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Sample records for bull spermatozoa express

  1. Protamine 3 expressions in crossbred bull spermatozoa may not be ...

    African Journals Online (AJOL)

    SAM

    2014-05-14

    May 14, 2014 ... To differentiate finally into spermatozoa, haploid sperma- tids undergo complex morphological and physiological changes during spermatogenesis. These changes include chromatin remodeling and condensation mediated through the replacement of somatic histones by transition proteins and protamines ...

  2. Protamine 3 expressions in crossbred bull spermatozoa may not be ...

    African Journals Online (AJOL)

    Protamines are short and highly basic sperm-specific nuclear proteins. Though the ... The mRNA expression of PRM3 gene among two groups was evaluated by real time quantitative polymerase chain reaction (PCR) using TaqMan chemistry, where peptidylprolyl isomerase A (PPIA) was used as an internal control.

  3. Comparative proteomic analysis of Taurine, Indicine, and crossbred (Bos taurus × Bos indicus) bull spermatozoa for identification of proteins related to sperm malfunctions and subfertility in crossbred bulls.

    Science.gov (United States)

    Muhammad Aslam, Munchakkal Kather; Kumaresan, Arumugam; Rajak, Shailendra Kumar; Tajmul, Md; Datta, Tirtha Kumar; Mohanty, Tushar Kumar; Srinivasan, Alagiri; Yadav, Savita

    2015-09-01

    Subfertility is one of the most common problems observed among Taurine × Indicine crossbred bulls in tropical countries; however, the etiology remain unknown in most of the cases. In present study, we compared the proteomic profile of spermatozoa from crossbred bulls (Bos taurus × Bos indicus) against their purebred parent lines (Holstein Friesian [Taurine] and Tharparkar [Indicine]) to find out alteration in expressions of proteins, if any. The proteomic profiles of freshly ejaculated spermatozoa from these breeds were compared by two-dimensional difference gel electrophoresis, and differentially expressed proteins were identified through mass spectrometry. It was observed that compared to Holstein Friesian, nine proteins were underexpressed and eight proteins were overexpressed (P < 0.05) in the spermatozoa of crossbred bulls. Similarly, four proteins were overexpressed and four proteins were underexpressed (P < 0.05) in the spermatozoa of crossbred bulls compared to Tharparkar bulls. In concurrent three breed comparison, 14 proteins were found to be differentially expressed (P < 0.05) between these breeds. From the findings of the study, it is apparent that the expression levels of several functionally significant proteins are either upregulated or downregulated in spermatozoa of crossbred bulls, which might be related to high incidence of subfertility in these bulls. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. RELATIONSHIP BETWEEN MOTILITY AND VIABILITY PARAMETERS OF FROZEN-THAWED BULL SPERMATOZOA

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    Eliška Špaleková

    2013-02-01

    Full Text Available The aim of this study was to determine relationship between parameters of spermatozoa motility (total motility - TM and progressive movement - PM and viability of bull frozen-thawed spermatozoa (dead spermatozoa ratio, apoptotic spermatozoa ratio and plasma membrane integrity. Motility parameters were evaluated using computer-assisted semen analysis (CASA. Parameters of spermatozoa viability were analysed using fluorescent dyes PNA-FITC (plasma membrane, Yo-Pro-1 and propidium iodide (PI. All bulls (n=6 were divided into two groups. First group (n=3 A – better bulls with total motility after thawing over 40% and the second group (n=3 B – with total motility lower than 40%. It was observed significantly (P<0.001 higher TM and PM in group A. No significant differences in velocity parameters and ALH between the group A and B were detected. Occurrence of spermatozoa with disrupted membranes, dead/necrotic spermatozoa and apoptotic spermatozoa was significantly lower in the group A. Bulls in the group A showed significantly higher cleavage rate of embryos. These motility and viability characteristics are associated with a higher embryo cleavage rate in in vitro fertilizatioThe aim of this study was to determine relationship between parameters of spermatozoa motility (total motility - TM and progressive movement - PM and viability of bull frozen-thawed spermatozoa (dead spermatozoa ratio, apoptotic spermatozoa ratio and plasma membrane integrity. Motility parameters were evaluated using computer-assisted semen analysis (CASA. Parameters of spermatozoa viability were analysed using fluorescent dyes PNA-FITC (plasma membrane, Yo-Pro-1 and propidium iodide (PI. All bulls (n=6 were divided into two groups. First group (n=3 A – better bulls with total motility after thawing over 40% and the second group (n=3 B – with total motility lower than 40%. It was observed significantly (P<0.001 higher TM and PM in group A. No significant differences in

  5. Light-scattering studies of bull spermatozoa. II. Interaction and concentration effects.

    OpenAIRE

    Woolford, M.W.; Harvey, J D

    1982-01-01

    The complete autocorrelation function of the intensity fluctuations of laser light scattered from motile bull spermatozoa is shown to depend upon several factors not previously considered. Samples of bull spermatozoa generally contain a substantial proportion of dead cells, which give rise to slowly decaying components of the autocorrelation function. Whereas previous work has concentrated on the form of the fast decaying autocorrelation component, we are concerned here with the relative ampl...

  6. Dynamics of microRNAs in bull spermatozoa

    OpenAIRE

    Govindaraju Aruna; Uzun Alper; Robertson LaShonda; Atli Mehmet O; Kaya Abdullah; Topper Einko; Crate Elizabeth A; Padbury James; Perkins Andy; Memili Erdogan

    2012-01-01

    Abstract Background MicroRNAs are small non-coding RNAs that regulate gene expression and thus play important roles in mammalian development. However, the comprehensive lists of microRNAs, as well as, molecular mechanisms by which microRNAs regulate gene expression during gamete and embryo development are poorly defined. The objectives of this study were to determine microRNAs in bull sperm and predict their functions. Methods To accomplish our objectives we isolated miRNAs from sperm of high...

  7. Identification of cryodamage on plasma membrane integrity in bull spermatozoa and its relationship with field fertility

    OpenAIRE

    Quintero-Moreno, Armando; González-Villalobos, Decio; Gutiérrez, Juan Carlos; Madrid-Bury, Ninoska; Garde López-Brea, José julián; Rubio-Guillén, Jorge

    2011-01-01

    The aim of this study was to investigate the effect of cryopreservation on the membrane integrity (MI) in the bull sperm, and determine the relationship between intact cryopreserved sperm and field fertility (FF). The eosin nigrosin exclusion and the hypo-osmotic swelling test were combined in a single test (HOS-EN test) to identify the spermatozoa with four types of MI. After cryopreservation, there was a marked decline (21.3%) in the percentage of spermatozoa with head membrane intact/ t...

  8. Microencapsulation of bull spermatozoa: Its viability in alginate-egg yolk media

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    Kusumaningrum DA

    2015-03-01

    Full Text Available Microencapsulation of spermatozoa is a process to entrap a number of spermatozoa in microcapsule. Alginate, as a natural polymer polysaccharide is commonly used in cell microencapsulation. Tris Yolk Citrate buffer is a good buffer for spermatozoa dilution, therefore this experiment aimed to determine optimal concentration of alginate and egg yolk to sperm quality in bull spermatozoa microencapsulation. Concentration of egg yolk and alginate in media of encapsulation were determined in applications of sperm microencapsulation. Four bulls were used as semen source and only semen with good quality were used in this study. Poolled semen was diluted using the medium to get final concentration 100 x 106 cell/ ml. The first study was conducted to determine the effect of concentration of alginate (0, 1, and 1.5% on viability of spermatozoa. The second study to determine the effect of alginate concentration, egg yolk and its interaction was done by comparing two levels of alginate (1 and 1.5% with four levels of egg yolk (5, 10, 15 and 20%. Viability of spermatozoa, motility (M, live spermatozoa (L and Intact Apical Ridge (IAR were observed at 0, 1, 2 and 3 h incubation at room temperature. Results indicated that alginate concentration increased the osmolality and viscosity but did not affect pH of the medium. The osmolality and viscosity of medium were 275, 325, 425 and 1.12, 26.62, 47.98 for concentration of alginate 0, 1 and 1.5% respectively. Percentage of motility is significantly lower (P<0.05 in alginate medium than those of control, and 1.5% alginate could produce more uniform beads. Concentration of alginate, egg yolk and its interaction did not significantly affect viability of sperm. It is concluded that the combination of 1.5% alginate with 5, 10, 15 or 20% egg yolk can be used as media for sperm encapsulation.

  9. Anandamide capacitates bull spermatozoa through CB1 and TRPV1 activation.

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    María Gracia Gervasi

    2011-02-01

    Full Text Available Anandamide (AEA, a major endocannabinoid, binds to cannabinoid and vanilloid receptors (CB1, CB2 and TRPV1 and affects many reproductive functions. Nanomolar levels of anandamide are found in reproductive fluids including mid-cycle oviductal fluid. Previously, we found that R(+-methanandamide, an anandamide analogue, induces sperm releasing from bovine oviductal epithelium and the CB1 antagonist, SR141716A, reversed this effect. Since sperm detachment may be due to surface remodeling brought about by capacitation, the aim of this paper was to investigate whether anandamide at physiological concentrations could act as a capacitating agent in bull spermatozoa. We demonstrated that at nanomolar concentrations R(+-methanandamide or anandamide induced bull sperm capacitation, whereas SR141716A and capsazepine (a TRPV1 antagonist inhibited this induction. Previous studies indicate that mammalian spermatozoa possess the enzymatic machinery to produce and degrade their own AEA via the actions of the AEA-synthesizing phospholipase D and the fatty acid amide hydrolase (FAAH respectively. Our results indicated that, URB597, a potent inhibitor of the FAAH, produced effects on bovine sperm capacitation similar to those elicited by exogenous AEA suggesting that this process is normally regulated by an endogenous tone. We also investigated whether anandamide is involved in bovine heparin-capacitated spermatozoa, since heparin is a known capacitating agent of bovine sperm. When the spermatozoa were incubated in the presence of R(+-methanandamide and heparin, the percentage of capacitated spermatozoa was similar to that in the presence of R(+-methanandamide alone. The pre-incubation with CB1 or TRPV1 antagonists inhibited heparin-induced sperm capacitation; moreover the activity of FAAH was 30% lower in heparin-capacitated spermatozoa as compared to control conditions. This suggests that heparin may increase endogenous anandamide levels. Our findings indicate

  10. EFFECT OF MALTOSE CONCENTRATION IN TRIS DILUTION ON EPIDIDYMAL SPERMATOZOA QUALITY OF BALI BULL PRESERVED AT 50C

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    J. Wattimena

    2014-10-01

    Full Text Available The objective of research was to evaluate the effect of maltose concentration in Tris dilution onepididymal spermatozoa quality of Bali bull that preserved at 50C. Five testis of Bali bull collected fromslaughter house were used in this study. Epididymal spermatozoa were collected through slicing andflushing methods, pressing cauda epididymal was conducted in NaCl physiology (NaCl 0.9% emulsion.Spermatozoa which collected were divided into three reaction tube and each diluted by Tris dilutioncontaining: Tris dilution + 20% of yolk (control; Tris dilution + 20% of yolk + 0.3 g of maltose/100ml(M0.3; and Tris dilution + 20% of yolk + 0.6 g maltose/100 ml (M0.6. Spermatozoa qualities observedwere motile spermatozoa (MS, live-spermatozoa (LS and intact-plasma membrane (IPM thatevaluated daily in refrigerator at temperature of 5oC. Completely Randomized Design with threetreatments and five replications was used in this study. Data was analyzed by analysis of variance.Examination on fresh spermatozoa showed that spermatozoa concentration was 11,222.5 million cell/ml,motile spermatozoa was 75.00%, live-sperm was 86.75%, abnormal spermatozoa was 10.50%,cytoplasmic droplet was 14.00% and IPM was 86.75%. At the seventh day of preservation, thepercentages of MS, LS and IPM in M0.3 were 37.0 %, 49.2% and 50.4%, respectively, and M0.6 were38.05%; 51.8 % and 52.0%, respectively that were significantly higher (P<0.05 than control (29.0%;41.8% and 42.4%, respectively. It was concluded that maltose added into Tris dilution could lengthenepididymal spermatozoa quality of Bali bull which persevered at 50C.

  11. Semen quality parameters as fertility predictors of water buffalo bull spermatozoa during low-breeding season.

    Science.gov (United States)

    Ahmed, Hussain; Andrabi, S Murtaza Hassan; Jahan, Sarwat

    2016-10-01

    The present study was carried out to assess various postthaw semen quality parameters for the prediction of fertility in buffalo bull during low-breeding season. Semen (30 ejaculates) was collected from five adult buffalo bulls with artificial vagina (42 °C). Sperm motility parameters, velocity distribution, motion kinematics, and subpopulations were analyzed by computer-aided sperm motion analyzer (CASA). Moreover, sperm visual motility, supravital plasma membrane integrity, viability/acrosome integrity, viability/mitochondrial transmembrane potential, DNA fragmentation/integrity, and morphology were analyzed by phase-contrast microscope, supravital hypoosmotic swelling test, Trypan blue/Giemsa staining, propidium iodide/"5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide" (JC-1) fluorochromes, neutral comet assay/acridine orange assay and wet mount technique, respectively. Outcome of 528 inseminations was analyzed for in vivo fertility. Pearson's correlation coefficients revealed that sperm progressive motility (%), rapid velocity (%), average path velocity (μm/s), straight line velocity (μm/s), subpopulation one (most rapid, and progressive) of motile spermatozoa (%), supravital plasma membrane integrity (%), and viable spermatozoa with intact acrosome (%) were significantly correlated with in vivo fertility (r = 0.64, P comet length (μm) of neutral comet assay were negatively associated with in vivo fertility (r = -0.79, r = -0.75, P sperm structural and functional parameters, that is, integrity of plasma membrane and acrosome, and transmembrane potential of mitochondria were able to predict the in vivo fertility of water buffalo bull during low-breeding season. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Novel insights into the role of cell-free seminal mRNAs on semen quality and cryotolerance of spermatozoa in bulls (Bos taurus).

    Science.gov (United States)

    Shilpa, Munivenkatappa; Selvaraju, Sellappan; GirishKumar, Venkataswamy; Parthipan, Sivashanmugam; Binsila, Krishnan B; Arangasamy, Arunachalam; Ravindra, Janivara Parameshwaraiah

    2017-11-01

    The aim of the present study was to ascertain the effectiveness of seminal plasma mRNAs as markers to assess the reproductive performance of bulls. Semen samples (33 ejaculates) from 11 bulls were evaluated for sperm kinematic and functional parameters. Total RNA was isolated from cell-free seminal (cfs) using TRIzol LS reagent and the concentration of cfs-RNA was 24.4±2.3µgmL-1 seminal plasma. The cfs-RNA was fragmented to a size of 25-500bp. Of the cfs-mRNAs screened using real time PCR, expression of protamine 1 (PRM1) was positively (P2.5μm and straightness >85%) in raw semen was positively (P<0.05) associated with bone morphogenetic protein 2 (BMP2), ubiquitin conjugating enzyme E2D3 (UBE2D3), tumour-associated necrotic factor-associated death domain (TRADD) and caspase-3 (CASP3) expression. Nerve growth factor (NGF) expression was positively (P<0.05) associated with the maintenance of post-thaw functional membrane integrity in spermatozoa and could be used to assess the cryotolerance of bull semen. In conclusion, the expression of cfs mRNAs can be used to assess the reproductive performance of males and to predict the sensitivity of spermatozoa to cryoinjury.

  13. Soya-lecithin in extender improves the freezability and fertility of buffalo (Bubalus bubalis) bull spermatozoa.

    Science.gov (United States)

    Akhter, S; Ansari, M S; Andrabi, S M H; Rakha, B A; Ullah, N; Khalid, M

    2012-10-01

    Egg yolk is routinely used as a cryoprotectant in semen extenders. However, it may contain cryoprotective antagonists, and there are hygienic risks associated with its use. Proteins of plant origin, like soya-lecithin, lack these hazards. The aim of this study was to use soya-lecithin as a cryoprotectant in extender and to investigate its effects on in vitro quality and in vivo fertility of buffalo semen. Semen from three buffalo bulls was frozen in tris-citric extender containing 5.0%, 10% or 15% soya-lecithin or 20% egg yolk. Sperm motility, plasma membrane integrity and viability were assessed post-dilution, pre-freezing and post-thaw. In Post-dilution and pre-freezing, the values for motility, plasma membrane integrity and viability remained higher (p ≤ 0.05) in extenders containing 10% soya-lecithin and control compared with extender containing 5% and 15% soya-lecithin. However, motility, plasma membrane integrity and viability were higher (p lecithin compared with control and extenders containing 5% and 15% soya-lecithin. Semen from two buffalo bulls was frozen in tris-citric extender containing either 10% soya-lecithin or 20% egg yolk. Higher (p lecithin (56%) compared with 20% egg yolk (41.5%). The results suggest that 10% soya-lecithin in extender improves the freezability and fertility of buffalo bull spermatozoa and can be used as an alternate to egg yolk in cryopreservation of buffalo semen. © 2011 Blackwell Verlag GmbH.

  14. Antioxidant effects of bovine serum albumin on kinetics, microscopic and oxidative characters of cryopreserved bull spermatozoa

    Directory of Open Access Journals (Sweden)

    I. Ashrafi

    2013-07-01

    Full Text Available The aim of this study was to determine the effects of bovine serum albumin (BSA as an antioxidant on post-thaw characters of bull spermatozoa such as motion variables, viability, plasma membrane integrity, morphology, lipid peroxidation, total antioxidant capacity (TAC, total thiols (TT and the enzymes activities. Ejaculates were collected from six proven bulls and diluted with a citrate-based extender supplemented with various concentrations of BSA (0, 0.5, 1, 1.5 and 2 g/100 mL. The results showed that the semen extender supplemented with various concentrations of BSA increased (p0.05 were detected in progressive motility, sperm track straightness, beat cross frequency, curvilinear velocity and malondialdehyde production between the experimental groups. The highest concentration of BSA (2 g/100 mL in the semen extender reduced (p<0.05 the sperm motion variables. The results indicated that the most effective concentration of BSA is 1 g/100 mL in the semen extender which is associated with an increase in the total motility, antioxidant enzymes activities, TT and TAC.

