12-01

    Strategy document and tentative schedule for testing of HyRAM, a software toolkit that integrates data and methods relevant to assessing the safety of hydrogen fueling and storage infrastructure. Because proposed and existing features in HyRAM that support testing are important factors in this discussion, relevant design considerations of HyRAM are also discussed. However, t his document does not cover all of HyRAM desig n, nor is the full HyRAM software development schedule included.

  4. Addition of superoxide dismutase mimics during cooling process prevents oxidative stress and improves semen quality parameters in frozen/thawed ram spermatozoa.

    Science.gov (United States)

    Santiani, Alexei; Evangelista, Shirley; Sepúlveda, Néstor; Risopatrón, Jennie; Villegas, Juana; Sánchez, Raúl

    2014-10-01

    High levels of reactive oxygen species (ROS), which may be related to reduced semen quality, are detected during semen cryopreservation in some species. The objectives of this study were to measure the oxidative stress during ram semen cryopreservation and to evaluate the effect of adding 2 antioxidant mimics of superoxide dismutase (Tempo and Tempol) during the cooling process on sperm motility, viability, acrosomal integrity, capacitation status, ROS levels, and lipid peroxidation in frozen and/or thawed ram spermatozoa. Measuring of ROS levels during the cooling process at 35, 25, 15, and 5 °C and after freezing and/or thawing showed a directly proportional increase (P ram spermatozoa are exposed to oxidative stress during the cooling process, specifically when maintained at 5 °C and that lipid peroxidation induced by high levels of ROS decreases sperm motility and induces premature sperm capacitation. In contrast, the addition of Tempo or Tempol at 0.5 to 1 mM during the cooling process (10 °C) protects ram spermatozoa from oxidative stress.

  5. THE EFFECT OF SEMEN WASHING AND SOYBEAN LECITHIN LEVEL ON MOTILITY AND VIABILITY OF RAM SPERMATOZOA STORED AT 5oC

    Directory of Open Access Journals (Sweden)

    Salmin

    2012-12-01

    Full Text Available The objectives of this reserarch were to study the effect of semen washing and soybean lecithin level on the motility and viability of ram spermatozoa stored at 5oC. Semen was collected from three mature rams by artificial vagina. Semen was evaluated by macroscopic and microscopic. Semen was divided into ten tubes, the semen of first five tubes was unwashed (W0 and diluted with extender 0%, 1%, 2%, 3% and 4% soybean lecithin (L0, L1, L2, L3 and L4. Semen of another five tubes were washed (W1 then each diluted with similar extender levels. The diluted semen samples were stored at 5oC and the sperm motility and viability were evaluated each day. The interaction of the semen washing and soybean lecithin levels were no significant differences (P>0.05 but the single factor of soybean lecithin level was significant differeNces (P0.05 on progressive motility and viability of ram sperm.The best extender was 3% soybean lecithin (L3 with the percentage of ram sperm progressive motility was 63.18 ± 3.61% and viability was 71.76±2.32%.

  6. THE EFFECT OF SEMEN WASHING AND SOYBEAN LECITHIN LEVEL ON MOTILITY AND VIABILITY OF RAM SPERMATOZOA STORED AT 5⁰C

    Directory of Open Access Journals (Sweden)

    S. Salmin

    2014-10-01

    Full Text Available The objectives of this reserarch were to study the effect of semen washing and soybean lecithinlevel on the motility and viability of ram spermatozoa stored at 5⁰C. Semen was collected from threemature rams by artificial vagina. Semen was evaluated by macroscopic and microscopic. Semen wasdivided into ten tubes, the semen of first five tubes was unwashed (W0 and diluted with extender 0%,1%, 2%, 3% and 4% soybean lecithin (L0, L1, L2, L3 and L4. Semen of another five tubes werewashed (W1 then each diluted with similar extender levels. The diluted semen samples were stored at5⁰C and the sperm motility and viability were evaluated each day. The interaction of the semen washingand soybean lecithin levels were no significant differences (P>0.05 but the single factor of soybeanlecithin level was significant differeNces (P<0.05 and the single factor of semen washing were nosignificant differences (P>0.05 on progressive motility and viability of ram sperm.The best extenderwas 3% soybean lecithin (L3 with the percentage of ram sperm progressive motility was 63.18 ± 3.61%and viability was 71.76±2.32%.

  7. FPS-RAM: Fast Prefix Search RAM-Based Hardware for Forwarding Engine

    Science.gov (United States)

    Zaitsu, Kazuya; Yamamoto, Koji; Kuroda, Yasuto; Inoue, Kazunari; Ata, Shingo; Oka, Ikuo

    Ternary content addressable memory (TCAM) is becoming very popular for designing high-throughput forwarding engines on routers. However, TCAM has potential problems in terms of hardware and power costs, which limits its ability to deploy large amounts of capacity in IP routers. In this paper, we propose new hardware architecture for fast forwarding engines, called fast prefix search RAM-based hardware (FPS-RAM). We designed FPS-RAM hardware with the intent of maintaining the same search performance and physical user interface as TCAM because our objective is to replace the TCAM in the market. Our RAM-based hardware architecture is completely different from that of TCAM and has dramatically reduced the costs and power consumption to 62% and 52%, respectively. We implemented FPS-RAM on an FPGA to examine its lookup operation.

  8. Effect of nutritional state on semen characteristics, testicular size and serum testosterone concentration in Sanjabi ram lambs during the natural breeding season.

    Science.gov (United States)

    Ghorbankhani, F; Souri, M; Moeini, M M; Mirmahmoudi, R

    2015-02-01

    This experiment was conducted to investigate the effect of plane of nutrition on body weight, average daily gain, dry mater intake, semen characteristics, serum testosterone concentration and testicular circumference of Sanjabi ram lambs during the natural breeding season. Sanjabi ram lambs (n = 20) that were 8 months of age were penned under natural photoperiod at latitude 34°18'N for a period of 9 months. The control group (C, n = 10) received a diet consisting of 80% alfalfa and 20% concentrate, providing 2.18 Mcal metabolizable energy and 130.0 g /kg DM crude protein, whereas, treatment group (T, n = 10) was fed with 65% alfalfa and 35% concentrate, providing 2.34 Mcal metabolizable energy and 160.0 g/kg DM crude protein. Body weight, additive daily gain and feed intake in T group were significantly greater than those obtained in C group. Body weight and testicular circumference increased at a steady rate throughout the experiment. All semen variables (except percentage of abnormal sperm and semen pH), serum testosterone concentration and testicular circumference were positively influenced by nutritional state (P nutritional state with season was found for semen volume, sperm concentration and abnormal sperm, but there was no interaction on the total sperm, progressive motility, live sperm, semen pH and semen index. It is concluded that the reproductive activity of growing Sanjabi ram lambs is affected by nutritional state. These results also demonstrated a monthly pattern in reproductive characteristics of Sanjabi ram lambs, independent of the nutritional state.

  9. Effect of the infection with the nematode Haemonchus contortus (Strongylida: Trichostrongylidae on the haematological, biochemical, clinical and reproductive traits in rams

    Directory of Open Access Journals (Sweden)

    Mariem Rouatbi

    2016-03-01

    Full Text Available This study aimed to investigate the effect of Haemonchus contortus infection on rams’ haematological, biochemical and clinical parameters and reproductive performances. A total number of 12 Barbarine rams (control and infected were included in the experiment. The infected group received 30 000 H. contortus third-stage larvae orally. Each ram’s ejaculate was immediately evaluated for volume, sperm cell concentration and mortality rate. At the end of the experiment (day 82 post-infection, which lasted 89 days, serial blood samples were collected in order to assess plasma testosterone and luteinising hormone (LH concentrations. There was an effect of time, infection and their interaction on haematological parameters (p < 0.001. In infected rams, haematocrit, red blood cell count and haemoglobin started to decrease from 21 days post-infection. There was an effect of time and infection for albumin. For total protein, only infection had a statistically significant effect. For glucose, only time had a statistically significant effect. Concentrations were significantly lower in infected rams compared to control animals. A significant effect of infection and time on sperm concentrations and sperm mortality was observed. The effect of infection appears in time for sperm concentrations at days 69 and 76 post-infection. Sperm mortality rate was significantly higher in infected animals at day 46 post-infection when compared to control group (p < 0.05. Finally, plasma testosterone traits (average concentration, cumulated levels during the sampling period and pulse frequency were depressed in infected rams when compared to control counterparts; none of these endocrine traits were affected for plasma LH.

  10. Cryopreservation of tench, Tinca tinca, sperm: Sperm motility and hatching success of embryos.

    Science.gov (United States)

    Rodina, M; Gela, D; Kocour, M; Alavi, S M Hadi; Hulak, M; Linhart, O

    2007-03-15

    samples were held 6min at -80 degrees C. Finally, samples were transferred into liquid N(2). The frozen spermatozoa were thawed in a water bath (40 degrees C) according to the frozen volume and checked for fertilization and hatching rates. Percentage of sperm motility and sperm velocity were measured using video recorded frames. ANOVA showed a significant influence of frozen and fresh sperm in all treatments. The hatching rates of 33.8% were obtained when sperm was equilibrated for 0h before freezing in IS of Kurokura 180 and frozen with a 10% of mixture 1:1 of DMSO and propanediol into straws of 5ml and cooled using program L1. The velocity of frozen-thawed spermatozoa ranged from 31 to 46microms(-1) and in post-thawed sperm was not significantly different according to frozen sperm volume, but a higher velocity was obtained when sperm was fast frozen using programme L1. A large volume of frozen sperm could reveal the best procedure for freezing, but also for simulating methods of artificial propagation for future practical use of frozen tench sperm at a large scale.

  11. Sperm length, sperm storage and mating system characteristics in bumblebees

    DEFF Research Database (Denmark)

    Baer, Boris; Schmid-Hempel, Paul; Høeg, Jens Thorvald

    2003-01-01

    of ejaculated sperm that was stored in a queen's spermatheca. Both longer sperm and shorter sperm could be preferentially stored, depending on the colony in which the males and queens were born and raised. These results indicate that the genotype of males may affect sperm length and that cryptic female choice...

  12. Reactive approach motivation (RAM) for religion.

    Science.gov (United States)

    McGregor, Ian; Nash, Kyle; Prentice, Mike

    2010-07-01

    In 3 experiments, participants reacted with religious zeal to anxious uncertainty threats that have caused reactive approach motivation (RAM) in past research (see McGregor, Nash, Mann, & Phills, 2010, for implicit, explicit, and neural evidence of RAM). In Study 1, results were specific to religious ideals and did not extend to merely superstitious beliefs. Effects were most pronounced among the most anxious and uncertainty-averse participants in Study 1 and among the most approach-motivated participants in Study 2 (i.e., with high Promotion Focus, Behavioral Activation, Action Orientation, and Self-Esteem Scale scores). In Studies 2 and 3, anxious uncertainty threats amplified even the most jingoistic and extreme aspects of religious zeal. In Study 3, reactive religious zeal occurred only among participants who reported feeling disempowered in their everyday goals in life. Results support a RAM view of empowered religious idealism for anxiety management (cf. Armstrong, 2000; Inzlicht, McGregor, Hirsch, & Nash, 2009).

  13. Transmission electron microscopic observations of flagellum abnormalities in impala (Aepyceros melampus sperm from the Kruger National Park

    Directory of Open Access Journals (Sweden)

    D.J. Ackerman

    1997-01-01

    Full Text Available Sperm must remain motile in order to reach and penetrate the ovum and defects in the ultrastructure of the tail can have an adverse influence on motility. Live spermatozoa were collected from the cauda epididymis of 64 impala rams in the Kruger National Park and studied by transmission electron microscopy to document sperm abnormalities. The following abnormalities of the flagellum were documented from micrographs: abnormal baseplate and neck attachments; neck vacuoles and displaced organelles; double or short flagella; bent flagella; principal-piece vacuoles; displaced axoneme and the Dag defect. The implications of these abnormalities for sperm motility are discussed.

  14. Effect of cryopreservation on the sperm DNA fragmentation dynamics of the bottlenose dolphin (Tursiops truncatus).

    Science.gov (United States)

    Sánchez-Calabuig, M J; López-Fernández, C; Johnston, S D; Blyde, D; Cooper, J; Harrison, K; de la Fuente, J; Gosálvez, J

    2015-04-01

    Sperm DNA fragmentation is one of the major causes of infertility; the sperm chromatin dispersion test (SCDt) evaluates this parameter and offers the advantage of species-specific validated protocol and ease of use under field conditions. The main purpose of this study was to evaluate sperm DNA fragmentation dynamics in both fresh and post-thaw bottlenose dolphin sperm using the SCDt following different cryopreservation protocols to gain new information about the post-thaw differential sperm DNA longevity in this species. Fresh and cryopreserved semen samples from five bottlenose dolphins were examined for sperm DNA fragmentation dynamics using the SCDt (Halomax(®)). Sperm DNA fragmentation was assessed immediately at collection and following cryopreservation (T0) and then after 0.5, 1, 4, 8, 24, 48 and 72 h incubation at 37°C. Serially collected ejaculates from four dolphins were frozen using different cryopreservation protocols in a TES-TRIS-fructose buffer (TTF), an egg-yolk-free vegetable lipid LP1 buffer (LP1) and human sperm preservation medium (HSPM). Fresh ejaculated spermatozoa initially showed low levels of DNA fragmentation for up to 48 h. Lower Sperm DNA fragmentation (SDF) was found in the second fresh ejaculate compared to the first when more than one sample was collected on the same day (p fragmentation after 24- and 48-h incubation than those frozen in TTF or HSPM. No correlation was found between any seminal characteristic and DNA fragmentation in either fresh and/or frozen-thawed samples. © 2015 Blackwell Verlag GmbH.

  15. Function of ram spermatozoa frozen in diluents supplemented with casein and vegetable oils.

    Science.gov (United States)

    Del Valle, I; Souter, A; Maxwell, W M C; Muiño-Blanco, T; Cebrián-Pérez, J A

    2013-05-01

    The aim of this study was to assess biologically safer components as alternatives to egg yolk for the frozen storage of ram semen using casein, coconut or palm oil in either Salamon's diluent (S) or a swim-up medium (SU). Ejaculates were frozen as pellets and sperm motility (subjectively) and acrosome integrity (FITC-PNA/PI) by flow cytometry were assessed at 0, 3 and 6h after thawing and incubation at 37°C. Three experiments were done: different concentrations of palm oil (5%, 10% and 20%); casein added as emulsifier and protective agent; and differences between egg yolk, coconut and palm oil in S and SU. 20% of oil added to SU accounted for a lesser percentage (Poil levels on viable cells. When casein was added to diluents containing 5% of palm oil, no differences were found between palm or casein (P>0.05). No differences were found when S and SU were compared neither as groups nor between S alone and containing coconut or palm oil; however, SU alone yielded less motility than SU 5% coconut. However, in both groups, S and SU, egg yolk accounted for the greatest values in both bases. These results indicate that none of biologically safer media components (casein, palm or coconut oil) used in this study maintained the function of ram spermatozoa after freeze-thawing better than S-containing egg yolk. The application of vegetable oils as substitutes for egg yolk in diluents for the cryopreservation of ram spermatozoa requires further research.

  16. Cleaved PARP-1, an Apoptotic Marker, can be Detected in Ram Spermatozoa.

    Science.gov (United States)

    Casao, A; Mata-Campuzano, M; Ordás, L; Cebrián-Pérez, J A; Muiño-Blanco, T; Martínez-Pastor, F

    2015-08-01

    The presence of apoptotic features in spermatozoa has been related to lower quality and functional impairment. Members of the poly-ADP-ribose polymerases (PARP) familyare involved in both DNA repair and apoptosis, playing important roles in spermatogenesis. Poly-ADP-ribose polymerase can be cleaved by caspases, and the presence of its cleavage product (cPARP) in spermatozoa has been related to chromatin remodelling during spermatogenesis and to the activation of apoptotic pathways. There are no reports on immunodetection of cPARP in ram spermatozoa; thus, we have tested a commercially available antibody for this purpose. cPARP was microscopically detected in the acrosomal ridge of some spermatozoa (indirect immunofluorescence). A preliminary study was carried out by flow cytometry (direct immunofluorescence, FITC). Ram semen was extended in TALP and incubated for 4 h with apoptosis inducers staurosporine (10 μm) or betulinic acid (200 μm). Both inducers and incubation caused a significant increase in cPARP spermatozoa (0 h, control: 21.4±3.3%, inducers: 44.3±1.4%; 4 h, control: 44.3±2.4%, inducers: 53.3±1.4%). In a second experiment, we compared the sperm fractions after density gradient separation (pellet and interface). The pellet yielded a slightly lower proportion of cPARP spermatozoa (28.5±1.2% vs 36.2±2.0% in the interface; p ram semen, although its presence in untreated samples was weakly related to worse quality (pellet/interface). We suggest to study the relationship of PARP and cPARP levels with between-male differences on sperm fertility.

  17. Sperm competition and the evolution of sperm design in mammals

    Directory of Open Access Journals (Sweden)

    Gomendio Montserrat

    2011-01-01

    Full Text Available Abstract Background The influence of sperm competition upon sperm size has been a controversial issue during the last 20 years which remains unresolved for mammals. The hypothesis that, when ejaculates compete with rival males, an increase in sperm size would make sperm more competitive because it would increase sperm swimming speed, has generated contradictory results from both theoretical and empirical studies. In addition, the debate has extended to which sperm components should increase in size: the midpiece to accommodate more mitochondria and produce more energy to fuel motility, or the principal piece to generate greater propulsion forces. Results In this study we examined the influence of sperm competition upon sperm design in mammals using a much larger data set (226 species than in previous analyses, and we corrected for phylogenetic effects by using a more complete and resolved phylogeny, and more robust phylogenetic control methods. Our results show that, as sperm competition increases, all sperm components increase in an integrated manner and sperm heads become more elongated. The increase in sperm length was found to be associated with enhanced swimming velocity, an adaptive trait under sperm competition. Conclusions We conclude that sperm competition has played an important role in the evolution of sperm design in mammals, and discuss why previous studies have failed to detect it.

  18. Sperm competition and the evolution of sperm design in mammals.

    Science.gov (United States)

    Tourmente, Maximiliano; Gomendio, Montserrat; Roldan, Eduardo R S

    2011-01-13

    The influence of sperm competition upon sperm size has been a controversial issue during the last 20 years which remains unresolved for mammals. The hypothesis that, when ejaculates compete with rival males, an increase in sperm size would make sperm more competitive because it would increase sperm swimming speed, has generated contradictory results from both theoretical and empirical studies. In addition, the debate has extended to which sperm components should increase in size: the midpiece to accommodate more mitochondria and produce more energy to fuel motility, or the principal piece to generate greater propulsion forces. In this study we examined the influence of sperm competition upon sperm design in mammals using a much larger data set (226 species) than in previous analyses, and we corrected for phylogenetic effects by using a more complete and resolved phylogeny, and more robust phylogenetic control methods. Our results show that, as sperm competition increases, all sperm components increase in an integrated manner and sperm heads become more elongated. The increase in sperm length was found to be associated with enhanced swimming velocity, an adaptive trait under sperm competition. We conclude that sperm competition has played an important role in the evolution of sperm design in mammals, and discuss why previous studies have failed to detect it.

  19. EFFECT OF SEASON ON BOAR SPERM MORPHOLOGY

    Directory of Open Access Journals (Sweden)

    Jan LIPENSKÝ

    2011-01-01

    Full Text Available The objective of this study was to analyze the influence of the year–season effect on semen production parameters in the fertile AI boars. The evaluation was especially focused on the morphologically abnormal spermatozoa (MAS incidence. It was microscopically evaluated after making fresh semen smears and staining on microscopic slides. MAS incidence 19.46 % was lower at first half-year than at second half-year 25.00 % (P<0.01. Spermatozoa with distal protoplasmic droplet were furthest participated in total MAS incidence. Its rate was the highest at fourth quarter in comparison with annual period (P<0.001. We found that season has the negative effect on sperm morphology and significantly affects boar sperm quality and subsequently AI dose quality.

  20. Giant panda (Ailuropoda melanoleuca) sperm morphometry and function after repeated freezing and thawing.

    Science.gov (United States)

    Santiago-Moreno, J; Esteso, M C; Pradiee, J; Castaño, C; Toledano-Díaz, A; O'Brien, E; Lopez-Sebastián, A; Martínez-Nevado, E; Delclaux, M; Fernández-Morán, J; Zhihe, Z

    2016-05-01

    This work examines the effects of subsequent cycles of freezing-thawing on giant panda (Ailuropoda melanoleuca) sperm morphometry and function, and assesses whether density-gradient centrifugation (DGC) can increase the number of freezing-thawing cycles this sperm can withstand. A sperm sample was collected by electroejaculation from a mature giant panda and subjected to five freezing-thawing cycles. Although repeated freezing-thawing negatively affected (P 60% of the sperm cells in both treatments showed acrosome integrity even after the fifth freezing cycle. In fresh semen, the sperm head length was 4.7 μm, the head width 3.6 μm, area 14.3 μm(2) and perimeter length 14.1 μm. The present results suggest that giant panda sperm trends to be resistant to repeated freezing-thawing, even without DGC selection. © 2015 Blackwell Verlag GmbH.

  1. Evaluating rammed earth walls: a case study

    Energy Technology Data Exchange (ETDEWEB)

    Taylor, P. [Deakin University, Geelong (Australia). Built Environment Research Group; La Trobe University, Wodonga (Australia); Luther, M.B. [Deakin University, Geelong (Australia). Built Environment Research Group

    2004-03-01

    The following research has been undertaken as a response to the recent controversy regarding the suitability of rammed earth wall construction as an effective building envelope in regard to its thermal performance. The R-value for rammed earth walls is low hence they might be expected to conduct heat into a building during summer. However the large mass of these walls and the associated thermal lag in heat transfer from outside to inside may result in the walls performing satisfactorily in a building which is only occupied during working hours. Internal rammed earth walls may act as moderators of large diurnal temperature swings helping to produce an even comfortable temperature within a building. Empirical (in situ) measurements of temperature and heat flux were taken on the walls of an existing rammed earth office building in New South Wales, Australia during the summer. An analysis was performed which established a methodology to measure the heat flow associated with the walls, floor, ceiling, windows and infiltration for one office during occupied hours and the net energy transferred between the office and these elements was established. During this time the earth walls performed well. External walls were found to transmit comparatively little heat to the office and the internal walls absorbed heat during this time. Diffuse sky radiation transmitted by the window and infiltration are both likely to be important factors in the summer heat load. (author)

  2. Perfectly Secure Oblivious RAM without Random Oracles

    DEFF Research Database (Denmark)

    Damgård, Ivan Bjerre; Meldgaard, Sigurd Torkel; Nielsen, Jesper Buus

    2011-01-01

    We present an algorithm for implementing a secure oblivious RAM where the access pattern is perfectly hidden in the information theoretic sense, without assuming that the CPU has access to a random oracle. In addition we prove a lower bound on the amount of randomness needed for implementing...

  3. Free-radical production after post-thaw incubation of ram spermatozoa is related to decreased in vivo fertility.

    Science.gov (United States)

    Del Olmo, Enrique; Bisbal, Alfonso; García-Álvarez, Olga; Maroto-Morales, Alejandro; Ramón, Manuel; Jiménez-Rabadán, Pilar; Anel-López, Luis; Soler, Ana J; Garde, J Julián; Fernández-Santos, María R

    2015-11-01

    The aim of the present study was to evaluate the effect of sperm reactive oxygen species (ROS) production and DNA changes on male fertility. For that purpose, six rams with significantly different pregnancy rates were used; these were classified as having high fertility, i.e. 59.4% average pregnancy rate, or low fertility, i.e. 23.1% average pregnancy rate. Sperm quality was assessed after a two-step process of sample thawing followed by an incubation of 2h, either in the freezing extender (37°C) or after dilution in synthetic oviductal fluid (SOF; 38°C, 5%CO2). Sperm viability (YO-PRO-1), ROS production (5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein acetyl ester (CM-H2DCFDA)) and undamaged chromatin (sperm chromatin structure assay, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling, chromomycin A3) were evaluated by flow cytometry. Although no significant differences in sperm viability were observed, our results showed increased ROS production during incubation in the freezing extender as well as in SOF medium. Comparison between fertility groups showed significant differences in ROS production after 2h of incubation for the two treatments. Regarding DNA integrity, our results showed no significant differences either between treatments and incubation times or fertility groups. Linear regression analysis showed that ROS production determined by CM-H2DCFDA was a good indicator parameter for in vivo male fertility of SOF-incubated samples, yielding a fair correlation between both parameters (r=-0.92). These results indicate that detection of ROS production by CM-H2DCFDA and flow cytometry after 2h of incubation in SOF could be a useful procedure for predicting fertility of ram spermatozoa.

  4. Influence of addition of different antibiotics in semen diluent on viable bacterial count and spermatozoal viability of Awassi ram semen

    Directory of Open Access Journals (Sweden)

    O I Azawi

    2012-04-01

    Full Text Available The objectives of the present study were to determine the effects of six different antibiotics in controlling the growth of semen contaminating bacteria and if these antibiotics have any adverse effect on Awassi ram spermatozoa. Semen samples from six mature Awassi rams were used in this study. A total number of 120 ejaculates were collected from the rams using an artificial vagina once a week. Semen ejaculates were evaluated for volume, sperm concentration, mass motility, individual motility, percentage live sperm, sperm abnormalities, and viable bacterial count. Semen samples were diluted by sodium citrate-fructose-egg yolk. The diluted semen sample was divided into 7 parts. Six types of antibiotics were added to the semen diluent parts including; penicillin G 1000 IU ml-1 with streptomycin 1 mg ml-1, gentamicin sulphate 250 mg ml-1, tetracycline 0.5 mg ml-1, lincomycin 1 mg ml-1, cefoperazone sodium 1mg ml-1, cefdinir 1 mg ml-1 and the seventh part considered as a control group without antibiotic addition. The diluted semen samples were cooled and preserved at 5 Co for 5 days. Cooled diluted semen samples were examined for individual motility, percent of live sperm, sperm abnormalities, acrosomal defects and bacterial count every 24 h until 5 days. Comparing with the control, all the antibiotics examined were effective in controlling bacterial growth (P<0.05 from 24 h to 96 h of preservation at 5 Co. Cefdinir and cefoperazone sodium proved to be significantly (P<0.05 effective than other antibiotics in controlling bacterial growth at 96 h of preservation as the bacterial count were 23.3 ± 3.7 x 103 / ml and 25.4 ± 6.2 x 103 / ml, respectively. Lincomycin, gentamicin sulphate and tetracycline proved ineffective in controlling bacterial growth at 96 h of preservation as the bacterial count were 57.1 ± 20.1 x 103 / ml, 52.5 ± 29.4 x 103 / ml and 46.5 ± 8.8 x 103 / ml, respectively. The addition of tetracycline to diluted ram semen

  5. Silymarin protects plasma membrane and acrosome integrity in sperm treated with sodium arsenite

    Directory of Open Access Journals (Sweden)

    Farzaneh Eskandari

    2016-01-01

    Full Text Available Background: Exposure to arsenic is associated with impairment of male reproductive function by inducing oxidative stress. Silymarin with an antioxidant property scavenges free radicals. Objective: The aim of this study was to investigate if silymarin can prevent the adverse effects of sodium arsenite on ram sperm plasma membrane and acrosome integrity. Materials and Methods: Ram epidydimal spermatozoa were divided into five groups: spermatozoa at 0 hr, spermatozoa at 180 min (control, spermatozoa treated with silymarin (20 μM + sodium arsenite (10 μM for 180 min, spermatozoa treated with sodium arsenite (10 μM for 180 min and spermatozoa treated with silymarin (20 μM for 180 min. Double staining of Hoechst and propidium iodide was performed to evaluate sperm plasma membrane integrity, whereas comassie brilliant blue staining was used to assess acrosome integrity. Results: Plasma membrane (p< 0.001 and acrosome integrity (p< 0.05 of the spermatozoa were significantly reduced in sodium arsenite group compared to the control. In silymarin + sodium arsenite group, silymarin was able to significantly (p< 0.001 ameliorate the adverse effects of sodium arsenite on these sperm parameters compared to sodium arsenite group. The incubation of sperm for 180 min (control group showed a significant (p< 0.001 decrease in acrosome integrity compared to the spermatozoa at 0 hour. The application of silymarin alone for 180 min could also significantly (p< 0.05 increase sperm acrosome integrity compared to the control. Conclusion: Silymarin as a potent antioxidant could compensate the adverse effects of sodium arsenite on the ram sperm plasma membrane and acrosome integrity.

  6. Direct visualization of sperm competition and sperm storage in Drosophila.

    Science.gov (United States)

    Civetta, A

    Drosophila females engage in multiple matings [1] [2] [3] [4] even though they can store sperm in specialized organs for most of their life [5]. The existence of sperm competition in Drosophila has been inferred from the proportion of progeny sired by the second male in double-mating experiments [6] [7] [8]. Investigators have used this approach to quantify genetic variation underlying sperm competition [8] [9] [10], to elucidate its genetic basis [11], to identify the dependence of different male competitive ability on the genotype of the females with which they mate [12] and to discern the potential role of sperm competition in species isolation [13] [14]. This approach assumes that the sperm from two males stored in a female compete to fertilize the eggs. The mechanism by which sperm competition is accomplished is still unknown, however. Here, fluorescence microscopy, cytometry, and differently labeled sperm were used to analyze the fate of sperm inside the female's sperm storage organs, to quantify sperm competition, and to assess how closely paternity success corresponds to the appearance and location of the sperm. The results show that the first male's sperm is retained for a shortened period if the female remates, and that the second males that sire more progeny either induce females to store and use more of their sperm or strongly displace resident sperm.

  7. Evaluation of sperm DNA damage in bulls by TUNEL assay as a parameter of semen quality.

    Science.gov (United States)

    Takeda, Kumiko; Uchiyama, Kyoko; Kinukawa, Masashi; Tagami, Takahiro; Kaneda, Masahiro; Watanabe, Shinya

    2015-01-01

    Sperm DNA damage affects the conception rate resulting from human assisted reproduction technology. The objective of this study was to adapt the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay to provide a quality parameter for bull semen based on the detection of sperm DNA damage. Fresh semen was collected from two Japanese Black bulls (A, B) several times over the course of a year, and the percentage of TUNEL-positive spermatozoa (sperm TUNEL index) was determined. Individual differences in semen were detected using the sperm TUNEL index in these bulls (P bulls with a conception rate lower than 10%) and Holstein (n = 34) bulls were analyzed. The average sperm TUNEL index and conception rate resulting from artificial insemination (AI) were 4.7% and 55.7% for Japanese Black, and 4.9% and 39.5% for Holstein, respectively. A weak negative correlation between sperm TUNEL index and conception rate was observed in Holstein bulls (P bulls with more than 10% sperm TUNEL index were studied, and these samples showed low sperm viability. However, semen resulting in a very low conception rate did not have a high sperm TUNEL index. Although it would be difficult to predict a low conception rate resulting from AI using the sperm TUNEL index alone, the index can be used as an additional parameter to provide a more comprehensive description of semen quality.

  8. Fertilization is not a new beginning: the relationship between sperm longevity and offspring performance.

    Directory of Open Access Journals (Sweden)

    Angela J Crean

    Full Text Available Sperm are the most diverse cell type known: varying not only among- and within- species, but also among- and within-ejaculates of a single male. Recently, the causes and consequences of variability in sperm phenotypes have received much attention, but the importance of within-ejaculate variability remains largely unknown. Correlative evidence suggests that reduced within-ejaculate variation in sperm phenotype increases a male's fertilization success in competitive conditions; but the transgenerational consequences of within-ejaculate variation in sperm phenotype remain relatively unexplored. Here we examine the relationship between sperm longevity and offspring performance in a marine invertebrate with external fertilization, Styela plicata. Offspring sired by longer-lived sperm had higher performance compared to offspring sired by freshly-extracted sperm of the same ejaculate, both in the laboratory and the field. This indicates that within-ejaculate differences in sperm longevity can influence offspring fitness - a source of variability in offspring phenotypes that has not previously been considered. Links between sperm phenotype and offspring performance may constrain responses to selection on either sperm or offspring traits, with broad ecological and evolutionary implications.

  9. Sperm cryopreservation in live-bearing Xiphophorus fishes: offspring production from Xiphophorus variatus and strategies for establishment of sperm repositories.

    Science.gov (United States)

    Yang, Huiping; Cuevas-Uribe, Rafael; Savage, Markita G; Walter, Ronald B; Tiersch, Terrence R

    2012-09-01

    Cryopreservation of sperm from Xiphophorus fishes has produced live young in three species: X. hellerii, X. couchianus, and X. maculatus. In this study, the goal was to establish protocols for sperm cryopreservation and artificial insemination to produce live young in X. variatus, and to identify needs for repository development. The objectives were to: 1) collect basic biological characteristics of males; 2) cryopreserve sperm from X. variatus, 3) harvest live young from cryopreserved sperm, and 4) discuss the requirements for establishment of sperm repositories. The 35 males used in this study had a body weight of 0.298±0.096 g (mean±SD), body length of 2.5±0.2 cm, and testis weight of 6.4±3.4 mg. The sperm production per gram of testis was 2.33±1.32×10(9) cells. After freezing, the post-thaw motility decreased significantly to 37%±17% (ranging from 5% to 70%) (p=0.000) from 57%±14% (40%-80%) of fresh sperm (N=20). Artificial insemination of post-thaw sperm produced confirmed offspring from females of X. hellerii and X. variatus. This research, taken together with previous studies, provides a foundation for development of strategies for sperm repositories of Xiphophorus fishes. This includes: 1) the need for breeding strategies for regeneration of target populations, 2) identification of minimum fertilization capacity of frozen samples, 3) identification of fish numbers necessary for sampling and their genetic relationships, 4) selection of packaging containers for labeling and biosecurity, 5) assurance of quality control and standardization of procedures, 6) information systems that can manage the data associated with cryopreserved samples, including the genetic data, 7) biological data of sampled fish, 8) inventory data associated with frozen samples, and 9) data linking germplasm samples with other related materials such as body tissues or cells saved for DNA and RNA analyses.

  10. Supplementation of soybean lecithin-based semen extender by antioxidants: complementary flowcytometric study on post-thawed ram spermatozoa.

    Science.gov (United States)

    Sharafi, Mohsen; Zhandi, Mahdi; Akbari Sharif, Abbas

    2015-06-01

    The purpose of the current study was to evaluate the effects of cysteine (C) and glutathione (G) on the post-thawed ram sperm quality. Collected semen samples from four mature rams were diluted with five soybean lecithin (SL)-based extenders containing: no antioxidant (SL-0), 5 mM cysteine (SL-C5), 10 mM cysteine (SL-C10), 5 mM glutathione (SL-G5) and 10 mM glutathione (SL-G10). After freeze-thawing process, motion and velocity parameters, plasma membrane integrity and functionality, morphological abnormality, lipid peroxidation, acrosomal status, mitochondria activity, and apoptosis status of post-thawed ram spermatozoa were assessed. The results showed that SL-C10 increased the total motility and plasma membrane integrity (p ram spermatozoa (55.86 ± 1.37 and 60.57 ± 1.34 %) compared to other extenders. Progressive motility was significantly higher in SL-C10 (24.71 ± 1.13 %) compared to SL-0 (20 ± 1.13 %) and SL-G10 (15 ± 1.13 %). Mitochondrial activity was significantly higher in SL-C10 (56.83 ± 2.29 %) compared to SL-G10 (38.75 ± 2.29 %). Capacitation and acrosomal status, lipid peroxidation, and the percentage of dead spermatozoa were not affected by different extenders. The percentage of live spermatozoa was higher in SL-C10 (56.33 ± 1.35 %) compared to other extenders. Also, SL-C10 resulted in a lower percentage of apoptotic spermatozoa (14.17 ± 0.53 %) compared to other extenders. The results of this study showed that supplementation of SL-based ram semen extender with 10 mM cysteine resulted in an improved quality of post-thawed ram spermatozoa.

  11. Chemotactic Maneuverability of Sperm

    CERN Document Server

    Guasto, Jeffrey S; Zimmer, Richard K; Stocker, Roman

    2011-01-01

    In this fluid mechanics video, we explore the kinematics of chemotaxing sperm cells (sea urchin, \\textit{Arbacia punctulata}) swimming in a chemoattractant gradient. We demonstrate that the complex swimming trajectories resulting in chemotactic behavior (`turn-and-run' motility) are comprised of several distinct flagellar maneuvers. These motility patterns likely play an important role optimizing chemotaxic motility and navigation, when the sperm cells are subjected external fluid flows.

  12. Is prnt a Pseudogene? Identification of Ram Prt in Testis and Ejaculated Spermatozoa

    Science.gov (United States)

    Pimenta, Jorge; Domingos, Ana; Santos, Pedro; Marques, Carla C.; Cantante, Cátia; Santos, Ana; Barbas, João P.; Baptista, Maria C.; Horta, António E. M.; Viegas, Aldino; Mesquita, Patrícia; Gonçalves, João; Fontes, Carlos A.; Prates, José A. M.; Pereira, Rosa M. L. N.

    2012-01-01

    A hallmark of prion diseases or transmissible spongiform encephalopaties is the conversion of the cellular prion protein (PrPC), expressed by the prion gene (prnp), into an abnormally folded isoform (PrPSc) with amyloid-like features that causes scrapie in sheep among other diseases. prnp together with prnd (which encodes a prion-like protein designated as Doppel), and prnt (that encodes the prion protein testis specific - Prt) with sprn (shadow of prion protein gene, that encodes Shadoo or Sho) genes, constitute the “prion gene complex”. Whereas a role for prnd in the proper functioning of male reproductive system has been confirmed, the function of prnt, a recently discovered prion family gene, comprises a conundrum leading to the assumption that ruminant prnt is a pseudogene with no protein expression. The main objective of the present study was to identify Prt localization in the ram reproductive system and simultaneously to elucidate if ovine prnt gene is transcribed into protein-coding RNA. Moreover, as Prt is a prnp-related protein, the amyloid propensity was also tested for ovine and caprine Prt. Recombinant Prt was used to immunize BALB/c mice, and the anti-Prt polyclonal antibody (APPA) immune response was evaluated by ELISA and Western Blot. When tested by indirect immunofluorescence, APPA showed high avidity to the ram sperm head apical ridge subdomain, before and after induced capacitation, but did not show the same behavior against goat spermatozoa, suggesting high antibody specificity against ovine-Prt. Prt was also found in the testis when assayed by immunohistochemistry during ram spermatogenesis, where spermatogonia, spermatocytes, spermatids and spermatozoa, stained positive. These observations strongly suggest ovine prnt to be a translated protein-coding gene, pointing to a role for Prt protein in the ram reproductive physiology. Besides, caprine Prt appears to exhibit a higher amyloid propensity than ovine Prt, mostly associated with its

  13. Cryopreservation of ram semen in extenders containing soybean lecithin as cryoprotectant and hyaluronic acid as antioxidant.

    Science.gov (United States)

    Najafi, A; Najafi, M H; Zanganeh, Z; Sharafi, M; Martinez-Pastor, F; Adeldust, H

    2014-12-01

    A soybean lecithin-based extender supplemented with hyaluronic acid (HA) was assayed for effectiveness to improve the quality of frozen-thawed ram semen. HA has not been tested yet in an extender containing soybean lecithin for freezing ram semen. Thus, the aim of this study was to analyse the effects of soybean lecithin at 1% or 1.5% along with HA at 0, 0.5 and 1 mg ml(-1) in a Tris-based extender on the motion characteristics, membrane integrity (HOST), viability, GSH peroxidase (GSH-PX) activity, lipid peroxidation and acrosomal status after freezing-thawing. Semen was collected from four Mehraban rams during the breeding season and frozen in the six lecithin×HA extenders. The extender containing 1.5% lecithin supplemented with no HA yielded higher total motility (52.5%±1.6), viability (55.8%±1.6) and membrane integrity (44.5%±1.7), but the effects of the lecithin concentration did not reach signification. Linearity-related parameters, ALH, BCF, lipid peroxidation, GSH-PX activity, morphology and acrosomal status were not affected by the extender composition. In general, adding HA significantly decreased sperm velocity (1 mg ml(-1) HA), total motility (only with 1.5% lecithin), viability (1 mg ml(-1) HA for 1% lecithin; both concentrations for 1.5% lecithin) and membrane integrity. In conclusion, adding HA to the freezing extender supplemented with soybean lecithin failed to improve quality-related variables in ram semen. Increasing the lecithin content could have a positive effect, but further studies are needed. © 2014 Blackwell Verlag GmbH.

  14. Is prnt a pseudogene? Identification of ram Prt in testis and ejaculated spermatozoa.

    Directory of Open Access Journals (Sweden)

    Jorge Pimenta

    Full Text Available A hallmark of prion diseases or transmissible spongiform encephalopaties is the conversion of the cellular prion protein (PrP(C, expressed by the prion gene (prnp, into an abnormally folded isoform (PrP(Sc with amyloid-like features that causes scrapie in sheep among other diseases. prnp together with prnd (which encodes a prion-like protein designated as Doppel, and prnt (that encodes the prion protein testis specific--Prt with sprn (shadow of prion protein gene, that encodes Shadoo or Sho genes, constitute the "prion gene complex". Whereas a role for prnd in the proper functioning of male reproductive system has been confirmed, the function of prnt, a recently discovered prion family gene, comprises a conundrum leading to the assumption that ruminant prnt is a pseudogene with no protein expression. The main objective of the present study was to identify Prt localization in the ram reproductive system and simultaneously to elucidate if ovine prnt gene is transcribed into protein-coding RNA. Moreover, as Prt is a prnp-related protein, the amyloid propensity was also tested for ovine and caprine Prt. Recombinant Prt was used to immunize BALB/c mice, and the anti-Prt polyclonal antibody (APPA immune response was evaluated by ELISA and Western Blot. When tested by indirect immunofluorescence, APPA showed high avidity to the ram sperm head apical ridge subdomain, before and after induced capacitation, but did not show the same behavior against goat spermatozoa, suggesting high antibody specificity against ovine-Prt. Prt was also found in the testis when assayed by immunohistochemistry during ram spermatogenesis, where spermatogonia, spermatocytes, spermatids and spermatozoa, stained positive. These observations strongly suggest ovine prnt to be a translated protein-coding gene, pointing to a role for Prt protein in the ram reproductive physiology. Besides, caprine Prt appears to exhibit a higher amyloid propensity than ovine Prt, mostly associated

  15. Accessory sperm: a biomonitor of boar sperm fertilization capacity.

    Science.gov (United States)

    Ardón, Florencia; Evert, Meike; Beyerbach, Martin; Weitze, Karl-Fritz; Waberski, Dagmar

    2005-04-15

    The number of accessory sperm found in the zona pellucida of porcine embryos was correlated to their individual quality and to the embryo quality range found within a single sow. Our goal was to determine whether accessory sperm counts provide semen evaluation with additional, useful information. Accessory sperm count was highest when only normal embryos were found in a given sow and diminished if oocytes or degenerated embryos were present (P<0.01). Within a given sow, normal embryos had higher (P<0.05) accessory sperm counts than degenerated embryos, although not when oocytes were also present. Fertilization capacity of sperm is optimal when only normal embryos are found in a given sow; this capacity is indicated by high accessory sperm counts. A decrease in fertilization capacity is reflected in diminishing accessory sperm counts. The boar had a significant effect (P<0.01) on accessory sperm count, but not on the percentage of normal embryos; this suggests that accessory sperm may be more sensitive indicators of the fertilization capacity of sperm than the percentage of normal embryos. We conclude that accessory sperm count can be used for the detection of compensable defects in sperm and is a valid parameter for assessing sperm fertilization capacity.

  16. Sperm-egg penetration assay assessment of the contraceptive effects of glycerol and egg yolk in rooster sperm diluents.

    Science.gov (United States)

    Abouelezz, F M K; Castaño, C; Toledano-Díaz, A; Esteso, M C; López-Sebastián, A; Campo, J L; Santiago-Moreno, J

    2015-06-01

    Glycerol (GLY) and egg yolk (EY) are good cryoprotectants of avian and mammalian sperm, but in birds, they strongly inhibit the eventual fertilization of ova. Using the sperm penetration (SP-holes) assay and fertility trials, the present study investigates (1) the possible mechanism by which this contraceptive effect occurs in chickens and (2) the maximum concentrations of GLY and EY tolerated by fresh rooster sperm. Seventy Black-Barred Andaluza hens (five per treatment) were inseminated four times (twice per week) with 0.1 mL of fresh semen from roosters of the same breed diluted 1:1 (v:v) with Lake and Ravie medium containing different concentrations of GLY or EY. No adverse effects on acrosome integrity, sperm motility, or viability were seen with any concentration of GLY or EY. The number of SP-holes on perivitelline layer samples taken from above the germinal disc became progressively lower at GLY concentrations of 1.5% or greater (P > 0.05). No holes caused by sperms were seen in unfertilized eggs. The corresponding fertility results showed similar reductions when the GLY concentration was 1.5% or greater. No changes in the number of SP-holes were seen with increasing EY concentrations (0%-7.5%), nor were any differences in fertility observed, except for a reduction when 15% EY was used. The results therefore reveal that GLY affects the transit of sperms through the oviduct in their attempt to reach the infundibulum area, limiting their access to the ovum perivitelline layer. Egg yolk had no such effect, nor did it influence acrosome reaction capacity; its mechanism of contraceptive action therefore remains unknown. The maximum GLY and EY concentrations tolerated by the rooster sperm were 0.75% and 7.5%, respectively.

  17. Sperm DNA fragmentation index does not correlate with the sperm or embryo aneuploidy rate in recurrent miscarriage or implantation failure patients.

    Science.gov (United States)

    Bronet, F; Martínez, E; Gaytán, M; Liñán, A; Cernuda, D; Ariza, M; Nogales, M; Pacheco, A; San Celestino, M; Garcia-Velasco, J A

    2012-07-01

    The aneuploidy rate is higher in poor-quality sperm samples, which also have higher DNA fragmentation index values. The aim of this study was to assess the relationship between sperm DNA fragmentation in samples from infertile men belonging to couples with recurrent miscarriage or implantation failure and the aneuploidy rate in spermatozoa as well as in embryos from patients. This prospective study evaluated DNA damage and the aneuploidy rate in fresh and processed (density gradient centrifugation) ejaculated sperm as well as the aneuploidy rate in biopsied embryos from fertility cycles. Fluorescence in situ hybridization was used for the aneuploidy analysis. Results were compared using linear regression and analysis of variance. A total of 154 embryos were evaluated from 38 patients undergoing PGD cycles; 35.2% of the embryos were chromosomally normal. Analysis of the same sperm samples showed an increased DNA fragmentation after sperm preparation in 76% of the patients. There was no correlation between DNA fragmentation and the aneuploidy rate in embryos or in fresh or processed sperm samples. Sperm DNA fragmentation is not related to chromosomal anomalies in embryos from patients with recurrent miscarriage or implantation failure. However, we cannot rule out the possibility that a relationship between DNA fragmentation and aneuploidy exists for other causes of infertility. Furthermore, the different methods used to evaluate DNA fragmentation may produce different results.

  18. Ram pressure stripping of tilted galaxies

    CERN Document Server

    Jachym, P; Palous, J; Combes, F

    2009-01-01

    Ram pressure stripping of galaxies in clusters can yield gas deficient disks. Previous numerical simulations based on various approaches suggested that, except for near edge-on disk orientations, the amount of stripping depends very little on the inclination angle. Following our previous study of face-on stripping, we extend the set of parameters with the disk tilt angle and explore in detail the effects of the ram pressure on the interstellar content (ISM) of tilted galaxies that orbit in various environments of clusters, with compact or extended distributions of the intra-cluster medium (ICM). We further study how results of numerical simulations could be estimated analytically. A grid of numerical simulations with varying parameters is produced using the tree/SPH code GADGET with a modified method for calculating the ISM-ICM interaction. These SPH calculations extend the set of existing results obtained from different codes using various numerical techniques. The simulations confirm the general trend of le...

  19. Space and the Synchronic A-Ram

    CERN Document Server

    Berka, Alex V

    2010-01-01

    Space is a circuit oriented, spatial programming language designed to exploit the massive parallelism available in a novel formal model of computation called the Synchronic A-Ram, and physically related FPGA and reconfigurable architectures. Space expresses variable grained MIMD parallelism, is modular, strictly typed, and deterministic. Barring operations associated with memory allocation and compilation, modules cannot access global variables, and are referentially transparent. At a high level of abstraction, modules exhibit a small, sequential state transition system, aiding verification. Space deals with communication, scheduling, and resource contention issues in parallel computing, by resolving them explicitly in an incremental manner, module by module, whilst ascending the ladder of abstraction. Whilst the Synchronic A-Ram model was inspired by linguistic considerations, it is also put forward as a formal model for reconfigurable digital circuits. A programming environment has been developed, that inco...

  20. Subversive practices of sperm donation - globalizing Danish sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past two decades, Denmark has developed in to an important destination for fertility travellers in need of donor sperm. Furthermore, two of the largest sperm banks in Europe have been established in Denmark, exporting sperm globally. This development has taken place at the same time...

  1. Sperm-egg interaction.

    Science.gov (United States)

    Evans, Janice P

    2012-01-01

    A crucial step of fertilization is the sperm-egg interaction that allows the two gametes to fuse and create the zygote. In the mouse, CD9 on the egg and IZUMO1 on the sperm stand out as critical players, as Cd9(-/-) and Izumo1(-/-) mice are healthy but infertile or severely subfertile due to defective sperm-egg interaction. Moreover, work on several nonmammalian organisms has identified some of the most intriguing candidates implicated in sperm-egg interaction. Understanding of gamete membrane interactions is advancing through characterization of in vivo and in vitro fertilization phenotypes, including insights from less robust phenotypes that highlight potential supporting (albeit not absolutely essential) players. An emerging theme is that there are varied roles for gamete molecules that participate in sperm-egg interactions. Such roles include not only functioning as fusogens, or as adhesion molecules for the opposite gamete, but also functioning through interactions in cis with other proteins to regulate membrane order and functionality.