  15. EFFECT OF NON-ENZYMATIC ANTIOXIDANTS IN EXTENDER ON POST-THAW QUALITY OF BUFFALO (BUBALUS BUBALIS BULL SPERMATOZOA

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    S. M. H. ANDRABI, M. S. ANSARI, N. ULLAH AND M. AFZAL

    2008-12-01

    Full Text Available The objective of the present study was to determine the effect of non-enzymatic antioxidants (vitamin C or E in tris-citric acid buffer (TCA on post-thaw quality of buffalo (Bubalus bubalis bull spermatozoa. Split pooled buffalo bull ejaculates were diluted in TCA egg yolk glycerol extender containing either vitamin C (TCAC, vitamin E (TCAE or without antioxidant (TCAN at 37°C. Extended semen was cooled to 4C in 2 h and equilibrated for 4 h at 4C. Cooled semen was then filled in 0.5 ml straws at 4C and frozen in programmable cell freezer. Thawing of semen was performed at 37°C for 30 seconds. Sperm motility, plasma membrane integrity and sperm morphology (acrosome integrity, head, mid-piece and tail abnormalities of each semen sample were evaluated. Percentage of post-thaw spermatozoal motility assessed visually at 0 and 6 h and the post-thaw percentage of spermatozoa with intact plasma membranes at 0 h were higher (P0.05. In conclusion, non-enzymatic antioxidants, particularly vitamin E, in the tris citric acid extender may improve the quality of frozen-thawed buffalo bull spermatozoa.

  16. Freezability of water buffalo bull (Bubalus bubalis) spermatozoa is improved with the addition of curcumin (diferuoyl methane) in semen extender.

    Science.gov (United States)

    Shah, S A H; Andrabi, S M H; Qureshi, I Z

    2017-10-01

    Effects of curcumin as antioxidant in extender were evaluated on freezability of buffalo spermatozoa. Semen from each of the five bulls (n = 3 replicates, six ejaculates/bull, a total of 30 ejaculates) was diluted in Tris-citric acid extender containing curcumin (0.5, 1.0, 1.5 or 2.0 mM) or control. At pre-freezing and post-thawing, total antioxidant contents (μM/L) and lipid peroxidation levels (μM/ml) were higher (p straight-line velocity, μm/s; curved-line velocity, μm/s; straightness, %; linearity, %), in vitro longevity (%, PM and RV) and DNA integrity (%) at post-thawing were higher (p < .05) with 1.5 mM compared to control. At post-thawing, supravital plasma membrane integrity (%) and viable spermatozoa with intact acrosome (%) were higher with 1.5 compared to 2.0 mM curcumin and control. We concluded that freezability of water buffalo spermatozoa is improved with the addition of 1.5 mM curcumin in extender. © 2016 Blackwell Verlag GmbH.

  17. The effects of cholesterol on the viability and fertility of bull spermatozoa

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    Polmer Situmorang

    2002-12-01

    Full Text Available This study was conducted to evaluate the effect of cholesterol on the viability and fertility of chilled and deep-frozen bull spermatozoa. Semen was collected by means of artificial vagina, diluted in Tris-Citrat diluent and cooled to 50C for 60 minutes. Following an equiliberation for 4 hours, semen was frozen at 5 cm above surface of liquid nitrogen for 10 minutes. The experiment was 2 x 3 factorial designed with two level of egg yolk (10 and 20% v/v and 3 level of cholesterol (0; 0.5 and 1.0 mg/ml. The viability of spermatozoa was evaluated after the temperature reduced to 50C, stored at 50C for 1, 3 and 7 days and after thawing. For fertility test, cows were artificially inseminated (AI using chilled and frozen semen on the onset or 6 hours of oestrus. Rectal palpation was conducted 3 months after AI to determine the pregnancy. The percentages motile of chilled semen was higher in 0.5 mg/ml than those of 0.0 or 1.0 mg/ml cholesterol but this difference was not significant. After thawing, the effects of cholesterol on the percentage motile was significant (P<0.05. The mean percentage motile was 47.5; 51,5 and 56.0 for 0.0; 0.5 and 1.0 mg/ml cholesterol respectively. The percentage of live sperm and intact apical ridge was higher in cholesterol however this effects was not significant. The effects level of egg yolk and its interaction with cholesterol on the viability was not significant. The percentage of pregnant was higher in 1.0 mg/ml and the mean percentage of pregnant was 45.8; 48.2 and 55.7 for 0.0; 0.5 and 1.0 mg/ml cholesterol respectively. Percentage of pregnant was higher for chilled semen than those of frozen semen (54.3 vs 45.5. In conclusion the addition of 1 mg/ml cholesterol increase the percentage of motile after thawing and pregnancy of cows inseminated with chilled and frozen semen.

  18. CHARACTERIZATION OF THE OLFACTORY RECEPTORS EXPRESSED IN HUMAN SPERMATOZOA

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    Caroline eFlegel

    2016-01-01

    Full Text Available The detection of external cues is fundamental for human spermatozoa to locate the oocyte in the female reproductive tract. This task requires a specific chemoreceptor repertoire that is expressed on the surface of human spermatozoa, which is not fully identified to date. Olfactory receptors (ORs are candidate molecules and have been attributed to be involved in sperm chemotaxis and chemokinesis, indicating an important role in mammalian spermatozoa. An increasing importance has been suggested for spermatozoal RNA, which led us to investigate the expression of all 387 OR genes. This study provides the first comprehensive analysis of OR transcripts in human spermatozoa of several individuals by RNA-Seq. We detected 91 different transcripts in the spermatozoa samples that could be aligned to annotated OR genes. Using stranded mRNA-Seq, we detected a class of these putative OR transcripts in an antisense orientation, indicating a different function, rather than coding for a functional OR protein. Nevertheless, we were able to detect OR proteins in various compartments of human spermatozoa, indicating distinct functions in human sperm. A panel of various OR ligands induced Ca2+ signals in human spermatozoa, which could be inhibited by mibefradil. This study indicated that a variety of ORs are expressed at the mRNA and protein level in human spermatozoa and demonstrates that ORs are involved in the physiological processes.

  19. Specific binding of the glycosaminoglycan /sup 3/H-heparin to bull, monkey, and rabbit spermatozoa in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Handrow, R.R.; Boehm, S.K.; Lenz, R.W.; Robinson, J.A.; Ax, R.L.

    In Vitro binding and some binding parameters of the glycosaminoglycan heparin to viable epididymal or ejaculated bull spermatozoa, ejaculated rabbit spermatozoa, and frozen-thawed rhesus monkey spermatozoa were investigated. Nonspecific binding was affected only by the concentration of /sup 3/H-heparin, whereas specific binding was saturable, reversible, and dependent on the pH, temperature, and calcium concentration of the incubation medium. Magnesium concentration dependence was observed in the presence of calcium but could not be detected in the absence of calcium. Bound /sup 3/H-heparin was displaced by several orders of magnitude greater concentrations of chondroitin sulfate. Scatchard plot analysis suggested multiple binding affinities for /sup 3/H-heparin to spermatozoa. /sup 3/H-heparin was shown to bind to sperm heads and flagella. Fluorescein-labeled heparin bound to acrosomal, postacrosomal, and flagellar membranes. It was concluded that the specific binding of heparin involved a proteinaceous component on, or intercalated with, spermatozoal membranes. Thus, glycosaminoglycans present in the female reproductive tract may contribute to sperm capacitation and enhance the likelihood of successful fertilization in mammals.

  20. Effects of the platelet-activating factor (PAF) supplementation on ATP content of cryopreserved bull spermatozoa (AI).

    Science.gov (United States)

    Lecewicz, M; Kordan, W; Kamiński, S; Majewska, A M; Strzeżek, R

    2017-03-01

    The aim of this study was to investigate the effect of PAF supplementation in semen extender on ATP content in cryopreserved bull spermatozoa used for artificial insemination at different time intervals. Cryopreserved semen was treated with different concentrations of PAF: 1×10-5M, 1×10-6M, 1×10-7M, 1×10-8M and 1×10-9M at 37°C. In the present work we showed that content of ATP in cryopreserved semen supplemented with 1×10-9M PAF was statistically significantly higher at 90 and 120 minutes of incubation in comparison to the control group (p≤0.05). Present study indicates the potential influence of PAF on ATP content in male spermatozoa via it's protective role towards mitochondria metabolic activity.

  1. Impact of proximal cytoplasmic droplets on quality traits and in-vitro embryo production efficiency of cryopreserved bull spermatozoa

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    Carreira Janaina T

    2012-01-01

    Full Text Available Abstract Background Proximal cytoplasmic droplets (PCDs, a remnant of germ cell cytoplasm, are common non-specific morphological defects in bovine semen. This study evaluated the effect of higher percentages of PCDs on the quality of frozen-thawed bovine semen, embryo production and early embryo development. Methods Three ejaculates from each of five (group 1: PCD ≤ 1%, control and eight adult Bos indicus bulls (group 2: PCD ≥ 24% were analysed. Semen samples were examined for: post-thaw motility, vigour of movement, concentration, sperm morphology, slow thermoresistance test (STT, membrane integrity, acrosome status, mitochondrial function using fluorescent probes association (FITC-PSA, PI and JC-1 and sperm chromatin integrity using acridine orange assay. Two bulls from group 2, with 28.5% and 48.5% PCD, respectively, and three bulls from the control group, each with 0% PCD, were selected for IVF (in vitro fertilisation. Results Semen analyses revealed a significant correlation (P Conclusion Higher PCD levels influenced spermatozoa quality traits. IVF and embryo development data showed that cleavage, blastocyst formation and blastocyst hatching may have been influenced by the interaction of morphology traits and individual bull effects.

  2. Viability of Bali bull epididymal spermatozoa preserved at 3–5oC in Tris extender with different lactose concentrations

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    Muhammad Rizal

    2009-06-01

    Full Text Available Cauda epididymal spermatozoa could be used as an alternative source of gamete in the application of various reproductive technologies, because the spermatozoa is motile and has ability for fertilizing the oocyte. The objective of this research was to examine the effectivity of lactose in maintaining viability of Bali bull epididymal spermatozoa preserved at 3–5oC. Five testis with epididymides of Bali bulls were obtained from slaughterhouse. Epididymal spermatozoa was collected by the combination of slicing, flushing and tissues pressure of cauda epididymides with physiological saline (0.9% NaCl. Collected-spermatozoa was divided in equal volume into three tubes and diluted with Tris extender containing 20% egg yolk (control, Tris extender + 0.3 g lactose/100 ml (L0.3, and Tris extender + 0.6 g lactose/100 ml (L0.6, respectively. Diluted-spermatozoa was stored in refrigerator at 3–5oC. Quality of diluted-spermatozoa including percentages of motile spermatozoa (MS, live spermatozoa (LS, and intact plasma membrane (IPM were evaluated every day during storage at 3–5oC for six days. Data were analyzed using completely randomized design with three treatments and five replications. Results of this study showed that mean spermatozoa concentration, percentage of MS, percentage of LS, percentage of abnormal spermatozoa, percentage of cytoplasmic droplet, and percentage of IPM of Bali bull fresh epididymal spermatozoa were 11,222.5 million cell/ml, 75, 86.75, 10.5, 14, and 86.75%, respectively. At day-7 storage, percentages of MS, LS, and IPM for L0.3 (39, 50.6, and 51.6% and L0.6 (39, 51.4, and 51.8% were significantly (P<0.05 higher than control (29, 41.8, and 42.4%. In conclusion, addition of lactose in Tris extender extended viability of Bali bull epididymal spermatozoa preserved at 3–5oC.

  3. Cholesterol loaded cyclodextrin increases freezability of buffalo bull (Bubalus bubalis spermatozoa by increasing cholesterol to phospholipid ratio

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    J. S. Rajoriya

    2014-09-01

    Full Text Available Aim: The study was conducted to investigate the effect of cholesterol loaded cyclodextrin (CLC on freezability of buffalo spermatozoa. Materials and Methods: Murrah buffalo bull semen samples with progressive motility of 70% and greater were used. After the evaluation of motility and livability, four equal fractions of semen samples were made. Group I was kept as control and diluted with Tris, whereas Group II, III and IV were treated with CLC solution at the rate of 2.0, 3.0 and 4.0 mg/ml respectively to obtain 120 × 106 sperm/ml as final spermatozoa concentration. The aliquots of all the groups were incubated for action of CLC, followed by dilution and freezing. Evaluation at pre-freeze and post-thaw stage of progressive motility, viability and level of cholesterol and phospholipid was done. Results: The mean cholesterol content (μg/100 × 106 spermatozoa of Group I, II, III and IV at pre-freeze stage was 21.55±0.63, 49.56±1.38, 55.67±0.45 and 47.79±1.01 and at post-thaw stage were 13.18±0.45, 34.27±0.71, 36.21±0.48 and 33.68±0.56, respectively. At pre-freeze stage, cholesterol content was significantly (p<0.01 higher in Group III in comparison to other groups. The mean cholesterol and phospholipids content of fresh sperm was 24.14±0.58 and 51.13±0.66 μg/100 × 106 sperm cells, respectively, and C/P ratio of spermatozoa at fresh stage was 0.47±0.067. Conclusion: CLC treatment maintains the C/P ratio and plays an important role in maintaining membrane architecture of spermatozoa. Hence, addition of CLC may be helpful in increasing freezability of buffalo spermatozoa by increasing the C/P ratio of spermatozoa.

  4. During capacitation in bull spermatozoa, actin and PLC-ζ undergo dynamic interactions.

    Science.gov (United States)

    Mejía-Flores, Itzayana; Chiquete-Félix, Natalia; Palma-Lara, Icela; Uribe-Carvajal, Salvador; de Lourdes Juárez-Mosqueda, María

    2017-10-01

    The migration pattern of sperm-specific phospholipase C-ζ (PLC-ζ) was followed and the role of this migration in actin cytoskeleton dynamics was determined. We investigated whether PLC-ζ exits sperm, opening the possibility that PLC-ζ is the 'spermatozoidal activator factor' (SOAF). As capacitation progresses, the highly dynamic actin cytoskeleton bound different proteins to regulate their location and activity. PLC-ζ participation at the start of fertilization was established. In non-capacitated spermatozoa, PLC-ζ is in the perinuclear theca (PT) and in the flagellum, therefore it was decided to determine whether bovine sperm actin interacts with PLC-ζ to direct its relocation as it progresses from non-capacitated (NC) to capacitated (C) and to acrosome-reacted (AR) spermatozoa. PLC-ζ interacted with actin in NC spermatozoa (100%), PLC-ζ levels decreased in C spermatozoa to 32% and in AR spermatozoa to 57% (P PLC-ζ interaction was twice as high in G-actin (P PLC-ζ was partially released from the cell. It was concluded that actin cytoskeleton dynamics control the migration of PLC-ζ during capacitation and leads to its partial release at AR spermatozoa. It is suggested that liberated PLC-ζ could reach the egg and favour fertilization.

  5. Comparative studies on bull and stallion seminal DNase activity and interaction with semen extender and spermatozoa.

    Science.gov (United States)

    Alghamdi, Abdorrahman S; Funnell, Bethany J; Bird, Scott L; Lamb, G Cliff; Rendahl, Aaron K; Taube, Patrick C; Foster, Douglas N

    2010-09-01

    We performed a series of comparative studies of bull and stallion seminal plasma (SP) and its role on sperm-neutrophil binding as well as the interaction between semen extender and seminal DNase. Because of contrasting roles of SP on sperm-neutrophil binding between horses and cattle, it was suspected there were some species-specific differences on sperm interaction with SP proteins due to the variations in the natural location of semen deposition (uterus compared to vagina). Bull frozen-thawed sperm removed from egg yolk extender showed similar results to fresh sperm, but this also caused extensive sperm agglutination unless SP or egg yolk was included. If similar agglutination occurs after AI with frozen bull semen, it could interfere with sperm transport or sperm functions. Commonly used bull semen extenders were poor media for seminal DNase activity on plasmid DNA degradation, raising the prospect that the same may be true with other SP factors important to fertility. DNase activity per mg SP protein of bulls was less than that of horses (Pfertility. Modifications of semen extender and/or semen processing should be examined to allow sperm cells a maximum potential for fertilization. 2010 Elsevier B.V. All rights reserved.

  6. Preservability of bull spermatozoa in Tris-egg yolk extender enriched with different concentrations of butylated hydroxytoluene

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    El-Sheshtawy RI

    2017-02-01

    Full Text Available Objective: To explore the effect of BHT on cattle spermatozoa during cooling and cryopreservation. Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk (TCFY diluent considered as control (0 BHT and different concentrations of BHT (1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 mM were prepared in ethanol in prewarmed (37 ℃ test tubes. The ethanol was allowed to evaporate so that, a thin crystallized layer of BHT was deposited on the inner surface of the tubes. Then extended semen was added into the tubes and incubated at 37 ℃ for 5 min to allow uptake of BHT by spermatozoa. The tubes were cooled slowly (approximately for 2 h up to 5 ℃ and equilibrated for 4 h. After equilibration, semen freezing process was carried out. Extended semen was subjected to evaluation (motility, alive sperm, intact sperm membrane (HOST % and acrosome integrity in both cooled and cryopreserved semen. Results: The result revealed that sperm motility of post-cooled spermatozoa improved (P<0.05 by the use of BHT concentrations (1, 2 and 3 mM in Tris semen extender if compared to the control (85.00±1.09, (83.33± 0.63, (81.67± 0.63 and (78.33± 0.63, respectively. Alive sperm percent was significantly higher in all concentrations of BHT. Sperm abnormalities percent were significantly lower in concentrations of BHT 1 and 2 (11.2±0.2, (11.8±0.2and (13.4±0.4, respectively. Sperm membrane integrity were significantly higher in BHT concentrations (1, 2, 3, 4 and 5 mM. It is exhibited that improved sperm motility in post-thawed frozen semen in the concentrations of BHT (1, 2, 3 and 4 mM if compared to the control. The sperm membrane integrity were significantly improved at all concentrations of BHT. Acrosome integrity was significantly higher at BHT concentration 1 mM (81.80±0.57 and (76.00±2.05, respectively. Conclusions: It could be concluded that some concentrations of BHT improved bull semen quality post-cooling and post-freezing.

  7. Combinations of glycerol percent, glycerol equilibration time, and thawing rate upon freezability of bull spermatozoa in plastic straws.

    Science.gov (United States)

    Wiggin, H B; Almquist, J O

    1975-03-01

    Twelve ejaculates were used in a central composite experiment to test 15 combinations of glycerol (7, 9, 11, 13, or 15%), glycerol equilibration times (1, 2, 4, 8, or 16 h) and thawing rates (water at 35 C for 15 s, 50 C for 13 s, 65 C for 11 s, 80 C for 9 s, or 95 C for 7 s). Semen was diluted in heated skim milk-glycerol, packaged in .3-ml. Continental U.S. straws and frozen in liquid nitrogen vapor. Based on post-thaw progressive sperm motility after storage at -196 C for 9 to 11 days, estimated optima from multiple regression were 10.7% for glycerol, 2.0 h for glycerol equilibration time, and 76 C for thawing bath temperature. Only the linear effect for each variable was significant. Much faster thawing rates and shorter glycerol equilibration times than those for freezing bull spermatozoa in glass ampules should be used for maximum post-thaw sperm motility in straws.

  8. Sperm function during incubation with oestrus oviductal fluid differs in bulls with different fertility.

    Science.gov (United States)

    Kumaresan, A; Johannisson, A; Bergqvist, A-S

    2017-06-01

    Spermatozoa undergo several modifications in the oviduct before acquiring fertilising capacity. Although spermatozoa are exposed to similar conditions in the oviduct, the speed of the response varies with the male and the state of the spermatozoa. We hypothesised that spermatozoa from bulls with different fertility may differ in their ability to respond to oviductal fluid (ODF). Frozen-thawed spermatozoa from four bulls were incubated with oestrus oviductal fluid (OODF) for 6h. Sperm kinematics, tyrosine phosphorylation, phosphorylation patterns, capacitation and acrosome reaction were analysed at hourly intervals. The amplitude of lateral head displacement (ALH) and straightness coefficient (STR) were higher (P<0.05) in bulls with higher fertility compared with those with lower fertility, at 1-4h of incubation. At 4h of incubation and onwards, spermatozoa from bulls with higher fertility showed a lower degree (P<0.05) of tyrosine phosphorylation and higher degree of capacitation and acrosome reaction. At least five tyrosine-phosphorylated sperm proteins were detected in all bulls. However, the expression of two phosphorylated sperm proteins (183 and 109 kDa) was upregulated in bulls with lower fertility. It may be concluded that cryopreserved spermatozoa from high- and low- fertile bulls differ in their ability to respond to OODF. This may help in developing tools for assessing fertility of bulls, once validated in more animals.

  9. ESR (electron spin resonance)-determined osmotic behavior of bull spermatozoa

    Energy Technology Data Exchange (ETDEWEB)

    Du, J.; Kleinhans, F.W.; Spitzer, V.J.; Critser, J.K. (Methodist Hospital, Indianapolis, IN (USA). Dept. of Medical Research); Horstman, L. (Purdue Univ., Lafayette, IN (USA). School of Veterinary Medicine); Mazur, P. (Oak Ridge National Lab., TN (USA))

    1990-01-01

    Our laboratories are pursuing a fundamental approach to the problems of semen cryopreservation. For many cell types (human red cells, yeast, HeLa) it has been demonstrated that there is an optimum cooling rate for cryopreservation. Faster rates allow insufficient time for cell dehydration and result in intracellular ice formation and cell death. It is possible to predict this optimal rate provided that the cell acts as an ideal osmometer and several other cell parameters are known such as the membrane hydraulic conductivity. It is the purpose of this work to examine the osmotic response of bull sperm to sucrose and NaCl utilizing electron spin resonance (ESR) to measure cell volume. For calibration purposes we also measured the ESR response of human red cells (RBC), the osmotic response of which is well documented with other methods. 15 refs., 1 fig.

  10. Evaluation of cytotoxic action of antihistamines – desloratadine and loratadine – using bulls spermatozoa as a test object

    Directory of Open Access Journals (Sweden)

    O. Kuzminov

    2014-02-01

    Full Text Available Antihistamines with active ingredients of loratadine and desloratadine are produced by Ukrainian pharmaceutical industry. According to the law, ther are assessed for their potential danger to human health and the environment, including alternative test objects. Evaluation has been carried out with regard to cytotoxic effect of pharmacological substances (loratadine and desloratadine using the bull sperm suspension as test objects, standardized and highly sensitive to toxic substances. Sperm was divided into the control sample (dissolved by phosphate-buffered saline and the investigated sample. Loratadine was added to phosphate-buffered saline in doses of 1/500 LD50 (12.3 mg, 1/100 LD50 (61.5 mg and LD50 (6150 mg. Desloratadine doses were 1/500 LD50 (1.25 mg, 1/100 LD50 (6.25 mg and LD50 (625 mg. Survival of spermatozoa was defined until termination of rectilinear forward movement in sperm intacted at +2…+5 °C. Respiratory activity (ex tempore was defined in 1.0 ml thermostated cell (temperature of 38.5 °C by polarography with the automatic registration of process flow by potentiometer; restorative activity was defined potentiometrically, using the open microelectrodes that were inserted in thermostated polarographic cell. Survival of spermatozoa in the sperm under the impact of loratadine in doses of 1/500 LD50 and 1/100 LD50 is respectively 136.0 ± 26.2 hours and 144.0 ± 19.6 hours. Adding LD50 dose of loratadine reduced survival to 112.0 ± 26.2 hours, which is lower than the control (160.0 ± 26.1 hours, respectively, by 10.0–15.0 and 30.0%. Loratadine reduces the respiratory activity of sperm: in the dose of 1/500 LD50 by 20.5%, in the dose of 1/100 LD50 – by 43.6%, and that of 100 LD50 – by 61.5% compared to the control. Restorative sperm activity under the impact of the loratadine reduced by 84.0% (dose of 1/500 LD50, 98.0% (dose of 1/100 LD50, 80.0% (dose LD50 compared to controls. The survival of spermatozoa in the sperm

  11. Morphometry and subpopulation structure of Holstein bull spermatozoa: variations in ejaculates and cryopreservation straws

    Science.gov (United States)

    Valverde, Anthony; Arenán, Héctor; Sancho, María; Contell, Jesús; Yániz, Jesús; Fernández, Alejandro; Soler, Carles

    2016-01-01

    Sperm quality is evaluated for the calculation of sperm dosage in artificial reproductive programs. The most common parameter used is motility, but morphology has a higher potential as a predictor of genetic quality. Morphometry calculations from CASA-Morph technology improve morphological evaluation and allow mathematical approaches to the problem. Semen from 28 Holstein bulls was collected by artificial vagina, and several ejaculates were studied. After general evaluation, samples were diluted, packaged in 0.25 ml straws, and stored in liquid nitrogen. Two straws per sample were thawed, and slides were processed and stained with Diff-Quik. Samples were analyzed by a CASA-Morph system for eight morphometric parameters. In addition to the “classical” statistical approach, based on variance analysis (revealing differences between animals, ejaculates, and straws), principal component (PC) analysis showed that the variables were grouped into PC1, related to size, and PC2 to shape. Subpopulation structure analysis showed four groups, namely, big, small, short, and narrow from their dominant characteristics, representing 31.0%, 27.3%, 24.1%, and 17.7% of the total population, respectively. The distributions varied between animals and ejaculates, but between straws, there were no differences in only four animals. This modern approach of considering an ejaculate sperm population as divided into subpopulations reflecting quantifiable parameters generated by CASA-Morph systems technology opens a new view on sperm function. This is the first study applying this approach to evaluate different ejaculates and straws from the same individual. More work must be done to improve seminal dose calculations in assisted reproductive programs. PMID:27678464

  12. EFFECTS OF A NEW ANTIBIOTIC COMBINATION ON POST -THAW MOTION CHRACTERISTICS AND MEMBRANE INTEGRITY OF BUFFALO AND SAHIWAL BULL SPERMATOZOA AND ON THE BACTERIOLOGICAL QUALITY OF THEIR SEMEN

    Directory of Open Access Journals (Sweden)

    S. Hasan, S. M. H. Andrabi, R. Muneer, M. Anzar and N. Ahmad

    2001-01-01

    Full Text Available In this study the effects of a new antibiotic combination, i.e., gentamycin, tylosin and linco-spectin (STLS on post-thaw motion characteristics, plasma membrane integrity, sperm morphology and the total aerobic bacterial counts (TABC in buffalo and Sahiwal bull semen were investigated. Ten ejaculates, five each from a buffalo and a Sahiwal bull, possessing more than 60% sperm motility were used. These ejaculates were diluted in Tris-citric acid (TCA extender (at 37 °C; 50 X 106 spermatozoa/mi, containing either GTLS (gentamycin 500 g/ml, tylosin 100 g/ml and linco-spectin 300/600 g/ml, streptomycin 1000 g/ml and penicillin 1000 IU/ml (SP, or negative control with no antibiotics (NCON. Samples were cooled to 4°C in 2 hours, equilibrated at 4°C for 4 hours, filled in 0.5 ml straws, frozen in a controlled rate cell freezer and plunged into liquid nitrogen. Frozen semen was thawed at 37°C for 15 seconds. Post-thaw sperm motion characteristics, plasma membrane integrity and sperm morphology were determined. Total aerobic bacterial counts and the frequency of appearance of bacterial genera were determined in neat semen, after dilution, and after freezing and thawing. Mean motilities (visual; computer-assisted, linear and circular, velocities (straight-line, average path and curvilinear and lateral head displacement (LHD in post- thaw semen samples did not differ due to antibiotics or species. Same was true for sperm plasma membrane integrity. Morphologically abnormal spermatozoa were lower (P<0.05 in GTLS and SP than in NCON. Sperm cells possessing normal acrosomes were higher (P<0.01 in GTLS and SP than in NCON. Total aerobic bacterial counts in post-thaw samples were lower (P<0.05 in GTLS than in SP or NCON. Staphylococcus and micrococcus were lower in samples treated with GTLS than that of SP or NCON. Pseudomonas and E.coli were more frequent in buffaloes than Sahiwal bull samples. Proteus and corynebacteria were scarcely present

  13. The effects of inclusion of exogenous phospholipid in Tris diluent with different level of egg yolk on the viability of bull spermatozoa

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    P Situmorang

    2002-10-01

    Full Text Available This study has been conducted to evaluate the effect of inclusion of phospholipid in Tris-diluent containing different level of egg yolk on the viability of bull spermatozoa after chilling and freezing. Semen was collected by means of artificial vagina, diluted in Tris-diluent to get a final concentration 100 x 106 spermatozoa/ml. Diluted semen was cooled to 50C for 45-60 minute, equiliberated in those temperature for 4 hours and frozen by placing the straw 5 cm above surface of liquid nitrogen for 10 minutes. The experiments was factorial designed with two level of phospholipid (0 and 0.5 mm and 4 concentration of egg yolk (0, 5, 10 and 20% v/v. The viability of spermatozoa was assessed by evaluating the percentage of motile, live and condition of apical ridge after the temperature reduced to 50C, stored at those temperature for 3 and 7 days and after thawing. Inclusion of phospholipid and level of egg yolk in Tris diluent significantly increased the viability of spermatozoa for both chilling and deepfreezing. The mean percentage of motile, live and intact apical ridge for 3 and 7 days of storage times at 50C, were significantly higher (P<0.05 in diluent containing phospholipid than without phospholipid. After thawing, the mean percentage motile, live and intact apical ridges were significantly higher (P<0.05 in a diluent containing phospholipid (49.9; 60.2 and 60.0 than those without phospholipid (39.1; 54.1 and 51.5. The effects of interaction between phospholipid and level of egg yolk on the viability of spermatozoa was not significant for both chilling and freezing. Level of egg yolk significantly (P<0.05 affected the viability of spermatozoa where the optimal level of egg yolk was 10% v/v for chilled semen and the higher level (20% v/v was needed for frozen semen. In conclusion, Tris diluent containing 0,5 mM phosphatidyl coline with 10 or 20% egg yolk gave a best protection for chilled and frozen semen respectively.

  14. Expression, Localization of SUMO-1, and Analyses of Potential SUMOylated Proteins in Bubalus bubalis Spermatozoa

    OpenAIRE

    Rahim Dad Brohi; Li Wang; Najla Ben Hassine; Jing Cao; Hira Sajjad Talpur; Di Wu; Chun-Jie Huang; Zia-Ur Rehman; Dinesh Bhattarai; Li-Jun Huo

    2017-01-01

    Mature spermatozoa have highly condensed DNA that is essentially silent both transcriptionally and translationally. Therefore, post translational modifications are very important for regulating sperm motility, morphology, and for male fertility in general. Protein sumoylation was recently demonstrated in human and rodent spermatozoa, with potential consequences for sperm motility and DNA integrity. We examined the expression and localization of small ubiquitin-related modifier-1 (SUMO-1) in t...

  15. Retrotransposon expression and incorporation of cloned human and mouse retroelements in human spermatozoa.

    Science.gov (United States)

    Lazaros, Leandros; Kitsou, Chrysoula; Kostoulas, Charilaos; Bellou, Sofia; Hatzi, Elissavet; Ladias, Paris; Stefos, Theodoros; Markoula, Sofia; Galani, Vasiliki; Vartholomatos, Georgios; Tzavaras, Theodore; Georgiou, Ioannis

    2017-03-01

    To investigate the expression of long interspersed element (LINE) 1, human endogenous retrovirus (HERV) K10, and short interspersed element-VNTR-Alu element (SVA) retrotransposons in ejaculated human spermatozoa by means of reverse-transcription (RT) polymerase chain reaction (PCR) analysis as well as the potential incorporation of cloned human and mouse active retroelements in human sperm cell genome. Laboratory study. University research laboratories and academic hospital. Normozoospermic and oligozoospermic white men. RT-PCR analysis was performed to confirm the retrotransposon expression in human spermatozoa. Exogenous retroelements were tagged with a plasmid containing a green fluorescence (EGFP) retrotransposition cassette, and the de novo retrotransposition events were tested with the use of PCR, fluorescence-activated cell sorting analysis, and confocal microscopy. Retroelement expression in human spermatozoa, incorporation of cloned human and mouse active retroelements in human sperm genome, and de novo retrotransposition events in human spermatozoa. RT-PCR products of expressed human LINE-1, HERV-K10, and SVA retrotransposons were observed in ejaculated human sperm samples. The incubation of human spermatozoa with either retrotransposition-active human LINE-1 and HERV-K10 or mouse reverse transcriptase-deficient VL30 retrotransposons tagged with an EGFP-based retrotransposition cassette led to EGFP-positive spermatozo; 16.67% of the samples were positive for retrotransposition. The respective retrotransposition frequencies for the LINE-1, HERV-K10, and VL30 retrotransposons in the positive samples were 0.34 ± 0.13%, 0.37 ± 0.17%, and 0.30 ± 0.14% per sample of 10,000 spermatozoa. Our results show that: 1) LINE-1, HERV-K10, and SVA retrotransposons are transcriptionally expressed in human spermatozoa; 2) cloned active retroelements of human and mammalian origin can be incorporated in human sperm genome; 3) active reverse transcriptases exist in human

  16. Melatonin receptors MT1 and MT2 are expressed in spermatozoa from several seasonal and nonseasonal breeder species.

    Science.gov (United States)

    González-Arto, Marta; Vicente-Carrillo, Alejandro; Martínez-Pastor, Felipe; Fernández-Alegre, Estela; Roca, Jordi; Miró, Jordi; Rigau, Teresa; Rodríguez-Gil, Joan E; Pérez-Pé, Rosaura; Muiño-Blanco, Teresa; Cebrián-Pérez, José A; Casao, Adriana

    2016-11-01

    Melatonin is a ubiquitous and multipurpose molecule, and one of its roles is to regulate reproduction in some seasonal mammals. Our group has previously reported the variation in the melatonin levels in ram seminal plasma along the year and identified MT1 and MT2 receptors in ram spermatozoa. The objective of this study was to elucidate whether the presence of melatonin receptors (MT1 and MT2) in the sperm plasma membrane, and melatonin in the seminal plasma is related to seasonal breeding. For this purpose, the presence of melatonin receptors and the levels of melatonin in seminal plasma have been examined in several species: donkey and stallion as long-day breeders; red deer as a wild, short-day, highly seasonal breeder (epididymal spermatozoa); bull as a conventional nonseasonal breeder; boar as a seasonal breeder under management techniques; and dog as possible a seasonal breeder not regulated by melatonin. We have detected measurable levels of melatonin in the seminal plasma of all ejaculated semen samples (from donkey, stallion, boar, bull, and dog). Also, and for the first time, we have demonstrated the presence of MT1 and MT2 melatonin receptors in the spermatozoa of all these species, regardless their type of reproduction or sperm source (ejaculated or epididymal), using indirect immunofluorescence techniques and Western blotting. Our findings suggest that melatonin and melatonin receptors may be universally distributed in the reproductive system of mammals and that the sperm melatonin receptors cells may not be necessarily related with seasonal reproduction. Furthermore, the presence of MT1 at the cytoplasmic droplet in immature ejaculated stallion spermatozoa found in one sample and epididymal red deer spermatozoa suggests that melatonin may be involved in specific functions during spermatogenesis and sperm maturation, like protecting spermatozoa from oxidative damage, this activity being mediated through these receptors. Copyright © 2016 Elsevier Inc

  17. Nicotine Effects and Receptor Expression on Human Spermatozoa: Possible Neuroendocrine Mechanism

    Science.gov (United States)

    Condorelli, Rosita A.; La Vignera, Sandro; Giacone, Filippo; Iacoviello, Linda; Mongioì, Laura M.; Li Volti, Giovanni; Barbagallo, Ignazio; Avola, Roberto; Calogero, Aldo E.