  2. Sperm Motility in Flow

    Science.gov (United States)

    Guasto, Jeffrey; Juarez, Gabriel; Stocker, Roman

    2012-11-01

    A wide variety of plants and animals reproduce sexually by releasing motile sperm that seek out a conspecific egg, for example in the reproductive tract for mammals or in the water column for externally fertilizing organisms. Sperm are aided in their quest by chemical cues, but must also contend with hydrodynamic forces, resulting from laminar flows in reproductive tracts or turbulence in aquatic habitats. To understand how velocity gradients affect motility, we subjected swimming sperm to a range of highly-controlled straining flows using a cross-flow microfluidic device. The motion of the cell body and flagellum were captured through high-speed video microscopy. The effects of flow on swimming are twofold. For moderate velocity gradients, flow simply advects and reorients cells, quenching their ability to cross streamlines. For high velocity gradients, fluid stresses hinder the internal bending of the flagellum, directly inhibiting motility. The transition between the two regimes is governed by the Sperm number, which compares the external viscous stresses with the internal elastic stresses. Ultimately, unraveling the role of flow in sperm motility will lead to a better understanding of population dynamics among aquatic organisms and infertility problems in humans.

  3. Comparison of different methods for assessment of sperm concentration and membrane integrity with bull semen.

    Science.gov (United States)

    Anzar, Muhammad; Kroetsch, Tom; Buhr, Mary M

    2009-01-01

    Assessing semen quality is crucially important for the exploitation of genetically superior sires in an artificial insemination (AI) program. In this study, we compare modern and conventional techniques to estimate bovine sperm concentration and membrane integrity. First, the NucleoCounter SP-100 was validated for sperm concentration and provided statistically reliable and repeatable estimates among aliquots and replicates of 25 fresh ejaculates. Sperm concentrations in 78 ejaculates were then determined with hemacytometer, flow cytometer, and NucleoCounter SP-100 and were significantly correlated (P spectrophotometers, comparing optical density against counts drawn by hemacytometer and NucleoCounter SP-100 (n = 94 fresh ejaculates) showed different (P calibration of spectrophotometers. Flow cytometer and NucleoCounter SP-100 can be used with equal confidence to estimate sperm concentration and membrane integrity in domestic animals and human semen.

  4. Sorting of Sperm by Morphology

    Science.gov (United States)

    Koh, James; Marcos, Marcos

    2016-11-01

    Many studies have proven that the percentage of morphologically normal sperm is a significant factor in determining the success of assisted reproduction. The velocity of sperm in a microchannel with shear flow subjected to an external field will be explored theoretically. The difference in response between morphologically normal and abnormal sperm will be computed from a statistical approach, to study the feasibility and effectiveness of sorting by an external field to remove abnormal sperm. The full name of this author is Marcos.

  5. Sperm cryopreservation of the Indian major carp, Labeo calbasu: effects of cryoprotectants, cooling rates and thawing rates on egg fertilization.

    Science.gov (United States)

    Nahiduzzaman, Md; Hassan, Md Mahbubul; Roy, Pankoz Kumar; Hossain, Md Akhtar; Hossain, Mostafa Ali Reza; Tiersch, Terrence R

    2012-12-01

    A sperm cryopreservation protocol for the Indian major carp, Labeo calbasu, was developed for long-term preservation and artificial fertilization. Milt collected from mature male fish were placed in Alsever's solution (296mOsmolkg(-1)) to immobilize the sperm. Cryoprotectant toxicity was evaluated by motility assessment with dimethyl sulfoxide (DMSO) and methanol at 5, 10 and 15% concentrations. DMSO was more toxic at higher concentrations than methanol, and consequently 15% DMSO was excluded from further study. A one-step cooling protocol (from 5 to 80°C) with two cooling rates (5 and 10°C/min) was carried out in a computer-controlled freezer (FREEZE CONTROL(®) CL-3300; Australia). Based on post-thaw motility, the 10°C/min cooling rate with either 10% DMSO or 10% methanol yielded significantly higher (P=0.011) post-thaw motility than the other rate and cryoprotectant concentrations. Sperm thawed at 40°C for 15s and fresh sperm were used to fertilize freshly collected L. calbasu eggs and significant differences were observed (P=0.001) in percent fertilization between cryopreserved and fresh sperm as well as among different sperm-to-egg ratios (P=0.001). The highest fertilization and hatching rates were observed for thawed sperm at a sperm-to-egg ratio of 4.1×10(5):1. The cryopreservation protocol developed can facilitate hatchery operations and long-term conservation of genetic resources of L. calbasu.

  6. Correlation of frequency of spermatozoa morphological alterations with sperm concentration in ejaculates of Polish Landrace boars

    Directory of Open Access Journals (Sweden)

    Kondracki S.

    2013-01-01

    Full Text Available The experiments were performed on 448 ejaculates obtained from 41 Polish Landrace boars. Ejaculates collected from each boar at one-month intervals for approximately 10 months were analysed. Sperm morphometric measurements were taken from each boar and assessment of semen morphology was done on the basis of examination under a microscope of preparations made from fresh ejaculates. The ejaculates were classified based on the criterion of sperm concentration and divided into three groups. An attempt was made in the present study to assess the correlation of ejaculate parameters, morphological sperm alteration incidence and morphometric sperm parameters with the sperm concentration in ejaculates of Polish Landrace boars. It should be stated that morphometric traits of spermatozoa are related to sperm concentration. The spermatozoa in concentrated ejaculates had smaller heads than the spermatozoa in the ejaculates with lower sperm concentrations. This can mean that the high fertility of males that produce highly concentrated semen does not only result from a high sperm concentration, but also from the fact that the spermatozoa in such ejaculates have smaller heads. The highest frequency of morphologically well-formed spermatozoa was identified in ejaculates with the sperm concentration ranging from 400 to 500 thousand/mm3.

  7. Testicular sperm retrieval at the time of bilateral radical orchiectomy

    Directory of Open Access Journals (Sweden)

    Nassir Anmar

    2009-01-01

    Full Text Available A new indication for testicular tissue cryopreservation is discussed here. We evaluate the feasibility of testicular sperm extraction during bilateral orchiectomy for testicular cancer. A 26-year-old man with a history of right orchidopexy presented with primary infertility. Tests revealed left varicocele and oligospermia. He underwent varicocelectomy but was lost to follow-up. Nine months later, he presented with right-sided solid testicular swelling. Ultrasound examination confirmed the finding of possible neoplasm and revealed contralateral intratesticular lesion. Since the patient turned azoospermic, no semen sample could be cryopreserved. Permanent histopathological sections revealed bilateral classical seminomas. On processing the fresh healthy looking specimen, normal-looking viable sperm were obtained. Future attempts at in-vitro fertilization with intracytoplasmic sperm injection should be possible. This method has been described in metachronous testicular cancer, but to our knowledge, we are the first to use it in synchronous bilateral testicular tumors.

  8. Subversive practices of sperm donation - globalizing Danish sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past two decades, Denmark has developed in to an important destination for fertility travellers in need of donor sperm. Furthermore, two of the largest sperm banks in Europe have been established in Denmark, exporting sperm globally. This development has taken place at the same time...... into how the bending of boundaries by “inappropriate parents”, fertility travellers, private sperm banks and fertility clinics have been part in negotiating the changes of the legislation in practice, and thus been part of developing a Danish industry of sperm banking. The presentation is based on a multi...

  9. Sperm recovery and IVF after testicular sperm extraction (TESE: effect of male diagnosis and use of off-site surgical centers on sperm recovery and IVF.

    Directory of Open Access Journals (Sweden)

    Kenan Omurtag

    Full Text Available OBJECTIVE: Determine whether testicular sperm extractions and pregnancy outcomes are influenced by male and female infertility diagnoses, location of surgical center and time to cryopreservation. PATIENTS: One hundred and thirty men undergoing testicular sperm extraction and 76 couples undergoing 123 in vitro fertilization cycles with testicular sperm. OUTCOME MEASURES: Successful sperm recovery defined as 1-2 sperm/0.5 mL by diagnosis including obstructive azoospermia (n = 60, non-obstructive azoospermia (n = 39, cancer (n = 14, paralysis (n = 7 and other (n = 10. Obstructive azoospermia was analyzed as congenital absence of the vas deferens (n = 22, vasectomy or failed vasectomy reversal (n = 37 and "other"(n = 1. Sperm recovery was also evaluated by surgical site including infertility clinic (n = 54, hospital operating room (n = 67 and physician's office (n = 11. Treatment cycles were evaluated for number of oocytes, fertilization, embryo quality, implantation rate and clinical/ongoing pregnancies as related to male diagnosis, female diagnosis, and use of fresh or cryopreserved testicular sperm. RESULTS: Testicular sperm recovery from azoospermic males with all diagnoses was high (70 to 100% except non-obstructive azoospermia (31% and was not influenced by distance from surgical center to laboratory. Following in vitro fertilization, rate of fertilization was significantly lower with non-obstructive azoospermia (43%, p = <0.0001 compared to other male diagnoses (66%, p = <0.0001, 59% p = 0.015. No differences were noted in clinical pregnancy rate by male diagnosis; however, the delivery rate per cycle was significantly higher with obstructive azoospermia (38% p = 0.0371 compared to diagnoses of cancer, paralysis or other (16.7%. Women diagnosed with diminished ovarian reserve had a reduced clinical pregnancy rate (7.4% p = 0.007 compared to those with other diagnoses (44

  10. Cholesterol-Loaded Cyclodextrin Increases the Cholesterol Content of Goat Sperm to Improve Cold and Osmotic Resistance and Maintain Sperm Function after Cryopreservation.

    Science.gov (United States)

    Salmon, Vianney M; Leclerc, Pierre; Bailey, Janice L

    2016-04-01

    The success of semen cryopreservation depends on sperm membrane integrity and function after thawing. Cholesterol-loaded cyclodextrin (CLC) is used for in vitro incorporation of cholesterol to protect cells against cold temperatures. We hypothesized that CLC treatment also enhances sperm cholesterol content to increase tolerance to osmotic shock and cryoresistance, thereby improving fertility. We confirmed the fact that treatment of goat semen with 3 mg/ml CLC increases sperm cholesterol content using both the Liebermann-Burchard approach and filipin III labeling of membrane cholesterol. Sperm were then treated with or without CLC and cryopreserved. After thawing, sperm cholesterol dramatically fell, even in the presence of CLC, which explains the mechanism of cryocapacitation. CLC treatment, however, maintained a normal prefreeze cholesterol level in sperm after cryopreservation. Furthermore, fresh sperm treated with CLC and subjected to either cold shock or incubated in hypo-, iso-, and hyperosmotic media, designed to mimic stresses associated with freezing/thawing, displayed increased temperature and osmotic tolerance. CLC treatment also improved sperm viability, motility, and acrosome integrity after thawing. Furthermore, CLC treatment did not affect the sperm's ability to undergo in vitro capacitation according to chlortetracycline fluorescence and protein tyrosine phosphorylation. A pilot field trial demonstrated that artificial insemination with sperm that underwent increased cholesterol levels following CLC treatment yielded higher fertility ( ITALIC! P< 0.1) and proliferation ( ITALIC! P< 0.05) rates in vivo than untreated semen from the same ejaculate samples. These observations suggest that CLC treatment could be used to improve cryoprotection during the freezing and thawing of goat sperm.

  11. Subversive practices of sperm donation - globalizing Danish sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past two decades, Denmark has developed in to an important destination for fertility travellers in need of donor sperm. Furthermore, two of the largest sperm banks in Europe have been established in Denmark, exporting sperm globally. This development has taken place at the same time...... as the use of donated sperm continuously has been debated as an ethical issue, and increasingly been regulated. In this presentation I will discuss how Denmark became a destination for fertility travelling (sperm donation) as a result of various subversive strategies of family making. The article inquires...... into how the bending of boundaries by “inappropriate parents”, fertility travellers, private sperm banks and fertility clinics have been part in negotiating the changes of the legislation in practice, and thus been part of developing a Danish industry of sperm banking. The presentation is based on a multi...

  12. Subversive practices of sperm donation - globalizing Danish sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past two decades, Denmark has developed in to an important destination for fertility travellers in need of donor sperm. Furthermore, two of the largest sperm banks in Europe have been established in Denmark, exporting sperm globally. This development has taken place at the same time...... as the use of donated sperm continuously has been debated as an ethical issue, and increasingly been regulated. In this presentation I will discuss how Denmark became a destination for fertility travelling (sperm donation) as a result of various subversive strategies of family making. The article inquires...... into how the bending of boundaries by “inappropriate parents”, fertility travellers, private sperm banks and fertility clinics have been part in negotiating the changes of the legislation in practice, and thus been part of developing a Danish industry of sperm banking. The presentation is based on a multi...

  13. Comparative analysis of boar seminal plasma proteome from different freezability ejaculates and identification of Fibronectin 1 as sperm freezability marker.

    Science.gov (United States)

    Vilagran, I; Yeste, M; Sancho, S; Castillo, J; Oliva, R; Bonet, S

    2015-03-01

    Variation in boar sperm freezability (i.e. capacity to withstand cryopreservation) between ejaculates is a limitation largely reported in the literature. Prediction of sperm freezability and classification of boar ejaculates into good (GFEs) and poor freezability ejaculates (PFEs) before cryopreservation takes place may increase the use of frozen-thawed spermatozoa. While markers of boar sperm freezability have been found from sperm cell extracts, little attention has been paid to seminal plasma. On this basis, the present study compared the fresh seminal plasma proteome of 9 GFEs and 9 PFEs through two-dimensional difference gel electrophoresis (2D-DIGE) and liquid chromatography mass spectrometry (LC-MS/MS). The ejaculates were previously classified as GFE or PFE upon their sperm viability and progressive motility assessments at 30 and 240 min post thawing. From a total of 51 spots, four were found to significantly (p sperm quality parameters. Results confirmed that FN1 is a reliable marker of boar sperm freezability, because GFEs presented significantly (p boar sperm freezability marker. We can thus conclude that levels of FN1 in fresh seminal plasma from boar semen may be used as a sperm freezability marker, thereby facilitating the use of frozen-thawed boar spermatozoa.

  14. Changeability of sperm chromatin structure during liquid storage of ovine semen in milk-egg yolk- and soybean lecithin-based extenders and their relationships to field-fertility.

    Science.gov (United States)

    Khalifa, Tarek; Lymberopoulos, Aristotelis

    2013-12-01

    The aim of this experiment was to study the effect of semen extender on sperm chromatin structure and to correlate chromatin integrity with field-fertility of preserved ram semen. Ejaculates of at least 2 × 10(9) sperm/ml and 70 % progressive motility were collected using an artificial vagina from Chios rams (n = 11, 4-6 years old), split-diluted to 1 × 10(9) sperm/ml with milk-egg yolk- and soybean lecithin (Ovixcell®)-based extenders, packaged in 0.5-ml straws and examined after 6, 24 and 48 h of storage at 5 ± 1 °C. Evaluation endpoints were computer-assisted sperm motion analysis, fluorescence-based analysis of chromatin structure by chromomycin A3 and acridine orange assays, and 65-day pregnancy rate (PR) of 34- to 36-h preserved semen after intra-cervical insemination of ewes (n = 154) in progestagen-synchronized estrus. Neither extender nor storage time had any influence on incidence of decondensed chromatin. Unlike Ovixcell® extender, deterioration of sperm motility (P egg yolk extender. Sperm motility accounted for 14.4-18.5 % of variations in chromatin integrity (P egg yolk-stored semen. Nevertheless, PR differed between rams (14.3-71.4 %; P egg yolk extender in preserving chromatin stability and motility. Chromatin defects are negatively associated with sperm fertility.

  15. Incorporation of RAM techniques into simulation modeling

    Energy Technology Data Exchange (ETDEWEB)

    Nelson, S.C. Jr.; Haire, M.J.; Schryver, J.C.

    1995-07-01

    This work concludes that reliability, availability, and maintainability (RAM) analytical techniques can be incorporated into computer network simulation modeling to yield an important new analytical tool. This paper describes the incorporation of failure and repair information into network simulation to build a stochastic computer model represents the RAM Performance of two vehicles being developed for the US Army: The Advanced Field Artillery System (AFAS) and the Future Armored Resupply Vehicle (FARV). The AFAS is the US Army`s next generation self-propelled cannon artillery system. The FARV is a resupply vehicle for the AFAS. Both vehicles utilize automation technologies to improve the operational performance of the vehicles and reduce manpower. The network simulation model used in this work is task based. The model programmed in this application requirements a typical battle mission and the failures and repairs that occur during that battle. Each task that the FARV performs--upload, travel to the AFAS, refuel, perform tactical/survivability moves, return to logistic resupply, etc.--is modeled. Such a model reproduces a model reproduces operational phenomena (e.g., failures and repairs) that are likely to occur in actual performance. Simulation tasks are modeled as discrete chronological steps; after the completion of each task decisions are programmed that determine the next path to be followed. The result is a complex logic diagram or network. The network simulation model is developed within a hierarchy of vehicle systems, subsystems, and equipment and includes failure management subnetworks. RAM information and other performance measures are collected which have impact on design requirements. Design changes are evaluated through ``what if`` questions, sensitivity studies, and battle scenario changes.

  16. Confocal Microscopy and Image Analysis Indicates a Region-Specific Relation between Active Caspases and Cytoplasm in Ejaculated and Epididymal Sperm

    Science.gov (United States)

    García Vazquez, Susana; Aragón Martínez, Andrés; Flores-Alonso, Juan Carlos

    2012-01-01

    Previously, it was suggested a relation between the presence of apoptosis markers with cytoplasm in mammalian sperm. In this work, flow cytometry, confocal microscopy and image analysis were used to analyze the relationship between active caspase-3 and -7 and intracellular esterases expression in ejaculated or epididymal ram sperm. Sperm obtained from ejaculates from the caput, corpus, or cauda of the epididymis were treated with an inhibitor of active caspase-3 and -7 and a marker of cytoplasmic esterases. Additionally, ejaculated sperm were incubated for one, two, or three hours before evaluation for active caspases. Sperm subpopulations positive for active caspases and/or intracellular esterases were detected by flow cytometry and confocal microscopy; however, image analysis of confocal images showed that the correlation between active caspases and cytoplasmic esterases in sperm is region-specific. Lower values of Spearman correlation coefficients were found when whole sperm or head sperm was analyzed; however, a high correlation was observed for midpiece sperm. Incubation of sperm for two or three hours promoted the autoactivation of caspases. It has been suggested that the presence of apoptotic markers in sperm are related with a process of abortive apoptosis and with errors during spermiogenesis. Our results permit us suggest that the origin of the relationship between active caspases and cytoplasmic esterases is due to differentiation errors occurring during spermiogenesis because the percentages of sperm with active caspases are not different in the caput, corpus, or cauda of the epididymis. In this study we demonstrate that existing sperm subpopulations can express active caspases and intracellular esterases and that the correlation between these molecules is high in midpiece sperm. PMID:22530029

  17. Confocal microscopy and image analysis indicates a region-specific relation between active caspases and cytoplasm in ejaculated and epididymal sperm.

    Directory of Open Access Journals (Sweden)

    Susana García Vazquez

    Full Text Available Previously, it was suggested a relation between the presence of apoptosis markers with cytoplasm in mammalian sperm. In this work, flow cytometry, confocal microscopy and image analysis were used to analyze the relationship between active caspase-3 and -7 and intracellular esterases expression in ejaculated or epididymal ram sperm. Sperm obtained from ejaculates from the caput, corpus, or cauda of the epididymis were treated with an inhibitor of active caspase-3 and -7 and a marker of cytoplasmic esterases. Additionally, ejaculated sperm were incubated for one, two, or three hours before evaluation for active caspases. Sperm subpopulations positive for active caspases and/or intracellular esterases were detected by flow cytometry and confocal microscopy; however, image analysis of confocal images showed that the correlation between active caspases and cytoplasmic esterases in sperm is region-specific. Lower values of Spearman correlation coefficients were found when whole sperm or head sperm was analyzed; however, a high correlation was observed for midpiece sperm. Incubation of sperm for two or three hours promoted the autoactivation of caspases. It has been suggested that the presence of apoptotic markers in sperm are related with a process of abortive apoptosis and with errors during spermiogenesis. Our results permit us suggest that the origin of the relationship between active caspases and cytoplasmic esterases is due to differentiation errors occurring during spermiogenesis because the percentages of sperm with active caspases are not different in the caput, corpus, or cauda of the epididymis. In this study we demonstrate that existing sperm subpopulations can express active caspases and intracellular esterases and that the correlation between these molecules is high in midpiece sperm.

  18. RAM-Efficient External Memory Sorting

    DEFF Research Database (Denmark)

    Arge, Lars; Thorup, Mikkel

    2013-01-01

    In recent years a large number of problems have been considered in external memory models of computation, where the complexity measure is the number of blocks of data that are moved between slow external memory and fast internal memory (also called I/Os). In practice, however, internal memory time...... often dominates the total running time once I/O-efficiency has been obtained. In this paper we study algorithms for fundamental problems that are simultaneously I/O-efficient and internal memory efficient in the RAM model of computation....

  19. Turbine ram-jet engine. Turbinen-Staustrahltriebwerk

    Energy Technology Data Exchange (ETDEWEB)

    Wildner, W.

    1990-10-25

    A turbine ram-jet engine with a ram pressure air duct surrounding the basic turbine engine as an annulus is described, in which with the ram jet switched off with simultaneous release of suction air into the basic engine, the supply of ram pressure air is shut off. A suction side air inflow corss section of the basic engine can be released or shut off by axial adjustment of a drop-shaped body relative to a flow divider. A ring slide axially adjustable opposite to this body, should be provided, which shuts off the ram pressure air duct with the incoming crossection of the basic engine open and opens it for the shut-off state of the incoming crossection. Further, the ram pressure air duct should be expanded at the level of shut-off zone formed by a body with local counter-surface contact and the ring slider in the upstream direction.

  20. Microfluidic single sperm analysis

    NARCIS (Netherlands)

    Wagenaar, de Bjorn

    2016-01-01

    Microfluidic technology has been occasionally used for in vitro analysis and separation of cells. The small dimensions of microfluidic chips are very suitable to study cells on the single cell level rather than in whole populations. Also sperm cells have been studied and manipulated using microfluid

  1. Development of a simple and low cost electro ejaculation equipment in ram

    Directory of Open Access Journals (Sweden)

    Gutiérrez E

    2016-12-01

    Full Text Available The objective of the present study was to development of a simple and low cost electro ejaculation equipment in ram to semen collection. The work was carried out in the Alto Public University laboratories. The development of the equipment was implemented with an electronic circuit designed with source generating electrics pulses of frequency between 5 to 100 Hertz (hz and 30 to 80 milliamps (mA with pauses between 3 to 5 seconds with adjustable function. Likewise, an annular device was used and through a probe provided with electrodes circulates the electric energy in the animal rectum provoking muscle stimulation at the level of the accessory glands that trigger the ejaculation. Mass motility (0 to 5 3.8 ± 0.6, individually motility 70.7 ± 3.4%, sperm concentration 3.3 ± 0.2 billons and vitality 76.6 ± 6.9%. The cost of building the equipment was150 $ USD. In conclusion, we development a simple and low cost electro ejaculation equipment in ram, allowed to obtain samples of semen of creole sheep of good quality, without affecting the reproductive capacity of the animals in study.

  2. Successful low dose insemination of flow cytometrically sorted ram spermatozoa in sheep.

    Science.gov (United States)

    de Graaf, S P; Evans, G; Maxwell, W M C; Downing, J A; O'Brien, J K

    2007-12-01

    The fertility of ram spermatozoa that had undergone flow cytometric sorting (MoFlo SX) and cryopreservation was assessed after low-dose insemination of synchronized Merino ewes. Oestrus was synchronized with progestagen-impregnated pessaries, PMSG and GnRH treatment. Ewes (n = 360) were inseminated with 1 x 10(6), 5 x 10(6) or 15 x 10(6) motile sorted frozen-thawed (S(1), S(5), or S(15) respectively) or non-sorted frozen-thawed (C(1), C(5) or C(15) respectively) spermatozoa from three rams. An additional group of ewes were inseminated with 50 x 10(6) motile non-sorted frozen-thawed spermatozoa (C(50)) to provide a commercial dose control. The percentage of ewes lambing after insemination was similar for C(50) (24/38, 63.2%), C(15) (37/54, 68.5%), S(15) (38/57, 66.7%), S(5) (37/56, 66.1%) and S(1) (32/52, 61.5%) groups (p > 0.05), but lower for C(5) (19/48, 39.6%) and C(1) (19/55, 34.5%) treatments (p sheep as a reduction in the minimum effective sperm number will allow a corresponding decrease in the associated cost per dose.

  3. RAmM Algorithm(Simplex)

    CERN Document Server

    Manimala, Jose Mathew

    2010-01-01

    The evolution of encryption algorithms have led to the development of very complicated and highly versatile algorithms that sacrifice efficiency for better and harder to decrypt results. But by the application of a genetic schema to the encryption of data, a new structure can be created. Genetic methods and procedures are lethal in the way they handle and manipulate data. The RAmM algorithm uses four genetic operations that have been developed specifically for encryption of data. The operations are Replication, Augmentation, Mutation and Multiplication. The proper application of these methods according to the rules that have been found to be the best for getting optimal and correct results produces a "fingerprint" that is unique to a pair of . This means that every single data entry can only be decrypted by using the correct set of key. The application of the RAmM algorithm is in the field of image encryption and restoration. The boundary and the pixel values are separately encrypted to produce a very genuine...

  4. The rams horn in western history

    Science.gov (United States)

    Lubman, David

    2003-10-01

    The shofar or rams horn-one of the most ancient of surviving aerophones-may have originated with early Neolithic herders. The shofar is mentioned frequently and importantly in the Hebrew bible and in later biblical and post-biblical literature. Despite its long history, contemporary ritual uses, and profound symbolic significance to western religion, no documentation of shofar acoustical properties was found. Since ancient times, shepherds of many cultures have fashioned sound instruments from the horns of herd animals for practical and musical uses. Shepherd horns of other cultures exhibit an evolution of form and technology (e.g., the inclusion of finger holes). The shofar is unique in having retained its primitive form. It is suggested that after centuries of practical use, the shofar became emblematic of the shepherd culture. Ritual use then developed, which froze its form. A modern ritual rams horn played by an experienced blower was examined. This rather short horn was determined to have a source strength of 92 dB (A) at 1 m, a fundamental frequency near 420 Hz, and maximum power output between 1.2 and 1.8 kHz. Sample sounds and detection range estimates are provided.

  5. Wakes of ram pressure stripped disc galaxies

    CERN Document Server

    Roediger, E; Hoeft, M

    2006-01-01

    Spiral galaxies that move through the intracluster medium lose a substantial amount of their gas discs due to ram pressure stripping. The recent observations of NGC 4388 by Oosterloo & van Gorkom 2005 reveal a tail of stripped gas of ~ 100 kpc behind the source galaxy. We present first 3D hydrodynamical simulations of the evolution of such ram pressure stripped tails. We find that if the ICM wind does not vary significantly over a period of a few 100 Myr, subsonic galaxies produce a tail with regular features similar to a von-Karman vortex street. In this case, the tail widens systematically by about 45 kpc per 100 kpc distance behind the source galaxy. The widening rate is independent of the galaxy's inclination for a large range of inclinations. For supersonic galaxies, the tail is more irregular than for subsonic ones. The tail observed for NGC 4388 is narrower than the tails in our simulations. Reasons for this difference may be additional physical processes such as heat conduction or viscosity. In ad...

  6. The evolution of sperm morphometry in pheasants.

    Science.gov (United States)

    Immler, S; Saint-Jalme, M; Lesobre, L; Sorci, G; Roman, Y; Birkhead, T R

    2007-05-01

    Post-copulatory sexual selection is thought to be a potent evolutionary force driving the diversification of sperm shape and function across species. In birds, insemination and fertilization are separated in time and sperm storage increases the duration of sperm-female interaction and hence the opportunity for sperm competition and cryptic female choice. We performed a comparative study of 24 pheasant species (Phasianidae, Galliformes) to establish the relative importance of sperm competition and the duration of sperm storage for the evolution of sperm morphometry (i.e. size of different sperm traits). We found that sperm size traits were negatively associated with the duration of sperm storage but were independent of the risk of sperm competition estimated from relative testis mass. Our study emphasizes the importance of female reproductive biology for the evolution of sperm morphometry particularly in sperm-storing taxa.

  7. Slow oocyte freezing and thawing in couples with no sperm or an insufficient number of sperm on the day of in vitro fertilization

    Directory of Open Access Journals (Sweden)

    Tomazevic Tomaz

    2011-02-01

    Full Text Available Abstract Background The clinical results of in vitro fertilization of slowly frozen-thawed oocytes are known to be significantly worse than those obtained by fresh oocytes. Little is known about the factors affecting the clinical outcome of frozen-thawed oocytes. The aim of this retrospective study was to explore the role of oocyte cryopreservation in the group of patients with no available sperm on the day of in vitro fertilization. Additionally, the effects of the female serum FSH level and sperm quality on the clinical outcome of frozen-thawed oocytes were evaluated. Methods Oocytes were slowly frozen and thawed in 22 infertile couples with no sperm or insufficient number of sperm on the day of in vitro fertilization (IVF. In 9 couples with severe azoospermia or oligoasthenoteratozoospermia frozen-thawed oocytes were fertilized by autologous sperm of bad quality when available (Group 1. In 13 couples with non-ejaculation due to psychological stress on the day of classical IVF or severe azoospermia frozen-thawed oocytes were fertilized by autologous or donated sperm of normal quality (Group 2. Oocytes were thawed in 23 cycles and microinjected by the autologous or donated sperm, when available. The clinical outcome of intracytoplasmic sperm injection - ICSI (fertilization, blastocyst, and pregnancy rates was compared to the outcome of fresh oocytes of the same group of patients; additionally, the female serum FSH level and the sperm quality were compared. Results In all couples, 70.5% of oocytes survived the freeze-thaw procedure. After ICSI, 61.5% of thawed oocytes were fertilized. Twenty one% of embryos developed to the blastocyst stage. The pregnancy rates per embryo transfer and freeze-thaw cycle were 33.3% and 17.4%, respectively. All pregnancies ended in the birth of a baby without congenital anomalies. In patients with severe azoospermia or oligoasthenoteratozoospermia there was no statistically significant difference in pregnancy rates

  8. Sperm counts and sperm sex ratio in male infertility patients

    Institute of Scientific and Technical Information of China (English)

    Michael L Eisenberg; Lata Murthy; Kathleen Hwang; Dolores J Lamb; Larry I Lipshultz

    2012-01-01

    In recent years,investigators have noted a trend toward a declining proportion of male births in many industrialized nations.While men bear the sex-determining chromosome,the role of the female partner as it pertains to fertilization or miscarriage may also alter the gender ratio.We attempted to determine a man's secondary sex ratio (F1 generation) by directly examining the sex chromosomes of his sperm.We examined our male infertility clinic database for all men who had undergone a semen fluorescence in situ hybridization (FISH).Patient demographic and semen parameters were recorded.Chi-squared analysis was used to compare gender ratios (Ychromosomes/total chromosomes).Multivariable logistic regression was used to predict the odds of possessing a Y-bearing sperm after accounting for demographic and semen parameters.A total of 185 men underwent sperm FISH.For the entire cohort,the proportion of Y chromosome-bearing sperm was 51.5%.Men with less than five million motile sperm had a significantly lower proportion of Y chromosome-bearing sperm (50.8%) compared to men with higher sperm counts (51.6%; P=0.02).After multivariable adjustment,a higher sperm concentration,total motile sperm count and semen volume significantly increased the odds of having a Y chromosome-bearing sperm (P<0.01).As a man's sperm production declines,so does the proportion of Y chromosome-bearing sperm.Thus,a man's reproductive potential may predict his ability to sire male offspring.

  9. Morphology, Structure of Dimorphic Sperm, and Reproduction in the Hermaphroditic Commensal Bivalve Pseudopythina tsurumaru (Galeommatoidea: Kellidae)

    DEFF Research Database (Denmark)

    Lützen, Jørgen; Jespersen, Åse; Takahashi, Tohru

    2004-01-01

    Galeommatoide, commensal bivalve, reproduction, dimorphic sperm, sperm ultrastructure, spermatozeugma......Galeommatoide, commensal bivalve, reproduction, dimorphic sperm, sperm ultrastructure, spermatozeugma...

  10. Effects of extender type, sperm volume, cryoprotectant concentration, cryopreservation and time duration on motility, survival and fertilisation rates of Mekong giant catfish sperm

    Directory of Open Access Journals (Sweden)

    Kriangsak Mengumphan

    2010-10-01

    Full Text Available The objectives of this study are to evaluate the effects of some basic factors, namely extender type, sperm volume, cryoprotectant concentration, cryopreservation and storage time, on the quality of Mekong giant catfish (MGC sperm. The following results are obtained from conducted experiments. The sperm kept in Hanks balanced salt solution (HBSS extender consistently produced good results in terms of motility. The highest motility grade (4.0 was observed after 12 hours of examination and still a very satisfactory grade (3.3 was observed after 48 hours. The percentage of live cells of the sperm kept in HBSS was also highest (45.3%. The optimal amount of cryoprotectant (DMSO prior to cryopreservation was 8%, which gave the best motility grade (4.0 up to the first 72 hours of observation while at 120 hours the motility grade was 3.3. The fertilisation rate of MGC fresh sperm in HBSS (2 ml and 1 gram eggs was 47.1% while that of cryopreserved sperm under the same conditions was 36.2%. When crossed with P. hypophthalmus, the fertilisation rates of a 2-week- and a 1-year-cryopreserved sperm sample were 36.2% and 30.9% respectively.

  11. Turbulence of swarming sperm

    Science.gov (United States)

    Creppy, Adama; Praud, Olivier; Druart, Xavier; Kohnke, Philippa L.; Plouraboué, Franck

    2015-09-01

    Collective motion of self-sustained swarming flows has recently provided examples of small-scale turbulence arising where viscous effects are dominant. We report the first observation of universal enstrophy cascade in concentrated swarming sperm consistent with a body of evidence built from various independent measurements. We found a well-defined k-3 power-law decay of a velocity field power spectrum and relative dispersion of small beads consistent with theoretical predictions in 2D turbulence. Concentrated living sperm displays long-range, correlated whirlpool structures of a size that provides an integral scale of turbulence. We propose a consistent explanation for this quasi-2D turbulence based on self-structured laminated flow forced by steric interactions and alignment, a state of active matter that we call "swarming liquid crystal." We develop scaling arguments consistent with this interpretation.

  12. Design and verification of a self-timed RAM

    DEFF Research Database (Denmark)

    Nielsen, Lars Skovby; Staunstrup, Jørgen

    1995-01-01

    This paper describes a self-timed static RAM. A single bit RAM is described in the design language SYNCHRONIZED TRANSITIONS and using the verification tools supporting this language, it is shown that the design is speed-independent. Furthermore, a transistor level implementation of the design is ...

  13. Classification of handwritten digits using a RAM neural net architecture

    DEFF Research Database (Denmark)

    Jørgensen, T.M.

    1997-01-01

    Results are reported on the task of recognizing handwritten digits without any advanced pre-processing. The result are obtained using a RAM-based neural network, making use of small receptive fields. Furthermore, a technique that introduces negative weights into the RAM net is reported. The results...

  14. Cytometry of mammalian sperm

    Energy Technology Data Exchange (ETDEWEB)

    Gledhill, B.L.

    1983-10-11

    Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. The accessibility of male cells makes them well suited for analytical cytology. We might automate the process of determining sperm morphology but should not do so solely for increased speed. Rather, richer tangible benefits will derive from cytometric evaluation through increased sensitivity, reduced subjectivity, standardization between investigators and laboratories, enhanced archival systems, and the benefits of easily exchanged standardized data. Inroads on the standardization of assays for motility and functional integrity are being made. Flow cytometric analysis of total DNA content of individual sperm is an insensitive means to detect exposure to reproductive toxins because of the small size and low frequency of the DNA content errors. Flow cytometry can be applied to determine the proportions of X- and Y-sperm in semen samples.

  15. Sperm function test

    Directory of Open Access Journals (Sweden)

    Pankaj Talwar

    2015-01-01

    Full Text Available With absolute normal semen analysis parameters it may not be necessary to shift to specialized tests early but in cases with borderline parameters or with history of fertilization failure in past it becomes necessary to do a battery of tests to evaluate different parameters of spermatozoa. Various sperm function tests are proposed and endorsed by different researchers in addition to the routine evaluation of fertility. These tests detect function of a certain part of spermatozoon and give insight on the events in fertilization of the oocyte. The sperms need to get nutrition from the seminal plasma in the form of fructose and citrate (this can be assessed by fructose qualitative and quantitative estimation, citrate estimation. They should be protected from the bad effects of pus cells and reactive oxygen species (ROS (leukocyte detection test, ROS estimation. Their number should be in sufficient in terms of (count, structure normal to be able to fertilize eggs (semen morphology. Sperms should have intact and functioning membrane to survive harsh environment of vagina and uterine fluids (vitality and hypo-osmotic swelling test, should have good mitochondrial function to be able to provide energy (mitochondrial activity index test. They should also have satisfactory acrosome function to be able to burrow a hole in zona pellucida (acrosome intactness test, zona penetration test. Finally, they should have properly packed DNA in the nucleus to be able to transfer the male genes (nuclear chromatic decondensation test to the oocyte during fertilization.

  16. Feasibility of Integrated Insulation in Rammed Earth

    Science.gov (United States)

    Stone, C.; Balintova, M.; Holub, M.

    2015-11-01

    Building Codes in Europe stipulate strict thermal performance criteria which any traditional rammed earth recipe cannot meet. This does not infer that the material itself is inferior; it has many other face saving attributes such as low embodied energy, high workability, sound insulation, fire resistance, aesthetics, high diffusivity and thermal accumulation properties. Integrated insulation is experimented with, to try achieve a 0.22 [W/(m2.K)] overall coefficient of heat transfer for walls required by 2015 Slovak standards, without using external insulation or using technologically complex interstitial insulation. This has the added aesthetic benefit of leaving the earth wall exposed to the external environment. Results evaluate the feasibility of this traditional approach.

  17. Ramón Cote Baraibar

    Directory of Open Access Journals (Sweden)

    Juan Gustavo Cobo Borda

    2005-05-01

    Full Text Available Hijo de poeta, como María Mercedes Carranza y Santiago Mutis, Ramón Cote nació en Cúcuta el 19 de mayo de 1963- Su primer libro fue publicado en 1984 por ediciones Arnao, de Madrid, donde se graduó en historia del arte en la Universidad Complutense. Al instalarse en Colombia, se dedicó a la publicidad. Ha publicado ensayos sobre poesía colombiana en revistas como ínsula, y en 1992, con el título de  |Diez de ultramar publicó una muestra de joven poesía latinoamericana que abarcaba nombres como José Luis Rivas, Coral Bracho, Raúl Zurita, Fabio Morabito, Yolanda Pantin y Eduardo Chirinos. Entre la nueva poesía colombiana, su voz es una de las más reconocibles. Vale la pena, entonces, repasar la totalidad de su trayectoria.

  18. Sperm Shape (Morphology): Does It Affect Fertility?

    Science.gov (United States)

    ... Website of the American Society for Reproductive Medicine Sperm morphology (shape): Does it affect fertility? How is ... semen analysis. This measures many features of the sperm and semen (the fluid in which the sperm ...

  19. Fire effects in Pinus uncinata Ram plantations

    Directory of Open Access Journals (Sweden)

    Adrián Cardil Forradellas

    2016-04-01

    Full Text Available Aim of study: Understanding fire ecology of main forest species is essential for a sound, scientifically based on managing of wildlands and also to assess likely implications due to changes in fire regime under a global change scenario. Few references can be found about fire ecology of Pinus uncinata Ram. (PU. PU species grows in the Central Pyrenees where large, severe wildland fires did not occur frequently in the past. However, several fires with extreme fire behavior have affected PU stands in last years and they might disturb other PU forest in the future.Area of study: Cabdella fire (February 2012, in Lleida province, is one of the several wildland fires occurred in 2012 (winter season in the Central Pyrenees. Fire affected a large PU plantation (102 ha located at 1.800-2,100 meters above the sea.Material and methods: We have analyzed first order fire effects in three fireline intensity thresholds along three years in terms of mortality ratio, scorched height, percentage of scorched crown volume and bark char height.Main results: PU seems to be a very tolerant species to low and medium fire line intensity but fire effects were very significant when fire line intensity was high. In medium fireline intensity sites, probability of mortality ranged from 15 to 30% and the dead trees had the highest values on scorched height and percentage of scorched crown volume.Research highlights: Results from this work supports that prescribed burning might be used to efficiently decrease fuel load and fuel vertical continuity while avoiding considerable PU mortality. It also displayed that when fuel management has been implemented, PU mortality might be limited even under extreme fire behavior.Abbreviations used: PU: Pinus uncinata Ram.

  20. Baryogensis in fresh inflation

    CERN Document Server

    Bellini, M

    2002-01-01

    I study the possibility of baryogenesis can take place in fresh inflation. I find that it is possible that violation of baryon number conservation can occur during the period out-of-equilibrium in this scenario. Indeed, baryogenesis could be possible before the thermal equilibrium is restored at the end of fresh inflation.

  1. Mecobalamin promotes mouse sperm maturation.

    Science.gov (United States)

    Oshio, S; Ozaki, S; Ohkawa, I; Tajima, T; Kaneko, S; Mohri, H

    1989-01-01

    The effect of Mecobalamin (alpha-(5,6-dimethyl benzimidazolyl)-Co-methyl-cobamide: Me-B 12) on sperm production in the oligozoospermic mice experimentally induced by the treatment with adriamycin (0.3 mg/kg, three times a week for 5 weeks) was evaluated quantitatively by means of equilibrium sedimentation in Percoll. After centrifugation, the distribution profile of the sperm showed two peaks, i.e. the first peak near the bottom consisting of mature sperm with good motility and the second peak containing immature and/or immotile sperm. By oral administration of Me B 12 (1.0 mg/kg/day) to the oligozoospermic mice for 10 weeks, the sperm count, sperm motility, motile sperm count, diameter of seminiferous tubules and the percentage of good motile sperm with higher apparent density were increased as compared with those of the control. These results suggest that Me-B 12 enhanced the testicular function, resulting in an increased output of mature sperm.

  2. Sperm storage in caecilian amphibians

    Directory of Open Access Journals (Sweden)

    Kuehnel Susanne

    2012-06-01

    Full Text Available Abstract Background Female sperm storage has evolved independently multiple times among vertebrates to control reproduction in response to the environment. In internally fertilising amphibians, female salamanders store sperm in cloacal spermathecae, whereas among anurans sperm storage in oviducts is known only in tailed frogs. Facilitated through extensive field sampling following historical observations we tested for sperm storing structures in the female urogenital tract of fossorial, tropical caecilian amphibians. Findings In the oviparous Ichthyophis cf. kohtaoensis, aggregated sperm were present in a distinct region of the posterior oviduct but not in the cloaca in six out of seven vitellogenic females prior to oviposition. Spermatozoa were found most abundantly between the mucosal folds. In relation to the reproductive status decreased amounts of sperm were present in gravid females compared to pre-ovulatory females. Sperm were absent in females past oviposition. Conclusions Our findings indicate short-term oviductal sperm storage in the oviparous Ichthyophis cf. kohtaoensis. We assume that in female caecilians exhibiting high levels of parental investment sperm storage has evolved in order to optimally coordinate reproductive events and to increase fitness.

  3. Postcoital Sperm Assessment Comparative Study.

    Science.gov (United States)

    Pelekanos, Michael J

    2015-11-01

    This postcoital sperm assessment study was performed over a 10 month time period (November 2014-August 2015). Fifteen couples enrolled in the study. The study was a non-blinded, non-randomized, single-center comparison study comparing The Stork® OTC (Rinovum Women's Health, Monroeville, PA) to natural intercourse (NI), using the subjects as their own control/baseline. This was an efficacy study designed to compare the number of sperm in the cervical mucus following the use of The Stork OTC conception aid with the number of sperm in the cervical mucus following natural intercourse. Subjects used both The Stork OTC conception system and the natural intercourse method to evaluate concentrations of sperm in the cervical mucus. Post-coital test (PCT) data was collected demonstrating higher concentrations of sperm within the cervical mucus with The Stork OTC conception system versus natural intercourse for 85% of test subjects in this study. Of the 15 couples enrolled in the study, 2 were lost to follow-up. Mean age for male subjects was 31.7 +/ 5.4 years of age and mean age for female subjects was 29.7+/- 5.4. The average sperm score value of the 85% of test subjects with higher sperm concentrations from The Stork OTC was 3.23 times the score value of sperm concentration compared to natural intercourse. The remaining 15% of test subjects showed no change in sperm score value between The Stork OTC and natural intercourse.

  4. Characterization of alkaline phosphatase activity in seminal plasma and in fresh and frozen-thawed stallion spermatozoa.

    Science.gov (United States)

    Bucci, Diego; Giaretta, Elisa; Spinaci, Marcella; Rizzato, Giovanni; Isani, Gloria; Mislei, Beatrice; Mari, Gaetano; Tamanini, Carlo; Galeati, Giovanna

    2016-01-15

    Alkaline phosphatase (AP) has been studied in several situations to elucidate its role in reproductive biology of the male from different mammalian species; at present, its role in horse sperm physiology is not clear. The aim of the present work was to measure AP activity in seminal plasma and sperm extracts from freshly ejaculated as well as in frozen-thawed stallion spermatozoa and to verify whether relationship exists between AP activity and sperm quality parameters. Our data on 40 freshly ejaculated samples from 10 different stallions demonstrate that the main source of AP activity is seminal plasma, whereas sperm extracts contribution is very low. In addition, we found that AP activity at physiological pH (7.0) is significantly lower than that observed at pH 8.0, including the optimal AP pH (pH 10.0). Alkaline phosphatase did not exert any effect on sperm-oocyte interaction assessed by heterologous oocyte binding assay. Additionally, we observed a thermal stability of seminal plasma AP, concluding that it is similar to that of bone isoforms. Positive correlations were found between seminal plasma AP activity and sperm concentration, whereas a negative correlation was present between both spermatozoa extracts and seminal plasma AP activity and seminal plasma protein content. A significant decrease in sperm extract AP activity was found in frozen-thawed samples compared with freshly ejaculated ones (n = 21), concomitantly with the decrease in sperm quality parameters. The positive correlation between seminal plasma AP activity measured at pH 10 and viability of frozen-thawed spermatozoa suggests that seminal plasma AP activity could be used as an additional predictive parameter for stallion sperm freezability. In conclusion, we provide some insights into AP activity in both seminal plasma and sperm extracts and describe a decrease in AP after freezing and thawing.

  5. The effect of chilled storage and cryopreservation on the sperm DNA fragmentation dynamics of a captive population of koalas.

    Science.gov (United States)

    Johnston, Stephen D; Zee, Yeng Peng; López-Fernández, Carmen; Gosálvez, Jamie

    2012-01-01

    This experiment documented the incidence and variability of sperm characteristics found in freshly collected and ex vivo-manipulated semen samples from a population of disease-free captive koalas with a special emphasis on the dynamic aspects of DNA fragmentation. These changes were analyzed in light of the putative negative effect of iatrogenic damage after chilled storage and cryopreservation with respect to different semen extender compositions to maximize sperm longevity. Sperm DNA fragmentation (SDF) dynamics (SDF assessment after a varying period of time) was investigated with the sperm chromatin dispersion assay after either dilution in tris-citrate media and chilled preservation at 4°C for upward of 16 days or cryopreservation in either glycerol or dimethylacetamide (DMA) tris-citrate-based cryoprotectant media; corresponding data on progressive sperm motility, plasma membrane integrity, and the proportion of koala sperm with relaxed chromatin were also recorded. SDF analysis of the captive koala population revealed a low mean (± SEM) basal level of only 6.7% ± 0.9%. The percentage of progressive sperm motility, percentage of intact sperm plasma membranes, and the percentage of relaxed chromatin did not correlate significantly with that of basal SDF. Moreover, despite the absence of cysteine residues in marsupial protamines, koala spermatozoa showed remarkable stability in terms of their DNA integrity after the incubation of either fresh, chilled, or frozen-thawed semen samples; observations of progressive motility (P koala spermatozoa declined after 4 days, whereas the incidence of relaxed chromatin increased after 8 days. Although koala SDF increased significantly (P koala sperm chromatin tended to decondense; however, the incidence of sperm DNA fragmentation was not correlated with the incidence of sperm chromatin relaxation after glycerol (R = .2) or DMA (R = -.04) cryopreservation.

  6. Functional integrity of Colossoma macropomum (Cuvier, 1816 sperm cryopreserved with enriched extender solutions

    Directory of Open Access Journals (Sweden)

    Raycon Roberto Freitas Garcia

    Full Text Available Cryoprotectant solutions are used to protect the sperm from alterations caused by the low temperature in the cryopreservation process. We evaluated the quality of Colossoma macropomum semen after freezing, using dimethyl sulfoxide (DMSO as a cryoprotectant, combined with two extender solutions (T1 - Solution 1: Glucose 90.0 g/L, Sodium Citrate 6.0 g/L, EDTA 1.5 g/L, Sodium Bicarbonate 1.5 g/L, Potassium Chloride 0.8 g/L, Gentamycin Sulphate 0.2 g/L, and T2 - Solution 2: Glucose 90.0 g/L, ACP(r-104 10.0 g/L. Motility rate and motility time did not differ between T1 and T2 and were lower than fresh semen. The number of normal sperm was significantly different in treatments T1 (15.1% and T2 (21.9%, and both showed a reduction in the percentage of normal sperm compared to fresh semen (57.4%. The values found for the rates of fertilization and hatching, mitochondrial functionality and sperm DNA, did not differ between the treatments (T1 and T2. Regarding membrane integrity, there was a higher percentage of spermatozoa with intact membranes in T1 (53.4% than T2 (43.7%. The extender solutions, combined with 10% DMSO, maintained the sperm DNA intact in almost all the C. macropomum sperm cells, however there was a loss in their functionality.

  7. Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrity.