    2017-01-01

    The aim of this experimental study was to investigate the mechanism by which nicotine (NIC) alters spermatozoa and to evaluate the expression of nicotinic receptors (nAChR) subunits in human spermatozoa. We analyzed 30 healthy normozoospermic men. Spermatozoa were incubated with NIC 100 ng/ml and the nAChR antagonist, hexamethonium (HEX) (0, 100, 1,000, 10,000 ng/ml) for 3 and 24 h. The following sperm parameters evaluated: (a) progressive motility; (b) mitochondrial membrane potential (MMP); (c) chromatin compactness; (d) externalization of phosphatidylserine (PS); (e) late apoptosis; (f) viability; (g) DNA fragmentation; (h) degree of lipid peroxidation (LP) by flow cytometry; (i) nAChR subunits expression by quantitative Real Time PCR and (j) protein expression evaluation by Western blot analysis. HEX fully antagonized the effects of NIC both after 3 and 24 h of incubation with significant improvement (p NIC in raw semen but this effect was fully antagonized (p NIC on sperm function are mediated by interaction with a specific nicotinic receptor. The presence of nAChR subunits suggests the presence of a neuroendocrine mechanism on human spermatozoa. PMID:28400736

  18. The human "prion-like" protein Doppel is expressed in both Sertoli cells and spermatozoa.

    Science.gov (United States)

    Peoc'h, Katell; Serres, Catherine; Frobert, Yveline; Martin, Caroline; Lehmann, Sylvain; Chasseigneaux, Stephanie; Sazdovitch, Veronique; Grassi, Jacques; Jouannet, Pierre; Launay, Jean-Marie; Laplanche, Jean-Louis

    2002-11-08

    The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. Experimental data suggest either direct or indirect interaction between the two proteins. In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. It was found on Sertoli cells in the testis, on the flagella of epididymal and mature spermatozoa, and in seminal plasma. Dpl coexists only with N-terminally truncated isoforms of PrP(c) on mature spermatozoa. The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. Finally, our data indicate that spermatozoa are thus an interesting model for studies of the potential interaction between Dpl and PrP(c).

  19. HSP90 expression correlation with the freezing resistance of bull sperm.

    Science.gov (United States)

    Wang, Peng; Wang, Yan-Feng; Wang, Hong; Wang, Chun-Wei; Zan, Lin-Sen; Hu, Jian-Hong; Li, Qing-Wang; Jia, Yong-Hong; Ma, Guo-Ji

    2014-05-01

    To date, there has been little improvement in cryopreservation of bull sperm due to lack of understanding of the freezing mechanisms. Therefore, this study set out to investigate expression levels of fertility-associated proteins in bull sperm, and in particular the relationship between the 90 kDa heat-shock protein (HSP90) and the sperm characteristics after freezing-thawing. Semen was collected from eight Holstein bulls by artificial vagina. Characteristics of these fresh semen, including sperm motility, morphology, viability and concentration, were evaluated. Sperm quality was also assessed after freezing-thawing. Eight ejaculates were divided into two groups based on freezing resistance and sperm motility. Sperm proteins were extracted and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and western blotting were performed. SDS-PAGE results showed that there was substantial diversity in 90 kDa proteins in the frozen-thawed sperm and HSP90 was confirmed as one of the 90 kDa proteins by western blot. This study indicated that HSP90 expression correlated positively with sperm quality. The amount of expressed 90 kDa proteins in the high freezing resistance (HFR) group was significantly higher than that in the low freezing resistance (LFR) group (P sperm found after freezing-thawing. Therefore, we concluded that level of HSP90 expression could be used to predict reliably and simply the freezing resistance of bull sperm.

  20. Functional insulin-like factor 3 (INSL3) hormone-receptor system in the testes and spermatozoa of domestic ruminants and its potential as a predictor of sire fertility.

    Science.gov (United States)

    Pitia, Ali M; Uchiyama, Kyoko; Sano, Hiroaki; Kinukawa, Masashi; Minato, Yoshiaki; Sasada, Hiroshi; Kohsaka, Tetsuya

    2017-04-01

    Insulin-like factor 3 (INSL3) is essential for fetal testis descent, and has been implicated in the testicular and sperm functions in adult males; however, similar functions in domestic ruminants remain largely unknown. This study investigated the functional INSL3 hormone-receptor system in adult ruminant testes and spermatozoa, and explored its potential to diagnose the fertility of sires. Testes and spermatozoa were obtained from fertile bulls, rams and he-goats, whereas subfertile testes and spermatozoa were obtained only from bulls. As expected, INSL3 was visualized in Leydig cells, while we clearly demonstrated that the functional receptor, relaxin family peptide receptor 2 (RXFP2), enabling INSL3 to bind was identified in testicular germ cells and in the sperm equatorial segment of bulls, rams and he-goats. In comparison to fertile bulls, the percentage of INSL3- and RXFP2-expressing cells and their expression levels per cell were significantly reduced in the testes of subfertile bulls. In addition, the population of INSL3-binding spermatozoa was also significantly reduced in the semen of subfertile bulls. These results provide evidence for a functional INSL3 hormone-receptor system operating in ruminant testes and spermatozoa, and its potential to predict subfertility in sires. © 2016 Japanese Society of Animal Science.

  1. Comprehensive proteomic analysis of bovine spermatozoa of varying fertility rates and identification of biomarkers associated with fertility

    Directory of Open Access Journals (Sweden)

    Kaya Abdullah

    2008-02-01

    Full Text Available Abstract Background Male infertility is a major problem for mammalian reproduction. However, molecular details including the underlying mechanisms of male fertility are still not known. A thorough understanding of these mechanisms is essential for obtaining consistently high reproductive efficiency and to ensure lower cost and time-loss by breeder. Results Using high and low fertility bull spermatozoa, here we employed differential detergent fractionation multidimensional protein identification technology (DDF-Mud PIT and identified 125 putative biomarkers of fertility. We next used quantitative Systems Biology modeling and canonical protein interaction pathways and networks to show that high fertility spermatozoa differ from low fertility spermatozoa in four main ways. Compared to sperm from low fertility bulls, sperm from high fertility bulls have higher expression of proteins involved in: energy metabolism, cell communication, spermatogenesis, and cell motility. Our data also suggests a hypothesis that low fertility sperm DNA integrity may be compromised because cell cycle: G2/M DNA damage checkpoint regulation was most significant signaling pathway identified in low fertility spermatozoa. Conclusion This is the first comprehensive description of the bovine spermatozoa proteome. Comparative proteomic analysis of high fertility and low fertility bulls, in the context of protein interaction networks identified putative molecular markers associated with high fertility phenotype.

  2. Expression, Localization of SUMO-1, and Analyses of Potential SUMOylated Proteins in Bubalus bubalis Spermatozoa

    Directory of Open Access Journals (Sweden)

    Rahim Dad Brohi

    2017-06-01

    Full Text Available Mature spermatozoa have highly condensed DNA that is essentially silent both transcriptionally and translationally. Therefore, post translational modifications are very important for regulating sperm motility, morphology, and for male fertility in general. Protein sumoylation was recently demonstrated in human and rodent spermatozoa, with potential consequences for sperm motility and DNA integrity. We examined the expression and localization of small ubiquitin-related modifier-1 (SUMO-1 in the sperm of water buffalo (Bubalus bubalis using immunofluorescence analysis. We confirmed the expression of SUMO-1 in the acrosome. We further found that SUMO-1 was lost if the acrosome reaction was induced by calcium ionophore A23187. Proteins modified or conjugated by SUMO-1 in water buffalo sperm were pulled down and analyzed by mass spectrometry. Sixty proteins were identified, including proteins important for sperm morphology and motility, such as relaxin receptors and cytoskeletal proteins, including tubulin chains, actins, and dyneins. Forty-six proteins were predicted as potential sumoylation targets. The expression of SUMO-1 in the acrosome region of water buffalo sperm and the identification of potentially SUMOylated proteins important for sperm function implicates sumoylation as a crucial PTM related to sperm function.

  3. Expression, Localization of SUMO-1, and Analyses of Potential SUMOylated Proteins in Bubalus bubalis Spermatozoa.

    Science.gov (United States)

    Brohi, Rahim Dad; Wang, Li; Hassine, Najla Ben; Cao, Jing; Talpur, Hira Sajjad; Wu, Di; Huang, Chun-Jie; Rehman, Zia-Ur; Bhattarai, Dinesh; Huo, Li-Jun

    2017-01-01

    Mature spermatozoa have highly condensed DNA that is essentially silent both transcriptionally and translationally. Therefore, post translational modifications are very important for regulating sperm motility, morphology, and for male fertility in general. Protein sumoylation was recently demonstrated in human and rodent spermatozoa, with potential consequences for sperm motility and DNA integrity. We examined the expression and localization of small ubiquitin-related modifier-1 (SUMO-1) in the sperm of water buffalo (Bubalus bubalis) using immunofluorescence analysis. We confirmed the expression of SUMO-1 in the acrosome. We further found that SUMO-1 was lost if the acrosome reaction was induced by calcium ionophore A23187. Proteins modified or conjugated by SUMO-1 in water buffalo sperm were pulled down and analyzed by mass spectrometry. Sixty proteins were identified, including proteins important for sperm morphology and motility, such as relaxin receptors and cytoskeletal proteins, including tubulin chains, actins, and dyneins. Forty-six proteins were predicted as potential sumoylation targets. The expression of SUMO-1 in the acrosome region of water buffalo sperm and the identification of potentially SUMOylated proteins important for sperm function implicates sumoylation as a crucial PTM related to sperm function.

  4. Cystic fibrosis transmembrane conductance regulator protein expression rate in healthy spermatozoa is not correlated with ovum fertilisation rate.

    Science.gov (United States)

    Li, H-G; Xu, C-M; Chen, W-Y; Shi, Q-X; Ni, Y

    2012-05-01

    Our previous studies have shown that the cystic fibrosis transmembrane conductance regulator (CFTR) was important for capacitation and fertilisation in mouse, guinea pig and human spermatozoa. However, it is unclear whether CFTR is correlated with ovum fertilisation rate. The present study was to test the possible relationship between spermatozoa CFTR protein expression rate in healthy men and ovum fertilisation rate during in vitro fertilisation. Ninety-four couples for female factor infertility for IVF-ET treatments were retrospectively studied. All the patients were divided into three groups based on the fertilisation rate of ovum in vitro. It was performed to explore whether there were differences in sperm CFTR protein expression rate among the three groups and the relevance between CFTR protein expression rate and ovum fertilisation rate. Our study showed that there was no significant differences in sperm CFTR protein expression rate among the three groups (F = 0.614, P = 0.544), and the relevance between spermatozoa CFTR protein expression rate and ovum fertilisation rate was not significantly different (r = 0.013, P = 0.904). These results further suggest that CFTR protein expression rate in healthy men spermatozoa was not associated with ovum fertilisation rate and thus we cannot predict ovum fertilisation results by sperm CFTR protein expression rate. © 2011 Blackwell Verlag GmbH.

  5. Effects of Hormones on the Expression of Matrix Metalloproteinases and Their Inhibitors in Bovine Spermatozoa

    Directory of Open Access Journals (Sweden)

    Sang-Hwan Kim

    2013-03-01

    Full Text Available Proteases and protease inhibitors play key roles in most physiological processes, including cell migration, cell signaling, and cell surface and tissue remodeling. Among these, the matrix metalloproteinase (MMPs pathway is one of the most efficient biosynthetic pathways for controlling the activation of enzymes responsible for protein degradation. This also indicates the association of MMPs with the maturation of spermatozoa. In an attempt to investigate the effect of MMP activation and inhibitors in cultures with various hormones during sperm capacitation, we examined and monitored the localization and expression of MMPs (MMP-2 and MMP-9, tissue inhibitors of metalloproteinases (TIMP-2 and TIMP-3, as well as their expression profiles. Matured spermatozoa were collected from cultures with follicle-stimulating hormone (FSH, luteinizing hormone (LH, and Lutalyse at 1 h, 6 h, 18 h, and 24 h. ELISA detected the expression of MMP-2, MMP-9, TIMP-2, and TIMP-3 in all culture media, regardless of medium type (FSH-supplemented fertilization Brackett-Oliphant medium (FFBO, LH-supplemented FBO (LFBO, or Lutalyse-supplemented FBO (LuFBO. TIMP-2 and TIMP-3 expression patterns decreased in LFBO and LuFBO. MMP-2 and MMP-9 activity in FBO and FFBO progressively increased from 1 h to 24 h but was not detected in LFBO and LuFBO. The localization and expression of TIMP-2 and TIMP-3 in sperm heads was also measured by immunofluorescence analysis. However, MMPs were not detected in the sperm heads. MMP and TIMP expression patterns differed according to the effect of various hormones. These findings suggest that MMPs have a role in sperm viability during capacitation. In conjunction with hormones, MMPs play a role in maintaining capacitation and fertilization by controlling extracellular matrix inhibitors of sperm.

  6. Testicular cytology indicates differences in Sertoli cell counts between "good freezer" and "poor freezer" bulls.

    Science.gov (United States)

    Rajak, Shailendra Kumar; Thippeswamy, Vijetha Bajjalli; Kumaresan, Arumugam; Layek, Siddhartha Shankar; Mohanty, Tushar Kumar; Gaurav, Mukesh Kumar; Chakravarty, Atish Kumar; Datta, Tirtha Kumar; Manimaran, Ayyasamy; Prasad, Shiv

    2016-01-01

    In artificial insemination, poor quality of semen unsuitable for cryopreservation and susceptibility of spermatozoa to cryodamage in crossbred bulls have been a matter of concern. Present study was designed to identify the testicular cytology indices that might be used to predict the semen quality and cryotolerance of spermatozoa in bulls. Based on the ejaculate rejection rate and sperm cryotolerance, bulls (Holstein Friesian X Tharparkar crossbred) were classified into either good (producing good quality semen with spermatozoa having good cryotolerance; n = 4) or poor (producing poor quality semen with spermatozoa having poor cryotolerance; n = 4). Testicular cytology was studied in all the 8 bulls using fine needle aspiration technique. Testicular cytology of good bulls and poor bulls differed significantly. The proportion of Sertoli cells was significantly higher in good bulls (25.3 ± 1.6) compared to poor bulls (11.0 ± 0.8). The Sertoli cell index was 46.1 ± 5.0 in good bulls while it was only 13.8 ± 1.3 in poor bulls. The cut off values, as determined using Receiver Operating Characteristics analysis, indicate that the bulls having testicular cytogram comprising of 4.0 spermatogenic cells to Sertoli cell ratio might be a poor bull in terms of semen quality and cryotolerance of spermatozoa. The proportion of Sertoli cells in the testicular cytology had positive (P < 0.05) relationship with semen quality and cryotolerance of spermatozoa.

  7. Epididymosomes transfer epididymal sperm binding protein 1 (ELSPBP1) to dead spermatozoa during epididymal transit in bovine.

    Science.gov (United States)

    D'Amours, Olivier; Frenette, Gilles; Bordeleau, Louis-Jean; Allard, Nancy; Leclerc, Pierre; Blondin, Patrick; Sullivan, Robert

    2012-10-01

    Previously, we showed that epididymal sperm binding protein 1 (ELSPBP1) characterizes spermatozoa already dead before ejaculation in bovine. In this study, we investigated the presence of ELSPBP1 in bull genital tract as well as its acquisition by spermatozoa during epididymal transit. As assessed by real-time RT-PCR, ELSPBP1 was highly expressed in the caput and the corpus epididymis but was present in lower expression levels in the testis and the cauda epididymis. Immunohistochemistry revealed the same expression pattern. However, Western blot on tissue homogenates showed some discrepancies, as ELSPBP1 was found in a comparable concentration all along the epididymis. This difference was due to the presence of ELSPBP1 in the epididymal fluid. In both caput and cauda epididymal fluid, ELSPBP1 was associated with the epididymosomes, small membranous vesicles secreted by epithelial cells of the epididymis and implicated in the transfer of proteins to spermatozoa. As assessed by immunocytometry, ELSPBP1 was found on a subset of dead spermatozoa in caput epididymis but was found on all dead spermatozoa in cauda epididymis. To assess ELSPBP1 acquisition by spermatozoa, caput epididymal spermatozoa were incubated with cauda epididymosomes under various conditions. ELSPBP1 detection by immunocytometry assay revealed that only spermatozoa already dead before incubation were receptive to ELSPBP1 transfer by epididymosomes. This receptivity was enhanced by the presence of zinc in the incubation medium. This specificity for a sperm subpopulation suggests that an underlying mechanism is involved and that ELSPBP1 could be a tag for the recognition of dead spermatozoa during epididymal transit.

  8. Viability of bull semen extended with commercial semen extender ...

    African Journals Online (AJOL)

    Andrea Raseona

    Abstract. The aim of this study was to evaluate the viability of bull spermatozoa diluted with commercial semen extender and two culture media stored at controlled room temperature (24 °C) for 72 hours. Two Nguni bulls were used for semen collection with the aid of an electro-ejaculator. After macroscopic evaluation ...