    Science.gov (United States)

    Rahiminia, Tahereh; Hosseini, Akram; Anvari, Morteza; Ghasemi-Esmailabad, Saeed; Talebi, Ali Reza

    2017-08-01

    Presence of vitrification method in sperm freezing and the introduction of solid surface vitrification beside rapid freezing in vapour, opens an easy and safe way to help infertility centres. While the effects of cryopreservation on motility, morphology and viability of sperm are documented, the question of the probable alteration of sperm DNA, chromatin and acrosome integrity after freezing and thawing procedures in different methods is still controversial. Normal sample were collected according to WHO strict criteria. Sperm suspensions were mixed 1:1 with 0.5 M sucrose and divided into four equal aliquots for freezing: fresh, nitrogen direct immersion vitrification (Vit), solid surface vitrification (SSV) and in vapour (Vapour). Sperm suspensions were transferred into a 0.25 ml sterile plastic. Then straw was inserted inside the 0.5 ml straw. For thawing, the straws were immersed in a 42 °C water bath. Beside the sperm parameters, we assessed the acrosome reaction by double staining, chromatin integrity by toluidine blue (Tb) and chromomycin A3 (CMA3) and DNA integrity by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) respectively. In progressive motility, the highest rate occurred in Vit (39.9 ± 13.3). Moreover, the lowest rate of immotile sperm was in Vit (32.7 ± 16.3). In normal morphology, the group Vit was similar to the fresh, while SSV and Vapour were significantly different from the fresh. The percentage of acrosome-reacted sperms was more in Vit (81.3 ± 10.2) than the fresh group. TUNEL+ results showed that DNA fragmentation was significantly increased in Vit (p-value = 0.025). While in SSV and Vapour results were comparable to fresh. There was a significant correlation between TUNEL+ and normal morphology, TB, CMA3 and presence of intact acrosome. Sperm in Vapour was healthier in terms of DNA, chromatin and acrosome integrity. In contrast of higher motility and normal morphology; DNA, chromatin and acrosome

  8. In vitro fertilization/intracytoplasmic sperm injection for male infertility

    Directory of Open Access Journals (Sweden)

    Rubina Merchant

    2011-01-01

    Full Text Available Progress in the field of assisted reproduction, and particularly micromanipulation, now heralds a new era in the management of severe male factor infertility, not amenable to medical or surgical correction. By overcoming natural barriers to conception, in vitro fertilization and embryo transfer (IVF-ET, subzonal sperm insemination, partial zona dissection, and intracytoplasmatic injection of sperm (ICSI now offer couples considered irreversibly infertile, the option of parenting a genetically related child. However, unlike IVF, which necessitates an optimal sperm number and function to successfully complete the sequence of events leading to fertilization, micromanipulation techniques, such as ICSI, involving the direct injection of a spermatozoon into the oocyte, obviate all these requirements and may be used to alleviate severe male factor infertility due to the lack of sperm in the ejaculate due to severely impaired spermatogenesis (non-obstructive azoospermia or non-reconstructable reproductive tract obstruction (obstructive azoospermia. ICSI may be performed with fresh or cryopreserved ejaculate sperm where available, microsurgically extracted epididymal or testicular sperm with satisfactory fertilization, clinical pregnancy, and ongoing pregnancy rates. However, despite a lack of consensus regarding the genetic implications of ICSI or the application and efficacy of preimplantation genetic diagnosis prior to assisted reproductive technology (ART, the widespread use of ICSI, increasing evidence of the involvement of genetic factors in male infertility and the potential risk of transmission of genetic disorders to the offspring, generate major concerns with regard to the safety of the technique, necessitating a thorough genetic evaluation of the couple, classification of infertility and adequate counseling of the implications and associated risks prior to embarking on the procedure. The objective of this review is to highlight the indications

  9. In vitro fertilization/intracytoplasmic sperm injection for male infertility.

    Science.gov (United States)

    Merchant, Rubina; Gandhi, Goral; Allahbadia, Gautam N

    2011-01-01

    Progress in the field of assisted reproduction, and particularly micromanipulation, now heralds a new era in the management of severe male factor infertility, not amenable to medical or surgical correction. By overcoming natural barriers to conception, in vitro fertilization and embryo transfer (IVF-ET), subzonal sperm insemination, partial zona dissection, and intracytoplasmatic injection of sperm (ICSI) now offer couples considered irreversibly infertile, the option of parenting a genetically related child. However, unlike IVF, which necessitates an optimal sperm number and function to successfully complete the sequence of events leading to fertilization, micromanipulation techniques, such as ICSI, involving the direct injection of a spermatozoon into the oocyte, obviate all these requirements and may be used to alleviate severe male factor infertility due to the lack of sperm in the ejaculate due to severely impaired spermatogenesis (non-obstructive azoospermia) or non-reconstructable reproductive tract obstruction (obstructive azoospermia). ICSI may be performed with fresh or cryopreserved ejaculate sperm where available, microsurgically extracted epididymal or testicular sperm with satisfactory fertilization, clinical pregnancy, and ongoing pregnancy rates. However, despite a lack of consensus regarding the genetic implications of ICSI or the application and efficacy of preimplantation genetic diagnosis prior to assisted reproductive technology (ART), the widespread use of ICSI, increasing evidence of the involvement of genetic factors in male infertility and the potential risk of transmission of genetic disorders to the offspring, generate major concerns with regard to the safety of the technique, necessitating a thorough genetic evaluation of the couple, classification of infertility and adequate counseling of the implications and associated risks prior to embarking on the procedure. The objective of this review is to highlight the indications, advantages

  10. Cu/Zn-superoxide dismutase from the fungal strain Humicola lutea 103 improves ram spermatozoa functions in vitro.

    Science.gov (United States)

    Stefanov, R; Angelova, M; Stefanova, T; Subev, M; Dolashka, P; Voelter, W; Zachariev, Z

    2004-04-01

    In this study we determined the effect of reactive oxygen species (ROS) generation during incubation in media at 39 degrees C on ram spermatozoa and the protection by exogenously added antioxidant enzyme, superoxide dismutase (SOD). A novel Cu/Zn-SOD, isolated from the fungal strain Humicola lutea 103 (HLSOD), was used. Our results point out that the levels of both, superoxide anion radicals (*O2-) and H2O2, increase approximately 8-10- and 2-3-fold, respectively, during incubation of spermatozoa. Enhanced ROS generation coincided with reduction of motility, independently of the type of diluted medium. Addition of HLSOD (30, 60 and 120 U ml(-1) sperm) improved sperm functions, maintaining almost initial percentages of motile spermatozoa and increasing the values of mean cytochemical coefficient. At the same time, a significant diminution of *O2- and H2O2 content in the presence of antioxidant enzyme was established. The results suggest that HLSOD is an effective *O2- scavenger in semen that leads to protection of sperm functions.

  11. Sperm donation in Israel.

    Science.gov (United States)

    Mor-Yosef, S; Schenker, J G

    1995-04-01

    Science and technology in the field of human reproduction present new legal, ethical and religious questions which do not always have immediate answers. The first step in the rapidly developed field of reproductive technology was the use of sperm donation (artificial insemination by donor, AID) and the establishment of sperm banks. The state of Israel faced these problems when the regulations for sperm donation were discussed. The fact that the main holy places for the three monotheistic religions are in Israel directly influences the make-up of the population constituents. Therefore, besides a majority of secular people, a high percentage of the population of Israel is very religious: Jews, Moslems and Christians. Thus any resolution relating to AID should take this demographic combination into account. The practice of AID is opposed by the different monotheistic religions. To avoid the conflict between secular and religious people, and between the different religions' perspectives, the legal problem of AID in Israel was solved not by laws but by regulations which were published by the Ministry of Health. The main idea behind this attitude is that the state and its authorities should not and do not deal with ethical or religious questions. Thus, the decision was left to the couples and to the donors. The regulations address technical requirements, health problems and confidential issues concerning the couple, the donor and the child. In this paper we present the different views relating to these problems as perceived by the different religions, and describe the solution that was accepted by the Israeli Ministry of Health.

  12. Sperm preparation for ART

    Directory of Open Access Journals (Sweden)

    Schill Wolf-Bernhard

    2003-11-01

    Full Text Available Abstract The onset of clinical assisted reproduction, a quarter of a century ago, required the isolation of motile spermatozoa. As the indication of assisted reproduction shifted from mere gynaecological indications to andrological indications during the years, this urged andrological research to understand the physiology of male germ cell better and develop more sophisticated techniques to separate functional spermatozoa from those that are immotile, have poor morphology or are not capable to fertilize oocytes. Initially, starting from simple washing of spermatozoa, separation techniques, based on different principles like migration, filtration or density gradient centrifugation evolved. The most simple and cheapest is the conventional swim-up procedure. A more sophisticated and most gentle migration method is migration-sedimentation. However, its yield is relatively small and the technique is therefore normally only limited to ejaculates with a high number of motile spermatozoa. Recently, however, the method was also successfully used to isolate spermatozoa for intracytoplasmic sperm injection (ICSI. Sperm separation methods that yield a higher number of motile spermatozoa are glass wool filtration or density gradient centrifugation with different media. Since Percoll® as a density medium was removed from the market in 1996 for clinical use in the human because of its risk of contamination with endotoxins, other media like IxaPrep®, Nycodenz, SilSelect®, PureSperm® or Isolate® were developed in order to replace Percoll®. Today, an array of different methods is available and the selection depends on the quality of the ejaculates, which also includes production of reactive oxygen species (ROS by spermatozoa and leukocytes. Ejaculates with ROS production should not be separated by means of conventional swim-up, as this can severely damage the spermatozoa. In order to protect the male germ cells from the influence of ROS and to stimulate

  13. RNA in human sperm

    Institute of Scientific and Technical Information of China (English)

    Rui Pires Martins; Stephen A. Krawetz

    2005-01-01

    We have yet to develop a fundamental understanding of the molecular complexities of human spermatozoa. This encompasses the unique packaging and structure of the sperm genome along with their paternally derived RNAs in preparation for their delivery to the egg. The diversity of these transcripts is vast, including several anti-sense molecules resembling known regulatory micro-RNAs. The field is still grasping with its delivery to the oocyte at fertilization and possible significance. It remains tempting to analogize them to maternally-derived transcripts active in early embryo patterning. Irrespective of their role in the embryo, their use as a means to assess male factor infertility is promising.

  14. Effects of the Czech Propolis on Sperm Mitochondrial Function

    Directory of Open Access Journals (Sweden)

    Miroslava Cedikova

    2014-01-01

    Full Text Available Propolis is a natural product that honeybees collect from various plants. It is known for its beneficial pharmacological effects. The aim of our study was to evaluate the impact of propolis on human sperm motility, mitochondrial respiratory activity, and membrane potential. Semen samples from 10 normozoospermic donors were processed according to the World Health Organization criteria. Propolis effects on the sperm motility and mitochondrial activity parameters were tested in the fresh ejaculate and purified spermatozoa. Propolis preserved progressive motility of spermatozoa in the native semen samples. Oxygen consumption determined in purified permeabilized spermatozoa by high-resolution respirometry in the presence of adenosine diphosphate and substrates of complex I and complex II (state OXPHOSI+II was significantly increased in the propolis-treated samples. Propolis also increased uncoupled respiration in the presence of rotenone (state ETSII and complex IV activity, but it did not influence state LEAK induced by oligomycin. Mitochondrial membrane potential was not affected by propolis. This study demonstrates that propolis maintains sperm motility in the native ejaculates and increases activities of mitochondrial respiratory complexes II and IV without affecting mitochondrial membrane potential. The data suggest that propolis improves the total mitochondrial respiratory efficiency in the human spermatozoa in vitro thereby having potential to improve sperm motility.

  15. Laboratory handling of epididymal and testicular spermatozoa: What can be done to improve sperm injections outcome

    Directory of Open Access Journals (Sweden)

    Sandro C Esteves

    2012-01-01

    Full Text Available Spermatozoa from azoospermic males can be retrieved from either the epididymis or the testis, depending on the type of azoospermia, using different surgical methods such as percutaneous epididymal sperm aspiration (PESA, testicular sperm aspiration (TESA, testicular sperm extraction (TESE, and microsurgical testicular sperm extraction (micro- TESE. After collecting the epididymal fluid or testicular tissue, laboratory techniques are used to remove contaminants, cellular debris, noxious microorganisms, and red blood cells. Processed spermatozoa may be used for intracytoplasmic sperm injection or eventually be cryopreserved. However, spermatozoa collected from either the epididymis or the testis are often compromised and more fragile than ejaculated ones. Therefore, sperm processing techniques should be used with great caution to avoid jeopardizing the sperm fertilizing potential in treatment cycles. In this review, we describe the current methods for processing surgically-retrieved specimens, either fresh or frozen- thawed, and provide the tips and pitfalls for facilitating the handling of such specimens. In addition, we present the available laboratory tools to aid in the identification of viable immotile spermatozoa to be used in conjunction with assisted reproductive techniques. Review of the literature was carried out using PubMed and Science Direct search engines.

  16. Effect of 655 nm laser different powers on dog sperm motility parameters

    Science.gov (United States)

    Corral-Baqués, M. I.; Rigau, T.; Rivera, M. M.; Rodríguez-Gil, J. E.; Rigau, J.

    2006-04-01

    Introduction: One of the most appreciated features of the sperm is its motility, which depends on a big energy consumption despite differences among species. Laser acts direct or indirectly on mitochondria increasing ATP production. Material and method: By means of a Computer Aided Sperm Analysis (CASA) we have studied the effects of a 655 nm continuous wave diode laser irradiation at different power outputs with a dose of 3.3418 J on sperm motility. After an eosine-nigrosine stain to establish its quality, the second fraction of fresh beagle dog sperm was divided into 5 groups, 1 control and four to be irradiated respectively with an average output power of 6.84 mW, 15.43 mW, 33.05 mW and 49.66 mW. At times 0 and 45 minutes from irradiation pictures were taken and analysed with the Sperm class Analyzer SCA2002 programme. The motility parameters of 4987 spermatozoa studied were: curvilinear velocity (VCL), progressive velocity (VSL), straightness (STR), wobble (WOB), average path velocity (VAP), linearity (LIN), mean amplitude of lateral head displacement (ALHmed), beat cross frequency (BCF) and the total motility (MT). At time 15 minutes after irradiation a hypoosmotic swelling test (HOST) was done. Results: Several motility parameters that affect the overall motile sperm subpopulation structure have been changed by different output powers of a 655 nm diode laser irradiation, and prevents the decrease of the sperm motility properties along time.

  17. Orthogonal Range Searching on the RAM, Revisited

    CERN Document Server

    Chan, Timothy M; Patrascu, Mihai

    2011-01-01

    We present several new results on one of the most extensively studied topics in computational geometry, orthogonal range searching. All our results are in the standard word RAM model for points in rank space: ** We present two data structures for 2-d orthogonal range emptiness. The first achieves O(n lglg n) space and O(lglg n) query time. This improves the previous results by Alstrup, Brodal, and Rauhe(FOCS'00), with O(n lg^eps n) space and O(lglg n) query time, or with O(nlglg n) space and O(lg^2 lg n) query time. Our second data structure uses O(n) space and answers queries in O(lg^eps n) time. The best previous O(n)-space data structure, due to Nekrich (WADS'07), answers queries in O(lg n/lglg n) time. ** For 3-d orthogonal range reporting, we obtain space O(n lg^{1+eps} n) and query time O(lglg n + k), for any constant eps>0. This improves previous results by Afshani (ESA'08), Karpinski and Nekrich (COCOON'09), and Chan (SODA'11), with O(n lg^3 n) space and O(lglg n + k) query time, or with O(n lg^{1+eps...

  18. Detonation Type Ram Accelerator: A Computational Investigation

    Directory of Open Access Journals (Sweden)

    Sunil Bhat

    2000-01-01

    Full Text Available An analytical model explaining the functional characteristics of detonation type ram accelerator is presented. Major flow processes, namely, (i supersonic flow over the cone of the projectile, (ii initiation ofconical shock wave and its reflection from the tube wall, (iii supersonic combustion, and (iv expansion wave and its reflection are modelled. Taylor-Maccoll approach is adopted for modellingthe flow over the cone of the projectile. Shock reflection is treated in accordance with wave angle theorytor flows over the wedge. Prandtl-Mayer analysis is used to model the expansion wave and its reflection.Steady one-dimensional flow with heat transfer along with Rayleigh line equation for perfect gases isused to model supersonic combustion. A computer code is developed to compute the thrust producedby combustion of gases. Ballistic parameters like thrust-pressure ratio and ballistic efficiency of the accelerator are evaluated and their maximum values are 0.032 and 0.068, respectively. The code indicates possibility ofachieving high velocity of 7 km/s on utilising gaseous mixture of 2H2+O2 in the operation.Velocity range suitable for operation of the accelerator lies between 3.8 - 7.0 km/s. Maximum thrust valueis 33721 N which corresponds to the projectile velocity of 5 km/s.

  19. A diphtheria outbreak in Buri Ram, Thailand.

    Science.gov (United States)

    Pantukosit, Pantavee; Arpornsuwan, Manote; Sookananta, Kanokporn

    2008-07-01

    In May 1996 there was an outbreak of diphtheria in Buri Ram, Thailand which infected 31 patients, 8 males and 23 females. The mean age of the patients was 8 +/- 5 years. Seventy-four percent had a history of childhood vaccinations. Common signs and symptoms included fever (100%) which was low grade in 61%, sore throat (90%), upper airway obstruction (3%), and hoarseness (10%). Pseudomembranes (seen in 100%) were located on the tonsils (71%), pharynx (22%), larynx (9.6%), and uvula (6%). The mean duration of symptoms prior to admission was 2 days with a range of 1 to 5 days. Complications included upper airway obstruction (10%) and cardiac complications (10%). There were no neurological complication or deaths. There were negative associations between cardiac complications, severity of disease and previous diphtheria vaccination. The ages varied from children to adults. Early recognition and prompt treatment decreased complications and mortality in this group of patients when compared with Chiang Mai and Queen Sirikit National Institute of Child Health (QSNICH) studies.

  20. Sperm membrane modulation by Sapindus mukorossi during sperm maturation

    Institute of Scientific and Technical Information of China (English)

    ManishNivsarkar; NeetaShrivastava

    2002-01-01

    Aim:TO observe the alterations in the biochemical and biophysical changes in the sperm membrane during sperm maturation in male rats treated with the water extract of the fruit pericarp of S.mukorossi.Methods:Adult male Sprague-Dawley rats were gavaged the aqueous extract of the fruit pericarp of S.mukorossi at a dose of 50mg/kg/dfor45days.On day46,the sperm parameters were observed in idfferent sections of the epididymis and the sperm superoxide dismutase and the lipid peroxidation was edtermined and compared with the controls.The testis and epidiymis were routinely prepared for histological examination under the light microscope.Results:No significant differences in the sperm number and morphology were observed between the control and reated groups.However,a significant inhibition(P<0.05-0.01)of sperm motility in the caput,corpus and cauda regions of the epididymis was seen in the treated group.No significant histopathological changes were found in the testis and epididymis.Theimportant finding was that in the treated animals,the spermatozoa showed an abnormal distribution of the superoxide dismutase activity,being minimum in the caput and maximum in the corpus,which was just opposite to that of the controls.Conclusion:The study provides a unique observation where the plant extract alters the sperm membrane physiology without change the testicular and epididymal morphology.

  1. Evaluation of sperm head shape at high magnification revealed correlation of sperm DNA fragmentation with aberrant head ellipticity and angularity.

    Science.gov (United States)

    Utsuno, Hiroki; Oka, Kenji; Yamamoto, Ayako; Shiozawa, Tanri

    2013-05-01

    To test for an association between DNA fragmentation and head shape at high magnification in fresh motile spermatozoa. Observational study. Academic tertiary care center. A total of 60 men in our assisted reproductive program. Quantifying sperm head shape using elliptic Fourier analysis, and detecting DNA fragmentation by use of a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Correlation between percentage of spermatozoa with abnormal head shape and percentage of DNA fragmentation. Elliptic Fourier analysis decomposed sperm head shapes into four quantitative parameters: ellipticity, anteroposterior (AP) symmetry, lateral symmetry, and angularity. The DNA fragmentation was statistically significantly correlated with abnormal angularity, and moderately with abnormal ellipticity but not with abnormal AP symmetry or lateral symmetry. Forward stepwise multiple logistic regression analysis revealed a statistically significantly higher percentage of DNA fragmentation in spermatozoa with abnormal ellipticity and abnormal angularity than in spermatozoa with normal-shaped head (6.1% and 5.4% vs. 2.8%). Spermatozoa with large nuclear vacuoles also correlated with sperm DNA fragmentation, and had a statistically significantly higher percentage of DNA fragmentation (4.7%). Among the morphologic features of the sperm head, abnormal ellipticity, angularity, and large nuclear vacuoles are associated with DNA fragmentation. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  2. Scheme of a WDM Time-Limited RAM for OPS

    Institute of Scientific and Technical Information of China (English)

    Shuguang Zhu; Anshi Xu; Hongxi Yin; Deming Wu; Xuzong Chen

    2003-01-01

    We propose a new idea of utilizing Multiple Electromagnetically Induced Transparency Concurrence within a Single Medium to fullfill optical packet buffering and theoretically design a Wavelength Division Multiplexing (WDM) Random Access Memory (RAM).

  3. Computational Examination of Parameters Influencing Practicability of Ram Accelerator

    Directory of Open Access Journals (Sweden)

    Sunil Bhat

    2004-07-01

    Full Text Available The problems concerning practicability aspects of a ram accelerator, such as intense in-bore projectile ablation, large accelerator tube length to achieve high projectile muzzle velocity, and high entry velocity of projectile in the accelerator tube for starting the accelerator have been examined. Computational models of the processes like phenomenon of projectile ablation, flow in the aero-window used as accelerator tube-end closure device in case of high drive gas filling pressure in the ram accelerator tube have been presented. New projectile design to minimise the starting velocity of the ram accelerator is discussed. Possibility of deployment of ram accelerator in the defence-oriented role has been investigated to utilise its high velocity potential.

  4. Ram pressure profiles in galaxy groups and clusters

    CERN Document Server

    Tecce, Tomás E; Tissera, Patricia B

    2011-01-01

    Using a hybrid method which combines non-radiative hydrodynamical simulations with a semi-analytic model of galaxy formation, we determine the ram pressure as a function of halocentric distance experienced by galaxies in haloes with virial masses 12.5 <= log (M_200 h/M_Sun) < 15.35, for redshifts 0 <= z <= 3. The ram pressure is calculated with a self-consistent method which uses the simulation gas particles to obtain the properties of the intergalactic medium. The ram pressure profiles obtained can be well described by beta profile models, with parameters that depend on redshift and halo virial mass in a simple fashion. The fitting formulae provided here will prove useful to include ram pressure effects into semi-analytic models based on methods which lack gas physics, such as dark matter-only simulations or the Press-Schechter formalism.

  5. Genetic parameters of testicular measurements in Merino rams and ...

    African Journals Online (AJOL)

    NARGA

    Genetic parameters of testicular measurements in Merino rams and the influence of scrotal ... firmly and evenly to the bottom of the scrotum until ventral skin folds were eliminated. The testes were then ..... Ph.D. (Agric) treatise. University of.

  6. Human Sperm Competition: A Comparative Evolutionary Analysis

    Directory of Open Access Journals (Sweden)

    Michael N. Pham

    2014-08-01

    Full Text Available Sperm competition occurs when a female copulates with two or more males within a sufficiently brief time period, resulting in sperm of the different males competing to fertilize ova. Sperm competition has been documented or inferred to occur across several species. We address the evidence for sperm competition in humans by reviewing literature indicating apparently convergent adaptations to sperm competition in humans and non-humans. We discuss future research directions, and conclude that the evidence for anatomical, biological, physiological, and behavioral adaptations to human sperm competition provides compelling evidence that sperm competition has been a recurrent feature of human evolutionary history.

  7. Historical rammed earth process description thanks to micromorphological analysis

    OpenAIRE

    HAMARD, Erwan; Cammas, Cécilia; Fabbri, Antonin; Razakamanantsoa, Andry; Cazacliu, Bogdan; MOREL, Jean Claude

    2016-01-01

    Rammed earth was traditionally used in western European countries before industrial building materials replace it during 20th Century. Construction strategies developed by former builders were dictated by locally available construction materials and engendered local constructive cultures. Unfortunately, this knowledge was orally transmitted and is lost today. The rediscovery of these cultures can provide answers to modern rammed earth construction processes. Micromorphological analysis of ear...

  8. EM Scattering from Conducting Flat Plates Coated with Thin RAM

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    According to the equivalence principles, high frequency approximation and boundary conditions, a method has been developed to deal with the EM scattering by a rectangular conducting flat plate coated with uniaxial anisotropic radar absorbing material (RAM). The simple and effective method is available to the system of RCS prediction in which the large complex targets modeled by facets and wedges. Numerical results show some properties of EM scattering by conducting plate coated with thin uniaxial anisotropic RAM.

  9. Comparative sperm ultrastructure in Nemertea.

    Science.gov (United States)

    von Döhren, J; Beckers, P; Vogeler, R; Bartolomaeus, T

    2010-07-01

    Although the monophyly of Nemertea is strongly supported by unique morphological characters and results of molecular phylogenetic studies, their ingroup relationships are largely unresolved. To contribute solving this problem we studied sperm ultrastructure of 12 nemertean species that belong to different subtaxa representing the commonly recognized major monophyletic groups. The study yielded a set of 26 characters with an unexpected variation among species of the same genus (Tubulanus and Procephalothrix species), whereas other species varied in metric values or only one character state (Ramphogordius). In some species, the sperm nucleus has grooves (Zygonemertes virescens, Amphiporus imparispinosus) that may be twisted and give a spiral shape to the sperm head (Paranemertes peregrina, Emplectonema gracile). To make the characters from sperm ultrastructure accessible for further phylogenetic analyses, they were coded in a character matrix. Published data for eight species turned out to be sufficiently detailed to be included. Comparative evaluation of available information on the sperm ultrastructure suggests that subtaxa of Heteronemertea and Hoplonemertea are supported as monophyletic by sperm morphology. However, the data do not provide information on the existing contradictions regarding the internal relationships of "Palaeonemertea." Nevertheless, our study provides evidence that sperm ultrastructure yields numerous potentially informative characters that will be included in upcoming phylogenetic analyses.

  10. Cigarette smoking impairs sperm bioenergetics

    Directory of Open Access Journals (Sweden)

    Kazim R. Chohan

    2010-02-01

    Full Text Available OBJECTIVE: The growing consensus on the negative impact of cigarette smoking on fertility prompted us to compare the rate of sperm respiration in smokers and non-smokers. MATERIALS AND METHODS: Semen samples from 20 smokers and 58 non-smokers consulting at the andrology laboratory for fertility evaluation were used. Smoking was defined as consumption of at least a half a pack per day. A phosphorescence analyzer that measures O2 concentration in sperm suspensions as function of time was used to determine the rate of respiration. In a sealed vial, the rate of sperm respiration (k was defined as -d[O2]/dt; where [O2] was obtained from the phosphorescence decay rate of a palladium phosphor. [O2] in solutions containing sperm and glucose declined linearly with time, showing the kinetics of O2 consumption was zero-order. Inhibition of O2 consumption by cyanide confirmed the oxidations that occurred in the sperm mitochondrial respiratory chain. RESULTS: There were no differences (p > 0.28 between smokers and non-smokers for ejaculate volume, motility, concentration, normal morphology, viability and hypo-osmotic swelling test. The rate (mean ± SD, in µM O2/min/108 sperm of sperm mitochondrial O2 consumption in the smokers was 0.96 ± 0.58 and in the non-smokers 1.39 ± 0.67 (p = 0.004. CONCLUSIONS: The rate of sperm respiration was significantly lower in smokers. This negative impact of cigarette smoking on sperm aerobic metabolism may, in part, explain the lower rate of fertility in smokers.

  11. Human sperm rheotaxis: a passive physical process

    OpenAIRE

    Zhuoran Zhang; Jun Liu; Jim Meriano; Changhai Ru; Shaorong Xie; Jun Luo; Yu Sun

    2016-01-01

    A long-standing question in natural reproduction is how mammalian sperm navigate inside female reproductive tract and finally reach the egg cell, or oocyte. Recently, fluid flow was proposed as a long–range guidance cue for sperm navigation. Coitus induces fluid flow from oviduct to uterus, and sperm align themselves against the flow direction and swim upstream, a phenomenon termed rheotaxis. Whether sperm rheotaxis is a passive process dominated by fluid mechanics, or sperm actively sense an...

  12. Fire effects in Pinus uncinata Ram plantations

    Energy Technology Data Exchange (ETDEWEB)

    Cardil Forradellas, A.; Molina Terrén, D.M.; Oliveres, J.; Castellnou, M.

    2016-07-01

    Aim of study: Understanding fire ecology of main forest species is essential for a sound, scientifically based on managing of wildlands and also to assess likely implications due to changes in fire regime under a global change scenario. Few references can be found about fire ecology of Pinus uncinata Ram. (PU). PU species grows in the Central Pyrenees where large, severe wildland fires did not occur frequently in the past. However, several fires with extreme fire behavior have affected PU stands in last years and they might disturb other PU forest in the future. Area of study: Cabdella fire (February 2012), in Lleida province, is one of the several wildland fires occurred in 2012 (winter season) in the Central Pyrenees. Fire affected a large PU plantation (102 ha) located at 1.800-2,100 meters above the sea. Material and methods: We have analyzed first order fire effects in three fireline intensity thresholds along three years in terms of mortality ratio, scorched height, percentage of scorched crown volume and bark char height. Main results: PU seems to be a very tolerant species to low and medium fire line intensity but fire effects were very significant when fire line intensity was high. In medium fireline intensity sites, probability of mortality ranged from 15 to 30% and the dead trees had the highest values on scorched height and percentage of scorched crown volume. Research highlights: Results from this work supports that prescribed burning might be used to efficiently decrease fuel load and fuel vertical continuity while avoiding considerable PU mortality. It also displayed that when fuel management has been implemented, PU mortality might be limited even under extreme fire behavior. (Author)

  13. 33 CFR 147.811 - Ram-Powell Tension Leg Platform safety zone.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Ram-Powell Tension Leg Platform... SECURITY (CONTINUED) OUTER CONTINENTAL SHELF ACTIVITIES SAFETY ZONES § 147.811 Ram-Powell Tension Leg Platform safety zone. (a) Description. The Ram-Powell Tension Leg Platform (Ram-Powell TLP) is located...

  14. Daily Sperm Production, Gonadal and Extra-Gonadal Sperm ...

    African Journals Online (AJOL)

    SH

    The control diet 1 had neither probiotic nor prebiotic while prebiotic, probiotic and symbiotic (prebiotic + ... live microbial feed supplement that has beneficial effects on ... Four diets were formulated including the control ... same trend. The sperm ...

  15. Effect of exposing rams to a female stimulus before semen collection on ram libido and semen quality1

    National Research Council Canada - National Science Library

    A G Fahey; P Duffy; S Fair

    2012-01-01

      Rams with strong libido and desirable semen characteristics can provide more insemination doses per ejaculate and produce more progeny, improving population genetic linkage to improve the accuracy of EBV...

  16. Effect of exposing rams to a female stimulus before semen collection on ram libido and semen quality

    National Research Council Canada - National Science Library

    Fahey, A G; Duffy, P; Fair, S

    2012-01-01

    Rams with strong libido and desirable semen characteristics can provide more insemination doses per ejaculate and produce more progeny, improving population genetic linkage to improve the accuracy of EBV...

  17. Sperm Dynamics in Spiders (Araneae): Ultrastructural Analysis of the Sperm Activation Process in the Garden Spider Argiope bruennichi (Scopoli, 1772)

    OpenAIRE

    Oliver Vöcking; Gabriele Uhl; Peter Michalik

    2013-01-01

    Storage of sperm inside the female genital tract is an integral phase of reproduction in many animal species. The sperm storage site constitutes the arena for sperm activation, sperm competition and female sperm choice. Consequently, to understand animal mating systems information on the processes that occur from sperm transfer to fertilization is required. Here, we focus on sperm activation in spiders. Male spiders produce sperm whose cell components are coiled within the sperm cell and that...

  18. Effects of Enterobacter cloacae on boar sperm quality during liquid storage at 17°C.

    Science.gov (United States)

    Prieto-Martínez, Noelia; Bussalleu, Eva; Garcia-Bonavila, Estela; Bonet, Sergi; Yeste, Marc

    2014-07-01

    Contamination of fresh and extended boar sperm often occurs in farms and artificial insemination (AI) centres during semen collection, processing and storage. The presence of bacteria produces detrimental effects on boar sperm quality, which may cause economic losses in reproductive centres. The present study has evaluated for the first time how the presence of Enterobacter cloacae affects the preservation of boar spermatozoa in liquid storage at 15-17 °C for an 11-day period. With this purpose, extended semen samples from seven healthy post-pubertal boars were artificially contaminated with different sperm:bacterium ratios (2:1; 1:1; 1:5 and 1:10) of E. cloacae. The 1:0 ratio (non-inoculated) served as a negative control. The most infective ratios (i.e. 1:5 and 1:10) significantly damaged sperm motility and membrane integrity, increased sperm agglutination, and decreased the osmotic resistance of spermatozoa. In contrast, the negative impact that the lowest bacterial concentration (2:1) had on boar sperm quality was clearly lower. In addition, other parameters such as pH were also more affected at the highest infective ratios (i.e. 1:5 and 1:10), despite no damage being observed on sperm morphology. In conclusion, the present work shows that damage inflicted by the presence of E. cloacae in boar sperm during liquid storage at 15-17 °C compromises the longevity and fertilising ability of seminal doses when bacterial concentration is higher than a 1:1 ratio. Further research is warranted to address by which mechanism E. cloacae impairs boar sperm quality.

  19. Zebrafish reproduction: revisiting in vitro fertilization to increase sperm cryopreservation success.

    Directory of Open Access Journals (Sweden)

    Mary Hagedorn

    Full Text Available Although conventional cryopreservation is a proven method for long-term, safe storage of genetic material, protocols used by the zebrafish community are not standardized and yield inconsistent results, thereby putting the security of many genotypes in individual laboratories and stock centers at risk. An important challenge for a successful zebrafish sperm cryopreservation program is the large variability in the post-thaw in vitro fertilization success (0 to 80%. But how much of this variability was due to the reproductive traits of the in vitro fertilization process, and not due to the cryopreservation process? These experiments only assessed the in vitro process with fresh sperm, but yielded the basic metrics needed for successful in vitro fertilization using cryopreserved sperm, as well. We analyzed the reproductive traits for zebrafish males with a strict body condition range. It did not correlate with sperm volume, or motility (P>0.05, but it did correlate with sperm concentration. Younger males produced more concentrated sperm (P0.05, but pooling eggs reduced it by approximately 30 to 50% (P<0.05. This reduction in fertilization success was due not to the pooling of the females' eggs, but to the type of tools used to handle the eggs. Recommendations to enhance the in vitro process for zebrafish include: 1 using males of a body condition closer to 1.5 for maximal sperm concentration; 2 minimizing sperm wastage by using a working sperm concentration of 10(6 motile cells/ml for in vitro fertilization; and 3 never using metal or sharp-edged tools to handle eggs prior to fertilization.

  20. Sperm fractions obtained following density gradient centrifugation in human ejaculates show differences in sperm DNA longevity

    Directory of Open Access Journals (Sweden)

    Jaime Gosálvez

    2014-06-01

    Conclusion: 1 Unnecessary incubation of spermatozoa prior to artificial insemination or in vitro fertilization, should be avoided, since sperm DNA longevity is significantly reduced after ex vivo sperm handling and 2 Although sperm selection by DCG significantly reduces the baseline levels of SDF of sperm in Fraction 3, sperm DNA longevity in this fraction was ultimately lower following 24 h incubation when compared to sperm recovered from non-centrifuged NSS.

  1. Intelligent Fish Freshness Assessment

    Directory of Open Access Journals (Sweden)

    Hamid Gholam Hosseini

    2008-01-01

    Full Text Available Fish species identification and automated fish freshness assessment play important roles in fishery industry applications. This paper describes a method based on support vector machines (SVMs to improve the performance of fish identification systems. The result is used for the assessment of fish freshness using artificial neural network (ANN. Identification of the fish species involves processing of the images of fish. The most efficient features were extracted and combined with the down-sampled version of the images to create a 1D input vector. Max-Win algorithm applied to the SVM-based classifiers has enhanced the reliability of sorting to 96.46%. The realisation of Cyranose 320 Electronic nose (E-nose, in order to evaluate the fish freshness in real-time, is experimented. Intelligent processing of the sensor patterns involves the use of a dedicated ANN for each species under study. The best estimation of freshness was provided by the most sensitive sensors. Data was collected from four selected species of fishes over a period of ten days. It was concluded that the performance can be increased using individual trained ANN for each specie. The proposed system has been successful in identifying the number of days after catching the fish with an accuracy of up to 91%.

  2. Evidence of melatonin synthesis in the ram reproductive tract.

    Science.gov (United States)

    Gonzalez-Arto, M; Hamilton, T R dos S; Gallego, M; Gaspar-Torrubia, E; Aguilar, D; Serrano-Blesa, E; Abecia, J A; Pérez-Pé, R; Muiño-Blanco, T; Cebrián-Pérez, J A; Casao, A

    2016-01-01

    Melatonin is a ubiquitous molecule found in a wide range of fluids, one of them being ram seminal plasma, in which it can reach higher concentrations than those found in blood, suggesting an extrapineal secretion by the reproductive tract. In order to identify the source of the melatonin found in ram seminal plasma, we first tried to determine whether the melatonin levels were maintained during the day. For this purpose, melatonin concentrations were measured in seminal plasma obtained from first ejaculates of six rams at 6:00 a.m. in total darkness, at 10:00 a.m. and at 14:00 p.m. The melatonin concentration was higher (p ram testes, epididymis, and accessory glands. The RT-PCR showed the presence of the m-RNA codifying both AANAT and ASTM in all the tissues under study, but the q-PCR and Western-blot revealed that gene expression of these enzymes was significantly higher in the testis (p ram seminal plasma, at least during the day.

  3. Modeling rammed earth wall using discrete element method

    Science.gov (United States)

    Bui, T.-T.; Bui, Q.-B.; Limam, A.; Morel, J.-C.

    2016-03-01

    Rammed earth is attracting renewed interest throughout the world thanks to its "green" characteristics in the context of sustainable development. Several research studies have thus recently been carried out to investigate this material. Some of them attempted to simulate the rammed earth's mechanical behavior by using analytical or numerical models. Most of these studies assumed that there was a perfect cohesion at the interface between earthen layers. This hypothesis proved to be acceptable for the case of vertical loading, but it could be questionable for horizontal loading. To address this problem, discrete element modeling seems to be relevant to simulate a rammed earth wall. To our knowledge, no research has been conducted thus far using discrete element modeling to study a rammed earth wall. This paper presents an assessment of the discrete element modeling's robustness for rammed earth walls. Firstly, a brief description of the discrete element modeling is presented. Then the parameters necessary for discrete element modeling of the material law of the earthen layers and their interfaces law following the Mohr-Coulomb model with a tension cut-off and post-peak softening were given. The relevance of the model and the material parameters were assessed by comparing them with experimental results from the literature. The results showed that, in the case of vertical loading, interfaces did not have an important effect. In the case of diagonal loading, model with interfaces produced better results. Interface characteristics can vary from 85 to 100% of the corresponding earthen layer's characteristics.

  4. The impact of bacteriospermia on boar sperm storage and reproductive performance.

    Science.gov (United States)

    Kuster, C E; Althouse, G C

    2016-01-01

    Bacteriospermia is a documented risk to reproductive performance when using extended boar semen for artificial insemination. A substantial list of bacteria have been recovered from boar semen attributed to fecal, preputial, skin, and hair microorganisms, with these and other environmental bacteria from processing areas identified in doses prepared for artificial insemination. Gram-negative bacteria are most commonly recovered from extended doses, including both Enterobacteriaceae and environmental contaminants, such as those that inhabit water purification systems. The method of processing, distributing, and storing fresh liquid boar semen before insemination plays a role in bacterial growth dynamics and the degree to which the bacteria may damage the sperm or affect the sow. Not all bacterial isolates or contamination levels have the same impact on sperm, with multiple factors governing if and when storage longevity will be reduced through sperm-to-sperm agglutination, impaired motility, acrosome disruption, or loss of membrane viability. Suboptimal reproductive performance can occur because of reduced fertilizing capacity of the sperm or induction of a uterine environment hostile to sperm and/or embryonic survival. Effective bacterial control strategies are necessary to minimize the risk of bacteria contaminating extended semen doses, including monitoring programs designed for quick detection and intervention, should the need arise.

  5. Effect of cryopreservation on mitochondrial activity in buffalo sperm Bubalus bubalis

    Directory of Open Access Journals (Sweden)

    O. Kandil

    2010-02-01

    Full Text Available Sperm mitochondrial activity is investigated and used as “in vitro” spermatozoa vitality indicator and about quality effectiveness of different sperm diluents. It was studied the cytochemically activity of NADPH-diaphorase and LDH-C4 in cryopreserved buffalo sperm. Low intensity of the enzyme reaction was established in all examined sperm samples in both enzymes, regardless from the used cryoprotectors. The main part of the enzyme reaction was localized in mitochondrial sheath and in a very small degree in the head base of spermatozoa. No increase of the enzymes activities or the spermatozoa motility has been found after the incubating with Sp-TALP medium although the established caffeine stimulating effect on the glycolysis and fresh spermatozoa motility. Established by us low sperm motility after cryopreservation may be due to low LDH and NADPH-diaphorase activity due to glycolisis disturbances and ATP synthesis. This method allows to estimate quality of buffalo semen and to find some different disturbances in mitochondrial sheath, which could not be found by routine morphological studies and could be used in practice ejaculates with high number of metabolic active sperm cells.

  6. Effect of Different Nano-Zinc Levels in Dietary on Semen Quality, Activities of Antioxidant Enzyme and Expression of Copper Zinc Superoxide in Epididymis of Ram Lambs%纳米锌水平对公羊精液品质、抗氧化酶活性及附睾 Cu-ZnSOD 表达的影响

    Institute of Scientific and Technical Information of China (English)

    张春香; 秦小伟; 郭丽娜; 张国林; 张建新; 任有蛇

    2015-01-01

    >附睾头>附睾尾;然而对照组的顺序为:附睾头>附睾体>附睾尾。50 mg·kg-1或100 mg·kg-1纳米锌组公羊附睾头和附睾体阳性信号的平均光密度显著高于对照组和150 mg·kg-1纳米锌(P<0.05)。精浆 Cu-ZnSOD 活性与精子密度、精子活力和精子质膜完整性呈正相关,与精子畸形率呈负相关。【结论】日粮添加50 mg·kg-1或100 mg·kg-1纳米锌可改善精液品质,提高精浆抗氧化能力,增加附睾中 Cu-ZnSOD 蛋白表达量。精浆 Cu-ZnSOD 活性可以作为检测精液品质优劣的指标在羊生产中应用。微量元素锌对精液品质影响调控的分子机理还需进一步深入研究。%[Objective] The objectives of this study were to investigate the effects of nano-zinc supplementation in dietary on the semen quality parameters, the activities of antioxidant enzyme in seminal plasma and the expression of copper zinc superoxide dismutase in epididymis of ram lambs, and to analyze the correlation between semen quality parameters and the activities of antioxidant enzyme.[Method]Sixteen 9-month-old Jinzhong ram lambs with good health and approximate weight were randomly divided into 4 groups, fed with a basal diet with supplementation of 0, 50, 100 and 150 mg·kg-1 DM nano-zinc, respectively. The experimental period was 90 d. Semen was collected on 78 d and 79 d in consecutive two days, samples of 100 µL fresh semen was used to analyze semen quality parameters, the rest of fresh semen sample was centrifuged at 2 000 r/min for 10 min, the supernatant (seminal plasma) was collected for measurement of the activities of copper zinc superoxide dismutase, glutathione peroxidase and total anti-oxidation competence. All rams were castrated for collection of epididymis caput, corpus and cauda at the end of experiment. Expression of Cu-ZnSOD protein in epididymis was detected and located by immunohistochemistry. The mean optical density was analyzed with Image Pro

  7. Active immunization against GnRH in ovine (Ovis aries: testicular morphometry, histopathology and endocrine responses in prepubertal ram lambs

    Directory of Open Access Journals (Sweden)

    Gutierrez-Reinoso MA

    2017-08-01

    Full Text Available The main objective of the present research was to assess the effects of an active immunization against GnRH on male reproductive function as an alternative to surgical sterilization evaluating testicular morphometry, histopathological alterations and plasma gonadotropin levels in prepubertal ram lambs. Dorper ram lambs (age= 2 months; control group, n= 5; treatment group, n=15 were immunised by using an anti-GnRH vaccine (two administrations spaced 15d developed by linking a GnRH homologous molecule to tetanus toxoid (clostridial toxoid with aluminum hydroxide as coadjuvant. To determine the effectiveness of the vaccination protocol, testicular morphometry was evaluated (length, width, mean volume and total volume together with histopathological alterations in testicular tissue samples (seminiferous tubule diameters, spermatogonia per testis and sperm presence and endocrine responses (ELISA from blood samples (FSH, LH, testosterone and cortisol plasma levels. Morphometric parameters (testicular length, width and volume were significantly reduced in vaccinated animals with respect to the control group (p0.05. In conclusion, prepubertal active immunization against GnRH led to marked differences on testes morphometry and activity in ovine species, however, it did not affect endocrine levels nor spermatogenesis in all individuals studied showing a partial vaccination effect in some of them. Thus, taking into account the heterogeneous dysfunctional responses obtained, different vaccination assays and strategies should be tested before active immunization against GnRH is considered as a viable alternative to conventional surgical sterilization.

  8. Ram pressure induced star formation in Abell 3266

    Science.gov (United States)

    Bonsall, Brittany

    An X-ray observation of the merging galaxy cluster Abell 3266 was obtained via the ROSAT PSPC. This information, along with spectroscopic data from the WIde-field Nearby Galaxy-clusters Survey (i.e. WINGS), were used to investigate whether ram pressure is a mechanism that influences star formation. Galaxies exhibiting ongoing star formation are identified by the presence of strong Balmer lines (Hbeta), known to correspond to early type stars. Older galaxies where a rapid increase in star formation has recently ceased, known as E+A galaxies, are identified by strong Hbeta absorption coupled with little to no [OII] emission. The correlation between recent star formation and "high" ram pressure, as defined by Kapferer et al. (2009) as ≥ 5 x 10-11 dyn cm-2, was tested and lead to a contradiction of the previously held belief that ram pressure influences star formation on the global cluster scale.

  9. Localization of CD9 Molecule on Bull Spermatozoa: Its Involvement in the Sperm-Egg Interaction.

    Science.gov (United States)

    Antalíková, J; Jankovičová, J; Simon, M; Cupperová, P; Michalková, K; Horovská, Ľ

    2015-06-01

    Tetraspanin CD9 is one of the egg membrane proteins known to be essential in fertilization process. The presence and localization of CD9 molecule in spermatozoa and its possible function in reproduction are still unclear. In our study, we describe the localization of CD9 on bull spermatozoa. In the immunofluorescence assay, the positive signal has been observed in the high proportion of sperm cells as a fine grains either on the apical part or through the entire anterior region of sperm head. CD9 recognized by monoclonal antibody IVA-50 was detected on freshly ejaculated (83.4 ± 3.7%) and frozen-thawed (84.3 ± 2.3%) sperm. The same reaction pattern was observed on sperm capacitated for 1 h, 2 h, 3 h and 4 h (83.6 ± 2.0%; 84.0 ± 1.5%; 85.7 ± 0.8%; 77.5 ± 10.8%). The presence of CD9 exclusively on plasma membrane of the bovine sperm has been detected by Western blot analysis of the protein fractions after the discontinuous sucrose gradient fractionation of the bull sperm. Moreover, probable role of the sperm CD9 molecule in fertilization process of cattle has been suggested as sperm treatment with anti-CD9 antibody significantly reduced (by 25%, p ≤ 0.001) the number of fertilized oocytes compared to control group in fertilization assay in vitro.

  10. La Palabra de Ramón Frediani: Las Retenciones

    Directory of Open Access Journals (Sweden)

    Marcelo Coser

    2008-04-01

    Full Text Available En el marco del programa "Prisma Económico" del Instituto de Economía y Finanzas (IEF, Marcelo Coser, su conductor, consulta al Profesor Ramón Osvaldo Frediani, docente de la asignatura “Economía Regional y Urbana” e investigadordel Instituto. Durante la nota, Ramón Frediani nos da sus conceptos sobre las debatidas retenciones y los propósitos que éstas persiguen, brindando su visión respecto al accionar del Gobierno Nacional.

  11. The sperm penetration test (P-test) can predict fecundability in the male partner from infertile couples

    DEFF Research Database (Denmark)

    Bostofte, E; Bagger, P; Michael, A;

    1992-01-01

    Three hundred and twenty-one consecutive couples were investigated for infertility at Hvidovre University Hospital in the period from November 1977 to June 1985. The male partners were evaluated in two ways: the classical semen analysis, and the ability of sperm to penetrate fresh hen egg white...

  12. What use is an infertile sperm? A comparative study of sperm-heteromorphic Drosophila

    DEFF Research Database (Denmark)

    Holman, Luke; Freckleton, Robert P; Snook, Rhonda R

    2007-01-01

    Sperm size and number are important determinants of male reproductive success. The genus Drosophila exhibits a remarkable diversity of sperm production strategies, including the production of multiple sperm morphs by individual males, a phenomenon called sperm heteromorphism. Sperm-heteromorphic ......Sperm size and number are important determinants of male reproductive success. The genus Drosophila exhibits a remarkable diversity of sperm production strategies, including the production of multiple sperm morphs by individual males, a phenomenon called sperm heteromorphism. Sperm......-heteromorphic Drosophila species in the obscura group produce large numbers of infertile "parasperm" in addition to fertile eusperm. Parasperm have been hypothesized to perform a number of roles in place of fertilization, predominantly focused on their potential function in postcopulatory sexual selection. However...

  13. Transmission electron microscopic observations of acrosome and head abnormalities in impala (Aepyceros melampus sperm from the Kruger National Park

    Directory of Open Access Journals (Sweden)

    D.J. Ackerman

    1997-01-01

    Full Text Available Sperm morphological features play an important role in semen evaluation. Exposure to a variety of chemical compounds, especially environmental endocrine disrupters, elicit abnormalities in sperm of certain species. Baseline data on ultrastructure of normal sperm as well as abnormalities observed concomitantly, are required before causal links between such substances and abnormalities can be established. Live spermatozoa were collected from the cauda epididymis of 64 impala rams in the Kruger National Park and studied by transmission electron microscopy to document normal sperm features and abnormalities. The following abnormalities of the acrosome and sperm head were documented from micrographs: Loose acrosome in various stages of disintegration, lip forming of the acrosome; bizarre head, crater defect, poor condensation of the nucleus and the Dag defect. The observed abnormalities were very similar to those reported for other members of the Bovidae. Different forms of a hollow sphere, formed by the nucleus and covered by an abnormal acrosome have not previously been described for other species.

  14. Predictors of sperm recovery after cryopreservation in testicular cancer

    Directory of Open Access Journals (Sweden)

    James M Hotaling

    2016-01-01

    Full Text Available Our objective was to identify predictors of improved postthaw semen quality in men with testicular cancer banking sperm for fertility preservation. We reviewed 173 individual semen samples provided by 67 men with testicular germ cell tumor (TGCT who cryopreserved sperm before gonadotoxic treatment between 1994 and 2010 at our tertiary university medical center. Our main outcomes measures were independent predictors for the greater postthaw total motile count (TMC in men with TGCT. Men with NSGCT were more likely to be younger (P median fresh TMC each had increased odds of a postthaw TMC greater than median postthaw TMC. Interestingly, age, advanced cancer stage (II or III, rapid freezing protocol, and motility enhancer did not show increased odds of improved postthaw TMC in our models. In conclusion, men with TGCT or poor fresh TMC should consider preserving additional vials (at least 15 vials before oncologic treatment. Density gradient purification should be routinely used to optimize postthaw TMC in men with TGCT. Larger, randomized studies evaluating cancer stage and various cryopreservation techniques are needed to assist in counseling men with TGCT regarding fertility preservation and optimizing cryosurvival.

  15. Computer-assisted Sperm Analysis System for Detection of in Boars Fresh Semen Quality%应用计算机辅助精子质量分析系统对猪常温精液的品质检验

    Institute of Scientific and Technical Information of China (English)

    张洁; 陈敏恒; 李宝红; 王均亮; 苏文昌; 彭国良; 李剑豪

    2015-01-01

    计算机辅助精子质量分析(computer-assisted sperm analysis,CASA)系统具有快速、客观、精确等特点,与传统人工分析法相比具有明显的优势.实验选择3个品种9头公猪的精液,采用计算机辅助分析法和人工分析法同时分析精液密度和活力,比较两种方法的差异.CASA采用伟力彩色精子质量检测系统(WLJY-9000),人工分析法采用《种猪常温精液》(GB23238-2009)规定的方法.结果表明,CASA法和人工分析法对精子密度的检测结果差异不显著(P>0.05),对杜洛克和大白公猪精子活力的检测差异不显著(P>0.05).CASA与人工分析法对精液密度和活力的检测结果较为一致,CASA更能体现精子的运动状态,对畸形率的观测更为清晰.

  16. In vitro sperm characterization and development of a sperm cryopreservation method using directional solidification in the killer whale (Orcinus orca).

    Science.gov (United States)

    Robeck, T R; Gearhart, S A; Steinman, K J; Katsumata, E; Loureiro, J D; O'Brien, J K

    2011-07-15

    Research was conducted to characterize seminal traits and to develop a sperm cryopreservation method using directional freezing (DF) for the killer whale (Orcinus orca). Experiments evaluated effects of: (i) freezing rate (SLOW, MED, FAST) by diluent (BF5F, Biladyl®, EYC) in 0.5 mL straws; and (ii) freezing method (straw or DF) by glycerol (3, 6, or 9% final concentration, v:v) on in vitro sperm quality. Fresh ejaculates (n = 161) were (mean ± SD) 7.8 ± 7.4 mL at 740 × 10(6) sperm/mL with 92.2 ± 6.3% total motility (TM), 85.4 ± 6.9% progressive motility (PM), 89.6 ± 9.0% viability and 89.8 ± 9.2% acrosome integrity. Samples frozen using straws by the MED or SLOW method were improved (P Directional freezing samples at 0 h and 3 h PT, at all glycerol concentrations, displayed higher (P < 0.001) TM, PM, RM, VAP, VSL, VCL and viability /intact acrosomes (PI/FITC-PNA) than straw. These data provided the first information on ejaculate characteristics and the development of a semen cryopreservation method using DF in the killer whale.

  17. Global Shortage of Fresh Water

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    <正>阅读下表。以Global Shortage of Fresh Water为题写一篇短文。词数:100—120学生习作:Global Shortage of Fresh Water Fresh water seems ineverywhere,in rivers,lakes,wells as well as rain,which make some people think that we can’t use up water.

  18. Bioenergetics of Mammalian Sperm Capacitation

    Directory of Open Access Journals (Sweden)

    Alessandra Ferramosca

    2014-01-01

    Full Text Available After ejaculation, the mammalian male gamete must undergo the capacitation process, which is a prerequisite for egg fertilization. The bioenergetics of sperm capacitation is poorly understood despite its fundamental role in sustaining the biochemical and molecular events occurring during gamete activation. Glycolysis and mitochondrial oxidative phosphorylation (OXPHOS are the two major metabolic pathways producing ATP which is the primary source of energy for spermatozoa. Since recent data suggest that spermatozoa have the ability to use different metabolic substrates, the main aim of this work is to present a broad overview of the current knowledge on the energy-producing metabolic pathways operating inside sperm mitochondria during capacitation in different mammalian species. Metabolism of glucose and of other energetic substrates, such as pyruvate, lactate, and citrate, is critically analyzed. Such knowledge, besides its obvious importance for basic science, could eventually translate into the development of novel strategies for treatment of male infertility, artificial reproduction, and sperm selection methods.

  19. Autoradiographic visualization of the mouse egg's sperm receptor bound to sperm

    OpenAIRE

    1986-01-01

    The extracellular coat, or zona pellucida, of mammalian eggs contains species-specific receptors to which sperm bind as a prelude to fertilization. In mice, ZP3, one of only three zona pellucida glycoproteins, serves as sperm receptor. Acrosome-intact, but not acrosome-reacted, mouse sperm recognize and interact with specific O- linked oligosaccharides of ZP3 resulting in sperm-egg binding. Binding, in turn, causes sperm to undergo the acrosome reaction; a membrane fusion event that results i...

  20. Microfluidic single sperm entrapment and analysis

    NARCIS (Netherlands)

    Wagenaar, B. de; Berendsen, J.T.; Bomer, J.G.; Olthuis, W.; Berg, A. van den; Segerink, L.I.

    2015-01-01

    Selection of healthy spermatozoa is of crucial importance for the success rates of assisted reproduction technologies (ART) such as in vitro fertilization and intra-cytoplasmic sperm injection. Although sperm selection for ART procedures is predominantly based on sperm motility, successful fertiliza

  1. Windsock memory conditioned RAM (Co-Ram) pressure effect: forced reconnection in the Earth's magnetotail

    CERN Document Server

    Vörös, Z; Khodachenko, M; Honkonen, I; Janhunen, P; Palmroth, M

    2014-01-01

    Magnetic reconnection (MR) is a key physical concept explaining the addition of magnetic flux to the magnetotail and closed flux lines back-motion to the dayside magnetosphere. This scenario elaborated by \\citet{dung63}, can explain many aspects of solar wind-magnetosphere interaction processes, including substorms. However, neither the Dungey model nor its numerous modifications were able to explain fully the onset conditions for MR in the tail. In this paper, we introduce new onset conditions for forced MR in the tail. We call our scenario the "windsock memory conditioned ram pressure effect". Our non-flux-transfer associated forcing is introduced by a combination of large-scale windsock motions exhibiting memory effects and solar wind dynamic pressure actions on the nightside magnetopause during northward oriented IMF. Using global MHD GUMICS-4 simulation results, upstream data from WIND, magnetosheath data from Cluster-1 and distant-tail data from the two-probe ARTEMIS mission, we show that the simultaneo...

  2. A cost for high levels of sperm competition in rodents: increased sperm DNA fragmentation.

    Science.gov (United States)

    delBarco-Trillo, Javier; García-Álvarez, Olga; Soler, Ana Josefa; Tourmente, Maximiliano; Garde, José Julián; Roldan, Eduardo R S

    2016-03-16

    Sperm competition, a prevalent evolutionary process in which the spermatozoa of two or more males compete for the fertilization of the same ovum, leads to morphological and physiological adaptations, including increases in energetic metabolism that may serve to propel sperm faster but that may have negative effects on DNA integrity. Sperm DNA damage is associated with reduced rates of fertilization, embryo and fetal loss, offspring mortality, and mutations leading to genetic disease. We tested whether high levels of sperm competition affect sperm DNA integrity. We evaluated sperm DNA integrity in 18 species of rodents that differ in their levels of sperm competition using the sperm chromatin structure assay. DNA integrity was assessed upon sperm collection, in response to incubation under capacitating or non-capacitating conditions, and after exposure to physical and chemical stressors. Sperm DNA was very resistant to physical and chemical stressors, whereas incubation in non-capacitating and capacitating conditions resulted in only a small increase in sperm DNA damage. Importantly, levels of sperm competition were positively associated with sperm DNA fragmentation across rodent species. This is the first evidence showing that high levels of sperm competition lead to an important cost in the form of increased sperm DNA damage. © 2016 The Author(s).

  3. Sedimentation properties in density gradients correspond with levels of sperm DNA fragmentation, chromatin compaction and binding affinity to hyaluronic acid.

    Science.gov (United States)

    Torabi, Forough; Binduraihem, Adel; Miller, David

    2017-03-01

    Mature spermatozoa bind hyaluronic acid in the extracellular matrix via hyaladherins. Immature spermatozoa may be unable to interact because they do not express the appropriate hyaladherins on their surface. Fresh human semen samples were fractionated using differential density gradient centrifugation (DDGC) and the ability of these fractions to bind hyaluronic acid was evaluated. The presence of sperm hyaladherins was also assessed. CD44 was located mainly on the acrosome and equatorial segment and became more restricted to the equatorial segment in capacitated spermatozoa. Hyaluronic acid-TRITC (hyaluronic acid conjugated with tetramethylrhodamine isothiocyanante), a generic hyaluronic-acid-binding reagent, labelled the membrane and the neck region, particularly after capacitation. Sperm populations obtained after DDGC or after interaction with hyaluronic acid were assessed for DNA fragmentation and chromatin maturity. Strong relationships between both measures and sperm sedimentation and hyaluronic-acid-binding profiles were revealed. Capacitation enhanced hyaluronic acid binding of both DDGC-pelleted sperm and sperm washed free of seminal fluid. In conclusion, hyaladherins were detected on human sperm and a higher capacity for sperm hyaluronic-acid-binding was shown to correspond with their DDGC sedimentation profiles and with lower levels of DNA fragmentation and better chromatin maturity. Capacitation induced changes in the distribution and presence of hyaladherins may enhance hyaluronic-acid-binding.

  4. Birth after human chorionic gonadotropin-primed oocyte in vitro maturation and fertilization with testicular sperm in a normo-ovulatory patient

    Directory of Open Access Journals (Sweden)

    Claudia González-Ortega

    2016-01-01

    Full Text Available In this report, we present a case of in vitro maturation (IVM with surgical retrieved testicular sperm in a normo-ovulatory female. Human chorionic gonadotropin-primed IVM, testicular biopsy for sperm retrieval and intracytoplasmic sperm injection with fresh sperm were performed. Fourteen cumulus-oocyte complexes were obtained in germinal vesicle or metaphase I stage, eight oocytes reached metaphase II, seven presumptive zygotes were obtained, and three cleavage stages embryos in day 2 were transferred producing a singleton pregnancy. A single healthy newborn was obtained. Our results suggest that IVM may be an alternative for in vitro fertilization in normo-ovulatory women even if surgical retrieval of sperm is needed. Further research is required to depict contributing factors to the success of IVM in indications different from polycystic ovaries syndrome and the role of male gamete.

  5. Birth after human chorionic gonadotropin-primed oocyte in vitro maturation and fertilization with testicular sperm in a normo-ovulatory patient

    Science.gov (United States)

    González-Ortega, Claudia; Piña-Aguilar, Raul Eduardo; Cancino-Villareal, Patricia; Gutiérrez-Gutiérrez, Antonio Martin

    2016-01-01

    In this report, we present a case of in vitro maturation (IVM) with surgical retrieved testicular sperm in a normo-ovulatory female. Human chorionic gonadotropin-primed IVM, testicular biopsy for sperm retrieval and intracytoplasmic sperm injection with fresh sperm were performed. Fourteen cumulus-oocyte complexes were obtained in germinal vesicle or metaphase I stage, eight oocytes reached metaphase II, seven presumptive zygotes were obtained, and three cleavage stages embryos in day 2 were transferred producing a singleton pregnancy. A single healthy newborn was obtained. Our results suggest that IVM may be an alternative for in vitro fertilization in normo-ovulatory women even if surgical retrieval of sperm is needed. Further research is required to depict contributing factors to the success of IVM in indications different from polycystic ovaries syndrome and the role of male gamete. PMID:27803591

  6. Minsk ja RAM 7 / Evelyn Müürsepp

    Index Scriptorium Estoniae

    Müürsepp, Evelyn

    2005-01-01

    Minskis toimunud RAM 7-st (Reapproaching New Media. Models of Collaboration / Ümberhinnates uut meediat. Koostöömudelid". Korraldajad Tatjana Dushina ja Denis Romanovski üritavad luua uue meedia keskust Valgevene kunstiakadeemiasse. Eestit esindasid Evelin Müürsepp ja John Grzinich (modereeris vabade ülikoolide kursust). Osalemisest fotomängus "FotoOhota"

  7. Embodied energy in cement stabilised rammed earth walls

    Energy Technology Data Exchange (ETDEWEB)

    Venkatarama Reddy, B.V.; Prasanna Kumar, P. [Department of Civil Engineering, Indian Institute of Science, Bangalore 560012 (India)

    2010-03-15

    Rammed earth walls are low carbon emission and energy efficient alternatives to load bearing walls. Large numbers of rammed earth buildings have been constructed in the recent past across the globe. This paper is focused on embodied energy in cement stabilised rammed earth (CSRE) walls. Influence of soil grading, density and cement content on compaction energy input has been monitored. A comparison between energy content of cement and energy in transportation of materials, with that of the actual energy input during rammed earth compaction in the actual field conditions and the laboratory has been made. Major conclusions of the investigations are (a) compaction energy increases with increase in clay fraction of the soil mix and it is sensitive to density of the CSRE wall, (b) compaction energy varies between 0.033 MJ/m{sup 3} and 0.36 MJ/m{sup 3} for the range of densities and cement contents attempted, (c) energy expenditure in the compaction process is negligible when compared to energy content of the cement and (d) total embodied energy in CSRE walls increases linearly with the increase in cement content and is in the range of 0.4-0.5 GJ/m{sup 3} for cement content in the rage of 6-8%. (author)

  8. Detecting danger labels with RAM-based neural networks

    DEFF Research Database (Denmark)

    Jørgensen, T.M.; Christensen, S.S.; Andersen, A.W.

    1996-01-01

    An image processing system for the automatic location of danger labels on the back of containers is presented. The system uses RAM-based neural networks to locate and classify labels after a pre-processing step involving specially designed non-linear edge filters and RGB-to-HSV conversion. Results...

  9. Vitrification is not superior to rapid freezing of normozoospermic spermatozoa: effects on sperm parameters, DNA fragmentation and hyaluronan binding.

    Science.gov (United States)

    Agha-Rahimi, Azam; Khalili, Mohammad Ali; Nabi, Ali; Ashourzadeh, Sareh

    2014-03-01

    Human sperm vitrification is a new cryopreservation method. This study compared the effects of rapid freezing and vitrification on various sperm parameters, hyaluronan-binding assay and DNA fragmentation and assessed the impact of cryoprotectant agents (CPA) with vitrification. A total of 30 normo-ejaculates were prepared by swim up and the motile sperm fraction was divided into four: fresh (control), rapid freezing, and two vitrification groups (a, lacking CPA; b, with CPA). For rapid freezing, a cryovial of sperm suspension was held just above the liquid nitrogen surface, and for vitrification, 30μl suspension was dropped directly into liquid nitrogen. Sperm parameters, including motility, viability and morphology, declined after cryopreservation in both groups. DNA fragmentation was not significantly higher in the vitrification (15.7±4.4%) or rapid freezing (16.6±5.6%) groups when compared with controls (11.6±4.5%). The rates of hyaluronan binding were similar between the control and cryopreserved groups. Moreover, addition of CPA for vitrification had a neutral effect on rates of sperm recovery. In conclusion, vitrification has great potential for human sperm cryopreservation and does not require CPA, with its possible toxicity. However, it is not superior to rapid cryopreservation regarding sperm recovery rate in normozoospermia. Human sperm vitrification is a new cryopreservation method that has been introduced recently. This study compared the effects of rapid freezing with vitrification on rates of sperm parameters, hyaluronan-binding assay and DNA fragmentation after thawing/warming and assessed the impact of cryoprotectant agent (CPA) on vitrification. The study was performed on 30 ejaculates prepared using the swim-up technique. Each motile sperm suspension was divided into four: control (fresh); rapid freezing; and two vitrification groups (a, lacking CPA; b, with CPA). For rapid freezing, a cryovial of sperm suspension was held above the surface of

  10. Sperm motility patterns in Andalusian donkey (Equus asinus) semen: effects of body weight, age, and semen quality.

    Science.gov (United States)

    Dorado, J; Acha, D; Gálvez, M J; Ortiz, I; Carrasco, J J; Díaz, B; Gómez-Arrones, V; Calero-Carretero, R; Hidalgo, M

    2013-04-15

    The aims of this study were to (1) identify sperm subpopulations with specific motion characteristics in fresh Andalusian donkey ejaculates; (2) evaluate the effects of individual donkey and ejaculates within the same donkey on the distribution of the subpopulations found; and (3) explore the relationship between the age and the body weight of donkey donors, the sperm quality parameters, and the sperm subpopulations structure. Sixty ejaculates from 12 Andalusian donkeys (five ejaculates per donkey), ranging in age from 4 to 15 years, were collected. Immediately after collection, sperm characteristics (volume, sperm concentration, objective sperm motility, and sperm morphology) were assessed. Donkeys were evaluated for body weight. Significant (P donkeys and the pH (r = -0.52), sperm motility (percentage of motile spermatozoa: r = -0.31; percentage of progressive motile spermatozoa: r = -0.34), and total sperm abnormalities (r = 0.38). The correlations of the age with the measures of semen quality were low and not significant (P > 0.05). A multivariate clustering procedure separated 65,342 motile spermatozoa into four subpopulations: subpopulation 1, consisting of slow and nonprogressive spermatozoa (15.4%), subpopulation 2, consisting of moderately slow but progressive spermatozoa (35.9%), subpopulation 3, consisting of highly active but nonprogressive spermatozoa (18.5%), and subpopulation 4, consisting of highly active and progressive spermatozoa (30.2%). The distribution of these subpopulations varied significantly (P donkey, the ejaculate of the same donkey, the total motility, and the overall sperm concentration. Our results show the existence of four well-defined motile sperm subpopulations in Andalusian donkey ejaculates, and suggest a high heterogeneity in the ejaculate structure in donkey. The relationship between the distribution of the sperm subpopulations and individual donkey, total motility, and sperm concentration shows that the spermatozoa of each

  11. BOARD-LEVEL BUILT-IN SELF-REPAIR METHOD OF RAM

    Institute of Scientific and Technical Information of China (English)

    Dou Yanjie; Zhan Huiqin; Chen Yakun; Shang Hongliang

    2012-01-01

    This paper describes the method of built-in self-repairing of RAM on board,designs hardware circuit,and logic for the RAM's faults self-repairing system based on FPGA.The key technology is that it utilizes FPGA to test RAM according to some algorithm to find out failure memory units and replace the faulty units with FPGA.Then it can build a memory that has no fault concern to external controller,and realizes the logic binding between external controller and RAM.Micro Controller Unit (MCU) can operate external RAM correctly even if RAM has some fault address units.Conventional MCS-51 is used to simulate the operation of MCU operating external memory.Simulation shows FPGA can complete the faulty address units' mapping and MCU can normally read and write external RAM.This design realizes the RAM's built-in self-repairing on board.

  12. No evidence for sperm priming responses under varying sperm competition risk or intensity in guppies

    Science.gov (United States)

    Evans, Jonathan P.

    2009-07-01

    Sperm competition theory predicts that males should tailor their investment in ejaculates according to the number of rival males competing to fertilize a female’s eggs. Research spanning several taxa supports this prediction by showing that males are often sensitive to the level of sperm competition and adjust their investment in sperm numbers accordingly. More recent work has revealed that males may also tailor the quality of sperm according to the number of males competing for fertilization. Here I test for both effects in guppies ( Poecilia reticulata) in an experiment that simultaneously evaluates the risk and intensity models of sperm competition. The experiment determined whether male guppies adjust the number (stripped ejaculate size) and quality (sperm velocity and viability) of sperm that are primed over a 3-day period according to experimental changes in the perceived level of sperm competition. A total of 136 focal males were initially stripped of all retrievable sperm and assayed for these sperm traits before being allocated at random to one of four treatments simulating different levels of sperm competition risk and intensity. During the 3-day treatment phase, focal males had visual and olfactory access to a sexually receptive (initially virgin) female maintained with different numbers of stimulus males to simulate variation in the risk and intensity of sperm competition. Following this, males were assayed again for the sperm traits. Contrary to predictions, there was no significant change in any of the measured variables among treatments, although qualitatively the patterns for sperm velocity and viability did conform to expectation. The lack of any trend for the number of sperm primed was unequivocal and future work examining the effects of sperm competition on sperm production should focus on whether males differentially allocate sperm numbers among matings that differ in the level of sperm competition.

  13. [Effects of some extenders and monoamines on sperm cryopreservation in tree shrews (Tupaia belangeri)].

    Science.gov (United States)

    Ping, Shu-Huang; Wang, Cai-Yun; Tang, Wen-Ru; Luo, Ying; Yang, Shi-Hua

    2012-02-01

    The tree shrew may be an important experimental animal for disease models in humans. The effects of some extenders and momamines on sperm cryopreservation will provide helpful data for experimentation of strains and conservation of genetic resources in tree shrews. Epididymal sperm were surgically harvested from male tree shrews captured around Kunming, China and sperm motility, acrosome integrity and fertility were assessed during cryopreservation. In Experiment 1 eight extenders (TTE, TCG, TCF, TTG, BWW, BTS, DM, and SR) supplemented with 0.4 mol/L DMSO were used to dilute the sperm: only TTE, DM and SR showed no differences in motility and acrosome integrity compared to fresh controls after equilibration. After freezing and thawing, sperm in any extender showed lower motility than fresh control and sperm in DM showed higher motility than other groups. However, BWW produced the lowest motility. For acrosome integrity, TTE and DM showed higher than BWW, BTS and SR after equilibration. The parameter in DM was higher than other groups (except TTE) after thawing. In Experiment 2 four penetrating cryoprotectant agents (CPA) [dimethyl-formamide (DF), formamide (F), dimethylacetamide (DA), and acetamide (A)] at 0.2 mol/L, 0.4 mol/L, 0.8 mol/L, and 1.2 mol/L, respectively were added to the DM extender. Motility showed no difference among CPA groups and non-CPA group (control) after equilibration, but all thawed sperm showed lower values in motility and acrosome integrity than pre-freezing groups. However, sperm in 0.8 mol/L DF and 0.4 mol/L DMSO showed higher values in both parameters than that in other CPA groups (P>0.05). In Experiment 3 the fertilization rate of oocytes inseminated with 0.4mol/L DMSO (50%) were higher than that with 0.8mol/L DF (16%). In conclusion, non-ion extenders supplemented with egg yolk may be better for sperm cryopreservation in tree shrews and cryoprotectant effects of monoamines agents should be further studied in this species.

  14. Operational advances in ring current modeling using RAM-SCB

    Energy Technology Data Exchange (ETDEWEB)

    Welling, Daniel T [Los Alamos National Laboratory; Jordanova, Vania K [Los Alamos National Laboratory; Zaharia, Sorin G [Los Alamos National Laboratory; Morley, Steven K [Los Alamos National Laboratory

    2010-12-03

    The Ring current Atmosphere interaction Model with Self-Consistently calculated 3D Magnetic field (RAM-SCB) combines a kinetic model of the ring current with a force-balanced model of the magnetospheric magnetic field to create an inner magnetospheric model that is magnetically self consistent. RAM-SCB produces a wealth of outputs that are valuable to space weather applications. For example, the anisotropic particle distribution of the KeV-energy population calculated by the code is key for predicting surface charging on spacecraft. Furthermore, radiation belt codes stand to benefit substantially from RAM-SCB calculated magnetic field values and plasma wave growth rates - both important for determining the evolution of relativistic electron populations. RAM-SCB is undergoing development to bring these benefits to the space weather community. Data-model validation efforts are underway to assess the performance of the system. 'Virtual Satellite' capability has been added to yield satellite-specific particle distribution and magnetic field output. The code's outer boundary is being expanded to 10 Earth Radii to encompass previously neglected geosynchronous orbits and allow the code to be driven completely by either empirical or first-principles based inputs. These advances are culminating towards a new, real-time version of the code, rtRAM-SCB, that can monitor the inner magnetosphere conditions on both a global and spacecraft-specific level. This paper summarizes these new features as well as the benefits they provide the space weather community.

  15. RAMS管理探究%Research of RAMS Management

    Institute of Scientific and Technical Information of China (English)

    张文泉

    2013-01-01

    Set out the basic meaning and interpret of resource-saved and environment-friendlysociety, RAMS(Reliability, Availability, Maintainability and Safety) and its news deviloping which is form section of Total Quality Management(TQM). It sho-uld stand height pay close attention tothe theory and method of Reliability, Availability, Maintainability and Safety(RAMS). Based on develops of recycle economy, low carbon economy, theoty, methods and management of RAMS has attracted broad attention with increase of serious environmental and climate deterioration. Thus based on the know and study of writer this paper introduces definition of Reliability Availability Maintainability and Safety, discusses the theoty, methods and management of RAMS, inquire into the utilize of theoty, methodsof RAMS in Engineering Management.%从建设资源节约型、环境友好型社会、发展低碳经济、循环经济的内涵与意义出发,RAMS理论方法与管理作为全面质量管理的组成部分及其新进展,大有用武之地,并引起高度重视。基于低碳经济、循环经济的不断发展, RAMS理论方法与管理更具理论和实际意义,从而受到广泛关注。为此,本文以作者的粗浅认识与初步研究为基础,首先介绍可靠性(Reliability)、可用性(Availability)、维修性(Maintainability)与安全性(Safety)的定义内涵及其密切关系,然后探讨RAMS管理的组成内容与实际应用。

  16. RAM: a Relativistic Adaptive Mesh Refinement Hydrodynamics Code

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Wei-Qun; /KIPAC, Menlo Park; MacFadyen, Andrew I.; /Princeton, Inst. Advanced Study

    2005-06-06

    The authors have developed a new computer code, RAM, to solve the conservative equations of special relativistic hydrodynamics (SRHD) using adaptive mesh refinement (AMR) on parallel computers. They have implemented a characteristic-wise, finite difference, weighted essentially non-oscillatory (WENO) scheme using the full characteristic decomposition of the SRHD equations to achieve fifth-order accuracy in space. For time integration they use the method of lines with a third-order total variation diminishing (TVD) Runge-Kutta scheme. They have also implemented fourth and fifth order Runge-Kutta time integration schemes for comparison. The implementation of AMR and parallelization is based on the FLASH code. RAM is modular and includes the capability to easily swap hydrodynamics solvers, reconstruction methods and physics modules. In addition to WENO they have implemented a finite volume module with the piecewise parabolic method (PPM) for reconstruction and the modified Marquina approximate Riemann solver to work with TVD Runge-Kutta time integration. They examine the difficulty of accurately simulating shear flows in numerical relativistic hydrodynamics codes. They show that under-resolved simulations of simple test problems with transverse velocity components produce incorrect results and demonstrate the ability of RAM to correctly solve these problems. RAM has been tested in one, two and three dimensions and in Cartesian, cylindrical and spherical coordinates. they have demonstrated fifth-order accuracy for WENO in one and two dimensions and performed detailed comparison with other schemes for which they show significantly lower convergence rates. Extensive testing is presented demonstrating the ability of RAM to address challenging open questions in relativistic astrophysics.

  17. Use of commercial extenders and alternatives to prevent sperm agglutination for cryopreservation of brown bear semen.

    Science.gov (United States)

    Gomes-Alves, S; Alvarez, M; Nicolas, M; Lopez-Urueña, E; Martínez-Rodríguez, C; Borragan, S; de Paz, P; Anel, L

    2014-08-01

    The objective of this study was to evaluate different bovine and canine commercial semen extenders for cryopreservation of brown bear ejaculates and the effect of semen collection directly into extender on sperm agglutination. Semen samples were obtained by electroejaculation from 13 adult males. In experiment 1, eleven ejaculates from eight bears were used to evaluate Bioxcell and Andromed as extenders, whereas in experiment 2, nine ejaculates from six bears were used to evaluate Triladyl canine, CaniPro, and Extender 2 as extenders. An extender specifically developed for brown bears (Test-Tris-fructose-egg yolk-glycerol, TTF-ULE/bear) served as a control extender in both experiments. After thawing, total and progressive sperm motility and sperm viability were greater (P agglutination in fresh samples (score 0.5 ± 0.2 vs. 1.8 ± 0.4 in diluted and control samples, respectively) with no effect on pre-freeze and post-thawing semen quality. In conclusion, TTF-ULE/bear is the most suitable extender for brown bear semen cryopreservation, but comparable results can be obtained with the commercial extender Andromed. In addition, collection of ejaculates directly in TTF-H extender decreases sperm agglutination in fresh samples.

  18. A comparative study of the morphometry of sperm head components in cattle, sheep, and pigs with a computer-assisted fluorescence method

    Directory of Open Access Journals (Sweden)

    Jesús L Yániz

    2016-01-01

    Full Text Available The aim of this study was to compare the sperm nuclear and acrosomal morphometry of three species of domestic artiodactyls; cattle (Bos taurus, sheep (Ovis aries, and pigs (Sus scrofa. Semen smears of twenty ejaculates from each species were fixed and labeled with a propidium iodide-Pisum sativum agglutinin (PI/PSA combination. Digital images of the sperm nucleus, acrosome, and whole sperm head were captured and analyzed. The use of the PI/PSA combination and CASA-Morph fluorescence-based method allowed the capture, morphometric analysis, and differentiation of most sperm nuclei, acrosomes and whole heads, and the assessment of acrosomal integrity with a high precision in the three species studied. For the size of the head and nuclear area, the relationship between the three species may be summarized as bull > ram > boar. However, for the other morphometric parameters (length, width, and perimeter, there were differences in the relationships between species for sperm nuclei and whole sperm heads. Bull sperm acrosomes were clearly smaller than those in the other species studied and covered a smaller proportion of the sperm head. The acrosomal morphology, small in the bull, large and broad in the sheep, and large, long, and with a pronounced equatorial segment curve in the boar, was species-characteristic. It was concluded that there are clear variations in the size and shape of the sperm head components between the three species studied, the acrosome being the structure showing the most variability, allowing a clear distinction of the spermatozoa of each species.

  19. A comparative study of the morphometry of sperm head components in cattle, sheep, and pigs with a computer-assisted fluorescence method

    Science.gov (United States)

    Yániz, Jesús L; Capistrós, Sara; Vicente-Fiel, Sandra; Hidalgo, Carlos O; Santolaria, Pilar

    2016-01-01

    The aim of this study was to compare the sperm nuclear and acrosomal morphometry of three species of domestic artiodactyls; cattle (Bos taurus), sheep (Ovis aries), and pigs (Sus scrofa). Semen smears of twenty ejaculates from each species were fixed and labeled with a propidium iodide-Pisum sativum agglutinin (PI/PSA) combination. Digital images of the sperm nucleus, acrosome, and whole sperm head were captured and analyzed. The use of the PI/PSA combination and CASA-Morph fluorescence-based method allowed the capture, morphometric analysis, and differentiation of most sperm nuclei, acrosomes and whole heads, and the assessment of acrosomal integrity with a high precision in the three species studied. For the size of the head and nuclear area, the relationship between the three species may be summarized as bull > ram > boar. However, for the other morphometric parameters (length, width, and perimeter), there were differences in the relationships between species for sperm nuclei and whole sperm heads. Bull sperm acrosomes were clearly smaller than those in the other species studied and covered a smaller proportion of the sperm head. The acrosomal morphology, small in the bull, large and broad in the sheep, and large, long, and with a pronounced equatorial segment curve in the boar, was species-characteristic. It was concluded that there are clear variations in the size and shape of the sperm head components between the three species studied, the acrosome being the structure showing the most variability, allowing a clear distinction of the spermatozoa of each species. PMID:27624987

  20. Effects of frozen and liquid hypothermic storage and extender type on calcium homeostasis in relation to viability and ATP content in striped bass (Morone saxatilis) sperm.

    Science.gov (United States)

    Guthrie, H D; Welch, G R; Woods, L C

    2014-05-01

    The effect of hypothermic storage on striped bass sperm calcium homeostasis was determined by Fluo-3 flow cytometry. Calcium homeostasis was defined as the ability of cells to maintain a low concentration of intracellular free calcium as measured by Fluo-3 fluorescence. Sperm were stored frozen in striped bass extender (SBE) and Tris-NaCl medium (T350) modified with 50 mM glycine and 7.5% dimethylsulfoxide and in nonfrozen form diluted 1:3 (vol/vol) in SBE and T350 for 1, 24, and 48 hours at 4 °C in an oxygen atmosphere. Fluo-3 fluorescence was detected in less than 5% of fresh viable sperm cells indicating maintenance of calcium homeostasis. In contrast to sperm in fresh semen, frozen-thawed and nonfrozen sperm cells lost to a considerable extent the ability to maintain low intracellular free calcium even in the absence of exogenous calcium; positive Fluo-3 fluorescence was found in 26% and 39% of thawed sperm frozen in SBE- and T350-based freezing diluents, respectively, and increased (P 0.05) in SBE after 48 hours of nonfrozen storage but decreased (P < 0.05) to 55.7% after 48 hours in T350. Energy status in terms of ATP content, determined by luciferin-luciferase bioluminescence assay, was higher (P < 0.05) in sperm frozen in SBE than in T350 during the first 5 minutes post-thaw and decreased to essentially zero by 15 minutes post-thaw and did not differ among nonfrozen storage treatments. In conclusion, sperm cells impervious to propidium iodide after frozen or nonfrozen storage were unable to maintain low intracellular calcium content. SBE is a better medium than T350 for frozen or nonfrozen storage of striped bass sperm. The inability to regulate intracellular calcium in striped bass sperm may be associated with poor activation of motility after 4 °C storage and cryopreservation. Published by Elsevier Inc.

  1. A Procedure-Spanning Analysis of Plasma Membrane Integrity for Assessment of Cell Viability in Sperm Cryopreservation of Zebrafish Danio rerio.

    Science.gov (United States)

    Yang, Huiping; Daly, Jonathan; Carmichael, Carrie; Matthews, Jen; Varga, Zoltan M; Tiersch, Terrence

    2016-04-01

    The goal of this study was to evaluate plasma membrane integrity and motility for zebrafish sperm quality assessment along the cryopreservation pathway-from sample collection through refrigerated storage, cryoprotectant equilibration, freezing, thawing, and fertilization. The objectives were to: (1) evaluate the effects of osmolality, extender, and refrigerated storage on sperm plasma membrane integrity and motility, and (2) compare cryopreservation of sperm from farm-raised and well-characterized research populations by evaluating motility and membrane integrity of fresh, post-equilibration (before freezing) and post-thaw sperm, and post-thaw fertility. Osmolality, extender, and storage time each influenced sperm motility and membrane integrity. Isotonic osmolality showed the best protection for motility and membrane integrity compared to hypotonic and hypertonic osmolalities. Of the four tested extenders, Hanks' balanced salt solution (HBSS) and Ca(2+)-free HBSS showed the best protection compared with NaCl and glucose, and sperm retained motility and membrane integrity for 24 h of refrigerated storage. Sperm cryopreservation of zebrafish from a farm population (n = 20) and an AB research line (n = 20) showed significant differences in post-thaw fertility (32% ± 18% vs. 73% ± 21%). No differences were found in post-thaw motility, although the farm-raised zebrafish possessed a larger body size, testis weight, and higher fresh motility. Correlation analysis of pooled data did not identify correlations among motility, flow cytometry analysis of membrane integrity and recognizable cells, and post-thaw sperm fertility (p ≥ 0.202). More research is needed to standardize the fertilization conditions especially sperm-to-egg ratio to avoid possible overabundance of sperm to obscure the differences.

  2. The relationship between sperm viability and DNA fragmentation rates

    OpenAIRE

    Mary K. Samplaski; Dimitromanolakis, Apostolos; Lo, Kirk C; Grober, Ethan D.; Mullen, Brendan; Garbens, Alaina; Jarvi, Keith A

    2015-01-01

    Background In humans, sperm DNA fragmentation rates have been correlated with sperm viability rates. Reduced sperm viability is associated with high sperm DNA fragmentation, while conversely high sperm viability is associated with low rates of sperm DNA fragmentation. Both elevated DNA fragmentation rates and poor viability are correlated with impaired male fertility, with a DNA fragmentation rate of > 30% indicating subfertility. We postulated that in some men, the sperm viability assay coul...

  3. Characterisation of Caenorhabditis elegans sperm transcriptome and proteome

    OpenAIRE

    Ma, Xuan; Zhu, Yingjie; Li, Chunfang; Xue, Peng; Zhao, Yanmei; Chen, Shilin; Yang, Fuquan; Miao, Long

    2014-01-01

    Background Although sperm is transcriptionally and translationally quiescent, complex populations of RNAs, including mRNAs and non-coding RNAs, exist in sperm. Previous microarray analysis of germ cell mutants identified hundreds of sperm genes in Caenorhabditis elegans. To take a more comprehensive view on C. elegans sperm genes, here, we isolate highly pure sperm cells and employ high-throughput technologies to obtain sperm transcriptome and proteome. Results First, sperm transcriptome cons...

  4. Etiologies of sperm oxidative stress

    Directory of Open Access Journals (Sweden)

    Parvin Sabeti

    2016-04-01

    Full Text Available Sperm is particularly susceptible to reactive oxygen species (ROS during critical phases of spermiogenesis. However, the level of seminal ROS is restricted by seminal antioxidants which have beneficial effects on sperm parameters and developmental potentials. Mitochondria and sperm plasma membrane are two major sites of ROS generation in sperm cells. Besides, leukocytes including polymer phonuclear (PMN leukocytes and macrophages produce broad category of molecules including oxygen free radicals, non-radical species and reactive nitrogen species. Physiological role of ROS increase the intracellular cAMP which then activate protein kinase in male reproductive system. This indicates that spermatozoa need small amounts of ROS to acquire the ability of nuclear maturation regulation and condensation to fertilize the oocyte. There is a long list of intrinsic and extrinsic factors which can induce oxidative stress to interact with lipids, proteins and DNA molecules. As a result, we have lipid peroxidation, DNA fragmentation, axonemal damage, denaturation of the enzymes, over generation of superoxide in the mitochondria, lower antioxidant activity and finally abnormal spermatogenesis. If oxidative stress is considered as one of the main cause of DNA damage in the germ cells, then there should be good reason for antioxidant therapy in these conditions

  5. Effect of male age on sperm traits and sperm competition success in the guppy (Poecilia reticulata).

    Science.gov (United States)

    Gasparini, C; Marino, I A M; Boschetto, C; Pilastro, A

    2010-01-01

    Deleterious mutations can accumulate in the germline with age, decreasing the genetic quality of sperm and imposing a cost on female fitness. If these mutations also affect sperm competition ability or sperm production, then females will benefit from polyandry as it incites sperm competition and, consequently, minimizes the mutational load in the offspring. We tested this hypothesis in the guppy (Poecilia reticulata), a species characterized by polyandry and intense sperm competition, by investigating whether age affects post-copulatory male traits and sperm competition success. Females did not discriminate between old and young males in a mate choice experiment. While old males produced longer and slower sperm with larger reserves of strippable sperm, compared to young males, artificial insemination did not reveal any effect of age on sperm competition success. Altogether, these results do not support the hypothesis that polyandry evolved in response to costs associated with mating with old males in the guppy.

  6. Boundaries of Ethics, Sperm on the Border –The Globalization of Danish Sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    as sperm donation in parliamentary debates, and in the media, often raises ethical questions. This presentation inquires into how the bending of boundaries by “inappropriate parents”, fertility travelers, private sperm banks and fertility clinics have been part in negotiating the changes of the legislation...... in practice, and thus been part of developing a Danish industry of sperm banking. In this presentation, I will show how Danish sperm has become a global commodity even though, or maybe because the technology has been regulated, and continuously has raised and troubled questions of kinship, sexuality...... of the reasons why Danish fertility clinics have enabled women from Denmark and abroad to turn to sperm donation is, that two of the largest sperm banks in Europe are Danish. In contrast to many other countries where sperm is in demand, the Danish sperm banks are able to provide enough sperm for the national...

  7. Sperm competition promotes diversity of sperm bundles in Ohomopterus ground beetles

    Science.gov (United States)

    Takami, Yasuoki; Sota, Teiji

    2007-07-01

    Diversification of sperm morphology has been investigated in the context of sperm competition, but the adaptive significance of sperm bundles is still unclear. In analyzing 10 taxa of the genus Carabus subgenus Ohomopterus and one related Carabus ground beetles, we found that dimorphic sperm bundles occurred in most species with varied degrees of bimodality, whereas sperm were generally monomorphic. Comparative analyses with phylogenetically independent contrasts revealed that the sizes of large and small sperm bundles evolved more rapidly than, and were not correlated with, the length of sperm, suggesting more intense selection on sperm bundle sizes and their independent responses to different evolutionary forces. The size of large sperm bundles was positively correlated with male genital morphology (pertinent to displacement of rival spermatophores) and postcopulatory guarding duration as well as male body length, suggesting that larger sperm bundles have been favored when the risk of spermatophore displacement is high. Larger sperm bundles may be advantageous because of their ability to migrate more rapidly into the spermatheca. In contrast, no clear association was detected between the small sperm bundle size and mating traits despite its rapid diversification. The present study provides the first record of heteromorphic sperm bundles, the diversity of which may be promoted by sperm competition.

  8. Why small males have big sperm: dimorphic squid sperm linked to alternative mating behaviours

    Directory of Open Access Journals (Sweden)

    Shiba Kogiku

    2011-08-01

    Full Text Available Abstract Background Sperm cells are the target of strong sexual selection that may drive changes in sperm structure and function to maximize fertilisation success. Sperm evolution is regarded to be one of the major consequences of sperm competition in polyandrous species, however it can also be driven by adaptation to the environmental conditions at the site of fertilization. Strong stabilizing selection limits intra-specific variation, and therefore polymorphism, among fertile sperm (eusperm. Here we analyzed reproductive morphology differences among males employing characteristic alternative mating behaviours, and so potentially different conditions of sperm competition and fertilization environment, in the squid Loligo bleekeri. Results Large consort males transfer smaller (average total length = 73 μm sperm to a female's internal sperm storage location, inside the oviduct; whereas small sneaker males transfer larger (99 μm sperm to an external location around the seminal receptacle near the mouth. No significant difference in swimming speed was observed between consort and sneaker sperm. Furthermore, sperm precedence in the seminal receptacle was not biased toward longer sperm, suggesting no evidence for large sperm being favoured in competition for space in the sperm storage organ among sneaker males. Conclusions Here we report the first case, in the squid Loligo bleekeri, where distinctly dimorphic eusperm are produced by different sized males that employ alternative mating behaviours. Our results found no evidence that the distinct sperm dimorphism was driven by between- and within-tactic sperm competition. We propose that presence of alternative fertilization environments with distinct characteristics (i.e. internal or external, whether or not in combination with the effects of sperm competition, can drive the disruptive evolution of sperm size.

  9. Influence of recovery method and centrifugation on epididymal sperm from collared peccaries (Pecari tajacu Linnaeus, 1758).

    Science.gov (United States)

    Bezerra, José Artur Brilhante; da Silva, Andréia Maria; Peixoto, Gislayne Christianne Xavier; da Silva, Mariana de Araújo; Franco de Oliveira, Moacir; Silva, Alexandre Rodrigues

    2014-05-01

    In order to establish protocols for gamete recovery from accidentally killed wild animals, or to take advantage of those slaughtered by captive breeders, we assess the influence of two methods on the recovery of epididymal sperm from collared peccaries, and verify the effect of centrifugation on such gametes. Genitalia from nine animals were used. For each animal, one epididymis was processed by flotation and the other was processed by retrograde flushing, both using a buffered media based on Tris. Following recovery, sperm were evaluated for motility, vigor, viability, functional membrane integrity, and morphology. A 1-mL aliquot of each sample was centrifuged, the supernatant removed, and the pellet suspended and evaluated as fresh samples. The sperm characteristics did not differ between the samples collected by flotation or retrograde flushing (P < 0.05). Centrifugation promoted an increase in head and tail defects, thus reducing the percentage of viable sperm (P < 0.05). No other parameter assessed for both methods was affected by centrifugation. In conclusion, epididymal sperm from collared peccaries can be efficiently collected through flotation or retrograde flushing, but not when either is followed by centrifugation.

  10. Effects of alpha-lipoic acids on sperm membrane integrity during liquid storage of boar semen

    Directory of Open Access Journals (Sweden)

    Laura Parlapan

    2015-05-01

    Full Text Available Preliminary studies have shown that sperm membrane from swine shows high sensitivity to cryopreservation process, causing a dramatic reduction in sperm quality. This has been attributed to the production of reactive oxygen species, that cause lipid peroxidation in sperm membranes. The aim of the present study was to minimize the oxidative attack by adding different concentration of alpha-lipoic acid into the sperm liquid storage at 17ºC for 7 days. Freshly ejaculated boar semen was diluted with Beltsville Thawing Solution (BTS and supplemented with 5 levels of alpha-lipoic  acid (0.015, 0.02, 0.05, 0.1, 0.15 mmol/ml. The membrane integrity was evaluated at days 0, 1, 3, 5 and 7 of liquid preservation, using flow cytometer FACSCanto II (BD Biociencias systems. The experiment indicate that supplementation of alpha-lipoic  acid to the semen liquid storage extender improve sperm membrane

  11. Controlled cooling versus rapid freezing of teratozoospermic semen samples: Impact on sperm chromatin integrity

    Directory of Open Access Journals (Sweden)

    Shivananda N Kalludi

    2011-01-01

    Full Text Available Aim: The present study evaluates the impact of controlled slow cooling and rapid freezing techniques on the sperm chromatin integrity in teratozoospermic and normozoospermic samples. Setting: The study was done in a university infertility clinic, which is a tertiary healthcare center serving the general population. Design: It was a prospective study designed in vitro. Materials and Methods: Semen samples from normozoospermic (N=16 and teratozoospermic (N=13 infertile men were cryopreserved using controlled cooling and rapid freezing techniques. The sperm chromatin integrity was analyzed in fresh and frozen-thawed samples. Statistical Analysis Used: Data were reported as mean and standard error (mean ± SEM of mean. The difference between two techniques was determined by a paired t-test. Results: The freeze-thaw induced chromatin denaturation was significantly (P<0.01 elevated in the post-thaw samples of normozoospermic and teratozoospermic groups. Compared to rapid freezing, there was no difference in the number of red sperms (with DNA damage by the controlled slow cooling method in both normozoospermic and teratozoospermic groups. Freeze-thaw induced sperm chromatin denaturation in teratozoospermic samples did not vary between controlled slow cooling and rapid freezing techniques. Conclusions: Since the controlled slow cooling technique involves the use of expensive instrument and is a time consuming protocol, rapid freezing can be a good alternative technique for teratozoospermic and normozoospermic samples when sperm DNA damage is a concern.

  12. Pregnancy rates in cattle with cryopreserved sexed sperm: effects of sperm numbers per inseminate and site of sperm deposition.

    Science.gov (United States)

    Seidel, G E; Schenk, J L

    2008-04-01

    In six field trials, doses between 1.0 and 6.0 x 10(6) total sexed, frozen-thawed sperm were inseminated into the uterine body or bilaterally into the uterine horns of heifers and nursing Angus cows 12 or 24h after observed estrus. Except for one comparison in one trial in which uterine body insemination was slightly superior (P0.1) difference between sites of semen deposition. Additionally, except for one small study with limited numbers, there was essentially no difference in pregnancy rates in the range between 1.5 and 6 x 10(6) sexed, frozen-thawed sperm per inseminate. Pregnancy rates with smaller doses of sexed sperm averaged about 75% of controls of 20 x 10(6) total frozen-thawed, unsexed sperm. While 1.0 x 10(6) sexed, frozen-thawed sperm per insemination dose resulted in decreased pregnancy rates compared to larger doses, the lesser fertility with sexed sperm could not be compensated by increasing sperm numbers in the range of 1.5-6 x 10(6) sperm per dose. Pregnancy rates with 2 x 10(6) sexed, frozen-thawed sperm per dose were not markedly less than control pregnancy rates with 20 x 10(6) frozen-thawed unsexed sperm/dose in well-managed herds.

  13. Control of sperm concentration is necessary for standardization of sperm cryopreservation in aquatic species: evidence from sperm agglutination in oysters.

    Science.gov (United States)

    Dong, Qiaoxiang; Huang, Changjiang; Tiersch, Terrence R

    2007-02-01

    A lack of standardization in sperm cryopreservation of aquatic organisms is one of the main reasons for inconsistency observed among various studies. In particular, there have been few attempts to standardize sperm concentration during procedural optimization. This study was intended to call attention to sperm concentration standardization through research of sperm agglutination in Pacific oysters Crassostrea gigas. Sperm agglutination after thawing is a relatively frequent phenomenon observed for various aquatic species, especially when sub-optimal cryopreservation protocols are used; however, no systematic attempts have been made to explain this phenomenon. The present study evaluated various factors affecting sperm agglutination of thawed samples from diploid and tetraploid Pacific oysters, and is the first detailed report addressing the sperm agglutination phenomenon of thawed samples from any aquatic organism. Agglutination of oyster sperm was classified into six levels with a scale ranging from 0 (homogenous suspension) to 5 (well-developed "noodles"). It was found that agglutination in thawed samples was mainly due to the lack of sufficient cryoprotectant for a specific sperm concentration. Interestingly, high levels of agglutination did not necessarily lead to low fertilization. On the contrary, some sperm cells appeared to gain protection from the formation of peripheral agglutination within 0.5-ml French straws. The exact mechanism of sperm agglutination remains unclear. However, morphological examination of cross sections of the noodles (agglutination level 5) indicated at least two forms of agglutination (formed with and without cryoprotectant) which could be used as a tool to understand the cryopreservation process within the micro-environment of the straw. Furthermore, the fact that the level of sperm agglutination was directly determined by sperm concentration, in addition to the type of cryoprotectant, cryoprotectant concentration, and cooling and

  14. Irradiation of fresh fish

    Science.gov (United States)

    Yueh-jen, Yen; Jin-lai, Zhou; Shao-chun, Lai

    Occasionally, in China, marine products can not be provided for the markets in good quality, for during the time when they are being transported from the sea port to inland towns or even at the time when they are unloaded from the ship, they are beginning to spoil. Obviously, it is very important that appropiate measures should be taken to prevent them from decay. Our study has proved that the shelf life of fresh Flatfish (Cynoglossue robustus) and Silvery pomfret (stromateoides argenteus), which, packed in sealed containers, are irradiated by 1.5 kGy, 2.2 kGy and 3.0 kGy, can be stored for about 13-26 days at 3° - 5° C.