  9. Sperm protamine-status correlates to the fertility of breeding bulls.

    Science.gov (United States)

    Dogan, Sule; Vargovic, Peter; Oliveira, Rodrigo; Belser, Lauren E; Kaya, Abdullah; Moura, Arlindo; Sutovsky, Peter; Parrish, John; Topper, Einko; Memili, Erdoğan

    2015-04-01

    During fertilization, spermatozoa make essential contributions to embryo development by providing oocyte activating factors, centrosomal components, and paternal chromosomes. Protamines are essential for proper packaging of sperm DNA; however, in contrast to the studies of oocyte-related female infertility, the influence of sperm chromatin structure on male infertility has not been evaluated extensively. The objective of this study was to determine the sperm chromatin content of bull spermatozoa by evaluating DNA fragmentation, chromatin maturity/protamination, PRM1 protein status, and nuclear shape in spermatozoa from bulls with different fertility. Relationships between protamine 1 (PRM1) and the chromatin integrity were ascertained in spermatozoa from Holstein bulls with varied (high vs. low) but acceptable fertility. Sperm DNA fragmentation and chromatin maturity (protamination) were tested using Halomax assay and toluidine blue staining, respectively. The PRM1 content was assayed using Western blotting and in-gel densitometry, flow cytometry, and immunocytochemistry. Fragmentation of DNA was increased and chromatin maturity significantly reduced in spermatozoa from low-fertility bulls compared to those from high-fertility bulls. Field fertility scores of the bulls were negatively correlated with the percentage of spermatozoa displaying reduced protamination and fragmented DNA using toluidine blue and Halomax, respectively. Bull fertility was also positively correlated with PRM1 content by Western blotting and flow cytometry. However, detection of PRM1 content by Western blotting alone was not predictive of bull fertility. In immunocytochemistry, abnormal spermatozoa showed either a lack of PRM1 or scattered localization in the apical/acrosomal region of the nuclei. The nuclear shape was distorted in spermatozoa from low-fertility bulls. In conclusion, we showed that inadequate amount and localization of PRM1 were associated with defects in sperm chromatin

  10. Gene expression profile of Musculus longissimus dorsi in bulls of a Charolais × Holstein F2-cross with divergent intramuscular fat content

    OpenAIRE

    Katrin Komolka; Siriluck Ponsuksili; Elke Albrecht; Christa Kühn; Klaus Wimmers; Steffen Maak

    2016-01-01

    Transcriptomes of Musculus longissimus dorsi (MLD) were compared between bulls from a F2-cross derived from Charolais and Holstein Friesian. Two groups of 10 bulls were selected which differed significantly in intramuscular fat (IMF) deposition despite standardized husbandry and feeding conditions and identical sires in both groups. Consequently, genetic factors underlying the different capability of IMF deposition should be identified. A total of 32 differentially expressed genes (DEGs) were...

  11. Impact of cryopreservation on bull () semen proteome.

    Science.gov (United States)

    Westfalewicz, B; Dietrich, M A; Ciereszko, A

    2015-11-01

    Cryopreservation of bull spermatozoa is a well-established technique, allowing artificial insemination of cattle on a commercial scale. However, the extent of proteome changes in seminal plasma and spermatozoa during cryopreservation are not yet fully known. The objective of this study was to compare the proteomes of fresh, equilibrated, and cryopreserved bull semen (spermatozoa and seminal plasma) to establish the changes in semen proteins during the cryopreservation process. Semen was collected from 6 mature Holstein Friesian bulls. After sample processing, comparative analysis and identification of proteins was performed using 2-dimensional difference in-gel electrophoresis coupled with matrix-assisted laser desorption/ionization mass spectrometry. Analysis of spermatozoa extracts revealed that 25 identified protein spots, representing 16 proteins, underwent significant ( cryopreservation. Eighteen protein spots decreased in abundance, 5 protein spots increased in abundance, and 2 protein spots showed different, specific patterns of abundance changes. Analysis of seminal fluid containing seminal plasma showed that 6 identified protein spots, representing 4 proteins, underwent significant ( cryopreservation. Two protein spots increased in abundance and 4 decreased in abundance. Semen extending and equilibration seems to be responsible for a significant portion of the proteome changes related to cryopreservation technology. Most sperm proteins affected by equilibration and cryopreservation are membrane bound, and loss of those proteins may reduce natural spermatozoa coating. Further research is needed to unravel the mechanisms of the particular protein changes described in this study and establish the relationship between those changes and sperm quality.

  12. Expression of Tas1 taste receptors in mammalian spermatozoa: functional role of Tas1r1 in regulating basal Ca²⁺ and cAMP concentrations in spermatozoa.

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    Dorke Meyer

    Full Text Available BACKGROUND: During their transit through the female genital tract, sperm have to recognize and discriminate numerous chemical compounds. However, our current knowledge of the molecular identity of appropriate chemosensory receptor proteins in sperm is still rudimentary. Considering that members of the Tas1r family of taste receptors are able to discriminate between a broad diversity of hydrophilic chemosensory substances, the expression of taste receptors in mammalian spermatozoa was examined. METHODOLOGY/PRINCIPAL FINDINGS: The present manuscript documents that Tas1r1 and Tas1r3, which form the functional receptor for monosodium glutamate (umami in taste buds on the tongue, are expressed in murine and human spermatozoa, where their localization is restricted to distinct segments of the flagellum and the acrosomal cap of the sperm head. Employing a Tas1r1-deficient mCherry reporter mouse strain, we found that Tas1r1 gene deletion resulted in spermatogenic abnormalities. In addition, a significant increase in spontaneous acrosomal reaction was observed in Tas1r1 null mutant sperm whereas acrosomal secretion triggered by isolated zona pellucida or the Ca²⁺ ionophore A23187 was not different from wild-type spermatozoa. Remarkably, cytosolic Ca²⁺ levels in freshly isolated Tas1r1-deficient sperm were significantly higher compared to wild-type cells. Moreover, a significantly higher basal cAMP concentration was detected in freshly isolated Tas1r1-deficient epididymal spermatozoa, whereas upon inhibition of phosphodiesterase or sperm capacitation, the amount of cAMP was not different between both genotypes. CONCLUSIONS/SIGNIFICANCE: Since Ca²⁺ and cAMP control fundamental processes during the sequential process of fertilization, we propose that the identified taste receptors and coupled signaling cascades keep sperm in a chronically quiescent state until they arrive in the vicinity of the egg - either by constitutive receptor activity and

  13. The effect of Ureaplasma urealyticum on the level of P34H expression, the activity of hyaluronidase, and DNA fragmentation in human spermatozoa.

    Science.gov (United States)

    Ma, Xiao-Ping; Gao, Xiao-Qin

    2017-01-01

    To study the effect of Ureaplasma urealyticum on the level of P34H expression, the activity of hyaluronidase, and the DNA fragmentation in human spermatozoa. Western blot was used to detect the level of P34H expression on spermatozoa.The localization of this protein on human spermatozoa was determined by indirect immunofluorescent and observed by laser scanning confocal microscope. The activity of hyaluronidase was examined by improved fixed-substrate film method. The DNA fragmentation was assayed with the use of TUNEL. There were significant differences in the level of P34H protein expression, the percentage of the P34H-positive rate, the activity of HYD, and the percentage of DNA fragmentation between each infertile group and the control (Pfragmentation. Ureaplasma urealyticum infection may affect the level of P34H protein expression, the percentage of the P34H-positive rate, the activity of HYD, and DNA fragmentation that influence fertility. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. Cryopreservation of stallion spermatozoa.

    Science.gov (United States)

    Graham, J K

    1996-04-01

    The main advantage to using frozen semen in any breeding program is faster genetic gain for the inherited trait desired. Milk production of dairy cows doubled (from 26,000 to 52,000 kg of milk/cow per year) between 1950 and 1980, because the dairy industry was using semen only from bulls with the greatest genetic potential for milk production. This genetic gain could have been achieved without the use of frozen semen; however, the time required to achieve that same genetic progress would have been lengthened exceedingly. Fertility rates using frozen stallion spermatozoa are not equal to that of fresh semen, but with careful management of both the stallion and the mare fertility rates of 40% to 60% can be achieved for a single mare reproductive cycle and fertility rates of 60% to 75% during the course of a breeding season can be expected for a number of stallions. Because pregnancy rates have increased and because many breed registries now condone the use of frozen stallion semen, more people in the equine industry are using frozen semen. It has been the attempt of this article to make the practitioner aware of some of the problems that can occur when processing, freezing, storing, and using frozen semen, and how to control those problems. The practitioner also should be aware that there is large variation in semen quality from stallions, and that spermatozoa from some of the problem stallions can be improved by altering a "standard protocol," but that semen from some stallions will not cryopreserve effectively, regardless of the efforts put forth.

  15. Expression of the vitamin D metabolizing enzyme CYP24A1 at the annulus of human spermatozoa may serve as a novel marker of semen quality

    DEFF Research Database (Denmark)

    Jensen, Martin Blomberg; Jørgensen, A; Nielsen, J E

    2012-01-01

    Vitamin D (VD) is important for male reproduction in mammals and the VD receptor (VDR) and VD-metabolizing enzymes are expressed in human spermatozoa. The VD-inactivating enzyme CYP24A1 titrates the cellular responsiveness to VD, is transcriptionally regulated by VD, and has a distinct expression......) detection of CYP24A1. Another 40 men (22 young, 18 subfertile) were tested for in vitro effects of 1,25(OH)(2) D(3) on intracellular calcium concentration ([Ca(2+) ](i) ) and sperm motility. Double ICC staining showed that CYP24A1 and VDR were either concomitantly expressed or absent in 80...

  16. Infertility of the breeding bull in insemination technology

    Directory of Open Access Journals (Sweden)

    Predojević Mirko R.

    2004-01-01

    Full Text Available In spite of very strict breeding bull selection, especialy for A.I programes their infertility is a very serius problem in everyday practice. Especially bull semen has been marked as the main factor for unsatisfied cow fertility in the A.I.programme. The reason could be the bull semen which really may play as the spreading factor of the specific or non-specific reproductive infective disoders – IBR, IPV, BVD, Campylobacter-Vibrio fetus, brucellosis leptospirosis, tuberculosis and other reproductive diseases. Secondarily, the percentage of vitality, motility, penetration abilities, and immonological properties of bull spermatozoa also have an important role in unsuccessful bovine fecundation. That is, why it is necessary to secure professional health care for breedig bull in AI centres, becase only healthy bulls can ensure good bovine genetic transmission and progress in cattle production for today's growing population.

  17. Gene expression profile of Musculus longissimus dorsi in bulls of a Charolais × Holstein F2-cross with divergent intramuscular fat content

    Directory of Open Access Journals (Sweden)

    Katrin Komolka

    2016-03-01

    Full Text Available Transcriptomes of Musculus longissimus dorsi (MLD were compared between bulls from a F2-cross derived from Charolais and Holstein Friesian. Two groups of 10 bulls were selected which differed significantly in intramuscular fat (IMF deposition despite standardized husbandry and feeding conditions and identical sires in both groups. Consequently, genetic factors underlying the different capability of IMF deposition should be identified. A total of 32 differentially expressed genes (DEGs were found of which 11 were up-regulated and 21 were down-regulated in the high IMF group. Ingenuity Pathway Analysis (IPA identified a gene network comprising DEGs with functions in carbohydrate metabolism, lipid metabolism and molecular transport. The data from this study were deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE75347. We provide here a dataset which is of potential value to dissect molecular pathways influencing differences in IMF deposition in crossbred cattle with standardized genetic background.

  18. Gene expression profile of Musculus longissimus dorsi in bulls of a Charolais × Holstein F2-cross with divergent intramuscular fat content.

    Science.gov (United States)

    Komolka, Katrin; Ponsuksili, Siriluck; Albrecht, Elke; Kühn, Christa; Wimmers, Klaus; Maak, Steffen

    2016-03-01

    Transcriptomes of Musculus longissimus dorsi (MLD) were compared between bulls from a F2-cross derived from Charolais and Holstein Friesian. Two groups of 10 bulls were selected which differed significantly in intramuscular fat (IMF) deposition despite standardized husbandry and feeding conditions and identical sires in both groups. Consequently, genetic factors underlying the different capability of IMF deposition should be identified. A total of 32 differentially expressed genes (DEGs) were found of which 11 were up-regulated and 21 were down-regulated in the high IMF group. Ingenuity Pathway Analysis (IPA) identified a gene network comprising DEGs with functions in carbohydrate metabolism, lipid metabolism and molecular transport. The data from this study were deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE75347. We provide here a dataset which is of potential value to dissect molecular pathways influencing differences in IMF deposition in crossbred cattle with standardized genetic background.

  19. Relationship of nonreturn rates of dairy bulls to binding affinity of heparin to sperm

    Energy Technology Data Exchange (ETDEWEB)

    Marks, J.L.; Ax, R.L.

    1985-08-01

    The binding of the glycosaminoglycan (3H) heparin to bull spermatozoa was compared with nonreturn rates of dairy bulls. Semen samples from five bulls above and five below an average 71% nonreturn rate were used. Samples consisted of first and second ejaculates on a single day collected 1 d/wk for up to 5 consecutive wk. Saturation binding assays using (TH) heparin were performed to quantitate the binding characteristics of each sample. Scatchard plot analyses indicated a significant difference in the binding affinity for (TH) heparin between bulls of high and low fertility. Dissociation constants were 69.0 and 119.3 pmol for bulls of high and low fertility, respectively. In contrast, the number of binding sites for (TH) heparin did not differ significantly among bulls. Differences in binding affinity of (TH) heparin to bull sperm might be used to predict relative fertility of dairy bulls.

  20. Expression of a2 Vacuolar ATPase in Spermatozoa is Associated with Semen Quality and Chemokine-Cytokine Profiles in Infertile Men

    Science.gov (United States)

    Ota, Kuniaki; Jaiswal, Mukesh Kumar; Ramu, Sivakumar; Jeyendran, Rajasinjham; Kwak-Kim, Joanne; Gilman-Sachs, Alice; Beaman, Kenneth D.

    2013-01-01

    Background A number of laboratory tests have been developed to determine properties of spermatozoa quality but few have been adopted into routine clinical use in place of the WHO semen analysis. We investigated whether Atp6v0a2 (a2 isoform of vacuolar ATPase) is associated with abnormal semen quality and changes in chemokine-cytokine profiles in infertile men. Patients and Methods Semen samples were collected from 35 healthy donors and 35 infertile men at the Andrology laboratory from August 2011 to June 2012. The levels of Atp6v0a2 mRNA and protein, and its localization in spermatozoa were determined. a2NTD (the N-terminal portion of Atp6v0a2) and secreted chemokine-cytokine profiles in seminal fluid were measured. Results Atp6v0a2 protein (P<0.05) and mRNA (P<0.05) in spermatozoa from infertile men were significantly lower than those from fertile men. Fluorescent microscopy revealed that Atp6v0a2 is mainly expressed in the acrosomal region. Infertile men’s seminal fluid had significantly lower G-CSF (P<0.01), GM-CSF (P<0.01), MCP-1 (P<0.05), MIP-1α (P<0.01) and TGF-β1 (P<0.01) levels when compared to the seminal fluid from fertile men. Seminal fluid a2NTD levels were significantly correlated with G-CSF (P<0.01), GM-CSF (P<0.01), MCP-1 (P<0.05), MIP-1α (P<0.01) and TGF-β1 (P<0.01) which are key molecules during the onset of pregnancy. Conclusion These results suggested that a critical level of Atp6v0a2 is required for the fertile spermatozoa and its decreased level in spermatozoa could be used to predict male infertility. This study provides a possibility that Atp6v0a2 could be potentially used as a diagnostic marker for the evaluation of male infertility. PMID:23936208

  1. Assessment of semen quality in Swamp Buffalo AI Bulls in Thailand

    Directory of Open Access Journals (Sweden)

    S. Koonjaenak

    2010-02-01

    Full Text Available Characteristic of Thai swamp buffalo bulls semen used for artificial insemination (AI in Thailand, aspects relevance in freezing and thawing of semen are review. Semen and sperm characteristics were evaluated included sperm count, motility (assessed subjectively and by CASA, morphology (using phase-contrast light microscopy and SEM, plasma membrane integrity (PMI (using a hypo-osmotic swelling test [HOST] and SYBR- 14/propidium iodide [PI], plasma membrane stability (PMS (using Annexin-V/PI and deoxyribonucleic acid (DNA integrity (using SCSA and flow cytometry [FCM]. The average ejaculate volume was about 3.0–4.0 mL, with good viability (PMI measured by the HOST and motility (>65% and >70%, respectively. Sperm concentration ranged from 1.1 to 1.2 billion/mL, being also affected by bull age. Whereas semen quality (including sperm output, pH and initial sperm motility did not differ between the seasons. Few spermatozoa (<15%/ ejaculate had abnormal morphology with abnormalities resembling those in other bovidae. In FT semen, PMI (using SYBR-14/PI and PMS were highest in winter. Across seasons, ~50% of post-thaw spermatozoa depicted linear motility, a proportion that decreased to ~35% during incubation (38oC for 60 minutes, without marking any seasonal difference. The sperm DNA was hardly damaged (with <3% fragmentation, expressed as DNA fragmentation index [DFI], among seasons.

  2. Testicle size as indicator of fertility in bulls

    Directory of Open Access Journals (Sweden)

    Prka Igor

    2014-01-01

    Full Text Available Male calves from the high value parents, bull fathers and bull dams, enter the selection for artificial insemination. After laboratory tests, the calves are taken to the center for artificial insemination, and after a stay in quarantine the are moved to a test station. At the age of twelve months they are measured for assessing the value of each calf exterior. One of the measures recorded was the testicle scope. On the basis of testicle size, it is possible to predict sperm production potential. For the determination of testicle size (testicular biometry, tapes or rulers were used. The aim of this work was to investigate the possible effect of testicle size on sperm production in young bulls used for artificial insemination. For that purpose there were used the data on circumference of testicles of one year old bulls just starting production of sperm, and then compared with certain semen quality parameters such as: volume of ejaculate and concentration and percentage of alive and progressively mobile spermatozoa. The investigation included all young bulls that started production in the period from 2010. to 2012., that is 36 bulls of various breeds (Simmental, Holstein Friesian, Montafon. After the testicle scope measuring in these bulls, there were observed the parameters of the sperm quality during the following one year period. The obtained results showed that the increased testicle size was followed by the increased average ejaculate quantity, in other words: 3.7 ml in group of bulls with testicle circumference below 30 cm, to 6.7 ml in bulls whose testicle circumference was over 40 cm. Also, the results showed that there was a correlation between the increased testicle size and the increased spermatozoa concentration. The values grow to testicle scope of 36 cm, and above that they were still high but with some oscillations. When it came to relation between testicle scope and the percentage of alive and progressively mobile spermatozoa, the

  3. Possibilities of using the European bison (Bison bonasus epididymal spermatozoa collected post-mortem for cryopreservation and artificial insemination: a pilot study

    Directory of Open Access Journals (Sweden)

    Dubiel Andrzej

    2011-03-01

    Full Text Available Abstract Background European bison is the largest mammal in Europe with the population of approximately 4000 individuals. However, there is no report of post-mortem spermatozoa collection and cryopreservation from this species and the aim of this study was to test if the epididymal spermatozoa collected post-mortem from European bison are suitable for cryopreservation and artificial insemination (AI. Methods Epididymides were collected post-mortem from two European bison bulls at age of 8 (bull 1 and 11 year (bull 2. Epididymal sperm was harvested by making multiple incisions in caudae epididymidis, which were then rinsed with extender. The left epididymis of bull 1 was rinsed with BioXcell (IMV, France, whereas the right epididymis of bull 1 and the right and left epididymides of bull 2 were rinsed with the extender based on Tris, citric acid, glucose, egg yolk, glycerol, antibiotics and distilled water (extender II. The diluted semen was cooled to 5 degrees C, and frozen in liquid nitrogen vapour. Then, properties of the frozen/thawed semen were examined with the use of computer-assisted semen analysis system, and thirty cows and nine heifers of domestic cattle were artificially inseminated. Results Motility of fresh spermatozoa collected from the right epididymis of bull 1 was 70% (spermatozoa diluted with extender II, and from the left one was 60% (spermatozoa diluted with BioXcell, whereas motility of fresh spermatozoa collected from bull 2 was 90% (spermatozoa diluted with extender II. Spermatozoa motility just after thawing were 11 and 13% in bull 1, respectively for spermatozoa collected from the left and right epididymis and 48% in bull 2. As a result of AI of domestic cows and heifers with the frozen/thawed European bison spermatozoa, two pregnancies were obtained in heifers. One pregnancy finished with a premature labour after 253 days of pregnancy, and the second one after 264 days of pregnancy. Conclusions This is the first report

  4. Relationship between bovine fertility and the number of spermatozoa penetrating the cervical mucus within straws.

    Science.gov (United States)

    Taş, Muzaffer; Bacinoglu, Suleyman; Cirit, Umüt; Ozdaş, Ozen Banu; Ak, Kemal

    2007-09-01

    In this study, by using a recently developed test technique, the relationship between the total spermatozoa number penetrating determined sites of bovine cervical mucus in straws and potential fertility of bulls, and other spermatological characteristics were investigated. Furthermore, we aimed to determine the effect on the test results, of two different incubation temperatures (37 and 41 degrees C) and two sperm penetration distance ranges (PDRs). Frozen semen samples of six Holstein bulls were used in the study. The bulls were divided into two fertility groups (high and low fertility) according to the "non-return rates" (NRR). For the penetration test, cervical mucus was drawn into transparent plastic straws and incubated with semen at 37 and 41 degrees C for 15 min. After the incubation, straws were frozen in liquid nitrogen vapour and stored at -20 degrees C. On the evaluation day, concentrations of spermatozoa penetrated to the PDRs, each of which was 2.5 mm, between 32.5 and 35 mm (first penetration distance range, PDR1), and 50 and 52.5 mm (second penetration distance range, PDR2) distance in the straws from the open end, were measured. When compared with the low fertility group, bulls from the high fertility group showed a higher number of spermatozoa at the determined PDRs, and a significant positive correlation was found between the total number of spermatozoa at the penetration distances and the NRR scores of the bulls.

  5. Comparative sperm protein profiling in bulls differing in fertility and identification of phosphatidylethanolamine-binding protein 4, a potential fertility marker.

    Science.gov (United States)

    Somashekar, L; Selvaraju, S; Parthipan, S; Patil, S K; Binsila, B K; Venkataswamy, M M; Karthik Bhat, S; Ravindra, J P

    2017-09-01

    This study aimed to identify sperm proteomic signatures regulating sperm functions and fertility by: (i) comparing the sperm electrophoretic protein profiles and identifying the differentially abundant proteins among breeding bulls differing in fertility status and (ii) elucidating the possible role of one of the identified novel proteins, PEBP4 on sperm function and fertility. The grouping of bulls as fertile (n = 6) and low fertile (n = 6) was performed based on bull fertility index and infertile (n = 6) based on semen rejection rate (>33%). The sperm motility, fructolysis index, acrosomal reaction, intracellular calcium levels, and seminal plasma fructose and calcium levels were studied among fertility groups. The differentially expressed sperm proteins observed in single- and two-dimensional gel electrophoresis (2DE) were identified using Nano-LC-MS/MS. In the fertile bulls, the expression levels of calmodulin (CALM1), spermadhesinZ13 (SPADH2), and phosphatidylethanolamine-binding protein 4 (PEBP4) were significantly (p sperm fructose uptake was observed. Further, PEBP4 was localized in elongated spermatids, Leydig cells, excurrent duct system, and principal piece of spermatozoa. These findings suggest a crucial role for the PEBP4 protein in spermiogenesis, epididymal sperm maturation, and sperm motility. This first study in bovine indicates the positive association of PEBP4 in regulating sperm maturation, functions, and fertility and could be a potential marker for predicting semen quality and fertility. © 2017 American Society of Andrology and European Academy of Andrology.

  6. DNA integrity in sexed bull sperm assessed by neutral Comet assay and sperm chromatin structure assay.

    Science.gov (United States)

    Boe-Hansen, Gry B; Morris, Ian D; Ersbøll, Annette K; Greve, Torben; Christensen, Preben

    2005-04-01

    During the production of sex-sorted spermatozoa from bull semen, the cells are exposed to a number of potential hazards including: dilution, centrifugation, incubation, exposure to DNA stains and laser light. These factors may affect the survival capacity and fertilization potential of the sperm. The objective of this study was to determine whether sex-sorted bull spermatozoa have more DNA damage than sperm from conventional processed bull semen. Two methods were used to determine DNA integrity: the neutral Comet assay (NCA) and the sperm chromatin structure assay (SCSA). The NCA showed that the conventional samples had a higher tail moment (TM) (P sperm and that cell sorting by flow cytometry improves the integrity of the sperm cell population. Additionally the results from the SCSA indicated that the sex-sorted sperm had less homogenous sperm chromatin. In the future assessment of sperm DNA integrity may be used to select bulls for sperm sex sorting and optimizing sperm sex sorting procedures.

  7. Expression, immunolocalization and processing of fertilins ADAM-1 and ADAM-2 in the boar (sus domesticus) spermatozoa during epididymal maturation

    Science.gov (United States)

    2011-01-01

    Fertilin alpha (ADAM-1) and beta (ADAM-2) are integral membrane proteins of the ADAM family that form a fertilin complex involved in key steps of the sperm-oocyte membrane interaction. In the present work, we analyzed the presence of ADAM-1 and ADAM-2 mRNAs, the spermatozoa proteins' processing and their sub-cellular localization in epididymal samples from adult boars. ADAM-1 and ADAM-2 mRNAs were highly produced in the testis, but also in the vas efferens and the epididymis. On immunoblots of sperm extracts, ADAM-1 subunit appeared as a main reactive band of ~50-55 kDa corresponding to occurrence of different isoforms throughout the epididymal duct, especially in the corpus region where isoforms ranged from acidic to basic pI. In contrast, ADAM-2 was detected as several bands of ~90 kDa, ~75 kDa, ~50-55 kDa and ~40 kDa. The intensity of high molecular mass bands decreased progressively in the distal corpus where lower bands were also transiently observed, and only the ~40 kDa was observed in the cauda. The presence of bands of different molecular weights likely results from a proteolytic processing occurring mainly in the testis for ADAM-1, and also throughout the caput epididymis for ADAM-2. Immunolocalization showed that fertilin migrates from the acrosomal region to the acrosomal ridge during the sperm transit from the distal corpus to the proximal cauda. This migration is accompanied by an important change in the extractability of a part of ADAM-1 from the sperm membrane. This suggests that the fertilin surface migration may be triggered by the biochemical changes induced by the epididymal post-translational processing of both ADAM1 and ADAM-2. Different patterns of fertilin immunolocalization then define several populations of spermatozoa in the cauda epididymis. Characterization of such fertilin complex maturation patterns is an important step to develop fertility markers based on epididymal maturation of surface membrane proteins in domestic mammals. PMID

  8. Splicing-related single nucleotide polymorphism of RAB, member of RAS oncogene family like 2B (RABL2B) jeopardises semen quality in Chinese Holstein bulls.

    Science.gov (United States)

    Wang, Xiuge; Cui, Xiaohui; Zhang, Yan; Hao, Haisheng; Ju, Zhihua; Liu, Deyu; Jiang, Qiang; Yang, Chunhong; Sun, Yan; Wang, Changfa; Huang, Jinming; Zhu, Huabin

    2017-11-01

    RAB, member of RAS oncogene family like 2B (RABL2B) is a member of a poorly characterised clade of the RAS GTPase superfamily, which plays an essential role in male fertility, sperm intraflagellar transport and tail assembly. In the present study, we identified a novel RABL2B splice variant in bovine testis and spermatozoa. This splice variant, designated RABL2B-TV, is characterised by exon 2 skipping. Moreover, a single nucleotide polymorphism (SNP), namely c.125G>A, was found within the exonic splicing enhancer (ESE) motif, indicating that the SNP caused the production of the RABL2B-TV aberrant splice variant. This was demonstrated by constructing a pSPL3 exon capturing vector with different genotypes and transfecting these vectors into murine Leydig tumour cell line (MLTC-1) cells. Expression of the RABL2B-TV transcript was lower in semen from high- versus low-performance bulls. Association analysis showed that sperm deformity rate was significantly lower in Chinese Holstein bulls with the GG or GA genotype than in bulls with the AA genotype (Pquality in bulls with different RABL2B genotypes is generated via an alternative splicing mechanism caused by a functional SNP within the ESE motif.

  9. Isolation of RNA striped bass Monrone saxatilis spermatozoa: Implications for teleost male fertility and beyond?

    Science.gov (United States)

    Vertebrate sperm has been shown to provide more than paternal genomic material to the oocyte. For example, specific transcripts have been identified in bull sperm associated with fertility and with motility in boar sperm. Very little is currently known about functional, residual RNA in spermatozoa a...

  10. Isolation of RNA from striped bass (Morone saxatilis) spermatozoa: implications for teleost male fertility and beyond?

    Science.gov (United States)

    Vertebrate sperm has been shown to provide more than paternal genomic material to the oocyte. For example, specific transcripts have been identified in bull sperm associated with fertility and with motility in boar sperm. Very little is currently known about functional, residual RNA in spermatozoa a...

  11. Proteomic comparison of detergent-extracted sperm proteins from bulls with different fertility indexes.

    Science.gov (United States)

    D'Amours, Olivier; Frenette, Gilles; Fortier, Marlène; Leclerc, Pierre; Sullivan, Robert

    2010-03-01

    Intrinsic factors such as proteins modulate the fertilising ability of male gametes. We compared detergent-extracted sperm protein composition of bulls with different fertility indexes in order to highlight putative fertility markers of sperm. Frozen semen from 23 Holstein bulls with documented fertility was used. According to their 'fertility solution' (SOL), as calculated by the Canadian dairy network, bulls were divided into four groups: high fertility (HF) (SOL>3.0; n=6), medium-HF (2.9>SOL>2.0; n=5), medium-low fertility (-2.8>SOL>-4.9; n=8) and low fertility (LF; SOL<-5.0; n=4), with a SOL=0 being the average. Triton X-100 protein extracts from ejaculated spermatozoa were subjected to two-dimensional difference gel electrophoresis, and polypeptide maps were quantitatively analysed by ImageMaster software. Nine protein spots showed significant differences between the HF and LF groups, and eight of these proteins were identified by liquid chromatography-tandem mass spectrometry. T-complex protein 1 subunits epsilon and (CCT5 and CCT8), two isoforms of epididymal sperm-binding protein E12 (ELSPBP1), proteasome subunit alpha type-6 and binder of sperm 1 (BSP1) were more expressed in the LF group than in the HF group. On the other hand, adenylate kinase isoenzyme 1 (AK1) and phosphatidylethanolamine-binding protein 1 (PEBP1) were more expressed in the HF group than in the LF group. The presence and expression level of ELSPBP1, BSP1, AK1 and PEBP1 were confirmed by western blot. A linear regression model established that CCT5 and AK1 explained 64% (P<0.001) of the fertility scores. The reported functions of these proteins are in agreement with a putative involvement in defective sperm physiology, where lower or higher levels can jeopardise sperm ability to reach and fertilise the oocyte.

  12. Toxic and photodynamic effects of toluidine blue on living bull spermatoza

    NARCIS (Netherlands)

    Duijn, C. van

    1. 1. The effects of vital staining with toluidine blue on the total number of bull spermatozoa moving normally and on their mean velocity and velocity frequency distributions have been investigated with Rikmenspoel's photoelectric method. 2. 2. A significant decrease of the mean half-life period

  13. Monitoring rumen environment in finishing Lidia bulls

    Directory of Open Access Journals (Sweden)

    Juan García G

    2016-05-01

    Full Text Available Objective. The aim of this work was to characterize the changes in rumen pH and temperature in finishing Lidia breed bulls reared on pasture and fed a total mixed ration (TMR. Materials and methods. Five 4-year-old Lidia bulls received approximately 10 kg of the TMR per animal and day in the morning. Bulls could move freely in a 17-ha fenced area and express normally their feeding behaviour. Internal wireless boluses were used to collect pH and temperature values every 10 minutes throughout the measurement period. Results. Average daily pH was 6.2. Average and maximum daily temperatures were not high enough to be indicative of disease (infections of other pathologies. Conclusions. When rations and feeding systems are appropriately managed, Lidia bulls can be supplemented with concentrates in the finishing stages of their productive cycle without impairing the rumen environment.

  14. Evaluation of post-acrosomal sheath WW domain binding protein expression in spermatozoa from infertile men with varicocele

    Directory of Open Access Journals (Sweden)

    Nasim Ghazavi-Khorasgani

    2017-09-01

    Conclusion: Expression of PAWP as a protein involved in spermatogenesis and fertilization process, has decreased in infertile men with varicocele. In addition, sperm DNA integrity was disrupted in these individuals that can be considered as a main etiology of infertility.

  15. Sexing mammalian spermatozoa and embryos--state of the art.

    Science.gov (United States)

    Seidel, G E

    1999-01-01

    Methods for sexing preimplantation embryos range from karyotyping to recording speed of development in vitro. The only method used routinely on a commercial scale is to biopsy embryos and amplify Y-chromosome-specific DNA using the polymerase chain reaction. This method is effective for more than 90% of embryos and is > 95% accurate. Within males, spermatozoa are essentially identical phenotypically due to: (1) connection of spermatogenic cells by intercellular bridges, (2) transcriptional inactivation of sex chromosomes during meiosis and spermiogenesis, (3) severe limitation of all gene expression during the later stages of spermiogenesis, and (4) coating all spermatozoa with common macromolecules during and after spermiogenesis. One consequence is that no convincing phenotypic difference has been detected between X- and Y-chromosome-bearing spermatozoa. The only consistently successful, nondestructive approach to sexing spermatozoa is to quantify DNA in spermatozoa using a fluorescing DNA-binding dye followed by flow cytometry and cell sorting. X-chromosome-bearing ruminant spermatozoa have about 4% more DNA compared with Y-chromosome-bearing spermatozoa; accuracy of sorting can exceed 90% routinely, and sorting rates currently exceed 10(3) live spermatozoa of each sex chromosome composition s-1. Hundreds of apparently normal offspring from a number of species have been produced from sexed semen, some via intrauterine artificial insemination.

  16. SPERM HY-LITER™ for the identification of spermatozoa from sexual assault evidence

    DEFF Research Database (Denmark)

    Westring, Christian Gustav; Wiuf, Morten; Nielsen, S Jock

    2014-01-01

    Accurate microscopic identification of human spermatozoa is important in sexual assault cases. We have compared the results of examinations with (1) a fluorescent microscopy method, SPERM HY-LITER™, and (2) Baecchi's method for identification of human spermatozoa. In 35 artificial, forensic type......-LITER™ detected spermatozoa when Baecchi's method did not (ts=6.567, df=1, P=0.048). This trend was also seen in selected compromised or degraded samples and in selected adjudicative samples. The reactions with spermatozoa from dog, horse, pig and bull were negative with SPERM HY-LITER™, whereas Baecchi's method...... was non-selective. Data from forensic casework samples in Copenhagen from two years (2008 and 2009) are presented. The samples from 2008 were investigated using Baecchi's method, while those from 2009 were investigated using SPERM HY-LITER™. The frequencies of positive results were similar between the two...

  17. EFFECT OF REDUCING SPERM NUMBERS PER INSEMINATION DOSE ON FERTILITY OF CRYOPRESERVED BUFFALO BULL SEMEN

    Directory of Open Access Journals (Sweden)

    S. M. H. ANDRABI, M. SIDDIQUE1, N. ULLAH AND L. A. KHAN

    2006-01-01

    Full Text Available The objective of this study was to evaluate the effect of reducing sperm numbers per insemination dose on fertility of cryopreserved buffalo bull semen. For this purpose, semen was collected at weekly intervals from a Nili-Ravi buffalo bull (Bubalus bubalis using an artificial vagina in two batches. The ejaculates were split-sampled and diluted at 37°C with tris-citric acid extender having 15x106 or 30x106 motile spermatozoa/0.5 ml. After dilution, the semen was cooled to 4C, equilibrated for 4 hours, packaged in 0.5 ml straws and frozen in programmable cell freezer. Fertility test based on 75-days first service pregnancy rate was determined under field conditions. A total of 500 buffaloes were inseminated with frozen semen and out of these 431 could be followed, 209 for semen straws packaged with 15x106 spermatozoa/straw and 222 for doses filled with 30x106 spermatozoa/straw. The inseminations were performed in two batches and each batch was spread over a period of three months. The fertility rate for sperm concentration of 15x106 spermatozoa/0.5 ml vs. 30x106 spermatozoa/0.5 ml (49.28 vs. 56.75% was similar (P>0.05. The fertility rates were also similar (P>0.05 in the first and second batch of inseminations performed with 15x106 or 30x106 spermatozoa/0.5 ml straw of cryopreserved semen. In conclusion, reduction of sperm number from 30x106 to 15x106 spermatozoa/0.5 ml dose of insemination did not affect fertility of cryopreserved buffalo bull semen.