  15. Effect of gibberellic acid on the quality of sperm and in vitro fertilization outcome in adult male rats

    Directory of Open Access Journals (Sweden)

    Mohammadreza Hosseinchi

    2014-12-01

    Full Text Available Gibberellic acid (GA3 is a group of plant hormones identified in various plants. The aim of this study was to determine the effects of GA3 on sperm parameters and in vitro fertilization (IVF. Fifty six adult male rats were divided into seven groups as, control, treatment and sham. Following 15, 30 and 45 days of GA3 and methanol alcohol (MA administration, rats were euthanized and epididymis tail was transferred to human tubular fluid (HTF medium containing 4 mg mL-1 bovine serum albumin (BSA .Total number of sperms, the percentage of live sperms, immature sperms and sperms with damaged chromatin and IVF were examined. The oocytes were obtained from immature rats after the injection of pregnant mare's serum (PMSG and human chorionic gonadotropin (HCG hormones. Human tubular fluid was used as the fertilization medium and zygotes transferred to fresh 1-cell rat embryos culture medium (mR1ECM to reach the blastocyst stage. This study showed that GA3 could decrease the number of total sperms on days 30 and 45 in treated group comparison with the control and sham groups. Additionally, GA3 increased the immature sperms and sperms with damaged chromatin. The percentage of fertilization, two-cell embryos and blastocyst resulting from the treatment group on days 30 and 45 also decreased and showed significant differences with the control and sham groups (p < 0.05. The results obtained from this study indicated that the oral use of GA3 could reduce the fertility in rats by influencing the sperm number and the quality of sperm’s chromatins.

  16. Etiology and Evaluation of Sperm Chromatin Anomalies

    Directory of Open Access Journals (Sweden)

    Marziyeh Tavalaee

    2008-01-01

    Full Text Available Evidence suggests that human sperm chromatin anomalies adversely affect reproductive outcomesand infertile men possess substantially amount of sperm with chromatin anomalies than fertilemen.Routine semen analysis evaluates parameters such as sperm motility and morphology, but doesnot examine the nuclear DNA integrity of spermatozoa. It has been suggested that altered nuclearchromatin structure or damaged DNA in spermatozoa could modify the special cellular functionsof human spermatozoa, and thereby affect the fertility potential. Intra-cytoplasmic sperm injection(ICSI bypass the barriers to fertilization for such a sperm, then the effect of chromatin anomalies onthe development remains a concern. Therefore, it is essential to develop and use accurate diagnostictests, which may provide better prognostic capabilities than the standard sperm assessments. Thisreview discusses our current understanding of the structure and organization of sperm DNA,the different procedures for assessment of sperm chromatin anomalies including comet assay,Chromomycin A3 (CMA3, sperm chromatin structure assay (SCSA, acridine orange test (AOT,terminal TdT-mediated dUTP-nick-end labelling (TUNEL assay, aniline blue and sperm chromatindispersion (SCD test and the impact of chromatin anomalies on reproductive outcome.

  17. Treatment by testicular sperm extraction and intracytoplasmic sperm injection of 65 azoospermic patients with non-mosaic Klinefelter syndrome with birth of 17 healthy children.

    Science.gov (United States)

    Madureira, C; Cunha, M; Sousa, M; Neto, A P; Pinho, M J; Viana, P; Gonçalves, A; Silva, J; Teixeira da Silva, J; Oliveira, C; Ferraz, L; Dória, S; Carvalho, F; Barros, A

    2014-07-01

    The aim of this work was to present the clinical and embryological outcomes of 65 azoospermic patients with non-mosaic Klinefelter syndrome (KS), treated by testicular sperm extraction (TESE), followed by intracytoplasmic sperm injection (ICSI), either with fresh or cryopreserved testicular spermatozoa. In total, spermatozoa were recovered in 25/65 (38.5%) of the cases. Of the 48 patients who choose to perform TESE followed by ICSI using fresh testicular spermatozoa (treatment TESE), spermatozoa was recovered in 19 patients (40%), with birth of 12 newborn. Of the 17 patients who choose to perform TESE followed by testicular sperm cryopreservation, spermatozoa were recovered in six patients (35%), with birth of one child. Of the patients who performed treatment TESE, nine went for a new cycle using cryopreserved spermatozoa. Of these, five patients had a previous failed treatment cycle (two patients, three newborn) and four with a previous success went for a new cycle (one patient, one newborn). Overall, the embryological and clinical rates were as follows: 52% of fertilization, 41% of blastocyst, 27% of implantation, 39% of live birth delivery and 47% of newborn. Of the 16 clinical pregnancies, 14 had a successful delivery (12 girls and 5 boys). The 17 newborns had a mean gestation time of 37.2 weeks (35.3% pre-term) and a mean newborn weight of 2781.3 g (37.5% low weight). Comparisons between cycles with fresh and frozen-thaw spermatozoa revealed higher fertilization and clinical pregnancy rates with fresh spermatozoa, with no differences regarding implantation or newborn rates. Of the 17 newborns, no abnormal karyotypes (n = 3) or numerical abnormalities in chromosomes 13, 18, 21, X and Y (n = 14) as evaluated by Multiplex Ligation-dependent Probe Amplification were observed. In conclusion, this study presents further data that reassures that men with KS have no increased risk of transmitting their genetic problem to the offspring.

  18. Sperm DNA assays and their relationship to sperm motility and morphology in bulls (Bos Taurus).

    Science.gov (United States)

    Serafini, Rosanna; Romano, Juan E; Varner, Dickson D; Di Palo, Rossella; Love, Charles C

    2015-08-01

    The relationship among sperm DNA assays in bulls with different sperm motility and morphology measures has not been reported. The objectives of the present study were to (1) describe Comet assay measures and examine their repeatability (inter- and intra-assay); (2) compare sperm DNA quality assays (i.e., Sperm Chromatin Structure Assay-SCSA; alkaline and neutral Comet assays and Sperm Bos Halomax assay-SBH) in two groups of bulls selected on either greater and lesser sperm motility and morphology (greater compared with lesser); (3) determine the relationship among DNA assays and sperm motility and morphology values. Inter-assay repeatability was greater for the neutral Comet assay as compared to the alkaline Comet assay. Intra-assay repeatability was greater than inter-assay repeatability for both Comet assays. Comet assay dimension measures and percentage tail DNA were the most repeatable for both Comet assays. Among sperm DNA quality assays, only SCSA measures and neutral Comet assay Ghosts (% Ghosts), head diameter and area, and comet area were different between greater and lesser sperm quality groups (P<0.05). The SCSA measures were inversely correlated with neutral Comet head measures (diameter, area, and intensity) and positively with percentage Ghosts (P<0.05). The % Ghosts and COMP-αt were correlated with some measures of sperm morphology and sperm motility. The neutral Comet assay was more appropriate for sperm evaluation than the alkaline Comet assay for distinguishing among groups with different sperm quality.

  19. Sperm motility under exposure of hydrogen dioxide

    Directory of Open Access Journals (Sweden)

    V. V. Evdokimov

    2015-01-01

    Full Text Available The paper contains research data on the effect of low concentrations of hydrogen dioxide on human sperm motility and specific enzyme activity of sperms of glyceraldehyde-3-phosphate dehydrogenase. It is shown that incubation of sperms with hydrogen dioxide in a low concentration leads to a change and motility in sperm and activity of sperm enzyme. Intensity of observed effect depended on the concentration of hydrogen dioxide: active mobility increased by 17–19 % and the total mobility – 11 %. Motility changes in sperms were accompanied by increased activity of glyceraldehyde-3-phosphate dehydrogenase by 24 %, in normozoospermia response was higher than in pathozoospermia and also depended on the concentration of hydrogen dioxide. The use of sperm analyzer enabled revealing changes in the diapason of different speeds of the active fraction of sperm, which have been observed in the first 15 min of incubation with hydrogen dioxide. A possible mechanism of action of the detected effect is discussed. Reactive oxygen species easily oxidize enzyme for glyceraldehyde-3-phosphate dehydrogenase of sperms, which leads to a loss of sperm motility, for example, in varicocele. Initially low enzyme activity in varicocele (pathozoospermia may be associated with the suppression of sperm antioxidant defense. Addition of low concentrations of hydrogen dioxide into sperm samples leads to an increase in the concentration of reduced glutathione in a cell. Increase of sperm motility in this case can serve as an indicator of normal operation of the cellular antioxidant defense system. Obtained experimental results provide a background for their introduction into clinical practice in the program of assisted reproductive technologies. 

  20. Sperm motility under exposure of hydrogen dioxide

    Directory of Open Access Journals (Sweden)

    V. V. Evdokimov

    2015-04-01

    Full Text Available The paper contains research data on the effect of low concentrations of hydrogen dioxide on human sperm motility and specific enzyme activity of sperms of glyceraldehyde-3-phosphate dehydrogenase. It is shown that incubation of sperms with hydrogen dioxide in a low concentration leads to a change and motility in sperm and activity of sperm enzyme. Intensity of observed effect depended on the concentration of hydrogen dioxide: active mobility increased by 17–19 % and the total mobility – 11 %. Motility changes in sperms were accompanied by increased activity of glyceraldehyde-3-phosphate dehydrogenase by 24 %, in normozoospermia response was higher than in pathozoospermia and also depended on the concentration of hydrogen dioxide. The use of sperm analyzer enabled revealing changes in the diapason of different speeds of the active fraction of sperm, which have been observed in the first 15 min of incubation with hydrogen dioxide. A possible mechanism of action of the detected effect is discussed. Reactive oxygen species easily oxidize enzyme for glyceraldehyde-3-phosphate dehydrogenase of sperms, which leads to a loss of sperm motility, for example, in varicocele. Initially low enzyme activity in varicocele (pathozoospermia may be associated with the suppression of sperm antioxidant defense. Addition of low concentrations of hydrogen dioxide into sperm samples leads to an increase in the concentration of reduced glutathione in a cell. Increase of sperm motility in this case can serve as an indicator of normal operation of the cellular antioxidant defense system. Obtained experimental results provide a background for their introduction into clinical practice in the program of assisted reproductive technologies. 

  1. THE EFFECT OF INSULIN LIKE GROWTH FACTOR – I COMPLEX TO CORELATION MALONDIALDEHIDE CONCENTRATE WITH INTAC MEMBRAN PLASMA OF SPERM

    Directory of Open Access Journals (Sweden)

    Suherni Susilowati

    2008-12-01

    Full Text Available The aim of this research was study of Insulin Like Growth -I Complex to level of malondialdehide (MDA and percentage of intact membrane plasm. Report of several species suggest that seminal plasma contains factors that may influence sperm viability. Seminal plasma was reported to be important for maintaining spermatozoa motility in bull and ram. For improving ram sperm viability and for increasing the resistance of boar spermatozoa to cold shock damage. IGF-I has been identified in the testis, where secreted by Leydig and Sertoli cells. Collection of semen by using artificial vagina from male goats and then characterized of motility and viability of spermatozoa. Identification of Insulin Like Growth Factor –I Complex were done by Native-PAGE 12% techniques, after several bands were identified and was done isolate of first band protein Insulin Like Growth Factor-I Complex. Insulin Like Growth Factor – I (IGF-I Complex seminal plasma have molecular weight of 150,288 kDa, respectively. Sperm preparation for this research by centrifugation of semen with Bracket and Oliphan’s medium. These sperm preparation were characterized of motility, viability. Then after sperm preparation divided into three groups. The group I, added with Bracket and Oliphan’t medium, group II added with Bracket Oliphan’t and IGF – I Complex medium, group III added with IGF – I Complex medium and then were exposed 45 minutes for incubation and then percentage of MPU and level of MDA was evaluated. The result of this research showed that percentage of level MDA has significant different between group I,group II and group III, (p<0,05. The percentage of MPU sperm has significant different between groups I, group II and group III (p<0,05. The highest of percentage MPU was found in group III and the lower of level MDA was found in group III. Regresion test showed that negative correlation between level of MDA and percentage of MPU sperm.

  2. Modelling of composites materials and structures under hydrodynamic ram loading

    OpenAIRE

    Deletombe, E.

    2013-01-01

    The impact of high speed/high energy projectiles - possibly ballistic ones - and the occurrence of an hydraulic ram (HRAM) event in fuel tanks, constitutes a threat which is often legitimized if not always compulsory to consider for aircraft safety. To prevent from such an eventuality, the impact hardening through armouring the structure is an ultimate solution which is hardly acceptable in aeronautics for obvious mass penalty reasons. The reduction of fuel tanks vulnerability then turns to b...

  3. ON THE STRUCTURES OF RAM-BASED CHINESE CHARACTER LIBRARIES

    Institute of Scientific and Technical Information of China (English)

    钱培德

    1991-01-01

    The Chinese character library is one of the important data structures in the Chinese information Processing system.The behavior of the whole system depends directly on the reasonableness of design for its structure.This paper expounds the structures of RAM-based Chinese character libraries,static and dynamic ,The paper offers a descriptive method for this behavior and inquires into some algorithms related to the structures mentioned above.

  4. Tracing ram-pressure stripping with warm molecular hydrogen emission

    Energy Technology Data Exchange (ETDEWEB)

    Sivanandam, Suresh [Dunlap Institute for Astronomy and Astrophysics, University of Toronto, Rm 101, 50 St. George Street, Toronto, ON M5S 3H4 (Canada); Rieke, Marcia J.; Rieke, George H., E-mail: sivanandam@dunlap.utoronto.ca [Steward Observatory, University of Arizona, 933 North Cherry Avenue, Tucson, AZ 85721 (United States)

    2014-12-01

    We use the Spitzer Infrared Spectrograph to study four infalling cluster galaxies with signatures of ongoing ram-pressure stripping. H{sub 2} emission is detected in all four, and two show extraplanar H{sub 2} emission. The emission usually has a warm (T ∼ 115-160 K) and a hot (T ∼ 400-600 K) component that is approximately two orders of magnitude less massive than the warm one. The warm component column densities are typically 10{sup 19} to 10{sup 20} cm{sup –2} with masses of 10{sup 6} to 10{sup 8} M {sub ☉}. The warm H{sub 2} is anomalously bright compared with normal star-forming galaxies and therefore may be excited by ram-pressure. In the case of CGCG 97-073, the H{sub 2} is offset from the majority of star formation along the direction of the galaxy's motion in the cluster, suggesting that it is forming in the ram-pressure wake of the galaxy. Another galaxy, NGC 4522, exhibits a warm H{sub 2} tail approximately 4 kpc in length. These results support the hypothesis that H{sub 2} within these galaxies is shock-heated from the interaction with the intracluster medium. Stripping of dust is also a common feature of the galaxies. For NGC 4522, where the distribution of dust at 8 μm is well resolved, knots and ripples demonstrate the turbulent nature of the stripping process. The Hα and 24 μm luminosities show that most of the galaxies have star-formation rates comparable to similar mass counterparts in the field. Finally, we suggest a possible evolutionary sequence primarily related to the strength of ram-pressure that a galaxy experiences to explain the varied results observed in our sample.

  5. Ram-pressure feeding of supermassive black holes

    Science.gov (United States)

    Poggianti, Bianca M.; Jaffé, Yara L.; Moretti, Alessia; Gullieuszik, Marco; Radovich, Mario; Tonnesen, Stephanie; Fritz, Jacopo; Bettoni, Daniela; Vulcani, Benedetta; Fasano, Giovanni; Bellhouse, Callum; Hau, George; Omizzolo, Alessandro

    2017-08-01

    When a supermassive black hole at the centre of a galaxy accretes matter, it gives rise to a highly energetic phenomenon: an active galactic nucleus. Numerous physical processes have been proposed to account for the funnelling of gas towards the galactic centre to feed the black hole. There are also several physical processes that can remove gas from a galaxy, one of which is ram-pressure stripping by the hot gas that fills the space between galaxies in galaxy clusters. Here we report that six out of a sample of seven ‘jellyfish’ galaxies—galaxies with long ‘tentacles’ of material that extend for dozens of kiloparsecs beyond the galactic disks—host an active nucleus, and two of them also have galactic-scale ionization cones. The high incidence of nuclear activity among heavily stripped jellyfish galaxies may be due to ram pressure causing gas to flow towards the centre and triggering the activity, or to an enhancement of the stripping caused by energy injection from the active nucleus, or both. Our analysis of the galactic position and velocity relative to the cluster strongly supports the first hypothesis, and puts forward ram pressure as another possible mechanism for feeding the central supermassive black hole with gas.

  6. Measurement and analysis of the water hammer in ram pump

    Indian Academy of Sciences (India)

    W SOBIESKI; D GRYGO; S LIPINSKI

    2016-11-01

    This paper presents the results of experimental research of the phenomena occurring in water ram during a single cycle of its operation. Apart from a brief introduction and description of the test stand and data recording system, the work includes a broad interpretation of the obtained results. Based on the pressure waveforms recorded in two characteristic zones of the device and its detailed analysis, the single cycle of waterram is divided into three main stages: acceleration, pumping, and backflow. The waveforms of phenomena in each of these steps were considered separately. In discussion, some of the issues were supported with additional measurements, including Fourier analysis of signals from the electronic pressure transducers. The main topic ofdiscussion based on the results recorded for the impulse valve, is supplemented by the comments that take into account the results obtained for the impulse valve (flap check valve) and for the others two (self-made) impulse valves. In the final part, in a graphic form presented is the interpretation of the phenomena occurring during one work cycle of water ram. The motivation of this work was to supplement the knowledge concerning the water hammer waveform in ram pump.

  7. Tracing Ram-Pressure Stripping with Warm Molecular Hydrogen Emission

    CERN Document Server

    Sivanandam, Suresh; Rieke, George H

    2014-01-01

    We use the Spitzer Infrared Spectrograph (IRS) to study four infalling cluster galaxies with signatures of on-going ram-pressure stripping. H$_2$ emission is detected in all four; two show extraplanar H$_2$ emission. The emission usually has a warm (T $\\sim$ $115 - 160$K) and a hot (T $\\sim$ 400 $-$ 600K) component that is approximately two orders of magnitude less massive than the warm one. The warm component column densities are typically $10^{19} - 10^{20}$ cm$^{-2}$ with masses of $10^6 - 10^8 M_\\odot$. The warm H$_2$ is anomalously bright compared with normal star-forming galaxies and therefore may be excited by ram-pressure. In the case of CGCG 97-073, the H$_2$ is offset from the majority of star formation along the direction of the galaxy's motion in the cluster, suggesting it is forming in the ram-pressure wake of the galaxy. Another galaxy, NGC 4522, exhibits a warm H$_2$ tail approximately 4 kpc in length. These results support the hypothesis that H$_2$ within these galaxies is shock-heated from th...

  8. MapReduce Parallel Cuckoo Hashing and Oblivious RAM Simulations

    CERN Document Server

    Goodrich, Michael T

    2010-01-01

    We present an efficient algorithm for performing cuckoo hashing in the MapReduce parallel model of computation and we show how this result in turn leads to improved methods for performing data-oblivious RAM simulations. Our contributions involve a number of seemingly unrelated new results, including: a parallel MapReduce cuckoo hashing algorithm that runs in O(log n) time and uses O(n) total work, with very high probability a reduction of data-oblivious simulation of sparse-streaming MapReduce algorithms to oblivious sorting an external-memory data-oblivious sorting algorithm using O((N/B) log^2_(M/B) (N/B)) I/Os constant-memory data-oblivious RAM simulation with O(log^2 n) amortized time overhead, with very high probability, or with expected O(log2 n) amortized time overhead and better constant factors sublinear-memory data-oblivious RAM simulation with O(n^nu) private memory and O(log n) amortized time overhead, with very high probability, for constant nu > 0. This last result is, in fact, the main result o...

  9. Ram pressure statistics for bent tail radio galaxies

    CERN Document Server

    Mguda, Zolile; van der Heyden, Kurt; Gottlöber, Stefan; Cress, Catherine; Vaisanen, Petri; Yepes, Gustavo

    2014-01-01

    In this paper we use the MareNostrum Universe Simulation, a large scale, hydrodynamic, non-radiative simulation in combination with a simple abundance matching approach to determine the ram pressure statistics for bent radio sources (BRSs). The abundance matching approach allows us to determine the locations of all galaxies with stellar masses $> 10^{11} MSol$ in the simulation volume. Assuming ram pressure exceeding a critical value causes bent morphology, we compute the ratio of all galaxies exceeding the ram pressure limit (RPEX galaxies) relative to all galaxies in our sample. According to our model 50% of the RPEX galaxies at $z = 0$ are found in clusters with masses larger than $10^{14.5}MSol$ the other half resides in lower mass clusters. Therefore, the appearance of bent tail morphology alone does not put tight constraints on the host cluster mass. In low mass clusters, $M 10^{15}Msol$ they can be found at distances up to 1.5Mpc. Only clusters with masses $> 10^{15}MSol $ are likely to host more than...

  10. Boar seminal plasma exosomes maintain sperm function by infiltrating into the sperm membrane.

    Science.gov (United States)

    Du, Jian; Shen, Jian; Wang, Yuanxian; Pan, Chuanying; Pang, Weijun; Diao, Hua; Dong, Wuzi

    2016-09-13

    Seminal plasma ingredients are important for maintenance of sperm viability. This study focuses on the effect of boar seminal plasma exosomes on sperm function during long-term liquid storage. Boar seminal plasma exosomes had typical nano-structure morphology as measured by scanning electron microscopy (SEM) and molecular markers such as AWN, CD9 and CD63 by western blot analysis. The effect on sperm parameters of adding different ratio of boar seminal plasma exosomes to boar sperm preparations was analyzed. Compared to the diluent without exosomes, the diluent with four times or sixteen times exosomes compared to original semen had higher sperm motility, prolonged effective survival time, improved sperm plasma membrane integrity (p membrane of sperm head which could improve sperm plasma membrane integrity.

  11. Social imaginaries, sperm and whiteness

    DEFF Research Database (Denmark)

    Andreassen, Rikke

    2017-01-01

    often is associated with nineteenth-century racial science, can surface in contemporary times. Yet the article also reveals how colonial and racial legacies of whiteness tend to be erased from narratives of fertility, as the (neoliberal) focus on the individual as consumer frames reproduction......This article analyses narratives about so-called Viking babies and Viking sperm. Over the last few years an increasing number of British single women and lesbian couples have been creating families by becoming pregnant with Danish donor sperm, termed ‘Viking sperm’. Through analyses of British...... media coverage of these new families, this article explores contemporary constructions of race, especially whiteness, and gender, and shows how imaginaries of Vikings, genes and white superiority circulate in British media and among British mothers. The article illustrates how a racial discourse, which...

  12. Sperm navigation in complex environments

    Science.gov (United States)

    Olson, Sarah

    2016-11-01

    Sperm can swim in a variety of environments, interacting with chemicals and other proteins in the fluid. Some of these extra proteins or cells may act as friction, possibly preventing or enhancing forward progression of swimmers. The homogenized fluid flow is assumed to be governed by the incompressible Brinkman equation, where a friction term with a resistance parameter represents a sparse array of obstacles. Representing the swimmers with a centerline approximation, we employ regularized fundamental solutions to investigate swimming speeds, trajectories, and interactions of swimmers. Asymmetric waveforms due to an increase in flagellar calcium is known to be important for sperm to reach and fertilize the egg. The trajectories of hyperactivated swimmers are found to have a decreased path curvature. Although attraction of two swimmers is more efficient in the Stokes regime, we find that attraction does not occur for larger resistance. Additionally, we study interactions of swimmers in a channel. NSF DMS 1413110.

  13. Hypercholesterolemia Impaired Sperm Functionality in Rabbits

    Science.gov (United States)

    Monclus, Maria A.; Cabrillana, Maria E.; Clementi, Marisa A.; Espínola, Leandro S.; Cid Barría, Jose L.; Vincenti, Amanda E.; Santi, Analia G.; Fornés, Miguel W.

    2010-01-01

    Hypercholesterolemia represents a high risk factor for frequent diseases and it has also been associated with poor semen quality that may lead to male infertility. The aim of this study was to analyze semen and sperm function in diet-induced hypercholesterolemic rabbits. Twelve adult White New Zealand male rabbits were fed ad libitum a control diet or a diet supplemented with 0.05% cholesterol. Rabbits under cholesterol-enriched diet significantly increased total cholesterol level in the serum. Semen examination revealed a significant reduction in semen volume and sperm motility in hypercholesterolemic rabbits (HCR). Sperm cell morphology was seriously affected, displaying primarily a “folded head”-head fold along the major axe-, and the presence of cytoplasmic droplet on sperm flagellum. Cholesterol was particularly increased in acrosomal region when detected by filipin probe. The rise in cholesterol concentration in sperm cells was determined quantitatively by Gas chromatographic-mass spectrometric analyses. We also found a reduction of protein tyrosine phosphorylation in sperm incubated under capacitating conditions from HCR. Interestingly, the addition of Protein Kinase A pathway activators -dibutyryl-cyclic AMP and iso-butylmethylxanthine- to the medium restored sperm capacitation. Finally, it was also reported a significant decrease in the percentage of reacted sperm in the presence of progesterone. In conclusion, our data showed that diet-induced hypercholesterolemia adversely affects semen quality and sperm motility, capacitation and acrosomal reaction in rabbits; probably due to an increase in cellular cholesterol content that alters membrane related events. PMID:20976152

  14. Chicken sperm transcriptome profiling by microarray analysis.

    Science.gov (United States)

    Singh, R P; Shafeeque, C M; Sharma, S K; Singh, R; Mohan, J; Sastry, K V H; Saxena, V K; Azeez, P A

    2016-03-01

    It has been confirmed that mammalian sperm contain thousands of functional RNAs, and some of them have vital roles in fertilization and early embryonic development. Therefore, we attempted to characterize transcriptome of the sperm of fertile chickens using microarray analysis. Spermatozoal RNA was pooled from 10 fertile males and used for RNA preparation. Prior to performing the microarray, RNA quality was assessed using a bioanalyzer, and gDNA and somatic cell RNA contamination was assessed by CD4 and PTPRC gene amplification. The chicken sperm transcriptome was cross-examined by analysing sperm and testes RNA on a 4 × 44K chicken array, and results were verified by RT-PCR. Microarray analysis identified 21,639 predominantly nuclear-encoded transcripts in chicken sperm. The majority (66.55%) of the sperm transcripts were shared with the testes, while surprisingly, 33.45% transcripts were detected (raw signal intensity greater than 50) only in the sperm and not in the testes. The greatest proportion of up-regulated transcripts were responsible for signal transduction (63.20%) followed by embryonic development (56.76%) and cell structure (56.25%). Of the 20 most abundant transcripts, 18 remain uncharacterized, whereas the least abundant genes were mostly associated with the ribosome. These findings lay a foundation for more detailed investigations on sperm RNAs in chickens to identify sperm-based biomarkers for fertility.

  15. Nontransitivity of sperm precedence in Drosophila.

    Science.gov (United States)

    Clark, A G; Dermitzakis, E T; Civetta, A

    2000-06-01

    Sperm competition is an important component of fitness in Drosophila, but we still do not have a clear understanding of the unit of selection that is relevant to sperm competition. Here we demonstrate that sperm competitive ability is not a property of the sperm haplotype, but rather of the diploid male's genotype. Then we test whether the relative sperm competitive ability of males can be ranked on a linear array or whether competitive ability instead depends on particular pairwise contests among males. Sperm precedence of six chromosome-extracted lines was tested against three different visible marker lines (cn bw, bwD, and Cy), and the rank order of the six lines differed markedly among the mutant lines. Population genetic theory has shown that departures from transitivity of sperm precedence may be important to the maintenance of polymorphism for genes that influence sperm competitive ability. The nontransitivity seen in sperm precedence should theoretically increase the opportunity for polymorphism in genes that influence this phenotype.

  16. Hypercholesterolemia impaired sperm functionality in rabbits.

    Directory of Open Access Journals (Sweden)

    Tania E Saez Lancellotti

    Full Text Available Hypercholesterolemia represents a high risk factor for frequent diseases and it has also been associated with poor semen quality that may lead to male infertility. The aim of this study was to analyze semen and sperm function in diet-induced hypercholesterolemic rabbits. Twelve adult White New Zealand male rabbits were fed ad libitum a control diet or a diet supplemented with 0.05% cholesterol. Rabbits under cholesterol-enriched diet significantly increased total cholesterol level in the serum. Semen examination revealed a significant reduction in semen volume and sperm motility in hypercholesterolemic rabbits (HCR. Sperm cell morphology was seriously affected, displaying primarily a "folded head"-head fold along the major axe-, and the presence of cytoplasmic droplet on sperm flagellum. Cholesterol was particularly increased in acrosomal region when detected by filipin probe. The rise in cholesterol concentration in sperm cells was determined quantitatively by Gas chromatographic-mass spectrometric analyses. We also found a reduction of protein tyrosine phosphorylation in sperm incubated under capacitating conditions from HCR. Interestingly, the addition of Protein Kinase A pathway activators -dibutyryl-cyclic AMP and iso-butylmethylxanthine- to the medium restored sperm capacitation. Finally, it was also reported a significant decrease in the percentage of reacted sperm in the presence of progesterone. In conclusion, our data showed that diet-induced hypercholesterolemia adversely affects semen quality and sperm motility, capacitation and acrosomal reaction in rabbits; probably due to an increase in cellular cholesterol content that alters membrane related events.

  17. Microdissection testicular sperm extraction: an update

    Institute of Scientific and Technical Information of China (English)

    Ali A Dabaja; Peter N Schlegel

    2013-01-01

    Patients with non-obstructive azoospermia (NOA) were once considered to be infertile with few treatment options due to the absence of sperm in the ejaculate.In the last two decades,the advent of intracytoplasmic sperm injection (ICSI),and the application of various testicular sperm retrieval techniques,including fine needle aspiration (FNA),conventional testicular sperm extraction (TESE) and microdissection testicular sperm extraction (micro-TESE) have revolutionized treatment in this group of men.Because most men with NOA will have isolated regions of spermatogenesis within the testis,studies have illustrated that sperm can be retrieved in most men with NOA,including Klinefelter's syndrome (KS),prior history of chemotherapy and cryptorchidism.Micro-TESE,when compared with conventional TESE has a higher sperm retrieval rate (SRR) with fewer postoperative complications and negative effects on testicular function.In this article,we will compare the efficacy of the different procedures of sperm extraction,discuss the medical treatment and the role of testosterone optimization in men with NOA and describe the micro-TESE surgical technique.Furthermore,we will update our overall experience to allow counseling on the prognosis of sperm retrieval for the specific subsets of NOA.

  18. Intracellular pH in sperm physiology.

    Science.gov (United States)

    Nishigaki, Takuya; José, Omar; González-Cota, Ana Laura; Romero, Francisco; Treviño, Claudia L; Darszon, Alberto

    2014-08-01

    Intracellular pH (pHi) regulation is essential for cell function. Notably, several unique sperm ion transporters and enzymes whose elimination causes infertility are either pHi dependent or somehow related to pHi regulation. Amongst them are: CatSper, a Ca(2+) channel; Slo3, a K(+) channel; the sperm-specific Na(+)/H(+) exchanger and the soluble adenylyl cyclase. It is thus clear that pHi regulation is of the utmost importance for sperm physiology. This review briefly summarizes the key components involved in pHi regulation, their characteristics and participation in fundamental sperm functions such as motility, maturation and the acrosome reaction.

  19. SPERM DNA INTEGRITY IN BUFFALO, BULL AND STALLION

    OpenAIRE

    Serafini, Rosanna

    2015-01-01

    The interest in sperm DNA integrity evaluation and its relationship to subfertility and infertility loaded to development of several sperm DNA assays. The aim of this study was to compare several sperm DNA assays in buffaloes, bulls and stallions, and to identify the relationships between those DNA assays and traditional sperm features. In Italian Mediterranean Buffalo (IMB) bulls traditional sperm features (motility, viability, acrosome integrity and morphology), sperm DNA integrity (neutral...

  20. Current status and potential of morphometric sperm analysis

    OpenAIRE

    Alejandro Maroto-Morales; Olga García-Álvarez; Manuel Ramón; Felipe Martínez-Pastor; M Rocío Fernández-Santos; A Josefa Soler; José Julián Garde

    2016-01-01

    The spermatozoon is the most diverse cell type known and this diversity is considered to reflect differences in sperm function. How the diversity in sperm morphology arose during speciation and what role the different specializations play in sperm function, however, remain incompletely characterized. This work reviews the hypotheses proposed to explain sperm morphological evolution, with a focus on some aspects of sperm morphometric evaluation; the ability of morphometrics to predict sperm cr...

  1. Significance of peristaltic squeezing of sperm bundles in the silkworm, Bombyx mori: elimination of irregular eupyrene sperm nuclei of the triploid

    OpenAIRE

    Sahara, Ken; Kawamura, Naoko; Yamashiki, Naoko; Saitoh, Hiroshi

    2001-01-01

    Silkworm (Lepidoptera) males produce dimorphic sperm: nucleate eupyrene sperm and anucleate apyrene sperm. The eupyrene sperm are ordinary sperm to fertilise the eggs, while the function of apyrene sperm remains uncertain. After meiosis, 256 sperm cells are enclosed by a layer of cyst cells, forming a sperm bundle. We have previously documented that the nucleus of eupyrene sperm anchors to the head cyst cell, which locates at the anterior apex of the bundle, by an acrosome tubule-basal body a...

  2. Frozen-thawed spermatozoa from oligozoospermic ejaculates are susceptible to in situ DNA fragmentation in polyvinylpyrrolidone-based sperm-immobilization medium.

    Science.gov (United States)

    Salian, Sujit Raj; Kalthur, Guruprasad; Uppangala, Shubhashree; Kumar, Pratap; Adiga, Satish Kumar

    2012-08-01

    To elucidate the effect of sperm immobilization media that are and are not based on polyvinylpyrrolidone (PVP) on the DNA integrity of fresh and frozen-thawed spermatozoa during standard intracytoplasmic sperm injection (ICSI) conditions. Experimental prospective study. Embryology research laboratory. Forty-six ejaculates from normozoospermic and oligozoospermic men. Assessment of sperm DNA fragmentation by single-cell gel electrophoresis assay. DNA integrity of fresh and frozen-thawed spermatozoa from normozoospermic and oligozoospermic ejaculates exposed to PVP-based and non-PVP-based media. Exposure of fresh and frozen thawed spermatozoa from normozoospermic and oligozoospermic ejaculates to PVP-based medium in an ICSI dish for 30 minutes statistically significantly increased the DNA fragmentation. In contrast, the extent of DNA fragmentation in non-PVP-based medium did not statistically significantly differ from control. A PVP-based medium can induce a statistically significant amount of sperm DNA fragmentation in an ICSI dish, and frozen-thawed sperm from oligozoospermic ejaculates are more susceptible to in situ DNA fragmentation. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  3. COMPARATIVE CHARACTERISTICS OF SPERM FUNCTIONAL PARAMETERS OF SHEEP NORTH CAUCASIAN BREED WHEN YOU MAKE IT TO THE MEDIA FOR IN VITRO FERTILIZATION

    Directory of Open Access Journals (Sweden)

    Gvozdetskii N. A.

    2016-09-01

    Full Text Available The article presents the use of intensive methods of diet herd reproduction that will increase the productivity and profitability of the sheep industry. One of such methods is in vitro fertilization (IVF. For the success of in vitro fertilization procedure, an important requirement is the availability of highquality nutrient media, which help to preserve the genetic material and contribute to the further development of the zygote. The main requirement to the media for sperm is the ability of media not to cause their agglutination. The aim of our work was to search for new ways to reduce the agglutination of spermatozoa in the preparation of freshly prepared sperm in the process of production of embryos in vitro. To eliminate the agglutination of spermatozoa in the semen preparation stage, we used GCY medium, followed by demolition of seed in SOFw environment, which has resulted in a significant (almost 15 times! reduce of the number of bound sperm. In our opinion, a decrease in agglutination in GCY buffer was connected with a specific influence of the constituent components of sperm. Conclusion: Thus, our method of preparation of freshly prepared sperm for in vitro fertilization allows a sharp decline in sperm agglutination, which will improve the fertility of eggs during the production of embryos in vitro sheep

  4. Protective effect of Rhodiola rosea polysaccharides on cryopreserved boar sperm.

    Science.gov (United States)

    Yang, Shen-Min; Wang, Ting; Wen, Duan-Gai; Hou, Jian-Quan; Li, Hai-Bo

    2016-01-01

    Cryopreservation brings sublethal damage to sperm, resulting in reduced fertile life of sperm. Rhodiola rosea polysaccharides (RPs) have antiviral, antioxidant and antitumor activities. In the present study, the cryoprotective effect of RPs on boar sperm quality parameters after frozen-thawed process was investigated. Boar sperm was cryopreserved in the extender with RPs added at concentrations of 0 (used as control), 2, 4, 6, 8 and 10mg/L and their effects on the quality of frozen-thawed boar sperm were assessed. Addition of RPs significantly improved sperm motility, mitochondrial activity, acrosomal integrity, plasma membrane integrity, superoxide dismutase and glutathione peroxidase activity and decreased sperm malonaldehyde level (pboar sperm.

  5. Sperm chromatin structure and male fertility: biological and clinical aspects

    Institute of Scientific and Technical Information of China (English)

    J. Erenpreiss; M. Spano; J. Erenpreisa; M. Bungum; A. Giwercman

    2006-01-01

    Aim: Sperm chromatin/DNA integrity is essential for the accurate transmission of paternal genetic information, and normal sperm chromatin structure is important for sperm fertilizing ability. The routine examination of semen, which includes sperm concentration, motility and morphology, does not identify defects in sperm chromatin structure. The origin of sperm DNA damage and a variety of methods for its assessment are described. Evaluation of sperm DNA damage appears to be a useful tool for assessing male fertility potential both in vivo and in vitro. The possible impact of sperm DNA defects on the offspring is also discussed.

  6. Sperm dynamics in spiders (Araneae: ultrastructural analysis of the sperm activation process in the garden spider Argiope bruennichi (Scopoli, 1772.

    Directory of Open Access Journals (Sweden)

    Oliver Vöcking

    Full Text Available Storage of sperm inside the female genital tract is an integral phase of reproduction in many animal species. The sperm storage site constitutes the arena for sperm activation, sperm competition and female sperm choice. Consequently, to understand animal mating systems information on the processes that occur from sperm transfer to fertilization is required. Here, we focus on sperm activation in spiders. Male spiders produce sperm whose cell components are coiled within the sperm cell and that are surrounded by a proteinaceous sheath. These inactive and encapsulated sperm are transferred to the female spermathecae where they are stored for later fertilization. We analyzed the ultrastructural changes of sperm cells during residency time in the female genital system of the orb-web spider Argiope bruennichi. We found three clearly distinguishable sperm conditions: encapsulated sperm (secretion sheath present, decapsulated (secretion sheath absent and uncoiled sperm (cell components uncoiled, presumably activated. After insemination, sperm remain in the encapsulated condition for several days and become decapsulated after variable periods of time. A variable portion of the decapsulated sperm transforms rapidly to the uncoiled condition resulting in a simultaneous occurrence of decapsulated and uncoiled sperm. After oviposition, only decapsulated and uncoiled sperm are left in the spermathecae, strongly suggesting that the activation process is not reversible. Furthermore, we found four different types of secretion in the spermathecae which might play a role in the decapsulation and activation process.

  7. Effects of Cadmium on Rat Sperm Motility Evaluated With Computer Assisted Sperm Analysis

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective To study effects of cadmium on rat sperm motility evaluated with computer assisted sperm analysis. Methods  Different doses of cadmium chloride (0.2,0.4,0.8mg Cd/kg BW) were administrated ip to adult male Sprague-Dawley rats. Control animals received the same volume of 0.9% NaCl solution. After 7 days, the rats were sacrificed with their testes removed. A part of one testis was used for testicular sperm head counts and daily sperm production observation. The motility of spermatozoa obtained from cauda epididymides using the “diffusion”method was measured by computer assisted sperm analysis(CASA). Results  The sperm head counts and daily sperm production decreased significantly in the high dose group. The motility of spermatozoa in the middle dose group was reduced significantly. No motile sperm was found in the high dose group. The results suggest that germinal epithelium was impaired irreversibly in a short time to produce toxic effects on spermatogenesis at high cadmium doses. Conclusion  Cadmium may reduce sperm motility at a dose far below the dose affecting sperm production at this time point. The motility of sperm is an early and sensitive endpoint for the assessment of cadmium toxicity on male reproduction.

  8. Observation of sperm-head vacuoles and sperm morphology under light microscope.

    Science.gov (United States)

    Park, Yong-Seog; Park, Sol; Ko, Duck Sung; Park, Dong Wook; Seo, Ju Tae; Yang, Kwang Moon

    2014-09-01

    The presence of sperm-head vacuoles has been suspected to be deleterious to the outcomes of assisted reproductive technology (ART). It is difficult to accurately distinguish morphologically abnormal sperm with vacuoles under a light microscope. This study was performed to analyze the result of the observation of sperm-head vacuoles using Papanicolaou staining under a light microscope and whether the male partner's age affects these vacuoles. Sperm morphology with vacuoles was evaluated using Papanicolaou staining and observed under a light microscope (400×) in 980 men. The normal morphology was divided into three categories (group A, 14% of normal morphology). The criteria for the sperm-head vacuoles were those given in the World Health Organization manual. For the analysis of the age factor, the participants were divided into the following groups: 26-30 years, 31-35 years, 36-40 years, 41-45 years, and 46-50 years. The percentage of sperm-head vacuoles increased with normal sperm morphology (group A vs. groups B, C) (p<0.05). In the case of the age factor, a statistically significant difference was not observed across any of the age groups. A majority of the sperm-head vacuoles showed a statistically significant difference among normal morphology groups. Therefore, we should consider the probability of the percentage of sperm-head vacuoles not increasing with age but with abnormal sperm morphology. A further study is required to clarify the effect of the sperm-head vacuoles on ART outcomes.

  9. [Updated detection of the function of sperm plasma membrane].

    Science.gov (United States)

    Zhou, Xin; Xia, Xin-Yi; Huang, Yu-Feng

    2010-08-01

    The sperm plasma membrane is rich in polyunsaturated fatty acids and a variety of proteins, and its function is associated with sperm capacitation, acrosome reaction and sperm-egg fusion. Sperm fertilizability can be predicted by detecting the function of the sperm plasma membrane, which is performed mainly with the following five techniques: sperm hypoosmotic swelling test, Eosin gamma water test, sperm membrane lipid peroxidation determination, seminal plasma superoxide dismutase determination, and flow cytometry. The evaluation of the function of sperm plasma membrane can be applied in detecting semen quality, selecting semen centrifugation, assessing the quality and fertilizability of sex-sorted sperm, improving cryopreservation, and guiding the optimization of intracytoplasmic sperm injection. This review presents an update on the principles, methods and steps of the detection of sperm plasma membrane function, as well as an overview of its status quo and application.

  10. Design and Construction of a Hydraulic Ram Pump

    Directory of Open Access Journals (Sweden)

    Shuaibu Ndache MOHAMMED

    2007-09-01

    Full Text Available The Design and Fabrication of a Hydraulic Ram Pump (Hydram is undertaken. It is meant to lift water from a depth of 2m below the surface with no other external energy source required. Based on the design the volume flow rate in the derived pipe was 4.5238 × 10-5 m3/s (2.7 l/min, Power was 1.273 kW which results in an efficiency of 57.3%. The overall cost of fabrication of this hydram shows that the pump is relatively cheaper than the existing pumps.

  11. Software for creating an airspace in RAMS from Skyview2

    OpenAIRE

    2012-01-01

    In this project has been developed an application that creates an airspace defined by the user and creates the necessary files for RAMS Plus to simulate it. This application has been programed using the Ismael González's TFC named "Algoritmo de cálculo de rutas óptimas para aeronaves en el plano horizontal", therefore the application also allows to know the shortest route between two points of the ECAC airspace, if exists. In order to create the airspace the user can elect from 5 elements to ...

  12. The study of traumatic intracerebral hematoma at Buri Ram Hospital.

    Science.gov (United States)

    Nagabhand, A; Sangcham, K

    1993-07-01

    This report describes the study of traumatic intracerebral hematoma at Buri Ram Hospital. The total number was 71 cases. There were 26 cases with no investigation and were treated by exploratory burr hole with the mortality rate of 57.5 per cent, and 45 cases which were sent for CT scan before operation with the mortality rate of 37.5 per cent. CT scanning is a useful investigative tool for correct diagnosis and rapid treatment of head injury and the operation which helps to reduce the mortality rate.

  13. Ejercicio de lectura de Marcos Ramírez

    OpenAIRE

    2012-01-01

    Se hace un análisis de la novela Marcos Ramírez de Carlos Luis Fallas, señalándola como pilar de la tradición costarricense; enmarcándola en la novelística de la llamada Generación del 40, en la literatura nacional. Se analiza la instancia narrativa encontrando que esta novela posee un narrador autodiegético, esto quiere decir que ese plano es una instancia doble, un elemento único en el cual se reúnen dos aspectos diferentes: el personaje, que es agente o paciente de la acción y el narrador,...

  14. A model for the importance of zinc in the dynamics of human sperm chromatin stabilization after ejaculation in relation to sperm DNA vulnerability.

    Science.gov (United States)

    Björndahl, Lars; Kvist, Ulrik

    2011-02-01

    The focus of this review is the dual functions of the sperm chromatin stabilization and how external factors can interfere with these functions. Zinc depletion after ejaculation allows for rapid and total sperm chromatin decondensation without addition of exogenous disulfide cleaving agents. Zinc depletion without concomitant repulsion of chromatin fibers induces another type of stability that requires exogenous disulfide cleaving agents to allow decondensation. It is essential to extend the present concept, that the sperm chromatin stability is based on disulfide bridges only, to include also the functions of Zn(2+). It is suggested that the chromatin stability of the ejaculated human spermatozoon is rapidly reversible due to the dual function of Zn(2+) that stabilizes the structure and prevents the formation of excess disulfide bridges by a single mechanism: the formation of zinc bridges involving protamine thiols of cysteine and potentially also imidazole groups of histidine. Extraction of zinc from the freshly ejaculated spermatozoon allows two totally different biological results: (1) immediate decondensation if chromatin fibers concomitantly are induced to repel (e.g., through phosphorylation in the ooplasm) and (2) thiols freed from Zn(2+) are available to form disulfide bridges creating a superstabilized chromatin. Spermatozoa in the zinc rich prostatic fluid (in first ejaculated fraction) represent physiology. Extraction of chromatin zinc can be caused by unphysiological exposure of spermatozoa to the zinc chelating and oxidative seminal vesicular fluid, a situation common to most assisted reproductive techniques (ART) laboratories where the entire ejaculate is collected into a single container in which spermatozoa and secretions are mixed during at least 30 min. Some men in infertile couples have low content of sperm chromatin zinc due to loss of zinc during ejaculation and liquefaction. Tests for sperm DNA integrity may give false negative results due to

  15. The vitamin D receptor localization and mRNA expression in ram testis and epididymis.

    Science.gov (United States)

    Jin, Hui; Huang, Yang; Jin, Guang; Xue, Yanrong; Qin, Xiaowei; Yao, Xiaolei; Yue, Wenbing

    2015-02-01

    The objectives of present study were to investigate the presence of vitamin D receptor (VDR) in testis and epididymis of ram by polymerase chain reaction (PCR), to locate VDR in testis and epididymis by immunohistochemistry and to compare difference of VDR expression between testis and epididymis before and after sexual maturation by Real time-PCR and Western blot. The results showed that VDR exists in the testis and epididymis of ram while VDR protein in testis and epididymis was localized in Leydig cells, spermatogonial stem cells, spermatocytes, Sertoli cells and principal cells. For the adult ram, the amounts of VDR mRNA and VDR protein were less (p ram, the result showed the same trend (p 0.05) between adult and prepubertal. In conclusion, VDR exists in testis and epididymis of ram, suggesting 1α,25-(OH)(2)VD(3) may play a role in ram reproduction.

  16. Predominance of sperm motion in corners

    Science.gov (United States)

    Nosrati, Reza; Graham, Percival J.; Liu, Qiaozhi; Sinton, David

    2016-05-01

    Sperm migration through the female tract is crucial to fertilization, but the role of the complex and confined structure of the fallopian tube in sperm guidance remains unknown. Here, by confocal imaging microchannels head-on, we distinguish corner- vs. wall- vs. bulk-swimming bull sperm in confined geometries. Corner-swimming dominates with local areal concentrations as high as 200-fold that of the bulk. The relative degree of corner-swimming is strongest in small channels, decreases with increasing channel size, and plateaus for channels above 200 μm. Corner-swimming remains predominant across the physiologically-relevant range of viscosity and pH. Together, boundary-following sperm account for over 95% of the sperm distribution in small rectangular channels, which is similar to the percentage of wall swimmers in circular channels of similar size. We also demonstrate that wall-swimming sperm travel closer to walls in smaller channels (~100 μm), where the opposite wall is within the hydrodynamic interaction length-scale. The corner accumulation effect is more than the superposition of the influence of two walls, and over 5-fold stronger than that of a single wall. These findings suggest that folds and corners are dominant in sperm migration in the narrow (sub-mm) lumen of the fallopian tube and microchannel-based sperm selection devices.

  17. Children conceived after intracytoplasmic sperm injection (ICSI)

    DEFF Research Database (Denmark)

    Mau, C; Juul, A; Main, K M;

    2004-01-01

    The aim of the study was to evaluate current medical knowledge about children born after intracytoplasmic sperm injection (ICSI) with respect to congenital malformations, chromosome abnormalities and postnatal growth.......The aim of the study was to evaluate current medical knowledge about children born after intracytoplasmic sperm injection (ICSI) with respect to congenital malformations, chromosome abnormalities and postnatal growth....

  18. Pollen and sperm heteromorphism: convergence across kingdoms?

    Science.gov (United States)

    Till-Bottraud, I; Joly, D; Lachaise, D; Snook, R R

    2005-01-01

    Sperm competition theory predicts that males should produce many, similar sperm. However, in some species of animals and plants, males exhibit a heteromorphism that results in the production of at least two different types of sperm or pollen grains. In animals, sperm heteromorphism typically corresponds to the production of one fertile morph and one (or more) sterile morph(s), whereas in plants two or more pollen morphs (one of which can be either sterile or fertile) are produced in all flowers but sometimes in different anthers. Heteromorphism has arisen independently several times across phyla and at different phylogenetic levels. Here, we compare and contrast sperm and pollen heteromorphism and discuss the evolutionary hypotheses suggested to explain heteromorphism in these taxa. These hypotheses include facilitation, nutritive contribution, blocking, cheap filler, sperm flushing or killing for animals; outcrossing and precise cross-pollen transfer or bet-hedging strategy for plants; cryptic female choice for both. We conclude that heteromorphism in the two phyla is most likely linked to a general evolutionary response to sexual selection, either to increase one male's sperm or pollen success in competition with other males, or mediate male/female interactions. Therefore, although sperm and pollen are not homologous, we suggest that heteromorphism represents an example of convergence across kingdoms.

  19. HDAC6 deacetylates alpha tubulin in sperm and modulates sperm motility in Holtzman rat.

    Science.gov (United States)

    Parab, Sweta; Shetty, Omshree; Gaonkar, Reshma; Balasinor, Nafisa; Khole, Vrinda; Parte, Priyanka

    2015-02-01

    Histone deacetylase 6 (HDAC6) is an alpha (α)-tubulin deacetylase and its over-expression has been demonstrated to promote chemotactic cell movement. Motility in sperm is driven by the flagella, the cytoskeletal structure comprising the microtubules, which are heterodimers of α- and β-tubulins. We have hypothesized that HDAC6, by virtue of being an α-tubulin deacetylase, might modulate sperm motility. However, the presence of HDAC6 on sperm has hitherto not been reported. In this study, we have demonstrated, for the first time, the presence of HDAC6 transcript and protein in the testicular and caudal sperm of rat. We have observed a significantly overlapping expression of HDAC6 with acetyl α-tubulin (Ac α-tubulin) in the mid-piece and principal piece of sperm flagella, and the co-precipitation of α-tubulin and Ac α-tubulin together with HDAC6 and vice versa in sperm lysates. This indicates that HDAC6 interacts with α-tubulin. The HDAC6 activity of sperm, sperm motility and status of Ac α-tubulin investigated in the presence of HDAC inhibitors Trichostatin A, Tubastatin A and sodium butyrate demonstrate that HDAC6 in sperm is catalytically active and that inhibitors of HDAC6 increase acetylation and restrict sperm motility. Thus, we show that (1) active HDAC6 enzyme is present in sperm, (2) HDAC6 in sperm is able to deacetylate α-tubulin, (3) inhibition of HDAC6 results in increased Ac α-tubulin expression and (4) HDAC6 inhibition affects sperm motility. This evidence suggests that HDAC6 is involved in modulating sperm movement.

  20. Human semen cryopreservation: a sperm DNA fragmentation study with alkaline and neutral Comet assay.

    Science.gov (United States)

    Ribas-Maynou, J; Fernández-Encinas, A; García-Peiró, A; Prada, E; Abad, C; Amengual, M J; Navarro, J; Benet, J

    2014-01-01

    Sperm cryopreservation is widely used for both research and reproduction purposes, but its effect on sperm DNA damage remains controversial. Sperm DNA fragmentation (SDF) has become an important biomarker to assess male infertility. In particular, the differentiation between single- and double-stranded DNA fragmentation (ssSDF and dsSDF) has clinical implications for male infertility where ssSDF is associated with reduced fertility, whereas dsSDF is associated with increased risk of miscarriage. In this study, semen samples from 30 human males have been analysed in both fresh and cryopreserved using the alkaline and neutral Comet assays. Results show an increase of about 10% of ssSDF, assessed by the alkaline Comet assay, regardless of the male fertility status. Neutral Comet analysis of dsSDF does not show any statistical increase when comparing fresh and cryopreserved samples in any of the patient groups. Results support previous reports that oxidative stress is the major effector in DNA damage during sample cryopreservation, as, on one hand, ssSDF has previously been related to oxidative damage and, on the other hand, we have not found any effect on dsSDF. Therefore, there might be a slight risk of decreased fertility after using a freezed sample, but no evidence for increased miscarriage risk from cryopreserved spermatozoa should be expected. © 2013 American Society of Andrology and European Academy of Andrology.

  1. Sperm Whales Suffer the Bends

    Institute of Scientific and Technical Information of China (English)

    Michael; Hopkin; 席芳

    2005-01-01

    据原先推测,鲸鱼对由声纳系统导致的疾病具有免疫力,但是两位美国科学家发现,抹香鲸(Sperm whales)正遭受着骨坏死疾病的侵扰,这是由于声纳系统扰乱了鲸鱼的生活习性所致,这种疾病有可能导致它们的搁浅。鲸鱼亟需得到保护!

  2. Pressure Characteristics of a Diffuser in a Ram RDE Propulsive Device

    Science.gov (United States)

    2017-07-21

    Naval Research Laboratory Washington, DC 20375-5320 NRL/MR/6040--17-9699 Pressure Characteristics of a Diffuser in a Ram-RDE Propulsive Device July...RESPONSIBLE PERSON 19b. TELEPHONE NUMBER (include area code) b. ABSTRACT c. THIS PAGE 18. NUMBER OF PAGES 17. LIMITATION OF ABSTRACT Pressure ...representative ram RDE subsonic diffuser to study ram RDE compatibility with a supersonic inlet. Of particular interest is the propagation of pressure

  3. Effect of age on sperm fertility potential: oocyte donation as a model.

    Science.gov (United States)

    Gallardo, E; Simón, C; Levy, M; Guanes, P P; Remohí, J; Pellicer, A

    1996-08-01

    To determine the effect of age on sperm fecundability using oocyte donation as an in vivo model. Oocyte donation and IVF programs at the Instituto Valenciano de Infertilidad. Retrospective study in which four groups of oocyte donation cycles were established according to age of the male providing the semen sample: group 1 (n = 31) 51 years, the oldest being 64 years. All donated oocytes were obtained from patients < 35 years old. Male age, sperm characteristics (volume, concentration, motility, morphology), fertilization, embryo quality, pregnancy, implantation, and abortion rates among recipients. Similar sperm characteristics in fresh as well as after preparation for IVF were observed among males of different ages. Fertilization, embryo quality, pregnancy, and implantation were similar among the established groups. The mean age of the females included in each group significantly increased from group 1 to group 4. Age (up to 64 years) does not affect sperm characteristics or its ability to fertilize human eggs. Similarly, embryo development in vitro as well as implantation in recipient uteri are not affected by age of the male providing the semen sample.

  4. The influence of organophosphate and carbamate on sperm chromatin and reproductive hormones among pesticide sprayers.

    Science.gov (United States)

    Jamal, Farrukh; Haque, Quazi S; Singh, Sangram; Rastogi, S K

    2016-08-01

    This study is aimed at evaluating the association between occupational exposure to organophosphate (OP) and carbamate (CB) pesticides and semen quality as well as levels of reproductive and thyroid hormones of pesticide sprayers in Malihabad, Lucknow, Uttar Pradesh, India. Thirty-five healthy men (unexposed group) and 64 male pesticide sprayers (exposed group) were recruited for clinical evaluation of fertility status. Fresh semen samples were evaluated for sperm quality and analyzed for DNA fragmentation index (DFI) by flow cytometry. Pesticide exposure was assessed by measuring erythrocyte acetylcholinesterase and plasma butyrylcholinesterase (BuChE) with a Test-mate ChE field kit. Serum levels of total testosterone (Tt), prolactin (PRL), follicle-stimulating hormone (FSH), luteinizing hormone (LH), thyroid-stimulating hormone (TSH), and free thyroxine (FT4) were analyzed using enzyme immunoassay kits. Evidence of pesticide exposure was found in 88.5% of sprayers and significant increments were observed in sperm DFI with significant decrease in some semen parameters. DFI was negatively correlated with BuChE, sperm concentration, morphology, and vitality in these pesticide sprayers. The levels of Tt, PRL, FT4, and TSH appeared to be normal; however, there was a tendency for increased LH and FSH levels in exposed workers. The results confirm the potential impact of chronic occupational exposure to OP and CB pesticides on male reproductive function, which may cause damage to sperm chromatin, decrease semen quality, and produce alterations in reproductive hormones, leading to adverse reproductive health outcomes. © The Author(s) 2015.

  5. Differential gene expression by RamA in ciprofloxacin-resistant Salmonella Typhimurium.

    Directory of Open Access Journals (Sweden)

    Jie Zheng

    Full Text Available Overexpression of ramA has been implicated in resistance to multiple drugs in several enterobacterial pathogens. In the present study, Salmonella Typhimurium strain LTL with constitutive expression of ramA was compared to its ramA-deletion mutant by employing both DNA microarrays and phenotype microarrays (PM. The mutant strain with the disruption of ramA showed differential expression of at least 33 genes involved in 11 functional groups. The study confirmed at the transcriptional level that the constitutive expression of ramA was directly associated with increased expression of multidrug efflux pump AcrAB-TolC and decreased expression of porin protein OmpF, thereby conferring multiple drug resistance phenotype. Compared to the parent strain constitutively expressing ramA, the ramA mutant had increased susceptibility to over 70 antimicrobials and toxic compounds. The PM analysis also uncovered that the ramA mutant was better in utilization of 10 carbon sources and 5 phosphorus sources. This study suggested that the constitutive expression of ramA locus regulate not only multidrug efflux pump and accessory genes but also genes involved in carbon metabolic pathways.

  6. El pensamiento evolutivo de Ramón Margalef

    Directory of Open Access Journals (Sweden)

    J. Terradas

    2015-01-01

    Full Text Available El pensamiento evolutivo de Ramón Margalef. El ecólogo Ramón Margalef desarrolló, a lo largo de toda su vida, una visión global de la Biosfera, lo que le llevó, en muchos de sus trabajos, a hacer propuestas y sugerencias relacionadas con interpretaciones evolutivas. Para ello, se basó de modo especial en la física (termodinámica especialmente y en la teoría de la información, y buscó un puente entre las concepciones utilizadas por la ecología teórica y la teoría evolutiva en una serie de aspectos de la ecología, pero de modo destacado en la sucesión. Aquí trato de dar una síntesis de las aportaciones de Margalef en este terreno, aportaciones que creo que siguen siendo una fuente de inspiración para los trabajos en ecología y evolución y para forjar conexiones teóricas entre las dos disciplinas, hoy por hoy, si no inexistentes, ciertamente demasiado débiles.

  7. Body mass indices analysis in the German Giant Ram breed

    Directory of Open Access Journals (Sweden)

    Silvia Patruica

    2015-05-01

    Full Text Available The study of the main body mass indices in the giant German ram breed was performed between 6.06.2014 and 10.06.2014 in a rabbitry from Diniaș town, Timiș County. The biological material was represented by 16 adult rabbit females and 10 adult males, with the average body weight of 7 kg, raised in semi-intensive system. There have been performed the following body measurements: oblique and horizontal length of the trunk, the length of thorax, the length of head, the length of ears, of neck, and of tail, the height at weithers and rump, the height of thorax and stern, the width of chest, of thorax, of rump at hip and ischia, the wide width of forehead, the thoracic circumference and the circumference of shin. Giant German Ram males have registered higher values in the body size index, in the pelvic-thoracic index, and in the rump angle index compared to females from the same breed, which have registered higher values in the index of transverse body size, in the index of massiveness, in the pelvic-thoracic index, and in the cephalic index.

  8. Ram accelerator operating characteristics at elevated fill pressures

    Energy Technology Data Exchange (ETDEWEB)

    Bundy, C.; Knowlen, C.; Bruckner, A.P. [Washington Univ., Seattle, WA (United States). Aerospace and Energetics Research Program

    2000-11-01

    An experimental investigation of the starting and operational characteristics of a 38-mm bore ram accelerator with propellant fill pressures greater than 5 MPa is in progress. Successful starts with 60 - 124 gm projectiles have been achieved using methane/oxygen/nitrogen propellants at fill pressures ranging from 6 to 15 MPa. At fill pressures of 8.5 MPa and above, it was found that projectiles having the nominal throat diameter (29 mm) required a minimum entrance velocity of 1250 m/s, which is about 100 m/s faster than the minimum needed to successfully start the ram accelerator at fill pressures below 7.5 MPa. Projectiles with a reduced throat diameter (25 mm) and a solid magnesium nose cone were successfully started at fill pressures up to 15 MPa with entrance velocities around 1300 m/s. The average accelerations achieved using high pressure stages were in general less than predicted for thermally choked operation by the Boltzmann equation of state. (orig.)

  9. Flow over a Modern Ram-Air Parachute Canopy

    Science.gov (United States)

    Mohammadi, Mohammad; Johari, Hamid

    2010-11-01

    The flow field on the central section of a modern ram-air parachute canopy was examined numerically using a finite-volume flow solver coupled with the one equation Spalart-Allmaras turbulence model. Ram-air parachutes are used for guided airdrop applications, and the canopy resembles a wing with an open leading edge for inflation. The canopy surfaces were assumed to be impermeable and rigid. The flow field consisted of a vortex inside the leading edge opening which effectively closed off the canopy and diverted the flow around the leading edge. The flow experienced a rather bluff leading edge in contrast to the smooth leading of an airfoil, leading to a separation bubble on the lower lip of the canopy. The flow inside the canopy was stagnant beyond the halfway point. The section lift coefficient increased linearly with the angle of attack up to 8.5 and the lift curve slope was about 8% smaller than the baseline airfoil. The leading edge opening had a major effect on the drag prior to stall; the drag is at least twice the baseline airfoil drag. The minimum drag of the section occurs over the angle of attack range of 3 -- 7 .

  10. Flow over a Ram-Air Parachute Canopy

    Science.gov (United States)

    Eslambolchi, Ali; Johari, Hamid

    2012-11-01

    The flow field over a full-scale, ram-air personnel parachute canopy was investigated numerically using a finite-volume flow solver coupled with the Spalart-Allmaras turbulence model. Ram-air parachute canopies resemble wings with arc-anhedral, surface protuberances, and an open leading edge for inflation. The rectangular planform canopy had an aspect ratio of 2.2 and was assumed to be rigid and impermeable. The chord-based Reynolds number was 3.2 million. Results indicate that the oncoming flow barely penetrates the canopy opening, and creates a large separation bubble below the lower lip of canopy. A thick boundary layer exists over the entire lower surface of the canopy. The flow over the upper surface of the canopy remains attached for an extended fraction of the chord. Lift increases linearly with angle of attack up to about 12 degrees. To assess the capability of lifting-line theory in predicting the forces on the canopy, the lift and drag data from a two-dimensional simulation of the canopy profile were extended using finite-wing expressions and compared with the forces from the present simulations. The finite-wing predicted lift and drag trends compare poorly against the full-span simulation, and the maximum lift-to-drag ratio is over-predicted by 36%. Sponsored by the US Army NRDEC.

  11. Nondestructive Evaluation of Historic Hakka Rammed Earth Structures

    Directory of Open Access Journals (Sweden)

    Yanhao Li

    2013-01-01

    Full Text Available The in-service Hakka rammed earth buildings, in the Fujian Province of China, are unique in design and performance. Their UNESCO’s inscription as World Heritage sites recognizes their artistic, cultural, social and historic significance. Sponsored by the National Science Foundation of the United States, the authors have examined the engineering values of these buildings in terms of comfortable living at low energy consumption, sustainability and durability. The objective of this study was to better understand the thermo-mechanical and aging responses of Hakka earth buildings under thermal and earthquake loads through nondestructive field evaluation, including full-scale roof truss and floor testing, laboratory testing of field samples and finite element modeling. This paper presents our observations and findings from the field nondestructive evaluations with emphasis on the integrity of the rammed earth outer walls and inner wood structures, as well as the thermal comfort of living in these buildings, while a second paper presents the results from the material characterization of field samples and the structural responses of a representative building under earthquake induced loads through finite element analysis.

  12. Experimental and numerical investigation of ram extrusion of bread dough

    Science.gov (United States)

    Mohammed, M. A. P.; Wanigasooriya, L.; Charalambides, M. N.

    2016-10-01

    An experimental and numerical study on ram extrusion of bread dough was conducted. A laboratory ram extrusion rig was designed and manufactured, where dies with different angles and exit radii were employed. Rate dependent behaviour was observed from tests conducted at different extrusion speeds, and higher extrusion pressure was reported for dies with decreasing exit radius. A finite element simulation of extrusion was performed using the adaptive meshing technique in Abaqus. Simulations using a frictionless contact between the billet and die wall showed that the model underestimates the response at high entry angles. On the other hand, when the coefficient of friction value was set to 0.09 as measured from friction experiments, the dough response was overestimated, i.e. the model extrusion pressure was much higher than the experimentally measured values. When a critical shear stress limit, τmax, was used, the accuracy of the model predictions improved. The results showed that higher die angles require higher τmax values for the model and the experiments to agree.

  13. Simulating lightning into the RAMS model: implementation and preliminary results

    Directory of Open Access Journals (Sweden)

    S. Federico

    2014-05-01

    Full Text Available This paper shows the results of a tailored version of a previously published methodology, designed to simulate lightning activity, implemented into the Regional Atmospheric Modeling System (RAMS. The method gives the flash density at the resolution of the RAMS grid-scale allowing for a detailed analysis of the evolution of simulated lightning activity. The system is applied in detail to two case studies occurred over the Lazio Region, in Central Italy. Simulations are compared with the lightning activity detected by the LINET network. The cases refer to two thunderstorms of different intensity. Results show that the model predicts reasonably well both cases and that the lightning activity is well reproduced especially for the most intense case. However, there are errors in timing and positioning of the convection, whose magnitude depends on the case study, which mirrors in timing and positioning errors of the lightning distribution. To assess objectively the performance of the methodology, standard scores are presented for four additional case studies. Scores show the ability of the methodology to simulate the daily lightning activity for different spatial scales and for two different minimum thresholds of flash number density. The performance decreases at finer spatial scales and for higher thresholds. The comparison of simulated and observed lighting activity is an immediate and powerful tool to assess the model ability to reproduce the intensity and the evolution of the convection. This shows the importance of the use of computationally efficient lightning schemes, such as the one described in this paper, in forecast models.

  14. Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices

    Science.gov (United States)

    Matsuura, Koji; Huang, Han-Wei; Chen, Ming-Cheng; Chen, Yu; Cheng, Chao-Min

    2017-04-01

    Mammalian sperm motility has traditionally been analyzed to determine fertility using computer-assisted semen analysis (CASA) systems. To develop low-cost and robust male fertility diagnostics, we created a paper-based MTT assay and used it to estimate motile sperm concentration. When porcine sperm motility was inhibited using sperm enzyme inhibitors for sperm enzymes related to mitochondrial activity and glycolysis, we simultaneously recorded sperm motility and enzymatic reactivity using a portable motility analysis system (iSperm) and a paper-based MTT assay, respectively. When using our paper-based MTT-assay, we calculated the area mean value signal intensity (AMV) to evaluate enzymatic reactivity. Both sperm motility and AMV decreased following treatment with iodoacetamide (IODO) and 3-bromopyruvic acid (3BP), both of which are inhibitors of glycolytic enzymes including glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a correlation between recorded motility using iSperm and AMV from our paper-based assay (P Based on this inhibitor study, sperm motility can be estimated using our paper-based MTT-assay.

  15. Human Sperm Immotility Caused by Degeneration in the Epididymis

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective To investigate whether sperm immotility was caused by degeneration in the epididymisMethods Five patients with totally immotile sperm were selected in this study. Testic ular biopsy was used to obtain testicular sperm to evaluate sperm motility. The com bined hypoosmotic swelling-eosin Y exclusion test was carried out to determine the sperm head and tail membrane integrity for the ejaculated and the testicular sperm.The ultrastructure of ejaculated sperm was examined by transmission electron microscope.Results No motile sperm were found in the ejaculated semen samples from 5 patients,whereas 2% to 11% motile testicular sperm extracted from the testicular biopsy tissues were observed. The percentage of testicular sperm with intact head and tail membranes was higher than that of the ejaculated sperm (P< 0. 01). Ultrastructure of the ejacu lated sperm showed marked degenerative features. Seminal plasma from patients did not influence the motility of normal donor sperm.Conclusion Sperm could undergo degenerative changes during transit through and /or storage in the epididymis, which led to lose sperm motility in these patients. Using motile testicular sperm would benefit the treatment for such cases.

  16. Improvement of post-thawed sperm quality and fertility of Arian rooster by oral administration of d-aspartic acid.

    Science.gov (United States)

    Ansari, Mahdi; Zhandi, Mahdi; Kohram, Hamid; Zaghari, Mojtaba; Sadeghi, Mostafa; Sharafi, Mohsen

    2017-04-01

    This study was conducted to investigate the effect of d-Aspartic acid (D-Asp) on post-thawed sperm quality, fertility and hatchability outcomes in male broiler breeders. Twenty 55-week-old roosters were selected and equally split into four groups (n = 5 rooster/group). Different daily D-Asp doses including 0 (D-0), 100 (D-100), 200 (D-200) or 300 (D-300) mg/kg BW were capsulated and individually administered for 12 weeks to roosters in each group. Semen samples were weekly collected from 7th to 12th week of experiment. Sperm quality from 7th to 11th week was evaluated in both fresh (total and forward motility and plasma membrane functionality) and post-thawed (total and forward motility, plasma membrane functionality, apoptosis status and mitochondrial activity) conditions. Also, collected semen samples on the 12th week were frozen and artificially inseminated to evaluate fertility and hatchability. The results from fresh condition showed that total and forward motility and plasma membrane functionality were significantly higher in D-200 compared to other groups. Also, interaction effect of time and treatment was not significant for all assessed parameters in fresh condition. In post-thawed condition, D-200 showed significantly higher total and forward motility, fertility and hatchability compared to other groups. The higher value for plasma membrane functionality and mitochondrial activity was observed in D-200 compared to D-0 and D300 groups. However, the percentage of live, early apoptotic and dead spermatozoa were not significantly affected by applied treatment in the current study. No significant difference for time and treat interaction effect was observed for all assessed parameters except forward motility. In conclusion, it seems that D-Asp administration could improve fresh and post-thawed sperm quality and post-thawed sperm fertility in male broiler breeders.

  17. Prudent sperm use by leaf-cutter ant queens

    DEFF Research Database (Denmark)

    Den Boer, Susanne Petronella A; Baer, Boris; Dreier, Stéphanie Agnès Jeanine

    2009-01-01

    glandular secretions or deteriorating sperm-storage organs. We evaluate our findings with a heuristic model, which suggests that the average queen has sperm for almost 9 years of normal colony development. We discuss the extent to which leaf-cutter ant queens have been able to optimize their sperm...... expenditure and infer that our observed averages of sperm number, sperm robustness and sperm use are consistent with sperm depletion being a significant cause of mortality of mature colonies of Atta leaf-cutter ants.......In many species, females store sperm between copulation and egg fertilization, but the consequences of sperm storage and patterns of sperm use for female life history and reproductive success have not been investigated in great detail. In hymenopteran insect societies (ants, bees, wasps...

  18. Redox regulation of mammalian sperm capacitation

    Directory of Open Access Journals (Sweden)

    Cristian O′Flaherty

    2015-01-01

    Full Text Available Capacitation is a series of morphological and metabolic changes necessary for the spermatozoon to achieve fertilizing ability. One of the earlier happenings during mammalian sperm capacitation is the production of reactive oxygen species (ROS that will trigger and regulate a series of events including protein phosphorylation, in a time-dependent fashion. The identity of the sperm oxidase responsible for the production of ROS involved in capacitation is still elusive, and several candidates are discussed in this review. Interestingly, ROS-induced ROS formation has been described during human sperm capacitation. Redox signaling during capacitation is associated with changes in thiol groups of proteins located on the plasma membrane and subcellular compartments of the spermatozoon. Both, oxidation of thiols forming disulfide bridges and the increase on thiol content are necessary to regulate different sperm proteins associated with capacitation. Reducing equivalents such as NADH and NADPH are necessary to support capacitation in many species including humans. Lactate dehydrogenase, glucose-6-phospohate dehydrogenase, and isocitrate dehydrogenase are responsible in supplying NAD (P H for sperm capacitation. Peroxiredoxins (PRDXs are newly described enzymes with antioxidant properties that can protect mammalian spermatozoa; however, they are also candidates for assuring the regulation of redox signaling required for sperm capacitation. The dysregulation of PRDXs and of enzymes needed for their reactivation such as thioredoxin/thioredoxin reductase system and glutathione-S-transferases impairs sperm motility, capacitation, and promotes DNA damage in spermatozoa leading to male infertility.

  19. Sperm Preservation using Freeze-Drying Method

    Directory of Open Access Journals (Sweden)

    TAKDIR SAILI

    2005-03-01

    Full Text Available Since the discovery of cryopreservation method for bull semen, cryopreservation become an alternative method for maintaining gamet resources of certain animal which is threatened or near extinction. This technology was then applied to the preservation of embryo, oocyte, ovary and testis. The application of intracytoplasmic sperm injection (ICSI for which sperm motility is unnecessary had supported the effort to create simplified method such as freeze-drying for sperm preservation. Due to the benefit of ICSI over the conventional in vitro fertilization (IVF the spermatozoon could be mechanically driven to pass through the zona pellucida and entering the cytoplasm of oocytes prior to fertilization. The freeze-drying method is an alternative method in sperm preservation which ignored the motility of sperm. The sperm resulted from this technique is in drying state, therefore, it might be stored in room temperature or in refrigerator. Many reports have claimed that freeze-dried sperm which is not motile but has an intact DNA was able to fertilize oocytes, even produced offspring in mouse.

  20. Semen analysis and sperm function testing

    Institute of Scientific and Technical Information of China (English)

    Daniel R Franken; Sergio Oehninger

    2012-01-01

    Despite controversy regarding the clinical value of semen analysis,male fertility investigation still relies on a standardized analysis of the semen parameters.This is especially true for infertility clinics in both developing and developed countries.Other optional tests or sophisticated technologies have not been widely applied.The current review addresses important changes in the analysis of semen as described in the new World Health Organization (WHO) manual for semen analysis.The most important change in the manual is the use of evidence-based publications as references to determine cutoff values for normality.Apart from the above mentioned changes,the initial evaluation and handling methods remain,in most instances,the same as in previous editions.Furthermore,the review evaluates the importance of quality control in andrology with emphasis on the evaluation of sperm morphology.WHO sperm morphology training programmes for Sub-Saharan countries were initiated at Tygerberg Hospital in 1995.1he external quality control programme has ensured that the majority of participants have maintained their morphological reading skills acquired during initial training.This review reports on current sperm functional tests,such as the induced acrosome reaction,and sperm-zona pellucida binding assays,as well as the impact of sperm qualitv in terms of DNA integritv,and the relationship of sperm function tests to sperm morphologv.

  1. Tolerance to environmental desiccation in moss sperm.

    Science.gov (United States)

    Shortlidge, Erin E; Rosenstiel, Todd N; Eppley, Sarah M

    2012-05-01

    • Sexual reproduction in mosses requires that sperm be released freely into the environment before finding and fertilizing a receptive female. After release from the male plant, moss sperm may experience a range of abiotic stresses; however, few data are available examining stress tolerance of moss sperm and whether there is genetic variation for stress tolerance in this important life stage. • Here, we investigated the effects of environmental desiccation and recovery on the sperm cells of three moss species (Bryum argenteum, Campylopus introflexus, and Ceratodon purpureus). • We found that a fraction of sperm cells were tolerant to environmental desiccation for extended periods (d) and that tolerance did not vary among species. We found that this tolerance occurs irrespective of ambient dehydration conditions, and that the addition of sucrose during dry-down improved cell recovery. Although we observed no interspecific variation, significant variation among individuals within species in sperm cell tolerance to environmental desiccation was observed, suggesting selection could potentially act on this basic reproductive trait. • The observation of desiccation-tolerant sperm in multiple moss species has important implications for understanding bryophyte reproduction, suggesting the presence of a significant, uncharacterized complexity in the ecology of moss mating systems. © 2012 The Authors. New Phytologist © 2012 New Phytologist Trust.

  2. Measurement and significance of sperm morphology

    Institute of Scientific and Technical Information of China (English)

    Roelof Menkveld; Cas AG Holleboom; Johann PT Rhemrev

    2011-01-01

    The measurement or evaluation and clinical significance of human sperm morphology has always been and still is a controversial aspect of the semen analysis for the determination of a male's fertility potential.In this review the background of the development of the evaluation criteria for sperm morphology will be discussed.Aspects of criticism on the strict criteria definition and use of the criteria for sperm morphology evaluation will be discussed as well as possible reasons for the decline in normal sperm morphology values and how we can compromise for this phenomenon resulting in the very low normal reference value as published in the 2010 WHO manual for the Examination and Processing of Human Semen.One of the possible solutions may be to give more attention to a limited number of abnormal sperm morphology categories and the inclusion of sperm morphology patterns.It is concluded in this review that if done correctly and with care and with strict application of existing guidelines as outlined in the 2010 WHO manual,sperm morphology measurement still has a very important role to play in the clinical evaluation of male fertility potential.

  3. A C. elegans sperm TRP protein required for sperm-egg interactions during fertilization.

    Science.gov (United States)

    Xu, X-Z Shawn; Sternberg, Paul W

    2003-08-08

    Fertilization, a critical step in animal reproduction, is triggered by a series of specialized sperm-egg interactions. However, the molecular mechanisms underlying fertilization are not well understood. Here, we identify a sperm-enriched C. elegans TRPC homolog, TRP-3. Mutations in trp-3 lead to sterility in both hermaphrodites and males due to a defect in their sperm. trp-3 mutant sperm are motile, but fail to fertilize oocytes after gamete contact. TRP-3 is initially localized in intracellular vesicles, and then translocates to the plasma membrane during sperm activation. This translocation coincides with a marked increase in store-operated calcium entry, providing an in vivo mechanism for the regulation of TRP-3 activity. As C. elegans oocytes lack egg coats, our data suggest that some TRPC family channels might function to mediate calcium influx during sperm-egg plasma membrane interactions leading to fertilization.

  4. Boundaries of Ethics, Sperm on the Border –The Globalization of Danish Sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    market as well as clinics worldwide. Furthermore, one of the sperm banks market sperm for self-insemination and deliver it to private customers by UPS. Although sperm donation is an old and low technology compared to other reproductive technologies, this development has taken place at the same time...... as sperm donation in parliamentary debates, and in the media, often raises ethical questions. This presentation inquires into how the bending of boundaries by “inappropriate parents”, fertility travelers, private sperm banks and fertility clinics have been part in negotiating the changes of the legislation...... of “ethical” does to a technology and its users such as in the case of the globalization of Danish sperm, as well as I will question how understandings of normality, sexuality, race, age, gender and kinship become part of the negotiations of “the ethical”, or in the doing of ethics in practice...

  5. In cryptozoospermia or severe oligozoospermia is sperm freezing useful?

    OpenAIRE

    Bessonnat, Julien; Brouillet, Sophie; Sintzel, Sarah; Gillois, Pierre; Bergues, Ulrike; Boutte-Busquet, Caroline; Thomas-Cadi, Claire; Hennebicq, Sylviane

    2014-01-01

    Background Intracytoplasmic Sperm Injection (ICSI) is an Assisted Reproduction Technique (ART) which offers the chance to conceive to patients presenting very low sperm counts (cryptozoospermia/severe oligozoospermia). Sperm freezing before the oocyte pick-up, can prevent from a lack of spermatozoa on the day of the ICSI. It can avoid the cancellation of the ICSI or the use of TESE (Testicular sperm extraction). The objective of this study was to analyse the practice of sperm freezing for the...

  6. Sperm competition and the evolution of gamete morphology in frogs.

    OpenAIRE

    Byrne, Phillip G; Simmons, Leigh W.; Roberts, J. Dale

    2003-01-01

    Despite detailed knowledge of the ultrastructure of spermatozoa, there is a paucity of information on the selective pressures that influence sperm form and function. Theoretical models for both internal and external fertilizers predict that sperm competition could favour the evolution of longer sperm. Empirical tests of the external-fertilization model have been restricted to just one group, the fishes, and these tests have proved equivocal. We investigated how sperm competition affects sperm...

  7. Microscopic analysis of MTT stained boar sperm cells

    OpenAIRE

    B.M. van den Berg

    2015-01-01

    The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few ...

  8. Pengaruh Pencucian Sperma dengan Lama Waktu Sentrifugasi yang Berbeda Terhadap Kualitas Sperma Kambing Bligon (Effect of Sperm Washing with Different Centrifugation Duration on Sperm Quality of Bligon Buck

    Directory of Open Access Journals (Sweden)

    Sigit Bintara

    2012-02-01

    spermatozoa. It is concluded that washing Bligon buck sperm using SpermRinse with centrifugation at 1.500 rpm (r = 5.5 cm could increased sperm quality, of which 20 minutes was the best centrifugation time. (Key word: Bligon Buck, Sperm, SpermRinse, Sperm quality

  9. Sperm immobilization by dental focus microorganisms.

    Science.gov (United States)

    Linossier, A; Thumann, A; Bustos-Obregon, E

    1982-01-01

    Focal infections and their ability to produce alterations in different tissues have been in dispute for long time. The purpose of this work was to observe "in vitro" the effect of an Escherichia coli filtrate obtained from open pulpar necrosis on human sperm motility. It was observed that the E. coli filtrate produced a loss in sperm motility. The immobilizating factor was studied and characterized as a heat-stable, resistant to lyophilization and non-dializable substance, which could via blood stream reach the male reproductive system and affect sperm motility.

  10. Aquaporin3 is a sperm water channel essential for postcopulatory sperm osmoadaptation and migration

    Institute of Scientific and Technical Information of China (English)

    Qi Chen; Hongying Peng; Li Lei; Ying Zhang; Haibin Kuang; Yujing Cao; Qi-xian Shi; Tonghui Ma; Enkui Duan

    2011-01-01

    In the journey from the male to female reproductive tract,mammalian sperm experience a natural osmotic decrease (e.g.,in mouse,from ~415 mOsm in the cauda epididymis to ~310 mOsm in the uterine cavity). Sperm have evolved to utilize this hypotonic exposure for motility activation,meanwhile efficiently silence the negative impact of hypotonic cell swelling. Previous physiological and pharmacological studies have shown that ion channel-controlled water influx/efflux is actively involved in the process of sperm volume regulation; however,no specific sperm proteins have been found responsible for this rapid osmoadaptation. Here,we report that aquaporin3 (AQP3) is a sperm water channel in mice and humans. Aqp3-deficient sperm show normal motility activation in response to hypotonicity but display increased vulnerability to hypotonic cell swelling,characterized by increased tail bending after entering uterus. The sperm defect is a result of impaired sperm volume regulation and progressive cell swelling in response to physiological hypotonic stress during male-female reproductive tract transition. Time-lapse imaging revealed that the cell volume expansion begins at cytoplasmic droplet,forcing the tail to angulate and form a hairpin-like structure due to mechanical membrane stretch. The tail deformation hampered sperm migration into oviduct,resulting in impaired fertilization and reduced male fertility. These data suggest AQP3 as an essential membrane pathway for sperm regulatory volume decrease (RVD) that balances the "trade-off" between sperm motility and cell swelling upon physiological hypotonicity,thereby optimizing postcopulatory sperm behavior.

  11. Sperm Proteomics Reveals Intensified Selection on Mouse Sperm Membrane and Acrosome Genes

    OpenAIRE

    Dorus, Steve; Wasbrough, Elizabeth R.; Busby, Jennifer; Wilkin, Elaine C.; Karr, Timothy L.

    2010-01-01

    Spermatozoa are a focal point for the impact of sexual selection due to sperm competition and sperm–female interactions in a wide range of sexually reproducing organisms. In-depth molecular investigation of the ramifications of these selective regimes has been limited due to a lack of information concerning the molecular composition of sperm. In this study, we utilize three previously published proteomic data sets in conjunction with our whole mouse sperm proteomic analysis to delineate cellu...

  12. Ovarian fluid mediates the temporal decline in sperm viability in a fish with sperm storage.

    Directory of Open Access Journals (Sweden)

    Clelia Gasparini

    Full Text Available A loss of sperm viability and functionality during sperm transfer and storage within the female reproductive tract can have important fitness implications by disrupting fertilization and impairing offspring development and survival. Consequently, mechanisms that mitigate the temporal decline in sperm function are likely to be important targets of selection. In many species, ovarian fluid is known to regulate and maintain sperm quality. In this paper, we use the guppy Poecilia reticulata, a highly polyandrous freshwater fish exhibiting internal fertilization and sperm storage, to determine whether ovarian fluid (OF influences the decline in sperm viability (the proportion of live sperm in the ejaculate over time and whether any observed effects depend on male sexual ornamentation. To address these questions we used a paired experimental design in which ejaculates from individual males were tested in vitro both in presence and absence of OF. Our results revealed that the temporal decline in sperm viability was significantly reduced in the presence of OF compared to a saline control. This finding raises the intriguing possibility that OF may play a role in mediating the decline in sperm quality due to the deleterious effects of sperm ageing, although other possible explanations for this observation are discussed. Interestingly, we also show that the age-related decline in sperm viability was contingent on male sexual ornamentation; males with relatively high levels of iridescence (indicating higher sexual attractiveness exhibited a more pronounced decline in sperm viability over time than their less ornamented counterparts. This latter finding offers possible insights into the functional basis for the previously observed trade-off between these key components of pre- and postcopulatory sexual selection.

  13. Status specific tailoring of sperm behavior in an external fertilizer

    Directory of Open Access Journals (Sweden)

    Torvald Blikra Egeland

    2016-11-01

    Full Text Available Why dominant males experiencing intense sperm competition sometimes show low investments in sperm production is not always obvious. One well-documented example is that of the external fertilizing teleost, the Arctic charr (Salvelinus alpinus, where individuals becoming dominant reduce sperm production and sperm swimming speed in water compared to subordinates. Here we report how ovarian fluid differentially influences sperm velocity of dominant and subordinate male Arctic charr. That is, sperm from dominant males increase their velocity in water diluted ovarian fluid compared to that observed in water, while sperm from subordinates, on the other hand, decrease velocity in ovarian fluid compared to that observed in water. Thus, subordinates, who invest more resources in their sperm and usually show the highest sperm velocity in water, have lower gains from their investment than dominant males when sperm are swimming in ovarian fluid. In sum, our result suggests that ovarian fluid increase sperm velocity more in dominant males than in subordinate males. Although this finding could partly be caused by cryptic female choice exerted by the ovarian fluid for sperm from dominant males, an alternative and more parsimonious explanation is that sperm from dominant males may simply be better designed for swimming in ovarian fluid compared to sperm from subordinate males. Thus, sperm production in the two reproductive roles seems to be adaptively tailored to different external environments.

  14. Evaluation of Sperm Parameters of Infertile Men with Retrograde Ejaculation

    Institute of Scientific and Technical Information of China (English)

    Hong-xing ZHONG; Wei-jie ZHU; Jing LI

    2006-01-01

    Objective To investigate sperm parameters of infertile men with retrograde ejaculation.Methods Twelve infertile men with retrograde ejaculation (group A) were enrolled into this study. Sperm samples were obtained from the postejaculation urine. After sperm recovery and washing procedure, sperm parameters were assessed. Twelve semen samples from normospermic donors were used as the control (group B).Results In all retrograde cases, motile sperm with forward movement were observed in the medium. Motility of group A was significantly lower than that of group B (P<0. 01).In group A, sperm motility ranged from 11% to 56%, sperm with intact both head and tail membranes was 42.2 ± 12.3%, sperm count ranged (13-85)×106/ml, and the sperm survival time was highly shortened. Sperm with normal morphology and intact acrosome were observed in retrograde specimens.Conclusion Sperm parameters recovered from retrograde specimens were highly variable between subjects. The toxicity of urine caused deleterious to sperm functions.Motile sperm could be collected by sperm recovery procedure. Sperm parameters could meet the requirement for the use of assisted reproductive techniques for treating infertile men with retrograde ejaculation.

  15. Tactic-specific differences in seminal fluid influence sperm performance.

    Science.gov (United States)

    Locatello, Lisa; Poli, Federica; Rasotto, Maria B

    2013-03-22

    Seminal fluid often makes up a large part of an ejaculate, yet most empirical and theoretical studies on sperm competition have focused on how sperm characteristics (number and quality) affect fertilization success. However, seminal fluid influences own sperm performance and may potentially influence the outcome of sperm competition, by also affecting that of rivals. As a consequence males may be expected to allocate their investment in both sperm and seminal fluid in relation to the potential level of competition. Grass goby (Zosterisessor ophiocephalus) is an external fertilizer with guard-sneaker mating tactics, where sperm competition risk varies according to the tactic adopted. Here, we experimentally manipulated grass goby ejaculates by separately combining sperm and seminal fluid from territorial and sneaker males. While sperm of sneaker and territorial males did not differ in their performance when they interacted with their own seminal fluid only, sperm of sneakers increased their velocity and fertilization rate in the presence of territorial males' seminal fluid. By contrast, sneaker males' seminal fluid had a detrimental effect on the performance of territorial males' sperm. Sperm velocity was unaffected by the seminal fluid of males employing the same tactic, suggesting that seminal fluid's effect on rival-tactic sperm is not based on a self/non-self recognition mechanism. Our findings show that cross interactions of sperm and seminal fluid may influence the fertilization success of competing ejaculates with males investing in both sperm and seminal fluid in response to sperm competition risk.

  16. The RAM Scale: Development and Validation of the Revised Scale in Likert Format.

    Science.gov (United States)

    Wright, Claudia R.; And Others

    1983-01-01

    The development and validation of a revised form of the RAM Scale in Likert format are described. The RAM Scale measures student philosophical orientations in terms of relative, absolute, or mixed biases or preferences toward issues of knowledge, methods, and values. (Author/PN)

  17. Uudised : Hortus Musicus Moskvas. TPÜ naiskoor koorikonkursil. RAM samal laval Mark Knopfleriga. "Tuuleveskid. Kerkokell" / Joosep Sang

    Index Scriptorium Estoniae

    Sang, Joosep

    2000-01-01

    Hortus Musicus esines 23. ja 24. nov. Moskvas. TPÜnaiskoor võitis II preemia XVII rahvusvahelisel F.Schuberti nim. koorikonkursil Austrias. RAM esineb 8. dets. Helsingis Hartwall Areenas heategevuskontserdil. RAM esineb 6. dets. Estonia kontserdisaalis kavaga "Tuuleveskid. Kerkokell"

  18. Molecular basis of RNA guanine-7 methyltransferase (RNMT) activation by RAM.

    Science.gov (United States)

    Varshney, Dhaval; Petit, Alain-Pierre; Bueren-Calabuig, Juan A; Jansen, Chimed; Fletcher, Dan A; Peggie, Mark; Weidlich, Simone; Scullion, Paul; Pisliakov, Andrei V; Cowling, Victoria H

    2016-12-01

    Maturation and translation of mRNA in eukaryotes requires the addition of the 7-methylguanosine cap. In vertebrates, the cap methyltransferase, RNA guanine-7 methyltransferase (RNMT), has an activating subunit, RNMT-Activating Miniprotein (RAM). Here we report the first crystal structure of the human RNMT in complex with the activation domain of RAM. A relatively unstructured and negatively charged RAM binds to a positively charged surface groove on RNMT, distal to the active site. This results in stabilisation of a RNMT lobe structure which co-evolved with RAM and is required for RAM binding. Structure-guided mutagenesis and molecular dynamics simulations reveal that RAM stabilises the structure and positioning of the RNMT lobe and the adjacent α-helix hinge, resulting in optimal positioning of helix A which contacts substrates in the active site. Using biophysical and biochemical approaches, we observe that RAM increases the recruitment of the methyl donor, AdoMet (S-adenosyl methionine), to RNMT. Thus we report the mechanism by which RAM allosterically activates RNMT, allowing it to function as a molecular rheostat for mRNA cap methylation. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  19. Use of testicular sperm for intracytoplasmic sperm injection in men with high sperm DNA fragmentation: a SWOT analysis.

    Science.gov (United States)

    Esteves, Sandro C; Roque, Matheus; Garrido, Nicolás

    2017-04-18

    Spermatozoa retrieved from the testis of men with high levels of sperm DNA fragmentation (SDF) in the neat semen tend to have better DNA quality. Given the negative impact of SDF on the outcomes of Assisted Reproductive Technology (ART), an increased interest has emerged about the use of testicular sperm for intracytoplasmic sperm injection (Testi-ICSI). In this article, we used a SWOT (strengths, weaknesses, opportunities, and threats) analysis to summarize the advantages and drawbacks of this intervention. The rationale of Testi-ICSI is bypass posttesticular DNA fragmentation caused by oxidative stress during sperm transit through the epididymis. Hence, oocyte fertilization by genomically intact testicular spermatozoa may be optimized, thus increasing the chances of creating a normal embryonic genome and the likelihood of achieving a live birth, as recently demonstrated in men with high SDF. However, there is still limited evidence as regards the clinical efficacy of Testi-ICSI, thus creating opportunities for further confirmatory clinical research as well as investigation of Testi-ICSI in clinical scenarios other than high SDF. Furthermore, Testi-ICSI can be compared to other laboratory preparation methods for deselecting sperm with damaged DNA. At present, the available literature supports the use of testicular sperm when performing ICSI in infertile couples whose male partners have posttesticular SDF. Due to inherent risks of sperm retrieval, Testi-ICSI should be offered when less invasive treatments for alleviating DNA damage have failed. A call for continuous monitoring is nonetheless required concerning the health of generated offspring and the potential complications of sperm retrieval.

  20. Sperm nuclear histone H2B: correlation with sperm DNA denaturation and DNA stainability

    Institute of Scientific and Technical Information of China (English)

    Armand Zini; Xiaoyang Zhang; Maria San Gabriel

    2008-01-01

    Aim: To examine the relationship between sperm DNA damage and sperm nuclear histone (H2B) staining. Methods:We evaluated sperm samples from 14 consecutive asthenoteratozoospermic infertile men and six consecutive fertile controls. Sperm nuclear histone (H2B) staining and sperm chromatin integrity (assessed by sperm chromatin structure assay and expressed using the percentage of (I) DNA fragmentation index [%DFI] and (ii) high DNA stainability [%HDS)]) were evaluated. Results: Histone H2B immunocytochemistry demonstrated two nuclear staining patterns: (I) focal punctate staining; and (ii) diffuse staining. Infertile men had a higher mean percentage of spermatozoa exhibiting diffuse H2B staining than did fertile men (7.7% ± 4.6% vs. 1.6% ± 1.2%, respectively, P < 0.01). We observed significant relationships between the proportion of spermatozoa with diffuse nuclear histone staining and both sperm %DFI (r= 0.63, P < 0.01) and sperm %HDS (r= 0.63, P < 0.01). Conclusion: The data demonstrate that infertile men have a higher proportion of spermatozoa with diffuse histone H2B than do fertile men and suggest that sperm DNA damage might, at least in part, be due to abnormally high histone H2B levels.

  1. Spermatogenesis, the mature sperm, and sperm egg association in Nematospiroides dubius

    Science.gov (United States)

    Clark, W. H., Jr.

    1974-01-01

    Nematode spermatogenesis was investigated using the strongyloid Nematospiroides dubius. The primary spermatocytes, development of spermatids, and changes in the sperm in the female tract are described.

  2. Computer-assisted sperm analysis parameters in young fertile sperm donors and relationship with age.

    Science.gov (United States)

    Fréour, Thomas; Jean, Miguel; Mirallie, Sophie; Barriere, Paul

    2012-04-01

    Sperm parameter values have been shown to decline with age, according to conventional sperm analysis. However, the effect of age on sperm kinematic parameters has been rarely studied, especially in young fertile men. Here, we studied Computer-Assisted Sperm Analysis (CASA) parameters in a large cohort of men with proven fertility, in order to determine if there is a decline with age in this young fertile population. This retrospective analysis of CASA parameters was conducted on all donors included in the sperm donor programme in the Assisted Reproductive Techniques (ART) Centre of the University Hospital of Nantes between 2006 and 2009. Sperm concentration, motility, and kinetic parameters were recorded by a HTM-Ceros system and compared in 3 groups of sperm donors according to their age: donors were analyzed. Values for ALH, VCL, LIN, and STR significantly decreased with age. Sperm concentration, motile sperm proportion, and other kinetic parameters did not differ significantly among the groups. The use of CASA allowed the identification of ALH, VCL, LIN, and STR age-related decrease in young men with proven fertility.

  3. Protein restriction affects sperm number but not sperm viability in male ants.

    Science.gov (United States)

    Dávila, Francisco; Aron, Serge

    2017-07-01

    Sperm cells are costly to produce; diet should therefore affect sperm number and/or viability. In non-social insects and vertebrates, there is compelling evidence that diet influences sperm production. Less is known about this relationship in eusocial hymenopterans (all ants and some bees and wasps), whose mating systems impose unique selective pressures on sperm production. Males face physiological constraints: they acquire all of the resources they will use in future reproductive efforts as larvae and emerge from the pupal stage with a fixed, non-renewable amount of sperm. Furthermore, males die shortly after copulation, but their genetic material persists for years since their spermatozoa are stored in their mates' spermathecae. We examined the effects of protein restriction during larval development on sperm number and viability in the Argentine ant Linepithema humile. We also looked at its impact on male development, adult mass, and adult fluctuating asymmetry. We found that protein restriction during larval development significantly reduced sperm production, but not sperm viability. It did not affect the number of males reared, male mass, or male asymmetry. However, males from protein-restricted nests developed much more slowly than males from protein-supplemented nests. These results suggest investing in sperm quality and in somatic growth, which enhances a male's ability to disperse and find a mate, are critical to successful male reproduction. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Female sperm use and storage between fertilization events drive sperm competition and male ejaculate allocation.