  18. Identification of suitable combinations of in vitro sperm-function test for the prediction of fertility in buffalo bull.

    Science.gov (United States)

    Singh, Raushan K; Kumaresan, A; Chhillar, Shivani; Rajak, Shailendra K; Tripathi, Utkarsh K; Nayak, Samiksha; Datta, T K; Mohanty, T K; Malhotra, R

    2016-12-01

    The present study assessed sperm functional characteristics in the frozen-thawed semen of buffalo bulls and estimated their relationship with field fertility. Frozen semen samples from three different freezing operations each from nine Murrah buffalo bulls were used for the assessment of different sperm functions related to fertilizing potential. Bulls were classified into high (n = 2), medium (n = 5), and low (n = 2) fertile based on adjusted field fertility. The sperm functions estimated included membrane integrity using carboxyfluorescein diacetate-propidium iodide, acrosome reaction status using fluorescein isothiocyanate peanut agglutinine, status of apoptosis using Annexin-V, protamine deficiency using Chromomycin A3, membrane stability using Merocyanine 540 and lipid peroxidation status using 4, 4-difluoro-4-bora-3a, 4a-diaza-s-indacene. The relationship between the proportion of live acrosome-intact spermatozoa and fertility was positive and significant (r = 0.59; P = 0.001). The proportion of moribund spermatozoa showed a significantly negative correlation with fertility (r = -0.50; P = 0.008). Similarly, the relationship of spermatozoa with unstable membrane (r = -0.51; P = 0.007), necrotic (r = - 0.42; P = 0.028), early necrotic (r = -0.42; P = 0.031), and apoptotic spermatozoa (r = -0.39; P = 0.046) with bull fertility was negative and significant. The correlation between the protamine-deficient spermatozoa and fertility was negative, but not significant. Among different combinations of tests, live acrosome-intact spermatozoa and lipid peroxidation status of spermatozoa revealed high positive correlation with buffalo bull fertility (adjusted R(2) = 0.73, C[p] = 0.80). These preliminary findings may help in developing tools for assessing fertility of buffalo bulls, once validated in more animals. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Characterisation of haematological profiles and whole blood relative gene expression levels in Holstein-Friesian and Jersey bull calves undergoing gradual weaning

    National Research Council Canada - National Science Library

    Johnston, D; Kenny, D A; Kelly, A K; McCabe, M S; McGee, M; Waters, S M; Earley, B

    2016-01-01

    .... The objectives of the study were to examine (i) the effect of breed and plane of nutrition, on haematological profiles of artificially reared Holstein-Friesian and Jersey bull calves in response to gradual weaning, and (ii...

  20. Study on the reproductive capacity of bulls of the autochthonous Rhodope Shorthorn cattle breed

    Directory of Open Access Journals (Sweden)

    Radka Malinova

    2015-06-01

    Full Text Available The sperm production of bulls from the autochthonous Rhodope Shorthorn cattle breed was studied. The breed is among the smallest in Europe, the average weight of the cows ranging from 200 to 250 kg, and of the bulls from 330 to 370 kg. It was found that during the first 6 months from the start of exploitation, at the age of the bulls from 18 to 24 months, AI bulls had high reproductive capacity. The ejaculate volume was 1,74±0,09 ml in average (LS, the percentage of motile spermatozoa was 74,3±3,48% and the concentration 1268±13,1 x 106/ml. It was established that the bull had a significant impact on the reproductive performance, but the individual differences in the main characteristics were not high – motility 71,8-77,0%, concentration – 1222-1324 х 106/ml. The season also had a significant effect on the percentage of motile spermatozoa. Within the period from January to June, the highest reproductive capacity of the bulls was observed from February to May and the lowest in June.

  1. Advanced forensic validation for human spermatozoa identification using SPERM HY-LITER™ Express with quantitative image analysis.

    Science.gov (United States)

    Takamura, Ayari; Watanabe, Ken; Akutsu, Tomoko

    2017-07-01

    Identification of human semen is indispensable for the investigation of sexual assaults. Fluorescence staining methods using commercial kits, such as the series of SPERM HY-LITER™ kits, have been useful to detect human sperm via strong fluorescence. These kits have been examined from various forensic aspects. However, because of a lack of evaluation methods, these studies did not provide objective, or quantitative, descriptions of the results nor clear criteria for the decisions reached. In addition, the variety of validations was considerably limited. In this study, we conducted more advanced validations of SPERM HY-LITER™ Express using our established image analysis method. Use of this method enabled objective and specific identification of fluorescent sperm's spots and quantitative comparisons of the sperm detection performance under complex experimental conditions. For body fluid mixtures, we examined interference with the fluorescence staining from other body fluid components. Effects of sample decomposition were simulated in high humidity and high temperature conditions. Semen with quite low sperm concentrations, such as azoospermia and oligospermia samples, represented the most challenging cases in application of the kit. Finally, the tolerance of the kit against various acidic and basic environments was analyzed. The validations herein provide useful information for the practical applications of the SPERM HY-LITER™ Express kit, which were previously unobtainable. Moreover, the versatility of our image analysis method toward various complex cases was demonstrated.

  2. Bull Moose Tube Company

    Science.gov (United States)

    The EPA is providing notice of a proposed Administrative Penalty Assessment against the Bull Moose Tube Company, a business located at 1819 Clarkson Road, Chesterfield, MO, 63017, for alleged violations at the facility located at 406 East Industrial Drive,

  3. Testicular, semen and blood parameters in adapted and nonadapted Bos taurus bulls in the semi-arid tropics.

    Science.gov (United States)

    Wildeus, S; Hammond, A C

    1993-08-01

    Two experiments were conducted to evaluate differences in testicular, seminal and hematological characteristics in adapted (Senepol) and nonadapted (Holstein) Bos taurus bulls under the semi-arid environmental conditions of St. Croix, Virgin Islands (17 degrees N, 64 degrees W). In Experiment 1 mature, sexually-rested Senepol (n=10) and Holstein (n=9) bulls of similar age (37 months) and body weight (715 kg) and grazing on adjacent native pastures, were tested on the same day in July (28.8 degrees C mean ambient temperature, 81.5% humidity). Senepol bulls had lower (PSenepol compared to Holstein bulls. Ejaculates, obtained by electroejaculation, contained 3.2x10(9) more spermatozoa with fewer abnormal tails and detached acrosomes in Senepol than in Holstein bulls (PSenepol (n=42) and Holstein (n=30) bulls, representing 3 beef and 5 dairy farms, were evaluated during the summer (August/September) and winter (February/March). Again, scrotal circumference was larger (PSenepol than in Holstein bulls, with no effect of season. Seminal fructose was higher (PSenepol than in Holstein bulls and decreased (PSenepol bulls. The data point to differences between the adapted and nonadapted breed type in testicular and ejaculate characteristics, but also suggest that season has only a limited impact on bull reproductive function under the environmental conditions in St. Croix.

  4. Proteomic identification of cryostress in epididymal spermatozoa

    Directory of Open Access Journals (Sweden)

    Sung Jae Yoon

    2016-11-01

    Full Text Available Abstract Background Cryopreservation of epididymal spermatozoa is important in cases in which it is not possible to collect semen using normal methods, as the sudden death of an animal or a catastrophic injury. However, the freezing and thawing processes cause stress to spermatozoa, including cold shock, osmotic damage, and ice crystal formation, thereby reducing sperm quality. We assessed the motility (%, motion kinematics, capacitation status, and viability of spermatozoa using computer-assisted sperm analysis and Hoechst 33258/chlortetracycline fluorescence staining. Moreover, we identified proteins associated with cryostress using a proteomic approach and performed western blotting to validate two-dimensional electrophoresis (2-DE results using two commercial antibodies. Results Cryopreservation reduced viability (%, motility (%, straight-line velocity (VSL, average path velocity (VAP, amplitude of lateral head displacement (ALH, and capacitated spermatozoa, whereas straightness (STR and the acrosome reaction increased after cryopreservation (P 3 fold, P < 0.05 before and after cryopreservation. The proteins differentially expressed following cryopreservation are putatively related to several signaling pathways, including the ephrinR-actin pathway, the ROS metabolism pathway, actin cytoskeleton assembly, actin cytoskeleton regulation, and the guanylate cyclase pathway. Conclusion The results of the current study provide information on epididymal sperm proteome dynamics and possible protein markers of cryo-stress during cryopreservation. This information will further the basic understanding of cryopreservation and aid future studies aiming to identify the mechanism of cryostress responses.

  5. Kinetics data from bovine sex-specific embryo development from three different bulls

    Directory of Open Access Journals (Sweden)

    C.S. Oliveira

    2016-06-01

    Full Text Available Here we present kinetics data from bovine sex-specific embryo development. Embryos were originated using sex-sorted semen from three different Nelore bulls, and semen from the same batch was used for X-and Y-chromosome spermatozoa sorting. Data was obtained for six time points (24, 48, 96, 120, and 144 h.p.i.. Analyses for each bull׳s embryos (1, 2 and 3 is presented for female and male groups separately. Also, grouped data analysis, considering bull and sex interaction, is shown. For further interpretation and discussion, see "Cell death is involved in sexual dimorphism during preimplantation development" (Oliveira et al., 2015 [1].

  6. A COMPARATIVE STUDY WITH MURRAH BUFFALO AND INDIGENOUS GIR SPERMATOZOA TO HYPO-OSMOTIC SWELLING TEST

    Directory of Open Access Journals (Sweden)

    Sumit Chowdhury

    2013-12-01

    Full Text Available Hypo-Osmotic Swelling Test (HOST is a simple but authentic test to measure the integrity of sperm membrane. The present study was undertaken to evaluate the HOST value in Murrah buffalo and indigenous Gir bull in 150 mosm/l tri-sodium citrate and D-fructose HOST solution. Murrah buffalo was having more HOST (49±0.39 % reacted spermatozoa than indigenous Gir (42±0.57 % bulls suggesting lesser membrane damage during cryo-preservation and higher fertility rate in Murrah buffalo than Gir cattle.

  7. DNA fragmentation in spermatozoa

    DEFF Research Database (Denmark)

    Rex, A S; Aagaard, J.; Fedder, J

    2017-01-01

    Sperm DNA Fragmentation has been extensively studied for more than a decade. In the 1940s the uniqueness of the spermatozoa protein complex which stabilizes the DNA was discovered. In the fifties and sixties, the association between unstable chromatin structure and subfertility was investigated....... In the seventies, the impact of induced DNA damage was investigated. In the 1980s the concept of sperm DNA fragmentation as related to infertility was introduced as well as the first DNA fragmentation test: the Sperm Chromatin Structure Assay (SCSA). The terminal deoxynucleotidyl transferase nick end labelling...... (TUNEL) test followed by others was introduced in the nineties. The association between DNA fragmentation in spermatozoa and pregnancy loss has been extensively investigated spurring the need for a therapeutic tool for these patients. This gave rise to an increased interest in the aetiology of DNA damage...

  8. Mitochondria in teleost spermatozoa.

    Science.gov (United States)

    Ulloa-Rodríguez, Patricio; Figueroa, Elías; Díaz, Rommy; Lee-Estevez, Manuel; Short, Stefania; Farías, Jorge G

    2017-05-01

    There is an extraordinary diversity of reproductive modes in teleost and this variability is related to the phylogenetic relationships and adaption to very different biotopes. As in all vertebrates, sperm is produced as the end product of the process of spermatogenesis, and regarding teleost the spermatozoa lack an acrosome in almost all species and motility is activated as a response to osmolarity and ion content of the aquatic medium where the sperm is released. In this context, mitochondria possess a fundamental role for fish spermatozoa motility and integrity, hence, fertilizing potential; they are the energy supplier that allows flagellar movement and their dysfunction could play a main role in structural and functional damage to the spermatozoa. The ATP production through oxidative phosphorylation provides not only energy for cell activities, which includes Na+/K+ ATPase pump, endocytosis, protein synthesis and many other cell processes; but also produces reactive oxygen species, that under mitochondrial dysfunction causes oxidative stress. The assessment of mitochondrial function (e.g. through measurement of mitochondrial membrane potential) as well as ATP content (mostly supplied by mitochondrial respiration) can be useful as quality markers of fish spermatozoa. Also quantification of ROS and antioxidant status, strongly influenced by mitochondria, are used as complementary measurements. There is much information about sperm mitochondria and their function but studies of these aspects on fish reproduction are still required for applications in aquaculture. The real role of fish sperm mitochondria under short and long term storage and in vitro manipulation is not fully understood yet. Thus future research should focus on these matters. Copyright © 2017 Elsevier B.V. and Mitochondria Research Society. All rights reserved.

  9. [Mobilization of Ca2+ from intracellular stores of spermatozoa of Bos taurus depending on their functional status].

    Science.gov (United States)

    Denisenko, V Yu; Boytseva, E N; Kuzmina, T I

    2015-01-01

    On the basis of inhibitory analysis by using a fluorescence probe chlortetracycline, calcium transduction pathway in spermatozoa of Bos taurus has been examined. Additional release of Ca2+ from intracellular stores of sperm was found after combiened action of prolactin and GTP, which took place under influence of protein kinase C inhibitor (compound Ro 31-8220); the combined effect of theophylline and GDP also stimulated additional release of Ca2+ from intracellular stores, which was missing when adding inhibitor of protein kinase A, compound H-89. Using chlortetracycline test (analysis localization of chlortetracycline fluorescence in spermatozoa), we have shown the combined action of prolactin and GTP increases the number of sperm with acrosome reaction, which is reduced after influence of Ro 31-8220; the combined effect of theophylline and GDP increases the percentage of capacitated spermatozoa, which was decreased in the presence of H-89. According with the data obtained, we propose the hypothesis that the transduction of Ca2+ between intracellular stores in bull spermatozoa stimulated by the combined action of prolactin and GTP, or theophylline and GDP, determines the functional status of the spermatozoa. Namely: the transduction of Ca2+ between intracellular stores in bull spermatozoa stimulated by the combined action of prolactin and GTP is involved in the regulation of acrosomal processes, while sperm capacitation is mediated by the transduction of calcium between intracellular stores activated by the combined influence of theophylline and GDP.

  10. Tolerance of spermatozoa to hypotonic stress: role of membrane fluidity and correlation with cryosurvival.

    Science.gov (United States)

    Oldenhof, Harriëtte; Heutelbeck, Anna; Blässe, Anne-Kathrin; Bollwein, Heinrich; Martinsson, Gunilla; Wolkers, Willem F; Sieme, Harald

    2015-01-01

    The aim of this study was to evaluate inter-individual variability in osmotic properties of stallion spermatozoa and its correlation with cryosurvival. In addition, temperature dependency of hypo-osmotic tolerance and membrane fluidity were studied. Stallion sperm membranes exhibited good resistance towards hypotonic stress in the 15-30 °C temperature range, whereas membrane stability was found to be decreased at 4 and 37 °C. Bull spermatozoa showed greater hypo-osmotic tolerance compared with stallion spermatozoa, especially at temperatures above 30 °C, which coincided with decreased membrane fluidity of bovine spermatozoa in this temperature range. The critical osmolality at 22 °C, at which half of the sperm population survived exposure to hypotonic saline solution, was found to vary between 55 and 170 mOsm kg(-1) among different stallions. Clear correlations were found for pre- versus post-freeze sperm motility and membrane integrity. Pre-freeze percentages of membrane-intact spermatozoa after exposure to hypotonic stress showed a weak correlation with sperm motility after cryopreservation. This correlation, however, was not found when data were corrected for initial numbers of membrane-intact spermatozoa in the sample. We thus conclude that studies on pre-freeze tolerance towards hypotonic stress cannot be used to predict sperm cryosurvival rates for individual stallions.

  11. Reversal of motility loss in bongo antelope (Tragelaphus eurycerus isaaci) spermatozoa contaminated with hyposmotic urine during electroejaculation.

    Science.gov (United States)

    Wirtu, Gemechu; Pope, C Earle; MacLean, Robert A; Godke, Robert A; Paccamonti, Dale L; Dresser, Betsy L

    2008-01-30

    Semen collected by a combination of ampullary (rectal) massage and electroejaculation of a bongo bull was incidentally contaminated with urine (1:3.7). At 1.5h post-collection, progressive motility was 0% but some spermatozoa had intermittently twitching tails. Subsequent dilution with media and processing improved the progressive motility (up to 50%) and intact membranes (up to 71%) of spermatozoa. After thawing, the respective values were 35 and 70%. The osmolarity and pH of the contaminated supernatant was 151 mOsm and 7.45, respectively. Initial progressive motility in a non-contaminated portion of semen collected during the same procedure was 80%, and, after thawing, 60 and 90%, of the spermatozoa showed progressive motility and intact membranes, respectively. In conclusion, urine-contaminated bongo spermatozoa can regain progressive motility after dilution with isosmotic solutions and survive cryopreservation.