    Science.gov (United States)

    Requena, Gustavo S; Alonzo, Suzanne H

    2014-12-01

    Sperm competition theory has traditionally focused on how male allocation responds to female promiscuity, when males compete to fertilize a single clutch of eggs. Here, we develop a model to ask how female sperm use and storage across consecutive reproductive events affect male ejaculate allocation and patterns of mating and paternity. In our model, sperm use (a single parameter under female control) is the main determinant of sperm competition, which alters the effect of female promiscuity on male success and, ultimately, male reproductive allocation. Our theory reproduces the general pattern predicted by existing theory that increased sperm competition favors increased allocation to ejaculates. However, our model predicts a negative correlation between male ejaculate allocation and female promiscuity, challenging the generality of a prevailing expectation of sperm competition theory. Early models assumed that the energetic costs of precopulatory competition and the level of sperm competition are both determined by female promiscuity, which leads to an assumed covariation between these two processes. By modeling precopulatory costs and sperm competition independently, our theoretical framework allows us to examine how male allocation should respond independently to variation in sperm competition and energetic trade-offs in mating systems that have been overlooked in the past.

  5. Sperm morphology: consistency of assessment of the same sperm by different observers.

    Science.gov (United States)

    Baker, H W; Clarke, G N

    1987-01-01

    Photographic slides of 36 sperm were shown to a group of 28 observers with different lengths of experience in assessing sperm morphology. Experienced observers were generally consistent (10 or more of the 17 agreeing) in classifying sperm as normal, amorphous, small heads, lacking acrosomes, and having tail defects or cytoplasmic droplets but categorization was more variable for large, tapering and pyriform heads. This study highlights the need for more widespread agreement about definition of sperm shapes and the development of practical objective methods of assessment.

  6. Transport properties of anodic porous alumina for ReRAM

    Energy Technology Data Exchange (ETDEWEB)

    Kato, S; Nigo, S; Lee, J W; Mihalik, M; Kitazawa, H; Kido, G [National Institute for Materials Science, 1-2-1 Sengen, Tsukuba, Ibaraki, 305-0047 (Japan)], E-mail: KATO.Seiichi@nims.go.jp

    2008-03-15

    A voltage-induced bistable switching effect has been studied for M/AlO{sub x}/Al devices made of the anodic porous alumina with a top electrode of aluminium (or silver) to develop a next generation memory (AlO{sub x}-ReRAM). The resistance state of memory is switched between OFF-state (high resistance) and ON-state (low resistance), where the resistance ratio is higher than 10{sup 4}. In the thermally stimulated current (TSC) measurement, a narrow band was observed around 290 K, indicating the conduction mechanism comes from a kind of impurity band in the energy gap. An anomaly was also observed around 290 K in the temperature dependence of resistance at the ON-state.

  7. An introduction to RAM analysis of EPB tunnel boring machine

    Directory of Open Access Journals (Sweden)

    Hasel Amini Khoshalan

    2014-03-01

    Full Text Available Earth pressure balance tunnel boring machines (EPB - TBMs are favorably used in construction of tunnels in urban areas. These more expensive machines with high operational costs require an exclusive study and knowledge on these machine components and subsystems and their failure and downtimes for time planning, cost control and performance prediction with a high accuracy. Thus guidelines for reliability, availability and maintainability (RAM analysis of an EPB machine are completely expressed. Gathering and recording all daily failure and repair times and other maintenance tasks, dividing the machine into subsystems (including mechanical subsystem, electrical subsystem, hydraulic subsystem, pneumatic subsystem and water subsystem, graphical tests (trend test and correlation test, statistical tests and analysis are discussed in detail.

  8. First Exploratory Study on the Ageing of Rammed Earth Material

    Directory of Open Access Journals (Sweden)

    Quoc-Bao Bui

    2014-12-01

    Full Text Available Rammed earth (RE is attracting renewed interest throughout the world thanks to its “green” characteristics in the context of sustainable building. In this study, the ageing effects on RE material are studied on the walls which have been constructed and exposed for 22 years to natural weathering. First, mechanical characteristics of the “old” walls were determined by two approaches: in-situ dynamic measurements on the walls; laboratory tests on specimens which had been cut from the walls. Then, the walls’ soil was recycled and reused for manufacturing of new specimens which represented the initial state. Comparison between the compressive strength, the Young modulus of the walls after 22 years on site and that of the initial state enables to assess the ageing of the studied walls.

  9. First Exploratory Study on the Ageing of Rammed Earth Material.

    Science.gov (United States)

    Bui, Quoc-Bao; Morel, Jean-Claude

    2014-12-23

    Rammed earth (RE) is attracting renewed interest throughout the world thanks to its "green" characteristics in the context of sustainable building. In this study, the ageing effects on RE material are studied on the walls which have been constructed and exposed for 22 years to natural weathering. First, mechanical characteristics of the "old" walls were determined by two approaches: in-situ dynamic measurements on the walls; laboratory tests on specimens which had been cut from the walls. Then, the walls' soil was recycled and reused for manufacturing of new specimens which represented the initial state. Comparison between the compressive strength, the Young modulus of the walls after 22 years on site and that of the initial state enables to assess the ageing of the studied walls.

  10. Antioxidant activity of aqueous extract of noni in dilutent for ram semen cryopreservation

    Directory of Open Access Journals (Sweden)

    Ana Lauren Costa Nascimento

    2016-03-01

    Full Text Available Noni (Morinda citrifolia L. is a fruit consumed worldwide because of its nutritional and therapeutic properties resulting from the large amount of phenolic compounds, which has aroused interest of the scientific community. In order to identify new natural sources of antioxidants, the objective of this study was to evaluate the performance of noni in diluent for ram semen cryopreservation. A completely randomized design consisting of four treatments and three repetitions per treatment was used. The treatments differed in terms of the concentration of the aqueous extract of noni added to the diluent: control, no addition of the extract, and three concentrations (24, 72, and 120 µg/mL. The physical and chemical variables of the mature fruit were evaluated: total acidity (8.78, pH (4.12, and soluble solids (8.18%. The vitamin C content was 309.42 mg per 100 g fresh matter. The aqueous extract of noni was also evaluated regarding the quantity of total phenolic compounds, antioxidant activity, and lipid peroxidation inhibition capacity. The aqueous extract contained a moderate amount of phenolic compounds (47.96 ± 1.95 mg gallic acid equivalent/100 g extract. The concentrations of the aqueous extract of 72 and 120 µg/mL in diluent used for semen cryopreservation inhibited lipid peroxidation by 21.75% and 51.32%, respectively. There was no positive effect of the lowest concentration (24 µg/mL. The antioxidant activity index of noni was 33.33, corresponding to very strong antioxidant activity. The aqueous extract of noni exhibits very strong antioxidant activity and its addition to the diluent for semen cryopreservation at a concentration of 72 µg/mL is able to inhibit lipid peroxidation.

  11. Ram Pressure Stripping and Morphological Transformation in the Coma Cluster

    Science.gov (United States)

    Gregg, Michael; West, Michael

    2017-01-01

    The two largest spiral galaxies in the Coma cluster, NGC4911 and NGC4921, exhibit signs of being vigorously ram-pressure stripped by the hot intracluster medium. HST ACS and WFC3 images have revealed galactic scale shock fronts, giant "Pillars of Creation", rivulets of dust, and spatially coherent star formation in these grand design spirals. All evidence points to these galaxies being stressed by a global external source which can only be the hot intracluster medium (ICM). Inspired by these examples, we have obtained HST WFC3 imaging of five additional large spirals to search for and investigate the effects of ram pressure stripping across the wider Coma cluster. The results are equally spectacular as the first two examples. The geometry of the interactions in some cases allows us to estimate the various time scales involved, including gas flows out of the disk leading to creation of the ICM, and the attendant triggered star formation in the galaxy disks. The global star formation patterns and wholesale tidal stripping of matter yield insights into the spatial and temporal ISM-ICM interactions driving the evolution of galaxies in clusters and ultimately transforming their morphologies from spiral to S0. These processes, much more common in the early Universe, led to the wholesale morphological transformation of Hubble types during the assembly of rich clusters, when the intergalactic populations and hot ICM were first being created and laid down from such stripping and destruction of their member galaxies.We also report on two instrumental aspects of WFC3: 1) using the filter pair F350LP and F600LP to create an extremely broad pseudo Blue-Red color to achieve the greatest observing efficiency with HST, and 2) a WFC3 CCD effect which leads to apparent quantization of background counts, making automatic sky determination challenging when using drizzlepac routines.

  12. 21 CFR 101.95 - “Fresh,” “freshly frozen,” “fresh frozen,” “frozen fresh.”

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false âFresh,â âfreshly frozen,â âfresh frozen,â âfrozen... fresh,” when used on the label or in labeling of a food, mean that the food was quickly frozen while still fresh (i.e., the food had been recently harvested when frozen). Blanching of the food...

  13. Rat sperm motility analysis: methodologic considerations

    Science.gov (United States)

    The objective of these studies was to optimize conditions for computer-assisted sperm analysis (CASA) of rat epididymal spermatozoa. Methodologic issues addressed include sample collection technique, sampling region within the epididymis, type of diluent medium used, and sample c...

  14. Sperm macromolecules associated with bull fertility.

    Science.gov (United States)

    Kaya, Abdullah; Memili, Erdoğan

    2016-06-01

    Bull fertility, ability of the sperm to fertilize and activate the egg that sustain embryo development, is vitally important for effective and efficient production of cattle. Fertility is a complex trait with low heritability. Despite recent advances in genomic selection and possibility of enormous paternal benefits to profitable cattle production, there exist no reliable tests for evaluating semen quality and predicting bull fertility. This review focuses on sperm macromolecules such as transcripts, proteins and the epigenome, i.e., the functional genome that are associated with bull fertility. Generating new information in these systems is important beyond agriculture because such progress advances the fundamental science of the mammalian male gamete while at the same time introduces biotechnology into livestock production. Sperm macromolecules and epigenome markers associated with bull fertility can be used alone or in combination with the current SNP microarrays to determine sperm quality and to indicate bull fertility.

  15. Comparative evolutionary psychology of sperm competition.

    Science.gov (United States)

    Shackelford, Todd K; Goetz, Aaron T

    2006-05-01

    A comparative evolutionary psychological perspective predicts that species that recurrently faced similar adaptive problems may have evolved similar psychological mechanisms to solve these problems. Sperm competition provides an arena in which to assess the heuristic value of such a comparative evolutionary perspective. The sperm competition that results from female infidelity and polyandry presents a similar class of adaptive problems for individuals across many species. The authors first describe mechanisms of sperm competition in insects and in birds. They suggest that the adaptive problems and evolved solutions in these species provide insight into human anatomy, physiology, psychology, and behavior. The authors then review recent theoretical and empirical arguments for the existence of sperm competition in humans and discuss proposed adaptations in humans that have analogs in insects or birds. The authors conclude by highlighting the heuristic value of a comparative evolutionary psychological approach in this field.

  16. Mammalian Sperm Motility: Observation and Theory

    KAUST Repository

    Gaffney, E.A.

    2011-01-21

    Mammalian spermatozoa motility is a subject of growing importance because of rising human infertility and the possibility of improving animal breeding. We highlight opportunities for fluid and continuum dynamics to provide novel insights concerning the mechanics of these specialized cells, especially during their remarkable journey to the egg. The biological structure of the motile sperm appendage, the flagellum, is described and placed in the context of the mechanics underlying the migration of mammalian sperm through the numerous environments of the female reproductive tract. This process demands certain specific changes to flagellar movement and motility for which further mechanical insight would be valuable, although this requires improved modeling capabilities, particularly to increase our understanding of sperm progression in vivo. We summarize current theoretical studies, highlighting the synergistic combination of imaging and theory in exploring sperm motility, and discuss the challenges for future observational and theoretical studies in understanding the underlying mechanics. © 2011 by Annual Reviews. All rights reserved.

  17. Acrosomo-nuclear syndrome in canine sperm.

    Science.gov (United States)

    Hrudka, F

    1983-01-01

    An acrosomo-nuclear syndrome in sperm of an infertile semicryptorchid dog is described. Based on an EM study of developing and mature sperm the syndrome is defined by simultaneous occurrence of these symptoms: 1) intranuclear inclusions of acrosomal origin, 2) maldifferentiated apical segment of acrosome, 3) intraacrosomal inclusions of Sertoli cell origin, 4) characteristic change of nuclear shape and 5) retained cytoplasmic droplet. The cause of the syndrome and possibility of a transfer of somatic factors are discussed.

  18. Outdoor air pollution and sperm quality.

    Science.gov (United States)

    Lafuente, Rafael; García-Blàquez, Núria; Jacquemin, Bénédicte; Checa, Miguel Angel

    2016-09-15

    Exposure to air pollution has been clearly associated with a range of adverse health effects, including reproductive toxicity, but its effects on male semen quality are still unclear. We performed a systematic review (up to June 2016) to assess the impact of air pollutants on sperm quality. We included 17 semi-ecological, panel, and cohort studies, assessing outdoor air pollutants, such as PM2.5, PM10, NOx, SO2, and O3, and their effects on DNA fragmentation, sperm count, sperm motility, and sperm morphology. Thirteen studies assessed air pollution exposure measured environmentally, and six used biomarkers of air pollution exposure (two did both). We rated the studies using the Newcastle-Ottawa Scale and assessed with the exposure method. Taking into account these factors and the number of studies finding significant results (positive or negative), the evidence supporting an effect of air pollution on DNA fragmentation is weak but suggestive, on sperm motility is limited and probably inexistent, on lower sperm count is inconclusive, and on sperm morphology is very suggestive. Because of the diversity of air pollutants and sperm parameters, and the studies' designs, we were unable to perform a meta-analysis. In summary, most studies concluded that outdoor air pollution affects at least one of the four semen quality parameters included in the review. However, results lack consistency, and furthermore, studies were not comparable. Studies using standardized air pollution and semen measures are required to obtain more reliable conclusions. CRD42015007175. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  19. Sperm DNA oxidative damage and DNA adducts

    Science.gov (United States)

    Jeng, Hueiwang Anna; Pan, Chih-Hong; Chao, Mu-Rong; Lin, Wen-Yi

    2015-01-01

    The objective of this study was to investigate DNA damage and adducts in sperm from coke oven workers who have been exposed to polycyclic aromatic hydrocarbons. A longitudinal study was conducted with repeated measurements during spermatogenesis. Coke-oven workers (n=112) from a coke-oven plant served the PAH-exposed group, while administrators and security personnel (n=67) served the control. Routine semen parameters (concentration, motility, vitality, and morphology) were analyzed simultaneously; the assessment of sperm DNA integrity endpoints included DNA fragmentation, bulky DNA adducts, and 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxo-dGuo). The degree of sperm DNA fragmentation was measured using the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and sperm chromatin structure assay (SCSA). The PAH-exposed group had a significant increase in bulky DNA adducts and 8-oxo-dGuo compared to the control subjects (Ps = 0.002 and 0.045, respectively). Coke oven workers' percentages of DNA fragmentation and denaturation from the PAH-exposed group were not significantly different from those of the control subjects (Ps = 0.232 and 0.245, respectively). Routine semen parameters and DNA integrity endpoints were not correlated. Concentrations of 8-oxo-dGuo were positively correlated with percentages of DNA fragmentation measured by both TUNEL and SCSA (Ps = 0.045 and 0.034, respectively). However, the concentrations of 8-oxo-dGuo and percentages of DNA fragmentation did not correlate with concentrations of bulky DNA adducts. In summary, coke oven workers with chronic exposure to PAHs experienced decreased sperm DNA integrity. Oxidative stress could contribute to the degree of DNA fragmentation. Bulky DNA adducts may be independent of the formation of DNA fragmentation and oxidative adducts in sperm. Monitoring sperm DNA integrity is recommended as a part of the process of assessing the impact of occupational and environmental toxins on

  20. Sperm mobility determines the outcome of sperm competition in the domestic fowl.

    Science.gov (United States)

    Birkhead, T R; Martínez, J G; Burke, T; Froman, D P

    1999-01-01

    The aim of this study was to establish whether the mobility of sperm of the domestic fowl, as measured by an in vitro assay, predicted the outcome of sperm competition. Thirteen pairs of New Hampshire roosters, comprising one male categorized as having high-mobility sperm and the other as having average-mobility sperm, were used. Each male provided 25 x 10(6) sperm, which were mixed and artificially inseminated into between four and seven New Hampshire hens, each of which produced 2-11 offspring. The experiment was conducted twice, such that the same pair of males inseminated the same females. Paternity was assigned by using microsatellite markers. There was a clear effect of sperm-mobility phenotype on the outcome of sperm competition: in all 13 pairs the high-mobility male fathered the majority of offspring (75.3% overall; p < 0.0001). The proportion of offspring fathered by the high-mobility male within pairs varied significantly between male pairs (p < 0.0005). This effect was associated with the difference in sperm-mobility scores between males within pairs; there was a significant positive relationship between the proportion of offspring fathered by the high-mobility male and the ratio of mobility scores between males (p < 0.05). In addition, compared with their success predicted from the non-competitive situation, in the competitive situation high-mobility males were disproportionately successful in fertilizing eggs compared with average-mobility males. This may occur because female sperm storage is limited in some way and a greater proportion of high-mobility sperm gain access to the female's sperm storage tubules. There was no evidence that female effects accounted for any of the variation in paternity. PMID:10577160

  1. Lifestyle influences human sperm functional quality

    Institute of Scientific and Technical Information of China (English)

    Mnica Ferreira; Joana Vieira Silva; Vladimiro Silva; Antnio Barros; Margarida Fardilha

    2012-01-01

    Objective:To investigate the impact of acute lifestyle changes on human sperm functional quality.Methods:In the academic festivities week, young and apparently healthy male students who voluntarily submit themselves to acute lifestyle alterations(among the potentially important variations are increase in alcohol, caffeine, and tobacco consumption and circadian rhythm shifts) were used as a model system.Sperm samples were obtained before and after the academic week and compared by traditional semen analysis(n=54) and also tested for cleavedPolyADP-ribose polymerase(PARP) protein, an apoptotic marker(n=35).Results:Acute lifestyle changes that occurred during the academic week festivities(the study model) resulted both in a significant reduction in sperm quality, assessed by basic semen analysis(decrease in sperm concentration, total number of spermatozoa, progressive and non-progressive motility and increase in sperm morphological abnormalities) and by an increase in the expression of the apoptotic marker, cleavedPARP, in the ejaculate.Conclusions:Acute lifestyle changes have clear deleterious effects on sperm quality.We propose cleavedPARP as a novel molecular marker, valuable for assessing spermquality in parallel with the basic semen analysis method.

  2. Major regulatory mechanisms involved in sperm motility.

    Science.gov (United States)

    Pereira, Rute; Sá, Rosália; Barros, Alberto; Sousa, Mário

    2017-01-01

    The genetic bases and molecular mechanisms involved in the assembly and function of the flagellum components as well as in the regulation of the flagellar movement are not fully understood, especially in humans. There are several causes for sperm immotility, of which some can be avoided and corrected, whereas other are related to genetic defects and deserve full investigation to give a diagnosis to patients. This review was performed after an extensive literature search on the online databases PubMed, ScienceDirect, and Web of Science. Here, we review the involvement of regulatory pathways responsible for sperm motility, indicating possible causes for sperm immotility. These included the calcium pathway, the cAMP-dependent protein kinase pathway, the importance of kinases and phosphatases, the function of reactive oxygen species, and how the regulation of cell volume and osmolarity are also fundamental components. We then discuss main gene defects associated with specific morphological abnormalities. Finally, we slightly discuss some preventive and treatments approaches to avoid development of conditions that are associated with unspecified sperm immotility. We believe that in the near future, with the development of more powerful techniques, the genetic causes of sperm immotility and the regulatory mechanisms of sperm motility will be better understand, thus enabling to perform a full diagnosis and uncover new therapies.

  3. Role of the epididymis in sperm competition

    Institute of Scientific and Technical Information of China (English)

    Russell C. Jones; Jean-Louis Dacheux; Brett Nixon; Heath W. Ecroyd

    2007-01-01

    Although it is generally understood that the testes recruited kidney ducts for reproductive function during the evolution of vertebrates, little is understood of the biological significance of the adaptation. In the context of the evolution of the mammalian epididymis, this report provides evidence that a major role of the epididymis is to enhance a male's chance of achieving paternity in a competitive mating system. A unique example of sperm cooperation in monotremes is used as evidence that the epididymis produces sperm competition proteins to form groups of 100 sperm into bundles that have a forward motility nearly thrice that of individual spermatozoa. As it required 3-h incubation in vitro under capacitation conditions to release motile sperm from the bundles, it is suggested that the monotremes provide an example of capacitation that is quite different from capacitation in higher mammals. It is Suggested that variation between species in the intensity of sperm competition could explain the variation that occurs between species in the amount of post-testicular sperm maturation and storage in the epididymis, an explanation of why the human epididymis does not play as important a role in reproduction as the epididymis of most mammals.

  4. Sperm length evolution in the fungus-growing ants

    DEFF Research Database (Denmark)

    Baer, B.; Dijkstra, M. B.; Mueller, U. G.

    2009-01-01

    the evolution of sperm length. Sperm length does not decrease further in multiply mating leaf-cutting ants, despite substantial further increases in colony size. In a combined analysis, sexual dimorphism explained 63.1% of the variance in sperm length between species. As colony size was not a significant...... in phylogenetically independent contrasts. Some of the remaining variation was explained by the relative size of the sperm-storage organ, but only in the multiply mating leaf-cutting ants, suggesting that sperm-storage constraints become important for the evolution of sperm length in this derived group. Mate number...... affected sperm length to a minor extent, and only in interaction with other predictor variables, suggesting that sperm competition has not been a major selective force for sperm length evolution in these ants....

  5. Effect of short abstinence time on sperm motility parameters

    DEFF Research Database (Denmark)

    Alipour, Hiva; Dardmeh, Fereshteh; Van Der Horst, Gerhard

    2015-01-01

    Semen preparation medium have an important role in assisted reproduction techniques and their composition influences sperm binding and motility. Some studies have assessed the influence of pH on sperm kinetics. However, no study to date has assessed the effect of environmental pH on subtle...... differences in the details of the sperm movement (kinematics) of human sperm provided by computerized sperm analysis systems. This study was designed to assess the effect of two different media pH levels on kinematic parameters of the human sperm. Samples were prepared using the 40%/80% Pureception (Sage, USA......) density gradient and resuspended in customized sperm culture media with pH levels of 7.9 and 8.3 (Origio, Denmark). Kinematic parameters of the sperm in both groups were analyzed using the Sperm Class Analyzer (Microptic S.L., Spain) at 0, 6 and 24 hours following the addition of media. Results...

  6. Effect of environmental pH on sperm kinematic characteristics

    DEFF Research Database (Denmark)

    Alipour, Hiva; Dardmeh, Fereshteh; Dissing, Marianne Charlotte

    2015-01-01

    Semen preparation medium have an important role in assisted reproduction techniques and their composition influences sperm binding and motility. Some studies have assessed the influence of pH on sperm kinetics. However, no study to date has assessed the effect of environmental pH on subtle...... differences in the details of the sperm movement (kinematics) of human sperm provided by computerized sperm analysis systems. This study was designed to assess the effect of two different media pH levels on kinematic parameters of the human sperm. Samples were prepared using the 40%/80% Pureception (Sage, USA......) density gradient and resuspended in customized sperm culture media with pH levels of 7.9 and 8.3 (Origio, Denmark). Kinematic parameters of the sperm in both groups were analyzed using the Sperm Class Analyzer (Microptic S.L., Spain) at 0, 6 and 24 hours following the addition of media. Results...

  7. Evidence for the function of hyperactivated motility in sperm.

    Science.gov (United States)

    Suarez, S S; Katz, D F; Owen, D H; Andrew, J B; Powell, R L

    1991-02-01

    After insemination, mammalian sperm undergo a striking change in flagellar beat pattern, termed hyperactivation. In low-viscosity culture medium, nonhyperactivated sperm flagella generate relatively symmetrical, low-amplitude waves, while hyperactivated sperm flagella generate an asymetrical beating pattern that results in nonprogressive movement. Since sperm encounter highly viscous and viscoelastic fluids in the female reproductive tract, the progress of hyperactivated sperm was compared with that of nonhyperactivated and transitional sperm in media of increasing viscosity. Hamster sperm obtained from the caudal epididymis were incubated in a medium that promotes capacitation. After 0, 3, and 4 h of incubation, the majority of the sperm exhibited, respectively, activated, transitional, and hyperactivated motility. At each of these time points, aliquots of sperm were removed from incubation and added to solutions of 0, 5%, 10%, 20%, and 30% Ficoll in medium. Samples containing mostly hyperactivated sperm (4 h) maintained higher swimming and flagellar velocities and were able to generate greater forces in response to increased viscous loading than activated sperm (0 h). Transitional sperm (3 h) showed an intermediate response. The paths of hyperactivated sperm through solutions of 20% and 30% Ficoll were considerably straighter than those made through medium alone. This is the first demonstration that hyperactivation can confer a mechanical advantage upon sperm in the oviduct where they may encounter viscous oviductal fluid and a viscoelastic cumulus matrix.

  8. Phenotypic plasticity in sperm traits in scorpionflies (Mecoptera : Panorpidae): Consequences of larval history and seasonality on sperm length and sperm transfer

    NARCIS (Netherlands)

    Vermeulen, Andreas; Engels, Sierk; Engqvist, Leif; Sauer, Klaus Peter

    2009-01-01

    We examined effects of seasonality, larval food availability and larval rearing density on sperm length, sperm transfer rates and body size in the bivoltine scorpionfly Panorpa vulgaris. Males of the first annual generation were larger and had larger sperm. Comparing individuals of two summer genera

  9. Canine sperm vitrification with sucrose: effect on sperm function.

    Science.gov (United States)

    Sánchez, R; Risopatrón, J; Schulz, M; Villegas, J; Isachenko, V; Kreinberg, R; Isachenko, E

    2011-08-01

    The ability of sucrose to protect spermatozoa against mitochondrial damage, artificial acrosome reaction and DNA fragmentation during ultra-rapid cryopreservation in canine sperm was investigated. Swim-up selected spermatozoa of second-fraction semen were vitrified with different concentrations of sucrose (0.1, 0.25 and 0.4 m) in proportion 1 : 1 v/v with HTF-BSA 1%. From each group, 30-μl suspensions of cells were dropped directly into liquid nitrogen and stored for at least 24 h. Cells were thawed by submerging the spheres in HTF with 1% BSA at 37 °C. The number of progressively motile spermatozoa was significantly higher in the sucrose 0.25 m + HTF-BSA 1% (42.5 ± 2.3%, P HTF only (1.66 ± 0.3%). The same combination of sucrose 0.25 m + HTF-BSA 1% (42.7 ± 1.5%) had a stronger cryoprotective effect on the integrity of mitochondrial membrane potential (P HTF only (1.93 ± 0.6% and 5.6 ± 0.6% respectively). With respect to acrosome-reacted spermatozoa, no significant difference was found between the groups investigated (P > 0.05). It is concluded that sucrose, a nonpermeable cryoprotectant, can effectively preserve important physiological parameters such as mitochondrial membrane potential and DNA integrity during ultra-rapid cryopreservation. © 2011 Blackwell Verlag GmbH.

  10. Neonatal outcome and congenital malformations in children born after ICSI with testicular or epididymal sperm

    DEFF Research Database (Denmark)

    Fedder, Jens; Loft, A; Parner, Erik Thorlund;

    2013-01-01

    Does neonatal outcome including congenital malformations in children born after ICSI with epididymal and testicular sperm [testicular sperm extraction (TESE)/percutaneous epididymal sperm aspiration (PESA)/testicular sperm aspiration (TESA) (TPT)] differ from neonatal outcome in children born after...

  11. Sperm chromatin structure assay results in Nigerian men with unexplained infertility

    National Research Council Canada - National Science Library

    Faduola, Paul; Kolade, Charles Oluwabukunmi

    2015-01-01

    .... This study aimed to compare the extent of sperm DNA damage in sperm samples from Nigerian men with unexplained infertility and in sperm samples from a fertile group composed of sperm donors who had...

  12. Effects of dilution and centrifugation on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey semen.

    Science.gov (United States)

    Miró, J; Taberner, E; Rivera, M; Peña, A; Medrano, A; Rigau, T; Peñalba, A

    2009-11-01

    The aim of this work was to study the effects of dilution and centrifugation (i.e., two methods of reducing the influence of the seminal plasma) on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey (Equus asinus) semen. Fifty ejaculates from nine Catalonian jackasses were collected. Gel-free semen was diluted 1:1, 1:5 or 1:10 with Kenney extender. Another sample of semen was diluted 1:5, centrifuged, and then resuspended with Kenney extender until a final dilution of 25x10(6) sperm/ml was achieved (C). After 24 h, 48 h or 72 h of refrigerated storage at 5 degrees C, aliquots of these semen samples were incubated at 37 degrees C for 5 min. The percentage of viable sperm was determined by staining with eosin-nigrosin. The motility characteristics of the spermatozoa were examined using the CASA system (Microptic, Barcelona, Spain). At 24h, more surviving spermatozoa were seen in the more diluted and in the centrifuged semen samples (1:1 48.71%; 1:5 56.58%, 1:10 62.65%; C 72.40%). These differences were maintained at 48 h (1:1 34.31%, 1:5 40.56%, 1:10 48.52%, C 66.30%). After 72 h, only the C samples showed a survival rate of above 25%. The four known donkey motile sperm subpopulations were maintained by refrigeration. However, the percentage of motile sperms in each subpopulation changed with dilution. Only the centrifuged samples, and only at 24h, showed exactly the same motile sperm subpopulation proportions as recorded for fresh sperm. However, the 1:10 dilutions at 24 and 48 h, and the centrifuged semen at 48 h, showed few variations compared to fresh sperm. These results show that the elimination of seminal plasma increases the survival of spermatozoa and the maintenance of motility patterns. The initial sperm concentration had a significant (P<0.05) influence on centrifugation efficacy, but did not influence the number of spermatozoa damaged by centrifugation. In contrast, the percentage of live

  13. Ocean acidification impacts on sperm mitochondrial membrane potential bring sperm swimming behaviour near its tipping point.

    Science.gov (United States)

    Schlegel, Peter; Binet, Monique T; Havenhand, Jonathan N; Doyle, Christopher J; Williamson, Jane E

    2015-04-01

    Broadcast spawning marine invertebrates are susceptible to environmental stressors such as climate change, as their reproduction depends on the successful meeting and fertilization of gametes in the water column. Under near-future scenarios of ocean acidification, the swimming behaviour of marine invertebrate sperm is altered. We tested whether this was due to changes in sperm mitochondrial activity by investigating the effects of ocean acidification on sperm metabolism and swimming behaviour in the sea urchin Centrostephanus rodgersii. We used a fluorescent molecular probe (JC-1) and flow cytometry to visualize mitochondrial activity (measured as change in mitochondrial membrane potential, MMP). Sperm MMP was significantly reduced in ΔpH -0.3 (35% reduction) and ΔpH -0.5 (48% reduction) treatments, whereas sperm swimming behaviour was less sensitive with only slight changes (up to 11% decrease) observed overall. There was significant inter-individual variability in responses of sperm swimming behaviour and MMP to acidified seawater. We suggest it is likely that sperm exposed to these changes in pH are close to their tipping point in terms of physiological tolerance to acidity. Importantly, substantial inter-individual variation in responses of sperm swimming to ocean acidification may increase the scope for selection of resilient phenotypes, which, if heritable, could provide a basis for adaptation to future ocean acidification. © 2015. Published by The Company of Biologists Ltd.

  14. SMART USE OF COMPUTER-AIDED SPERM ANALYSIS (CASA) TO CHARACTERIZE SPERM MOTION

    Science.gov (United States)

    Computer-aided sperm analysis (CASA) has evolved over the past fifteen years to provide an objective, practical means of measuring and characterizing the velocity and parttern of sperm motion. CASA instruments use video frame-grabber boards to capture multiple images of spermato...

  15. MONTHLY VARIATION IN SPERM MOTILITY IN COMMON CARP ASSESSED USING COMPUTER-ASSISTED SPERM ANALYSIS (CASA)

    Science.gov (United States)

    Sperm motility variables from the milt of the common carp Cyprinus carpio were assessed using a computer-assisted sperm analysis (CASA) system across several months (March-August 1992) known to encompass the natural spawning period. Two-year-old pond-raised males obtained each mo...

  16. Sperm retrieval outcomes with microdissection testicular sperm extraction (micro-TESE) in men with cryptozoospermia.

    Science.gov (United States)

    Alrabeeah, K; Wachter, A; Phillips, S; Cohen, B; Al-Hathal, N; Zini, A

    2015-05-01

    Several studies support of the use of testicular rather than ejaculated spermatozoa for intracytoplasmic sperm injection (ICSI) in couples with virtual azoospermia or cryptozoospermia, although this approach remains controversial. We sought to evaluate sperm retrieval outcomes with microdissection testicular sperm extraction (micro-TESE) in men with cryptozoospermia. We conducted a retrospective study of 24 consecutive micro-TESEs in men with cryptozoospermia. We also evaluated the outcomes of seven consecutive TESAs (testicular sperm aspiration) in cryptozoospermic men during the same time period (January 2007 and September 2014). Micro-TESE and TESA were performed on the day prior to ICSI. Final assessment of sperm recovery (reported on the day of ICSI) was recorded as (i) successful (available spermatozoa for ICSI) or (ii) unsuccessful (no spermatozoa for ICSI). The decision to perform a unilateral or bilateral micro-TESE was guided by the intra-operative evaluation of sperm recovery from the first testicle. A unilateral procedure was performed in 87.5% (21/24) and 57% (4/7) of the micro-TESE and TESA cohorts, respectively. Sperm recovery was successful in 96% (23/24) of the men who underwent micro-TESE and 43% (3/7) of the men who underwent TESA (p cryptozoospermia and few of these men will require a bilateral procedure. Moreover, sperm retrieval rates are higher with micro-TESE than TESA in this group of men.

  17. MONTHLY VARIATION IN SPERM MOTILITY IN COMMON CARP ASSESSED USING COMPUTER-ASSISTED SPERM ANALYSIS (CASA)

    Science.gov (United States)

    Sperm motility variables from the milt of the common carp Cyprinus carpio were assessed using a computer-assisted sperm analysis (CASA) system across several months (March-August 1992) known to encompass the natural spawning period. Two-year-old pond-raised males obtained each mo...

  18. SMART USE OF COMPUTER-AIDED SPERM ANALYSIS (CASA) TO CHARACTERIZE SPERM MOTION

    Science.gov (United States)

    Computer-aided sperm analysis (CASA) has evolved over the past fifteen years to provide an objective, practical means of measuring and characterizing the velocity and parttern of sperm motion. CASA instruments use video frame-grabber boards to capture multiple images of spermato...

  19. Human sperm chromosome analysis after subzonal sperm insemination of hamster oocytes

    Energy Technology Data Exchange (ETDEWEB)

    Cozzi, J. [Medical School of Grenoble (France)

    1994-09-01

    Sperm microinjection techniques, subzonal sperm insemination (SUZI) and intracytoplasmic sperm injection (ICSI), have achieved a wide spread clinical application for the treatment of male infertility. To date, only one study has focused on sperm karyotypes after microinjection. Martin et al. reported a very high incidence of abnormal human sperm complements after ICSI into hamster oocytes. In the present study, are reported the first human sperm karyotypes after SUZI of hamster oocytes. Spermatozoa from two control donors were treated by calcium ionophore A23187 and injected under the zona of hamster eggs. The microinjected eggs were then cultured for cytogenetic analysis of the pronuclei. Out of 47 analyzed sperm chromosome metaphases, 5 (10.6%) were abnormal, 4 (8.5%) were hypohaploid and 1 (2.1%) had a structural abnormality. The sex ratio was not significantly different from the expected 1:1 ratio. Rates of chromosomal abnormalities in microinjected spermatozoa were similar to those observed in spermatozoa inseminated with zona free eggs, suggesting that SUZI procedure per se does not increase sperm chromosomal abnormalities.

  20. Factors influencing sperm retrieval following testicular sperm extraction in nonobstructive azoospermia patients.

    Science.gov (United States)

    Salehi, Peyman; Derakhshan-Horeh, Marzieh; Nadeali, Zakiye; Hosseinzadeh, Majid; Sadeghi, Erfan; Izadpanahi, Mohammad Hossein; Salehi, Mansour

    2017-03-01

    Azoospermia owing to testicular disorders is the most severe manifestation of male infertility. The main concern for patients with nonobstructive azoospermia (NOA) is the probability of successful sperm retrieval following testicular sperm extraction (TESE). Therefore, the goal of this study was to determine predictive factors correlated with sperm retrieval. We assessed the testicular histopathological patterns, the choice of TESE surgical procedure, hormone levels, and chromosomal abnormalities in patients with NOA (n=170). The histopathology specimens were analyzed based on the histopathological patterns of hypospermatogenesis, maturation arrest, and Sertoli cell-only syndrome. The mean rate of sperm retrieval was 48.8%. The rate of sperm retrieval was significantly higher in the hypospermatogenesis group than in the other groups (p<0.001). There was a positive correlation between micro-TESE (vs. conventional TESE) and the sperm retrieval rate (odds ratio, 8.077; p<0.01). A logistic regression model demonstrated that high levels of follicle-stimulating hormone (FSH) and small testicular volume were significantly associated with lower chances of successful sperm retrieval. Some parameters, including testicular histopathology patterns, FSH levels, testicular volume, and method of TESE surgery, may be able to predict the chances of obtaining spermatozoa in patients with NOA. However, despite the efficiency of some predictive models, the hope of retrieving any functioning spermatozoa may be sufficient to disregard predictive factors of the success of intracytoplasmic sperm injection in these patients.

  1. Spermatozoal traits and sperm competition in Atlantic salmon: relative sperm velocity is the primary determinant of fertilization success.

    Science.gov (United States)

    Gage, Matthew J G; Macfarlane, Christopher P; Yeates, Sarah; Ward, Richard G; Searle, Jeremy B; Parker, Geoffrey A

    2004-01-06

    Sperm competition occurs when sperm from more than one male compete for fertilizations. This form of post-copulatory sexual selection is recognized as a significant and widespread force in the evolution of male reproductive biology and as a key determinant of differential male reproductive success. Despite its importance, however, detailed mechanisms of sperm competition at the gamete level remain poorly understood. Here, we use natural variation in spermatozoal traits among wild Atlantic salmon (Salmo salar), a species naturally adapted to sperm competition, to examine how the relative influences of sperm (i) number, (ii) velocity, (iii) longevity, and (iv) total length determine sperm competition success. Atlantic salmon fertilize externally, and we were therefore able to conduct controlled in vitro fertilization competitions while concurrently measuring spermatozoal traits within the aqueous micro-environment to which salmon gametes are naturally adapted. Microsatellite DNA fingerprinting revealed that a male's relative sperm velocity was the primary determinant of sperm competition success. There was no significant relationship between fertilization success and either relative sperm number or total length; sperm longevity showed an inverse relationship with competition success. These relationships were consistent for two experimental repeats of the in vitro fertilization competitions. Our results therefore show, under the natural microenvironment for salmon gametes, that relative sperm velocity is a key spermatozoal component for sperm competition success. Atlantic salmon sperm can be considered to enter a competition analogous to a race in which the fastest sperm have the highest probability of success.

  2. Improved risk assessment by screening sperm parameters.

    Science.gov (United States)

    Plassmann, S; Urwyler, H

    2001-02-28

    The question of whether a 4 or 9 week premating treatment period is more suitable in studies for effects on fertility and early embryonic development, and the extent to which the screening of sperm parameters may contribute to the detection of effects, has been under discussion since the ICH guideline changed in 1994/1995. This study presents a comparison between 4 and 9 weeks treatment with known male reproductive toxicants with regard to sperm motility, count, morphology, abnormal movements and testicular and epididymal histopathology. Mating outcome was examined after 4 weeks treatment. Three compounds with different targets and mechanisms of action were chosen: two testicular toxicants, Pyridoxine and Adriamycin and the epididymal toxicant, alpha-Chlorohydrine. Sperm motility was reduced in males treated with Pyridoxine (markedly) and alpha-Chlorohydrine (slightly) after 4 weeks treatment and in males treated with Adriamycin after 9 weeks treatment. With Pyridoxine and Adriamycin, sperm count was significantly increased after 4 weeks. Histopathological examination after 4 weeks showed characteristic changes leading to marked testicular tubular atrophy at 8/9 weeks, which was confirmed by a significantly reduced sperm count at 8/9 weeks. With alpha-Chlorohydrine, sperm count was not affected and the results of the histopathological examination were equivocal. Changes in sperm morphology were observed after 4/9 weeks of treatment with Pyridoxine. Mating outcome after 4 weeks was markedly affected with both Pyridoxine and alpha-Chlorohydrine, but no effect was observed with Adriamycin. The results of this study indicate that the two testicular toxicants would have been detected as male reproductive toxicants in a 4-week general toxicity study with routine testicular histopathology and examination of sperm parameters, without the need for mating trials. For the epididymal toxicant, alpha-Chlorohydrine, there was slightly reduced sperm motility after 4 weeks

  3. Aspects of Toxoplasma Infection on the Reproductive System of Experimentally Infected Rams (Ovis Aries

    Directory of Open Access Journals (Sweden)

    Welber Daniel Zanetti Lopes

    2009-01-01

    Full Text Available Eight reproductive rams with no prior reproductive disease were distributed into three groups of infection with T. gondii: GI, 3 rams, 2.0×105 P strain oocysts; GII, 3 rams, 1.0×106 RH strain tachyzoites; GIII, 2 control rams. Clinical parameters were measured and serological evaluations (IIF were performed. Presence of the parasite in the semen was investigated by PCR and bioassay techniques. The rams presented clinical alterations (hyperthermia and apathy related to toxoplasmosis in both groups infected with Toxoplasma gondii. All the inoculated rams responded to antigenic stimulus, producing antibodies against T. gondii from postinoculation day 5 onwards. In ovine groups I and II, the greatest titers observed were 1 : 4096 and 1 : 8192, respectively. In semen samples collected from these two groups, the presence of T. gondii was detected by bioassay and PCR. This coccidian was isolated (bioassay and PCR in tissue pools (testicles, epididymis, seminal vesicle, and prostrate from two rams infected presenting oocysts and in one presenting tachyzoites.

  4. Do Pilea Microphylla Improve Sperm DNA Fragmentation and Sperm Parameters in Varicocelized Rats?

    Science.gov (United States)

    Heidari, Reza; Alizadeh, Rafieh; Abbasi, Niloufar; Pasbakhsh, Parichehr; Hedayatpour, Azim; Farajpour, Mostafa; Khaleghi, Mohammad Reza; Abbasi, Mehdi; Dehpour, Ahmad Reza

    2015-01-01

    Varicocele is one of the most common causes of primary male infertility. Pilea microphylla (PM) is being used as folk medicine. This study was aimed to investigate the effects of PM in a rat model of varicocele. A total of 30 male Wistar rats were divided into control, sham, varicocele, accessory varicocele and PM-treated groups. After 10 weeks of varicocele induction, sperm parameters and chromatin (Aniline blue, acridine orange and toluidine blue) were evaluated, except for the treated and accessory groups that received 50 mg/kg PM orally daily for 10 weeks and then were sacrificed. Sperm parameters significantly decreased in varicocele groups (P DNA fragmentation and sperm parameters in varicocelized rats. Administration of PM led to significantly increased sperm parameters and AO staining (P DNA fragmentation in varicocelized rats. PM can reduce the damage to sperm DNA but not chromatin condensation.

  5. The "Ram Effect": A "Non-Classical" Mechanism for Inducing LH Surges in Sheep.

    Science.gov (United States)

    Fabre-Nys, Claude; Chanvallon, Audrey; Dupont, Joëlle; Lardic, Lionel; Lomet, Didier; Martinet, Stéphanie; Scaramuzzi, Rex J

    2016-01-01

    During spring sheep do not normally ovulate but exposure to a ram can induce ovulation. In some ewes an LH surge is induced immediately after exposure to a ram thus raising questions about the control of this precocious LH surge. Our first aim was to determine the plasma concentrations of oestradiol (E2) E2 in anoestrous ewes before and after the "ram effect" in ewes that had a "precocious" LH surge (starting within 6 hours), a "normal" surge (between 6 and 28h) and "late» surge (not detected by 56h). In another experiment we tested if a small increase in circulating E2 could induce an LH surge in anoestrus ewes. The concentration of E2 significantly was not different at the time of ram introduction among ewes with the three types of LH surge. "Precocious" LH surges were not preceded by a large increase in E2 unlike "normal" surges and small elevations of circulating E2 alone were unable to induce LH surges. These results show that the "precocious" LH surge was not the result of E2 positive feedback. Our second aim was to test if noradrenaline (NA) is involved in the LH response to the "ram effect". Using double labelling for Fos and tyrosine hydroxylase (TH) we showed that exposure of anoestrous ewes to a ram induced a higher density of cells positive for both in the A1 nucleus and the Locus Coeruleus complex compared to unstimulated controls. Finally, the administration by retrodialysis into the preoptic area, of NA increased the proportion of ewes with an LH response to ram odor whereas treatment with the α1 antagonist Prazosin decreased the LH pulse frequency and amplitude induced by a sexually active ram. Collectively these results suggest that in anoestrous ewes NA is involved in ram-induced LH secretion as observed in other induced ovulators.

  6. Elucidation of the RamA regulon in Klebsiella pneumoniae reveals a role in LPS regulation.

    Directory of Open Access Journals (Sweden)

    Shyamasree De Majumdar

    2015-01-01

    Full Text Available Klebsiella pneumoniae is a significant human pathogen, in part due to high rates of multidrug resistance. RamA is an intrinsic regulator in K. pneumoniae established to be important for the bacterial response to antimicrobial challenge; however, little is known about its possible wider regulatory role in this organism during infection. In this work, we demonstrate that RamA is a global transcriptional regulator that significantly perturbs the transcriptional landscape of K. pneumoniae, resulting in altered microbe-drug or microbe-host response. This is largely due to the direct regulation of 68 genes associated with a myriad of cellular functions. Importantly, RamA directly binds and activates the lpxC, lpxL-2 and lpxO genes associated with lipid A biosynthesis, thus resulting in modifications within the lipid A moiety of the lipopolysaccharide. RamA-mediated alterations decrease susceptibility to colistin E, polymyxin B and human cationic antimicrobial peptide LL-37. Increased RamA levels reduce K. pneumoniae adhesion and uptake into macrophages, which is supported by in vivo infection studies, that demonstrate increased systemic dissemination of ramA overexpressing K. pneumoniae. These data establish that RamA-mediated regulation directly perturbs microbial surface properties, including lipid A biosynthesis, which facilitate evasion from the innate host response. This highlights RamA as a global regulator that confers pathoadaptive phenotypes with implications for our understanding of the pathogenesis of Enterobacter, Salmonella and Citrobacter spp. that express orthologous RamA proteins.

  7. Seminal plasma concentrations of Anti-Müllerian hormone and inhibin B predict motile sperm recovery from cryopreserved semen in asthenozoospermic men: a prospective cohort study.

    Science.gov (United States)

    Nery, S F; Vieira, M A F; Dela Cruz, C; Lobach, V N M; Del Puerto, H L; Torres, P B; Rocha, A L L; Reis, A B; Reis, F M

    2014-11-01

    The rate of motile sperm recovery after cryopreservation is very variable and difficult to predict. Anti-Müllerian hormone (AMH) and inhibin B are produced by Sertoli cells and released into the seminal plasma, where they could be functional markers of spermatogenesis and sperm resistance to thermal stress. The aim of this study was to evaluate whether seminal plasma levels of AMH and inhibin B predict sperm recovery after cryopreservation. The study included 153 men enrolled prospectively during a semen analysis. The cohort was stratified by the fresh semen characteristics into: normal (n = 52), high sperm count (n = 55), asthenozoospermia (n = 23), and oligozoospermia (n = 23). The main outcome measure was motile sperm recovery rate, defined as post-thaw total motile sperm count × 100/pre-freezing total motile sperm count. In men with asthenozoospermia there was a significant correlation between motile sperm recovery rate and the pre-freezing concentrations of AMH (r = 0.522, p < 0.05) and inhibin B (0.471, p < 0.05). In this group, the areas under the receiver operating characteristic curves of AMH and inhibin B for prediction of ≥50% motile sperm recovery after cryopreservation were, respectively, 0.808 and 0.638. AMH was particularly useful, with sensitivity of 0.85, specificity of 0.80, positive predictive value of 0.84 and negative predictive value of 0.80. The sensitivity, specificity, positive, and negative predictive values of inhibin B for the same outcome were, respectively, 0.62, 0.60, 0.67, and 0.55. The median motile sperm recovery rate was 83% when seminal plasma AMH concentration was ≥0.84 ng/mL, vs. 27% when AMH concentration was <0.84 ng/mL (p < 0.05). In other patient groups, there was no correlation between the two hormone levels in seminal plasma and the motile sperm recovery rate. In conclusion, seminal plasma AMH and inhibin B concentrations correlate with and can be used to predict motile sperm recovery after semen

  8. Different computer-assisted sperm analysis (CASA) systems highly influence sperm motility parameters.

    Science.gov (United States)

    Boryshpolets, S; Kowalski, R K; Dietrich, G J; Dzyuba, B; Ciereszko, A

    2013-10-15

    In this study, we examined different computer-assisted sperm analysis (CASA) systems (CRISMAS, Hobson Sperm Tracker, and Image J CASA) on the exact same video recordings to evaluate the differences in sperm motility parameters related to the specific CASA used. To cover a wide range of sperm motility parameters, we chose 12-second video recordings at 25 and 50 Hz frame rates after sperm motility activation using three taxonomically distinct fish species (sterlet: Acipenser ruthenus L.; common carp: Cyprinus carpio L.; and rainbow trout: Oncorhynchus mykiss Walbaum) that are characterized by essential differences in sperm behavior during motility. Systematically higher values of velocity and beat cross frequency (BCF) were observed in video recordings obtained at 50 Hz frame frequency compared with 25 Hz for all three systems. Motility parameters were affected by the CASA and species used for analyses. Image J and CRISMAS calculated higher curvilinear velocity (VCL) values for rainbow trout and common carp at 25 Hz frequency compared with the Hobson Sperm Tracker, whereas at 50 Hz, a significant difference was observed only for rainbow trout sperm recordings. No significant difference was observed between the CASA systems for sterlet sperm motility at 25 and 50 Hz. Additional analysis of 1-second segments taken at three time points (1, 6, and 12 seconds of the recording) revealed a dramatic decrease in common carp and rainbow trout sperm speed. The motility parameters of sterlet spermatozoa did not change significantly during the 12-second motility period and should be considered as a suitable model for longer motility analyses. Our results indicated that the CASA used can affect motility results even when the same motility recordings are used. These results could be critically altered by the recording quality, time of analysis, and frame rate of camera, and could result in erroneous conclusions. Copyright © 2013 Elsevier Inc. All rights reserved.

  9. Moonlighting proteins in sperm-egg interactions.

    Science.gov (United States)

    Petit, François M; Serres, Catherine; Auer, Jana

    2014-12-01

    Sperm-egg interaction is a highly species-specific step during the fertilization process. The first steps consist of recognition between proteins on the sperm head and zona pellucida (ZP) glycoproteins, the acellular coat that protects the oocyte. We aimed to determine which sperm head proteins interact with ZP2, ZP3 and ZP4 in humans. Two approaches were combined to identify these proteins: immunoblotting human spermatozoa targeted by antisperm antibodies (ASAs) from infertile men and far-Western blotting of human sperm proteins overlaid by each of the human recombinant ZP (hrZP) proteins. We used a proteomic approach with 2D electrophoretic separation of sperm protein revealed using either ASAs eluted from infertile patients or recombinant human ZP glycoproteins expressed in Chinese-hamster ovary (CHO) cells. Only spots highlighted by both methods were analysed by MALDI-MS/MS for identification. We identified proteins already described in human spermatozoa, but implicated in different metabolic pathways such as glycolytic enzymes [phosphokinase type 3 (PK3), enolase 1 (ENO1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), aldolase A (ALDOA) and triose phosphate isomerase (TPI)], detoxification enzymes [GST Mu (GSTM) and phospholipid hydroperoxide glutathione peroxidase (PHGPx) 4], ion channels [voltage-dependent anion channel 2 (VDAC2)] or structural proteins (outer dense fibre 2). Several proteins were localized on the sperm head by indirect immunofluorescence, and their interaction with ZP proteins was confirmed by co-precipitation experiments. These results confirm the complexity of the sperm-ZP recognition process in humans with the implication of different proteins interacting with the main three ZP glycoproteins. The multiple roles of these proteins suggest that they are multifaceted or moonlighting proteins.

  10. Thyroxin Is Useful to Improve Sperm Motility

    Directory of Open Access Journals (Sweden)

    Mendeluk Gabriela Ruth

    2016-07-01

    Full Text Available Background The aim of this study was to evaluate the non-genomic action of thyroxin on sperm kinetic and its probable use to improve sperm recovery after applying an en- richment method like “swim-up” in comparison with the available one, pentoxifylline. Materials and Methods This is an experimental study. A total of 50 patients were re- cruited, followed by infertility consultation. Conventional sperm assays were performed according to World Health Organization criteria-2010 (WHO-2010. A Computer Aided Semen Analysis System was employed to assess kinetic parameters and concentrations. Number of the motile sperm recovered after preparation technique was calculated. Results Addition of T4 (0.002 µg/ml to semen samples increased hypermotility at 20 minutes (control: 14.18 ± 5.1% vs. 17.66 ± 8.88%, P<0.03, data expressed as mean ± SD and remained unchanged after 40 minutes. Significant differences were found in the motile sperm recovered after swim-up (control: 8.93×106 ± 9.52× 06vs. 17.20×106 ± 21.16×106, P<0.03, achieving all of the tested samples a desirable threshold value for artificial insemination outcome, while adding pentoxifylline increased the number of recovered sperm after swim-up in 60% of the studied cases. No synergism between two treatments could be determined. Conclusion We propose a new physiological tool to artificially improve insemination. The discussion opens windows to investigate unknown pathways involved in sperm ca- pacitation and gives innovative arguments to better understand infertility mechanisms.

  11. Endocannabinoids and Human Sperm Cells

    Directory of Open Access Journals (Sweden)

    Giovanna Zolese

    2010-10-01

    Full Text Available N-acylethanolamides (NAEs are naturally occurring signaling lipids consisting of amides and esters of long-chain polyunsaturated fatty acids. Usually they are present in a very small amounts in many mammalian tissues and cells, including human reproductive tracts and fluids. Recently, the presence of N-arachidonoylethanolamide (anandamide, AEA, the most characterised member of endocannabinoids, and its congeners palmitoylethanolamide (PEA and oleylethanolamide (OEA in seminal plasma, oviductal fluid, and follicular fluids was demonstrated. AEA has been shown to bind not only type-1 (CB1 and type-2 (CB2 cannabinoid receptors, but also type-1 vanilloid receptor (TRPV1, while PEA and OEA are inactive with respect to classical cannabinoid CB1 and CB2 but activate TRPV1 or peroxisome proliferator activate receptors (PPARs. This review concerns the most recent experimental data on PEA and OEA, endocannabinoid-like molecules which appear to exert their action exclusively on sperm cells with altered features, such as membrane characteristics and kinematic parameters. Their beneficial effects on these cells could suggest a possible pharmacological use of PEA and OEA on patients affected by some forms of idiopathic infertility.

  12. Acrosin activity is a good predictor of boar sperm freezability.

    Science.gov (United States)

    Pinart, Elisabeth; Yeste, Marc; Bonet, Sergi

    2015-06-01

    The main aim of this study was to determine whether acrosin activity could predict boar sperm freezability. For this purpose, we characterized the changes in sperm quality and acrosin activity throughout the cryopreservation procedure of sperm samples from 30 Pietrain boars by analyzing four critical steps: step 1 (extended sperm at 15 °C), step 2 (cooled sperm at 5 °C), step 3 (30 minutes postthaw), and step 4 (240 minutes postthaw). Freezability ejaculate groups were set on the basis of sperm motility and membrane integrity after freeze-thawing. Results obtained highlighted the low predictive value in terms of freezability of sperm motility and kinematics and sperm membrane integrity, as no differences between good and poor freezability ejaculates were seen before cryopreservation. Significant differences (P sperm kinetic parameters, and after thawing for sperm motility and membrane integrity. In contrast, acrosin activity appeared as an indicator of boar sperm freezability because the differences (P sperm kinematics, membrane lipid disorder, intracellular calcium content, acrosome integrity, and acrosin activity throughout the cryopreservation procedure were indicative of a significant damage in spermatozoa during the cooling step in both ejaculate groups. In conclusion, the main finding of our study is that acrosin activity can be used as a reliable predictor of boar sperm freezability because it differs significantly between good and poor freezability ejaculates yet before freeze-thawing procedures took place, i.e., in the refrigeration step at 15 °C.

  13. Experimental evolution of sperm competitiveness in a mammal

    Directory of Open Access Journals (Sweden)

    Simmons Leigh W

    2011-01-01

    Full Text Available Abstract Background When females mate with multiple partners, sperm from rival males compete to fertilise the ova. Studies of experimental evolution have proven the selective action of sperm competition on male reproductive traits. However, while reproductive traits may evolve in response to sperm competition, this does not necessarily provide evidence that sperm competitive ability responds to selection. Indeed, a study of Drosophila failed to observe divergence in sperm competitive ability of males in lines selected for enhanced sperm offence and defence. Results Adopting the naturally polygamous house mouse (Mus domesticus as our vertebrate model, we performed an experimental evolution study and observed genetic divergence in sperm quality; males from the polygamous selection lines produced ejaculates with increased sperm numbers and greater sperm motility compared to males from the monogamous lines. Here, after 12 generations of experimental evolution, we conducted competitive matings between males from lineages evolving under sperm competition and males from lineages subject to relaxed selection. We reduced variation in paternity arising from embryo mortality by genotyping embryos in utero at 14 days gestation. Our microsatellite data revealed a significant paternity bias toward males that evolved under the selective regime of sperm competition. Conclusion We provide evidence that the sperm competitiveness phenotype can respond to selection, and show that improved sperm quality translates to greater competitive fertilisation success in house mice.

  14. Investigation of obturator and ignitor effects on low velocity starting of the ram accelerator

    Energy Technology Data Exchange (ETDEWEB)

    Knowlen, C.; Bundy, C.; Bruckner, A.P. [Washington Univ., Seattle, WA (United States). Aerospace and Energetics Research Program; Kruczynski, D. [Washington Univ., Seattle, WA (United States). Aerospace and Energetics Research Program; Utron Inc., Manassas, VA (United States)

    2000-11-01

    An experimental investigation of the effects of obturator geometry, propellant chemistry, and onboard pyrotechnic ignitor systems on the starting characteristics of the ram accelerator at low launch velocity has been conducted at the University of Washington 38-mm-bore facility. The ram accelerator was successfully started at entrance velocities as low as 760 m/s using stoichiometric methane/oxygen propellants with various levels of carbon dioxide dilution and obturator configurations at a fill pressure of 2.5 MPa. Experiments using an onboard pyrotechnic ignitor demonstrated that ignition of the propellant and starting of the ram accelerator could occur without the presence of the obturator-driven normal shock. (orig.)

  15. Administration of flutamide alters sperm ultrastructure, sperm plasma membrane integrity and its stability, and sperm mitochondrial oxidative capability in the boar: in vivo and in vitro approach.

    Science.gov (United States)

    Lydka, M; Piasecka, M; Gaczarzewicz, D; Koziorowski, M; Bilinska, B

    2012-08-01

    Our previous work has shown that an anti-androgen flutamide administered pre- and post-natally induced adverse effects on the epididymal morphology and function of adult boars. The present investigation is aimed to understand the effect of flutamide and its metabolite on changes in sperm plasma membrane integrity and its stability, changes in mitochondrial oxidative capability and frequency of abnormal sperm. In vivo effects of flutamide (50 mg/kg b.w.) on sperm ultrastructure were examined by electron microscopic observations. In vitro effects of 5, 50 and 100 μg/ml hydroxyflutamide, administered for 2 and 24 h, on sperm plasma membrane integrity were measured by LIVE/DEAD Sperm Vitality kit, while those on sperm membrane stability and mitochondrial oxidoreductive activity were investigated using Merocyanine 540 and NADH tests, respectively. The incidence of abnormal spermatozoa increased significantly (p boars compared with controls. In an in vitro approach, low dose of hydroxyflutamide in 2-h incubations appeared less effective in altering the sperm plasma membrane integrity and its stability than two higher doses used (p sperm membrane destabilization and mitochondrial oxidoreductive activity was strengthened after 24 h of hydroxyflutamide administration (p sperm parameters with regard to oxidative capability of mitochondria, plasma membrane changes and sperm ultrastructure provides novel data on the boar sperm sensitivity to anti-androgen action. Results indicate high sensitivity of boar spermatozoa to androgen withdrawal.

  16. A Minimum Leakage Quasi-Static RAM Bitcell

    Directory of Open Access Journals (Sweden)

    Adam Teman

    2011-05-01

    Full Text Available As SRAMs continue to grow and comprise larger percentages of the area and power consumption in advanced systems, the need to minimize static currents becomes essential. This brief presents a novel 9T Quasi-Static RAM Bitcell that provides aggressive leakage reduction and high write margins. The quasi-static operation method of this cell, based on internal feedback and leakage ratios, minimizes static power while maintaining sufficient, albeit depleted, noise margins. This paper presents the concept of the novel cell, and discusses the stability of the cell under hold, read and write operations. The cell was implemented in a low-power 40 nm TSMC process, showing as much as a 12× reduction in leakage current at typical conditions, as compared to a standard 6T or 8T bitcell at the same supply voltage. The implemented cell showed full functionality under global and local process variations at nominal and low voltages, as low as 300 mV.

  17. A ram-pressure threshold for star formation

    CERN Document Server

    Whitworth, A P

    2016-01-01

    In turbulent fragmentation, star formation occurs in condensations created by converging flows. The condensations must be sufficiently massive, dense and cool to be gravitationally unstable, so that they start to contract; {\\it and} they must then radiate away thermal energy fast enough for self-gravity to remain dominant, so that they continue to contract. For the metallicities and temperatures in local star forming clouds, this second requirement is only met robustly when the gas couples thermally to the dust, because this delivers the capacity to radiate across the full bandwidth of the continuum, rather than just in a few discrete spectral lines. This translates into a threshold for vigorous star formation, which can be written as a minimum ram-pressure Pcrit ~ 4 10^-11 dyne. Pcrit is independent of temperature, and corresponds to flows with molecular hydrogen number-density nH2 and velocity v satisfying nH2 v^2 > 800 cm^-3 (km/s)^2. This in turn corresponds to a minimum molecular hydrogen column-density ...

  18. Cardiac Safety of Diclofenac at a Single Dose in Ram

    Directory of Open Access Journals (Sweden)

    Ayse Er

    2013-01-01

    Full Text Available Nonsteroidal anti-inflammatory drugs are frequently prescribed drug group in human and veterinary medicine. However, diclofenac, a traditional nonsteroidal anti-inflammatory drug, related to cardiotoxicity is reported, and blood cardiac damage markers may increase within the first hours after damage. The aim of the current research was to determine the effect of diclofenac on the blood cardiac damage markers. Single dose of diclofenac (2.5 mg/kg, IM was injected to 6 rams. Blood samples were collected in before (0 hour, control and 6 hours after injection. Specific (troponin I, and creatine kinase-MB and nonspecific (lactate dehydrogenase, aspartate aminotransferase blood cardiac damage marker concentrations, routine biochemical (hepatic damage, renal damage, lipid metabolism, glucose, and phosphorus parameters, and hemogram values were measured. Diclofenac increased (P<0.05 specific (troponin I and nonspecific cardiac (lactate dehydrogenase, aspartate aminotransferase, hepatic (aspartate aminotransferase, alkaline phosphatase, and alanine aminotransferase, and muscular (creatine kinase damage markers and high density lipoprotein level, while it decreased (P<0.05 low density lipoprotein level. Moreover, diclofenac decreased (P<0.05 white blood cell counts and increased (P<0.05 red blood cell counts. In conclusion, it may be stated that diclofenac shows slight cardiotoxicity, whereas it may show potent hepatic and muscular damage effects at an intramuscularly single dose in sheep. Thereby, repeated injections of diclofenac may be more harmful in sheep.

  19. Temperature management during semen processing: Impact on boar sperm quality under laboratory and field conditions.

    Science.gov (United States)

    Schulze, M; Henning, H; Rüdiger, K; Wallner, U; Waberski, D

    2013-12-01

    Freshly collected boar spermatozoa are sensitive to a fast reduction in temperature because of lipid phase transition and phase separation processes. Temperature management during semen processing may determine the quality of stored samples. The aim of this study was to evaluate the influence of isothermic and hypothermic semen processing protocols on boar sperm quality under laboratory and field conditions. In the laboratory study, ejaculates (n = 12) were first diluted (1:1) with Beltsville Thawing Solution (BTS) at 32 °C, then processed either with isothermic (32 °C) or hypothermic (21 °C) BTS, stored at 17 °C, and assessed on days 1, 3, and 6. Temperature curves showed that 150 minutes after the first dilution, semen doses of both groups reached the same temperature. Two-step hypothermic processing resulted in lower sperm motility on days 1 and 6 (P sperm on days 3 and 6 (P boar semen compared with isothermic dilution and that the type of semen extender affects the outcomes.

  20. Effects of very rapid versus vapor phase freezing on human sperm parameters.

    Science.gov (United States)

    Darvishnia, Hamid; Lakpour, Niknam; Lahijani, Maryam Shams; Heidari-Vala, Hamed; Akhondi, Mohammad A; Zeraati, Hojjat; Sadeghi, Mohammad Reza

    2013-12-01

    The aim of the present study was to compare the effects of two freezing methods, vapor phase and very rapid freezing, with and without cryoprotectant on semen parameters in men with normal semen criteria. Cryopreservation was done on semen samples from 31 men by two methods of vapor phase freezing and very rapid freezing, with and without Test Yolk buffered glycerol (TYBG) as cryoprotectant. The motility, viability, acrosome and DNA integrity were evaluated on fresh and post-thaw samples. Post-thaw sperm progressive motility was significantly higher in the presence of TYBG in the vapor phase cryopreservation (%6.30 ± 3.74) compared with the very rapid freezing method (%2.2 ± 1.97 and %4.00 ± 2.42 in the presence and absence of TYBG, respectively). There was no significant difference in viability, acrosome status and DNA integrity between two methods in presence or absence of TYBG. The very rapid freezing method in the absence of TYBG showed better sperm motility but viability, acrosome and DNA integrity were similar to the presence of TYBG. The results show that cryopreservation of human spermatozoa together with seminal plasma by using vapor phase method is better than very rapid freezing method to preserve sperm progressive motility; however very rapid freezing method is quick and simple and do not require special cryoprotectant. It can be used for cryopreservation of small number of spermatozoa in IVF centers.

  1. Sperm Cryopreservation before Testicular Cancer Treatment and Its Subsequent Utilization for the Treatment of Infertility

    Directory of Open Access Journals (Sweden)

    Jana Žáková

    2014-01-01

    Full Text Available Aims. In this study we report our results with storage of cryopreserved semen intended for preservation and subsequent infertility treatment in men with testicular cancer during the last 18 years. Methods. Cryopreserved semen of 523 men with testicular cancer was collected between October 1995 and the end of December 2012. Semen of 34 men (6.5% was used for fertilization of their partners. They underwent 57 treatment cycles with cryopreserved, fresh, and/or donor sperm. Results. A total of 557 men have decided to freeze their semen before cancer treatment. Azoospermia was diagnosed in 34 men (6.1%, and semen was cryopreserved in 532 patients. Seminoma was diagnosed in 283 men (54.1% and nonseminomatous germ cell tumors in 240 men (45.9%. 34 patients who returned for infertility treatment underwent 46 treatment cycles with cryopreserved sperm. Totally 16 pregnancies were achieved, that is, 34.8% pregnancy rate. Conclusion. The testicular cancer survivors have a good chance of fathering a child by using sperm cryopreserved prior to the oncology treatment, even when it contains only limited number of spermatozoa.

  2. Inhibition of sperm motility does not affect live-dead separation of bull sperm by glass beads

    Institute of Scientific and Technical Information of China (English)

    Robert H. Foote

    2001-01-01

    Aim: This study was designed to explore factors which influence binding of dead versus live sperm to glass filters.Methods: Multiple semen collections from bulls were used to explore selective filtration of bull sperm as influenced by nonlethal inhibition of sperm motility with fluoride, killing of sperm by quick-freezing, alteration of the glass surface with silicone, and different intervals of sexual rest between semen collections. Results: A comparison of glass spheres 100, 200 and 390 μm in diameter indicated that 200μm spheres were optimal for selective filtration. Quantitative separation of live from dead sperm was demonstrated with a correlation between the percentage of motile sperm and retention of sperm by the filter of r =-0.87 (P < 0.05). Up to 0.02 mol/L NaFl did not alter the proportion of sperm retained by the filter despite inhibiting sperm motility during filtration, an inhibition which was reversible. Proportions of live-dead sperm, based upon eosin staining, were unaffected by fluoride. Coating the glass spheres with silicone greatly reduced selective filtration. Dead sperm adherence to glass wasreduced and resistance to NaFl inhibition was increased by daily ejaculation versus one-week intervals of sexual rest. Conclusion: These studies indicate that the adherence of sperm to glass is primarily due to some form of physico-chemical change accompanying death of the sperm cell independent of active sperm motility. This attraction between the sperm plasma membrane and glass is modified by the age of the ejaculated sperm. This information is useful in evaluating different clinical procedures used for sperm separation.

  3. Direct effect of macrophage migration inhibitory factor on sperm function: possible involvement in endometriosis-associated infertility.

    Science.gov (United States)

    Carli, Cédric; Leclerc, Pierre; Metz, Christine N; Akoum, Ali

    2007-10-01

    To evaluate the effect of macrophage migration inhibitory factor (MIF) on sperm capacitation, a maturational process that occurs in the female reproductive tract and enables spermatozoa to become fully competent at fertilizing the oocyte. Incubation of Percoll-washed spermatozoa with varying concentrations of human recombinant MIF or fetal cord serum (positive control). Human reproduction research laboratories. Fresh semen samples obtained from healthy volunteers after a minimum of 2 days of sexual abstinence. Protein tyrosine phosphorylation by Western blotting, the acrosomal status upon binding to the Pisum sativum agglutinin conjugated to fluorescein isothiocyanate, and sperm motility by computer-assisted sperm analysis. MIF displayed a dose-dependent effect on the phosphotyrosine content of p105 and p81, the two major tyrosine-phosphorylated proteins associated with human sperm capacitation. A significant induction of tyrosine phosphorylation was seen at 2 ng/mL of MIF for both p105 and p81, but a trend for a down-regulation of the basal tyrosine phosphorylation level was noted at elevated concentrations (12-24 ng/mL). MIF pretreatment of spermatozoa resulted in a dose-dependent change in the acrosome reaction induced by the Ca(2+) ionophore A23187. After being increased at 1-4 ng/mL MIF with a statistically significant effect observed at 4 ng/mL, the acrosome reaction gradually decreased and fell below the control levels at higher concentrations. Furthermore, a significant decrease in the motility of spermatozoa was observed after exposure to an elevated concentration of MIF (12 ng/mL). The present data indicate that MIF may play a physiological role in sperm capacitation but may have deleterious effects on sperm function at abnormal pathophysiological levels, which suggests a role in endometriosis-associated infertility.

  4. Intracytoplasmic sperm injection (ICSI) and chromosomally abnormal spermatozoa

    NARCIS (Netherlands)

    P.A. in 't Veld; F.J.M. Broekmans (Frank); H.F. de France; P.L. Pearson; M.H. Pieters; R.J. van Kooij

    1997-01-01

    textabstractAn infertile couple was referred for intracytoplasmic sperm injection (ICSI) because of primary infertility and oligoasthenoteratozoospermia (OAT) in the male. It was observed that although the sperm cells presented with an unusual head size and multiple tai

  5. Third Party Reproduction: Sperm, Egg, and Embryo Donation and Surrogacy

    Science.gov (United States)

    ... third-party reproduction” refers to the use of eggs , sperm , or embryos that have been donated by a ... the requisite screening and testing required of all egg and sperm donors. For embryos that are created specifically for ...

  6. Sperm viability staining in ecology and evolution: potential pitfalls

    DEFF Research Database (Denmark)

    Holman, Luke

    2009-01-01

    The causes and consequences of variation in sperm quality, survival and ageing are active areas of research in ecology and evolution. In order to address these topics, many recent studies have measured sperm viability using fluorescent staining. Although sperm viability staining has produced...... a number of interesting results, it has some potential pitfalls that have rarely been discussed. In the present paper, I review the major findings of ecology and evolution studies employing sperm viability staining and outline the method's principle limitations. The key problem is that the viability assay...... may itself kill sperm, which is likely to confound many common experimental designs in addition to producing artificially low estimates of sperm viability. I further suggest that sperm number should be routinely measured in sperm viability studies, as it may be an important but overlooked source...

  7. Gulf of Mexico sperm whale photo-ID catalog

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Photo-identification data on sperm whales occupying the north central Gulf of Mexico have been collected during vessel surveys. Photographs of sperm whales are taken...

  8. Sperm use economy of honeybee (Apis mellifera) queens

    DEFF Research Database (Denmark)

    Baer, Boris; Collins, Jason; Maalaps, Kristiina

    2016-01-01

    The queens of eusocial ants, bees, and wasps only mate during a very brief period early in life to acquire and store a lifetime supply of sperm. As sperm cannot be replenished, queens have to be highly economic when using stored sperm to fertilize eggs, especially in species with large and long......-lived colonies. However, queen fertility has not been studied in detail, so that we have little understanding of how economic sperm use is in different species, and whether queens are able to influence their sperm use. This is surprising given that sperm use is a key factor of eusocial life, as it determines...... the fecundity and longevity of queens and therefore colony fitness. We quantified the number of sperm that honeybee (Apis mellifera) queens use to fertilize eggs. We examined sperm use in naturally mated queens of different ages and in queens artificially inseminated with different volumes of semen. We found...

  9. Comparison of the major malformation rate of children conceived from cryopreserved embryos and fresh embryos

    Institute of Scientific and Technical Information of China (English)

    LI Hong-zhen; QIAO Jie; CHI Hong-bin; CHEN Xin-na; LIU Ping; MA Cai-hong

    2010-01-01

    Background Cryopreserved embryo transfer has become indispensable in reproductive technology. More and more children are conceived from frozen-thawed embryo transfer (FET). The risk of birth defects associated with frozen-thawed embryo transfer has been evaluated and conflict results are obtained. The aim of this study was to compare the rate of major malformations in children conceived from cryopreserved embryos with that of children from fresh embryos. Methods A retrospective analysis was performed on children conceived from frozen-thawed embryos and fresh embryos between January 2005 and December 2008 at the Reproduction Center of the Third Hospital, Peking University.The major malformation rates were compared between two groups for all children, as well as singletons or twins,separately. The frequencies of different subtypes of malformations classified according to different organ system were also compared.Results Thirty-four of 3125 children from cryopreserved embryos had a major malformation. The malformation rate was 1.09%, which was comparable to that for children after fresh embryos transfer (1.53%(55/3604), OR:0.71, 95% CI; 0.46-1.09). The malformation rate was also similar when the analysis was limited to children from cryopreserved embryos resulted from in vitro fertilization (IVF)(1.39%)and fresh IVF(1.3%). However, children from cryopreserved embryos resulted from intracytoplasmic sperm injections (ICSI) had much lower malformation rate than from fresh ICSI(0.63% vs.1.83%, OR: 0.34, 95% CI: 0.16-0.75). No difference was found in the incidence of major malformations in singletons from cryo ICSI (0.73%) and fresh ICS1(1.9%), or from cryo IVF(1.49%) and fresh IVF(1.67%). Similar malformation rate was found in multiples from cryo ICSI(0.52%) and fresh ICSI(1.76%), or cryo IVF(1.30%) and fresh IVF(0.90%). The distribution and risk of the subtype of malformations, such as cardiovascular, gastrointestinal, neural tube, urogenital, rnusculoskeletal and facial

  10. Activity of antioxidative enzymes in fresh and frozen thawed buffalo (Bubalus bubalis) spermatozoa in relation to lipid peroxidation and semen quality

    Institute of Scientific and Technical Information of China (English)

    G Kadirvel; SatishKumar; S K Ghosh; P Perumal

    2014-01-01

    Objective:To investigate the activity of antioxidative enzymes in fresh and frozen thawed spermatozoa in relation to lipid peroxidation and semen quality in buffalo(Bubalus bubalis) bulls.Methods:Forty two semen ejaculates from seven buffalo bulls were collected by artificial vagina method and were used for the study.Sperm motility, livability, plasma membrane and acrosomal integrity, buffalo cervical mucous penetration test were assessed in fresh and frozen thawed semen.Intracellular antioxidative enzymatic activity such as super oxide dismutase (SOD), catalase(CAT), glutathione peroxidase(GSHPx) and reduced glutathione(GSH), reactive oxygen species(ROS) and lipid peroxidation(LPO) were estimated in fresh and frozen thawed semen.Results:A significant(P<0.01) reduction in activity of antioxidative enzymes(SOD by 47.7%,GSHPx by62.7% andGSH by58.6%) in frozen thawed spermatozoa as compared to fresh spermatozoa was found.Although the catalase activity was varied from0 to3.8IU/109sperm in fresh semen, but after freezing and thawing this activity was not detectable.These enzyme activities had a strong positive association with sperm motility, membrane integrity and distance traveled by vanguard spermatozoa in buffalo cervical mucus and negative correlation withLPO andROS. However, no significant correlation with acrosomal integrity was found.Conclusion:It was concluded that loss of activity of intracellular antioxidative enzymes was evident after freezing and thawing and there was a strong association between the antioxidative enzyme activities,ROS, lipid peroxidation and sperm function in buffalo semen.

  11. Speract, a sea urchin egg peptide that regulates sperm motility, also stimulates sperm mitochondrial metabolism.

    Science.gov (United States)

    García-Rincón, Juan; Darszon, Alberto; Beltrán, Carmen

    2016-04-01

    Sea urchin sperm have only one mitochondrion, that in addition to being the main source of energy, may modulate intracellular Ca(2+) concentration ([Ca(2+)]i) to regulate their motility and possibly the acrosome reaction. Speract is a decapeptide from the outer jelly layer of the Strongylocentrotus purpuratus egg that upon binding to its receptor in the sperm, stimulates sperm motility, respiration and ion fluxes, among other physiological events. Altering the sea urchin sperm mitochondrial function with specific inhibitors of this organelle, increases [Ca(2+)]i in an external Ca(2+) concentration ([Ca(2+)]ext)-dependent manner (Ardón, et al., 2009. BBActa 1787: 15), suggesting that the mitochondrion is involved in sperm [Ca(2+)]i homeostasis. To further understand the interrelationship between the mitochondrion and the speract responses, we measured mitochondrial membrane potential (ΔΨ) and NADH levels. We found that the stimulation of sperm with speract depolarizes the mitochondrion and increases the levels of NADH. Surprisingly, these responses are independent of external Ca(2+) and are due to the increase in intracellular pH (pHi) induced by speract. Our findings indicate that speract, by regulating pHi, in addition to [Ca(2+)]i, may finely modulate mitochondrial metabolism to control motility and ensure that sperm reach the egg and fertilize it.

  12. Does breastfeeding influence future sperm quality and reproductive hormones?

    DEFF Research Database (Denmark)

    Laustsen, J M; Jensen, M S; Thulstrup, Ane Marie;

    2011-01-01

    was not statistically significantly associated with sperm concentration, total sperm count, sperm motility or morphology, oligozoospermia, follicle-stimulating hormone, inhibin B, luteinizing hormone, sex hormone-binding globulin (SHBG), the calculated level of free testosterone, free oestradiol, the free testosterone...... testosterone nor free oestradiol was different between the two groups. This study shows no association between breastfeeding and sperm quality or reproductive hormones and a strong association is unlikely. A larger study would be needed to detect more subtle effects....

  13. Integrated and independent evolution of heteromorphic sperm types

    OpenAIRE

    Moore, A.J.; Bacigalupe, L.D.; Snook, R.R.

    2013-01-01

    Sperm are a simple cell type with few components, yet they exhibit tremendous between-species morphological variation in those components thought to reflect selection in different fertilization environments. However, within a species, sperm components are expected to be selected to be functionally integrated for optimal fertilization of eggs. Here, we take advantage of within-species variation in sperm form and function to test whether sperm components are functionally and genetically integra...

  14. A single-center study examining the outcomes of percutaneous epididymal sperm aspiration in the treatment of obstructive azoospermia

    Directory of Open Access Journals (Sweden)

    Jason R Kovac

    2014-01-01

    Full Text Available Introduction: Obstructive azoospermia (OA is characterized by normal spermatogenesis and the absence of sperm in the ejaculate. Variable success rates have been reported using in-vitro fertilization (IVF combined with PESA in cases of men with OA. Aims: To determine fertilization and pregnancy outcomes from PESA-derived spermatozoa and to ascertain whether frozen spermatozoa yields similar outcomes compared to fresh specimens. Materials and Methods: The charts of 68 consecutive couples undergoing 68 cycles of sperm retrieval for OA over eight years (2002-2010 were retrospectively reviewed. Patients requiring testicular intervention were excluded ( n = 17. Results: Viable sperms were identified in 100% of men, and fresh spermatozoa were obtained in 40 patients (78.4% simultaneously with female egg retrieval. The average fertilization rate was 77.7% with five embryos not surviving to transfer (12.5%. Pregnancies were confirmed in 48.6% (17/35. Twin gestations occurred in 11.8% (2/17 of cases. Frozen-thawed spermatozoa were used in 11 patients (21.6%. In this subgroup, the average fertilization rate was 73.6% with pregnancies confirmed in 54.5% (6/11. No multiple gestations were generated, and no complications occurred. The use of fresh spermatozoa for PESA provided no significant improvements in outcomes over frozen specimens. Summary: PESA is a very effective, simple, and safe method of obtaining spermatozoa for IVF. Outcomes obtained using fresh and frozen PESA-derived spermatozoa were similar and as such, either could be used during the IVF process.

  15. Optimal Design of FPGA Switch Matrix with Ion Mobility Based Nonvolatile ReRAM

    Directory of Open Access Journals (Sweden)

    Peng Hai-yun

    2015-01-01

    Full Text Available There are high demands for research of new device with greater accessing speed and stability to replace the current SRAM storage cell. The resistive random access memory (ReRAM is a metal oxide which is based on nonvolatile memory device possessing the characteristics of high read/write speed, high storage density, low power, low cost, very small cell, being nonvolatile, and unlimited writing endurance. The device has extreme short erasing time and the stored charge cannot be destroyed after power-off. Therefore, the ReRAM device is a significant storage device for many applications in the next generation. In this paper, we first explored the mechanism of the ReRAM device based on ion mobility model and then applied this device to optimize the design of FPGA switching matrix. The results show that it is beneficial to enhance the FPGA performance to replace traditional SRAM cells with ReRAM cells for the switching matrix.

  16. Gas Loss by Ram Pressure Stripping and Internal Feedback From Low Mass Milky Way Satellites

    CERN Document Server

    Emerick, Andrew; Grcevich, Jana; Gatto, Andrea

    2016-01-01

    The evolution of dwarf satellites of the Milky Way is affected by the combination of ram pressure and tidal stripping, and internal feedback from massive stars. We investigate gas loss processes in the smallest satellites of the Milky Way using three-dimensional, high resolution, idealized wind tunnel simulations, accounting for gas loss through both ram pressure stripping and expulsion by supernova feedback. Using initial conditions appropriate for a dwarf galaxy like Leo T, we investigate whether or not environmental gas stripping and internal feedback can quench these low mass galaxies on the expected timescales, shorter than 2 Gyr. We find that supernova feedback contributes negligibly to the stripping rate for these low star formation rate galaxies. However, we also find that ram pressure stripping is less efficient than expected in the stripping scenarios we consider. Our work suggests that, although ram pressure stripping can eventually completely strip these galaxies, other physics is likely at play t...

  17. Investigation on MCU Clustering Methodologies for Cross-Section Estimation of RAMs

    CERN Document Server

    Bosser, A; Tsiligiannis, G; Javanainen, A; Kettunen, H; Puchner, H; Saigne, F; Virtanen, A; Wrobel, F; Dilillo, L

    2015-01-01

    During irradiation testing of RAMs, various failure scenarios may occur which may generate different characteristic Multiple Cell Upset (MCU) error patterns. This work proposes a method based on spatial and temporal criteria to identify them.

  18. Cryopreservation of ram semen with antioxidant supplemented soybean lecithin-based extenders and impacts on incubation resilience.

    Science.gov (United States)

    Toker, M Berk; Alcay, Selim; Gokce, Elif; Ustuner, Burcu

    2016-06-01

    The scope of this study was investigation the affects of various antioxidants on 1% soybean lecithin-based semen extenders for ram semen cryopreservation. Ejaculates, collected via electrically stimulated ejaculation, that have a thick consistency, rapid wave motion (3-5 on a 0-5 scale) and >75% initial motility were pooled. The pooled samples were split into four equal aliquots as 5 mM Methionine, 5 mM Cysteamine, 1 mM Cysteine and a sample of antioxidant-free control group. Each sample group was diluted to a ratio of 1/5 (semen/extender, v/v) as final concentration and two step dilution method was used for cryopreservation. Extender groups were assessed for sperm motility, plasma membrane functional integrity using hypoosmotic swelling test (HOST), damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) and DNA integrity using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Semen samples also incubated for 6 h in humidified air with 5% CO2 at 39 °C to evaluate post-thaw incubation resilience of semen characteristics. The results showed that freezing and thawing procedures had negative effects on motility (P < 0.05), plasma membrane integrity (P < 0.05) and acrosomal integrity (P < 0.05). After 6 h of incubation time, the Cysteine supplemented extender group yielded significantly higher results than other extender groups in terms of spermatological parameters. Furthermore MDA levels in the antioxidant groups were lower than control group (P < 0.05). Nevertheless, there were no significant differences among antioxidant groups. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Effect of dietary fish oil supplementation on ram semen freeze ability and fertility using soybean lecithin- and egg yolk-based extenders.

    Science.gov (United States)

    Masoudi, R; Sharafi, M; Zare Shahneh, A; Towhidi, A; Kohram, H; Zhandi, M; Esmaeili, V; Shahverdi, A

    2016-10-01

    Ram semen cryopreservation is not efficient for artificial insemination in commercial herds. Beneficial effects of dietary fish oil have been evaluated for cryopreservation of ram semen in soybean lecithin (SL) and egg yolk (EY)-based extenders. A factorial study (two diets × two extenders) was used to analyze the effects of two diets supplemented with fish oil (n-3 fatty acid) or palm oil (saturated fatty acids; [SFAs]) to freeze ram semen in two extenders containing SL or EY. Motility characteristics, membrane integrity, abnormal morphology, mitochondria activity, acrosome integrity, apoptotic status, and fertilizing ability were assessed after freeze-thawing. Although diet had significant (P ≤ 0.05) effects on the quality parameters of frozen-thawed sperm, effects of extenders on these traits were not significant (P > 0.05). The higher significant (P ≤ 0.05) percentage of total motility and progressive motility were observed in n-3/SL (44.83 ± 1.56 and 28.33 ± 1.4) and n-3/EY (43.33 ± 1.56 and 28.50 ± 1.4) than SFA/SL (32.16 ± 1.56 and 14.00 ± 1.4) and SFA/EY (31.66 ± 1.56 and 12.66 ± 1.4) groups. Moreover, n-3/SL and n-3/EY produced the higher significant (P ≤ 0.05) percentage of membrane integrity of sperm (39.83 ± 1.4 and 37.33 ± 1.4) than SFA/SL and SFA/EY (29.83 ± 1.4 and 28.5 ± 1.4). For viability results, the higher significant percentage of live sperm was observed in n-3/SL and n-3/EY (43.16 ± 1.38 and 45.66 ± 1.38) than SFA/SL and SFA/EY (28.66 ± 1.38 and 27.5 ± 1.38). For fertility trials, n-3-based diets (n-3/SL and n-3/EY) improved significantly (P ≤ 0.05) pregnancy rate (44% and 46%), parturition rate (42% and 42%), and lambing rate (46% and 44%) compared with the SFA-based diets (SFA/SL and SFA/EY). No interaction effects have been found between diets and extenders (P > 0.05). It seems that dietary fish oil can improve the semen performance after freezing-thawing process and

  20. Sperm dilution ratio affects post-thaw motility rate and velocity of Prochilodus lineatus (Characiformes) sperm.

    Science.gov (United States)

    Viveiros, Ana T M; Leal, Marcelo C

    2016-10-01

    There is a lack of standardization in sperm cryopreservation of aquatic organisms and, thus, a necessity of more accurate investigations in all steps of this process. In this study, the effects of sperm dilution ratio on post-thaw sperm quality of Prochilodus lineatus were evaluated. Sperm was diluted in a standard freezing medium (glucose and methyl glycol) at four different ratios (sperm to final volume = 1:5, 1:10, 1:50 or 1:100), frozen in a nitrogen vapour vessel at -170°C and then stored in liquid nitrogen vessel at -196°C. Post-thaw motility rate and velocities (curvilinear = VCL; average path = VAP; straight line = VSL) were determined using a Computer-Assisted Sperm Analyzer (CASA) at 10 and 40 s post-activation. The highest motility rates were observed when sperm was frozen at a ratio of 1:5 (76%) and 1:10 (75%). The highest VCL (225 μm/s) and VAP (203 μm/s) were observed at a ratio of 1:10, while VSL was similar among samples frozen at 1:5, 1:10 and 1:50 (97-124 μm/s). When those parameters were evaluated again 30 s later, motility decreased significantly in samples frozen at a ratio of 1:5 (57%) and 1:10 (61%), while velocities decreased significantly in all samples regardless of dilution ratio (75-85 μm/s of VCL, 38-53 μm/s of VAP and 25-39 μm/s of VSL). P. lineatus sperm should be frozen at a ratio of 1:10, where both the number of loaded sperm per straw and the post-thaw quality are maximized.

  1. Sperm preparation: state-of-the-art-physiological aspects and application of advanced sperm preparation methods

    Institute of Scientific and Technical Information of China (English)

    Ralf Henkel

    2012-01-01

    For assisted reproduction technologies (ART),numerous techniques were developed to isolate spermatozoa capable of fertilizing oocytes.While early methodologies only focused on isolating viable,motile spermatozoa,with progress of ART,particularly intracytoplasmic sperm injection (ICSI),it became clear that these parameters are insufficient for the identification of the most suitable spermatozoon for fertilization.Conventional sperm preparation techniques,namely,swim-up,density gradient centrifugation and glass wool filtration,are not efficient enough to produce sperm populations free of DNA damage,because these techniques are not physiological and not modeled on the stringent sperm selection processes taking place in the female genital tract.These processes only allow one male germ cell out of tens of millions to fuse with the oocyte.Sites of sperm selection in the female genital tract are the cervix,uterus,uterotubal junction,oviduct,cumulus oophorus and the zona pellucida.Newer strategies of sperm preparation are founded on:(i) morphological assessment by means of‘motile sperm organelle morphological examination (MSOME)'; (ii) electrical charge; and (iii) molecular binding characteristics of the sperm cell.Whereas separation methods based on electrical charge take advantage of the sperm's adherence to a test tube surface or separate in an electrophoresis,molecular binding techniques use Annexin V or hyaluronic acid (HA) as substrates.Techniques in this category are magnet-activated cell sorting,Annexin V-activated glass wool filtration,flow cytometry and picked spermatozoa for ICSI (PICSI) from HA-coated dishes and HA-containing media.Future developments may include Raman microspectrometry,confocal light absorption and scattering spectroscopic microscopy and polarization microscopy.

  2. Sperm fractions obtained following density gradient centrifugation in human ejaculates show differences in sperm DNA longevity

    Institute of Scientific and Technical Information of China (English)

    Jaime Goslvez; Stephen Johnston; Carmen Lpez-Fernndez; Altea Goslbez; Francisca Arroyo; Jose Lus Fernndez; Juan G lvarez

    2014-01-01

    Objective:To investigate the DNA longevity characteristics associated with each resultant fraction following density gradient centrifugation (DGC) in comparison to that of the original neat ejaculated sample. Methods:An aliquot of neat semen (NSS) collected from 7 patients was processed using DGC resulting in 3 fractions;Fraction 1:seminal plasma/40%gradient interface (GI);Fraction 2:40%GI/80%GI;Fraction 3:80%GI/pellet. An aliquot of each fraction and NSS was cryopreserved, thawed and incubated at 37 ℃for 24h;the increase of sperm DNA fragmentation was assessed using the Dyn-Halosperm assay following 0, 3, 6 and 24h of incubation. Results:While there was a significant reduction in the incidence of baseline sperm DNA fragmentation following DGC in Fraction 3, sperm DNA longevity was shown to be higher in the NSS than in any other sub-population following incubation. The highest levels of baseline DNA damage were found in Fractions 1 and 2;these fractions also showed the highest rate DNA fragmentation following incubation, subsequently exhibiting the lowest DNA longevity. Conclusion:1) Unnecessary incubation of spermatozoa prior to artificial insemination or in vitro fertilization, should be avoided, since sperm DNA longevity is significantly reduced after ex vivo sperm handling and 2) Although sperm selection by DCG significantly reduces the baseline levels of SDF of sperm in Fraction 3, sperm DNA longevity in this fraction was ultimately lower following 24 h incubation when compared to sperm recovered from non-centrifuged NSS.

  3. Unraveling the sperm proteome and post-genomic pathways associated with sperm nuclear DNA fragmentation.

    Science.gov (United States)

    Intasqui, Paula; Camargo, Mariana; Del Giudice, Paula T; Spaine, Deborah M; Carvalho, Valdemir M; Cardozo, Karina H M; Cedenho, Agnaldo P; Bertolla, Ricardo P

    2013-09-01

    Sperm DNA fragmentation has been suggested as a marker for infertility diagnosis and prognosis. Hence, understanding its impact on male physiology and post-genomic pathways would be clinically important. We performed the proteomics and functional enrichment analyses of viable spermatozoa from ejaculates with low and high sperm DNA fragmentation to identify protein expression and pathways altered in association with sperm DNA fragmentation. Sperm DNA fragmentation using the Comet assay and the Komet 6.0.1 software was assessed in raw samples from 89 subjects from a human reproduction service. The Low and High sperm DNA fragmentation groups were formed according to the Olive Tail Moment variable. Spermatozoa proteins from these groups were pooled and analyzed by a shotgun proteomic approach (2D nanoUPLC-ESI-MS(E)). Differentially expressed proteins were used for a functional enrichment study. Two hundred and fifty-seven proteins were identified or quantified in sperm from the Low and High sperm DNA fragmentation groups. Of these, seventy-one proteins were exclusively or overexpressed in the Low group, whereas twenty-three proteins were exclusively or overexpressed in the High group. One hundred and sixty-three proteins were conserved between these groups. We also functionally related the differentially expressed proteins in viable spermatozoa from the groups. Processes such as triacylglycerol metabolism, energy production, protein folding, response to unfolded proteins, and cellular detoxification were found to be altered in these cells. Sperm DNA fragmentation is associated with differential protein expression in viable spermatozoa. These proteins may potentially be used as biomarkers for sperm DNA integrity.

  4. Could using the zona pellucida bound sperm for intracytoplasmic sperm injection(ICSI)enhance the outcome of ICSI?

    Institute of Scientific and Technical Information of China (English)

    De-Yi Liu

    2011-01-01

    @@ In the recent literature, several interesting articles have been published using the zona pellucdia(ZP)-bound sperm for intra-cytoplasmic sperm injection(ICSI)to enhance embryo quality, implantation and clinical pregnancy rates.1-3

  5. Cryopreservation of mutton snapper ( Lutjanus analis) sperm.

    Science.gov (United States)

    Sanches, Eduardo G; Oliveira, Idili R; Serralheiro, Pedro C Da Silva; Cerqueira, Vinicius R

    2013-09-01

    This study aimed to develop a protocol of semen cryopreservation of the mutton snapper Lutjanus analis. The interaction between three extenders ( pH 6.1; 7.8 and 8.2) , two concentrations of dimethyl sulfoxide ( DMSO, 5 and 10%) and three cooling rates ( -90; -60 and -30°C.min-1) on the sperm motility rate and motility time were analyzed by a factorial experiment. A sample of 30 fishes ( 1,261 ± 449 g) collected in the nature was kept in floating net cages. The semen was frozen by using cryogenic straws, in nitrogen vapour and transferred, later, to liquid nitrogen. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The highest sperm motility rate and motility time ( P snapper.

  6. Trehalose enhances osmotic tolerance and suppresses lysophosphatidylcholine-induced acrosome reaction in ram spermatozoon.

    Science.gov (United States)

    Ahmad, E; Naseer, Z; Aksoy, M; Küçük, N; Uçan, U; Serin, I; Ceylan, A

    2015-09-01

    This study was aimed to investigate the influence of trehalose on osmotic tolerance and the ability of ram spermatozoon to undergo acrosome reaction induced by lysophosphatidylcholine (LPC). In experiment 1, the diluted ejaculates were exposed to anisosmotic fructose solutions (70, 500, 750 and 1000 mOsm l(-1) ) with or without 50 mm trehalose. The presence of trehalose in hyperosmotic conditions enhanced (P ram spermatozoon and suppresses their ability to undergo LPC and cryo-induced acrosome reaction.

  7. Radiation immune RAM semiconductor technology for the 80's. [Random Access Memory

    Science.gov (United States)

    Hanna, W. A.; Panagos, P.

    1983-01-01

    This paper presents current and short term future characteristics of RAM semiconductor technologies which were obtained by literature survey and discussions with cognizant Government and industry personnel. In particular, total ionizing dose tolerance and high energy particle susceptibility of the technologies are addressed. Technologies judged compatible with spacecraft applications are ranked to determine the best current and future technology for fast access (less than 60 ns), radiation tolerant RAM.

  8. Experimental Study of Cement Mortar-Steel Fiber Reinforced Rammed Earth Wall

    OpenAIRE

    Pang, Miao; Yang, Shuai; Zhang, Yongqiang

    2012-01-01

    Rammed earth construction is an ancient technique which has recently attracted renewed interest throughout the world. Although rammed earth is currently regarded as a promising material in the construction industry in the context of sustainable development, it is difficult to quantify its bearing capacity, mechanical performance, as well as retrofitting approach, which discourages people from large-scale application in architectural engineering. This paper is devoted to the study of these pro...

  9. A Report of Contemporary Rammed Earth Construction and Research in North America

    OpenAIRE

    Arno Schmidt; Bly Windstorm

    2013-01-01

    Contemporary stabilized rammed earth (SRE) draws upon traditional rammed earth (RE) methods and materials, often incorporating reinforcing steel and rigid insulation, enhancing the structural and energy performance of the walls while satisfying building codes. SRE structures are typically engineered by licensed Structural Engineers using the Concrete Building Code or the Masonry Building Code. The construction process of SRE creates structural walls of relatively high compressive strength app...

  10. Rammed earth walls in the late middle age castles in the actual province of Soria, Spain

    OpenAIRE

    Gil Crespo, Ignacio Javier

    2013-01-01

    This paper analyzed the building techniques based in the use of rammed earth in the Late Medieval fortifications in the province of Soria, Spain. The manorial castles of Serón de Nágima and Yanguas were built completely with rammed earth. However, these techinques are different. Through the study of the constructive signals, we can reconstruct the constructive process and made an hypothesis of the auxiliary scaffolding necessary for the construction. After, other four cases in which there is ...

  11. Evolution: sperm, cryptic choice, and the origin of species.

    Science.gov (United States)

    Chippindale, Adam K

    2013-10-07

    In two fruit fly species, in vivo observations of competing sperm reveal how differences in sperm size, female behavior and reproductive architecture promote retention of same-species sperm. Sexual selection continues after mating and may play an important role in speciation.

  12. 21 CFR 173.275 - Hydrogenated sperm oil.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Hydrogenated sperm oil. 173.275 Section 173.275... CONSUMPTION Solvents, Lubricants, Release Agents and Related Substances § 173.275 Hydrogenated sperm oil. The food additive hydrogenated sperm oil may be safely used in accordance with the following...