  12. STIMULATING AND PROTECTIVE EFFECTS OF VITAMIN E ON BOVINE SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    Eva Tvrdá

    2013-02-01

    Full Text Available As spermatozoa are particularly sensitive towards oxidative damage, the search for a potential antioxidant substance to preserve and protect them under in vitro conditions has recently attracted the attention of the scientific community. The aim of this study was to assess the dose- and time-dependent in vitro effects of vitamin E on bovine spermatozoa during three different time periods (Time 0 h, 12 h and 24 h. Semen samples were collected from 15 adult breeding bulls, and diluted in physiological saline solution containing 0.5% ethanol (96% together with 0, 10, 50, 100, 500 and 1000 µM of vitamin E. Spermatozoa motility was determined using the Sperm VisionTM and CASA (Computer Assisted Semen Analyzer system. Cell viability was measured using the metabolic activity MTT assay, the nitroblue-tetrazolium (NBT test was used to assess the intracellular superoxide formation. Spermatozoa motility differences between the control and groups A and B were the only one significant (P<0.05 and P<0.001, respectively at 0 h, however, increased motility parameters were observed in all experimental groups after 12 h, with significant differences (P<0.001 after 24 h. The MTT assay indicated that none of the vitamin E concentrations had a negative or cytotoxic effect on the spermatozoa mitochondrial activity and furthermore showed a significantly (P<0.001 improved cell viability in groups B, C and D at 24 h. The NBT test showed that the addition of 500, 100 and 50 µM of vitamin E had an instant positive effect on the spermatozoa protection against free radical production. This protection remained present with a significant impact at 12 h (P<0.001 and P<0.05, respectively. Furthermore, all vitamin E concentrations exhibited significant (P<0.05; P<0.001 protective effects on the spermatozoa free radical formation. The results indicate that the addition of vitamin E, especially in concentrations of 500 µM to 50 µM to the culture medium could be beneficial for

  13. Hydrodynamics of insect spermatozoa

    Science.gov (United States)

    Pak, On Shun; Lauga, Eric

    2010-11-01

    Microorganism motility plays important roles in many biological processes including reproduction. Many microorganisms propel themselves by propagating traveling waves along their flagella. Depending on the species, propagation of planar waves (e.g. Ceratium) and helical waves (e.g. Trichomonas) were observed in eukaryotic flagellar motion, and hydrodynamic models for both were proposed in the past. However, the motility of insect spermatozoa remains largely unexplored. An interesting morphological feature of such cells, first observed in Tenebrio molitor and Bacillus rossius, is the double helical deformation pattern along the flagella, which is characterized by the presence of two superimposed helical flagellar waves (one with a large amplitude and low frequency, and the other with a small amplitude and high frequency). Here we present the first hydrodynamic investigation of the locomotion of insect spermatozoa. The swimming kinematics, trajectories and hydrodynamic efficiency of the swimmer are computed based on the prescribed double helical deformation pattern. We then compare our theoretical predictions with experimental measurements, and explore the dependence of the swimming performance on the geometric and dynamical parameters.

  14. Exploration of Biological Markers of Feed Efficiency in Young Bulls.

    Science.gov (United States)

    Meale, Sarah J; Morgavi, Diego P; Cassar-Malek, Isabelle; Andueza, Donato; Ortigues-Marty, Isabelle; Robins, Richard J; Schiphorst, Anne-Marie; Laverroux, Sophie; Graulet, Benoit; Boudra, Hamid; Cantalapiedra-Hijar, Gonzalo

    2017-11-15

    The efficiency with which ruminants convert feed to desirable products is difficult to measure under normal commercial settings. We explored the use of potential biological markers from easily obtainable samples, that is, blood, hair, and feces, to characterize potential causes of divergent efficiency when considered as residual feed intake (RFI) or feed conversion efficiency (FCE). A total of 54 Charolais bulls, 20 in period 1 and 34 in period 2, were examined for individual dry matter intake (DMI) and growth. Bulls were offered a diet of 70:30 wrapped grass silage to concentrate for 99 d. At the conclusion of the test period, blood samples were collected for the determination of vitamins B2 and B6, and plasma used for the determination of metabolites, natural isotopic 15N abundance (15N NIA, expressed as δ15N ‰) and fractionation (Δ15Nplasma proteins-diet and Δ13Cplasma proteins-diet) and near-infrared spectroscopy (NIRS). Feces were analyzed by NIRS. Bulls were slaughtered at 15-17 months of age and carcass characteristics determined. Bulls were ranked according to RFI with extremes (SD ± 0.5; n = 31) classified as either efficient (Neg-RFI) or inefficient (Pos-RFI). Extreme bulls were then classified for FCE (high vs low FCE), changing the groups. Pos-RFI bulls consumed 14% more feed than Neg-RFI bulls for the same level of weight gain. Low FCE bulls tended to eat more, but had lower weight gains than high FCE bulls. No differences were detected in carcass conformation, fat scores, hot carcass weight, or dressing percentage. Yet, heart and bladder weights were heavier in Pos-RFI, and rumen weight tended to be heavier in Pos-RFI bulls. RFI did not affect bulk 15N or 13C fractionation. A negative correlation was observed between FCE and Δ15Nplasma proteins-diet. Inefficient bulls (Pos-RFI) had higher δ15N in glycine compared to Neg-RFI bulls. Similarly, metabolomic analysis showed a tendency for concentrations of glycine and sarcosine to be elevated in

  15. Evaluation of chromatin integrity of motile bovine spermatozoa capacitated in vitro.

    Science.gov (United States)

    Reckova, Z; Machatkova, M; Rybar, R; Horakova, J; Hulinska, P; Machal, L

    2008-08-01

    The efficiency of in vitro embryo production is highly variable amongst individual sires in cattle. To eliminate that this variability is not caused by sperm chromatin damage caused by separation or capacitacion, chromatin integrity was evaluated. Seventeen of AI bulls with good NRRs but variable embryo production efficiency were used. For each bull, motile spermatozoa were separated on a Percoll gradient, resuspended in IVF-TALP medium and capacitated with or incubated without heparin for 6 h. Samples before and after separation and after 3-h and 6-h capacitacion or incubation were evaluated by the Sperm Chromatin Structure Assay (SCSA) and the proportion of sperm with intact chromatin structure was calculated. Based on changes in the non-DFI-sperm proportion, the sires were categorized as DNA-unstable (DNA-us), DNA-stable (DNA-s) and DNA-most stable (DNA-ms) bulls (n=3, n=5 and n=9, respectively). In DNA-us bulls, separation produced a significant increase of the mean non-DFI-sperm proportion (p Capacitacion produced a significant decrease in the mean non-DFI-sperm proportion in H+ sperm (p capacitacion, the mean non-DFI-sperm proportion remained almost unchanged. In DNA-ms bulls, neither separation nor capacitacion had any effect on the mean non-DFI-sperm proportion. It can be concluded that, although separation and capacitacion may produce some changes in sperm chromatin integrity, these are not associated with different in vitro fertility of the bulls involved.

  16. Post Testicular Sperm Maturational Changes in the Bull: Important Role of the Epididymosomes and Prostasomes

    Directory of Open Access Journals (Sweden)

    Julieta Caballero

    2011-01-01

    Full Text Available After spermatogenesis, testicular spermatozoa are not able to fertilize an oocyte, they must undergo sequential maturational processes. Part of these essential processes occurs during the transit of the spermatozoa through the male reproductive tract. Since the sperm become silent in terms of translation and transcription at the testicular level, all the maturational changes that take place on them are dependent on the interaction of spermatozoa with epididymal and accessory gland fluids. During the last decades, reproductive biotechnologies applied to bovine species have advanced significantly. The knowledge of the bull reproductive physiology is really important for the improvement of these techniques and the development of new ones. This paper focuses on the importance of the sperm interaction with the male reproductive fluids to acquire the fertilizing ability, with special attention to the role of the membranous vesicles present in those fluids and the recent mechanisms of protein acquisition during sperm maturation.

  17. Growth, puberty, and carcass characteristics of Brahman-, Senepol-, and Tuli-sired F1 Angus bulls.

    Science.gov (United States)

    Chase, C C; Chenoweth, P J; Larsen, R E; Hammond, A C; Olson, T A; West, R L; Johnson, D D

    2001-08-01

    Postweaning growth, sexual development, libido, and carcass data were collected from two consecutive calf crops using 31 Brahman x Angus (B x A), 41 Senepol x Angus (S x A), and 38 Tuli x Angus (T x A) F1 bulls. Following weaning (by mid-September) and preconditioning, at the start of the study (late September) bulls were fed concentrate (three times each week at a rate equivalent to 4.5 kg/d) on bahiagrass pasture for approximately 250 d. At the start of the study and at 28-d intervals, BW, hip height, and scrotal circumference (SC) were measured. Concurrently at 28-d intervals, when the SC of a bull was > or = 23 cm, semen collection was attempted using electroejaculation. Ejaculates were evaluated for presence of first spermatozoa (FS), 50 x 10(6) sperm with at least 10% motility (PU), and 500 x 10(6) sperm with at least 50% motility (PP). After all bulls reached PP they were subjected to two libido tests. Carcass data were collected on all bulls (n = 110) and Warner-Bratzler shear (WBS) force values were assessed on a subset (n = 80). For both years, B x A bulls were heavier (P 0.10) gain in BW or hip height during the study. Scrotal circumference of T x A bulls was larger (P 0.10) of breed type by the end of the study. At PU and PP, B x A bulls were older (P 0.10) USDA quality grade. In conclusion, tropically adapted F1 bulls produced from Senepol (Bos taurus) and Tuli (Sanga) sires bred to Angus cows in Florida had lighter BW, shorter hip heights, and smaller carcasses than those from Brahman sires but reached puberty earlier and had higher libido scores and lower WBS force values.

  18. 29 CFR 1918.84 - Bulling cargo.

    Science.gov (United States)

    2010-07-01

    ... 29 Labor 7 2010-07-01 2010-07-01 false Bulling cargo. 1918.84 Section 1918.84 Labor Regulations...) SAFETY AND HEALTH REGULATIONS FOR LONGSHORING Handling Cargo § 1918.84 Bulling cargo. (a) Bulling cargo shall be done with the bull line led directly from the heel block. However, bulling may be done from the...

  19. Prediction of bull fertility.

    Science.gov (United States)

    Utt, Matthew D

    2016-06-01

    Prediction of male fertility is an often sought-after endeavor for many species of domestic animals. This review will primarily focus on providing some examples of dependent and independent variables to stimulate thought about the approach and methodology of identifying the most appropriate of those variables to predict bull (bovine) fertility. Although the list of variables will continue to grow with advancements in science, the principles behind making predictions will likely not change significantly. The basic principle of prediction requires identifying a dependent variable that is an estimate of fertility and an independent variable or variables that may be useful in predicting the fertility estimate. Fertility estimates vary in which parts of the process leading to conception that they infer about and the amount of variation that influences the estimate and the uncertainty thereof. The list of potential independent variables can be divided into competence of sperm based on their performance in bioassays or direct measurement of sperm attributes. A good prediction will use a sample population of bulls that is representative of the population to which an inference will be made. Both dependent and independent variables should have a dynamic range in their values. Careful selection of independent variables includes reasonable measurement repeatability and minimal correlation among variables. Proper estimation and having an appreciation of the degree of uncertainty of dependent and independent variables are crucial for using predictions to make decisions regarding bull fertility. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Nutritional composition analysis of meat from human lactoferrin transgenic bulls.

    Science.gov (United States)

    Zhao, Jie; Xu, Jianxiang; Wang, Jianwu; Li, Ning

    2013-01-01

    Transgenic technology has many potential advantages in food production. However, the transgenic technology process may influence the composition of food products derived from genetically engineered (GE) animals, which may be adverse to human health. Therefore, it is very important to research the compositions of GE animal products. Here, we analyzed the compositions of meat from the offspring of human lactoferrin (hLF) transgenic cows, which can express human lactoferrin proteins in their mammary gland. Six hLF transgenic bulls and three wide-type (WT) bulls, 10 months of age, were slaughtered for meat composition analysis. To determine the comparative health of hLF bulls for meat analysis, hematological analyses, organ/body weight analyses and pathology analyses were conducted. Results of the meat analysis show that there were no significant differences in the hematological parameters, organ/body weight ratios of hLF and WT bulls (P>0.05), and histopathological examination of the main organs of hLF bulls revealed no abnormalities. Nutrient parameters of meat compositions of hLF and WT bulls did not show any significant differences (P>0.05). All of these results suggest that the hLF transgene did not have an impact on the meat nutrient compositions of hLF bulls.

  1. [Effect of the E-50 Biostimulant on sperm production in young bulls].

    Science.gov (United States)

    Elezov, G; Marinkov, T; Krŭstev, M

    1986-01-01

    The Biostimulant E-50 preparation, a biogenic stumulant, was applied to 8 bulls divided into two groups--with normal and with disturbed semen production (lower qualitative indices of semen and higher number of discarded ejaculates). The stimulant was injected muscularly at the rate of 40 cu. cm, twice, at a seven-day interval, with one of the bulls it being used in a second series. Studied were a total of 587 ejaculates taken from the test bulls following treatment. It was found that the agent produced a favourable effect on semen production, the reaction being individual and better manifested with bulls having disturbances. Up to 13.7 per cent of the ejaculates of bulls with normal semen production were discarded, and up to 31.8 per cent of those showing signs of disturbed spermiogenesis. The spermogram indices with some bulls reliably rose (P less than 0.01 and 0.05) in terms of concentration and motility, the number of dead spermatozoa coming back to normal at negligible rise of the pH value.

  2. Correlation between hypoosmotic swelling test and breeding soundness evaluation of adult Nelore bulls

    Directory of Open Access Journals (Sweden)

    Tamires Miranda Neto

    2011-07-01

    Full Text Available This study aimed at evaluating the relationship between physical and morphological semen features with the hypoosmotic swelling (HOS test in raw semen of adult Nelore bulls classified as sound and unsound for breeding. Two hundred and six Nelore bulls aging from 3-10 years old were subjected to breeding soundness examination. After physical and morphological semen examination, HOS test was done. After the breeding soundness examination, 94.2% of the bulls were classified as sound for breeding. There was no difference between the average scrotal circumference of bulls classified as sound and unsound for breeding (P>0.05, but there was difference between all semen physical and morphological aspects of bulls classified as sound and unsound for breeding (P>0.05, but there was no difference in the mean percentage of reactive spermatozoa to HOS test results both for sound (38.4±17.9 and unsound animals (39.5±16.4; P>0.05, with no Pearson correlation between the HOS test and variables. According to these results HOS test can not be used alone to predict the reproductive potential of adult Nelore bulls.

  3. Proteomic Markers of Functional Sperm Population in Bovines: Comparison of Low- and High-Density Spermatozoa Following Cryopreservation.

    Science.gov (United States)

    D'Amours, Olivier; Frenette, Gilles; Bourassa, Sylvie; Calvo, Ézéchiel; Blondin, Patrick; Sullivan, Robert

    2018-01-05

    Mammalian semen contains a heterogeneous population of sperm cells. This heterogeneity results from variability in the complex processes of cell differentiation in the testis, biochemical modifications undergone by spermatozoa during transit along the male reproductive tract, interactions with secretions from accessory sex glands at ejaculation, and, in the context of reproductive technologies, in the ability of ejaculated spermatozoa to resist damage associated with freeze-thaw procedures. When submitted to density gradient centrifugation, ejaculated spermatozoa distribute themselves into two distinct populations: a low-density population characterized by low motility parameters, and a high-density population with high motility characteristics. To understand the origin of ejaculated spermatozoa heterogeneity, cryopreserved semen samples from bulls used by the artificial insemination (A.I.) industry were submitted to Percoll gradient centrifugation. Proteins from low and high density spermatozoa were then extracted with sodium deoxycholate and submitted to proteomic analysis using iTRAQ (isobaric tag for relative and absolute quantitation) methodologies. Quantification of selected sperm proteins was confirmed by multiple reaction monitoring (MRM). Overall, 31 different proteins were more abundant in low-density spermatozoa, while 80 different proteins were more abundant in the high-density subpopulation. Proteins enriched in high-density spermatozoa were markers of sperm functionality such as the glycolytic process, binding to the egg zona pellucida, and motility. Low-density spermatozoa were not solely characterized by loss of proteins and their associated functions. Chaperonin-containing TCP1s and chaperones are hallmarks of the low-density subpopulation. iTRAQ analysis revealed that other proteins such as binder of sperm proteins, histone, GPX5, ELSPBP1, and clusterin are overexpressed in low-density spermatozoa suggesting that these proteins represent defects

  4. ABCA17 mediates sterol efflux from mouse spermatozoa plasma membranes.

    Science.gov (United States)

    Morales, Carlos R; Ni, Xiaoyan; Smith, Charles E; Inagaki, Nobuya; Hermo, Louis

    2012-03-01

    Mammalian spermatozoa lose plasma membrane cholesterol during maturation in the epididymis and during capacitation in the female reproductive tract. While cholesterol acceptors such as high-density lipoproteins (HDL) and apolipoproteins A-I (apoA-I) and J (Apo J) have been found in male and female reproductive tracts, transporters that mediate cholesterol efflux from plasma membranes of spermatozoa to acceptors are not well defined. Candidates include members of the ATP-binding cassette (ABC) transporter superfamily including ABCA1, ABCA7, ABCA17, and ABCG1. In this study, we utilize immunocytochemistry on sections of adult mouse testis and epididymis and RT-PCR on isolated germ cells. The data reveal that ABCA17 is expressed by steps 12-16 elongated spermatids in the mouse in testis and by spermatozoa in the lumen of the epididymis where ABCA17 localizes to the sperm head and tail midpiece. It also localizes on these areas of mouse sperm isolated from the epididymis. Moreover, ABCA17 antibody interferes with cholesterol efflux from spermatozoa to lipid acceptors apoA-I. Taken together, these results suggest that ABCA17 plays an important role in the process of sterol efflux which renders spermatozoa capable of fertilizing an oocyte.

  5. Detection of spermatozoa following consensual sexual intercourse

    DEFF Research Database (Denmark)

    Astrup, Birgitte Schmidt; Thomsen, Jørgen Lange; Lauritsen, Jens

    2012-01-01

    examinations of sexual assault victims. For an objective evaluation of the findings, substantial knowledge of aspects regarding consensual sexual intercourse is crucial. The aim of this study was to examine detection frequencies and genital sampling sites of spermatozoa following consensual sexual intercourse...... that 88% of possible cases were positive for spermatozoa. The posterior fornix was significantly better than the other sites for detection of spermatozoa and the number of spermatozoa decreased significantly over time. In a large sub-group of women who reported that no intra-vaginal ejaculation had taken...... place during their latest intercourse, a significant number (14%) had spermatozoa in the vagina. CONCLUSION: Spermatozoa were best recovered from the posterior fornix, but spermatozoa were also present on swabs taken from the external genitalia. Detection of spermatozoa is thus possible in cases where...

  6. The Effect of Thawing Temperature on Sperm Quality of Friesian Holstein Bulls

    Directory of Open Access Journals (Sweden)

    Tri Utami

    2014-11-01

    Full Text Available The percentage of sperm motility and morphology are important criteria in evaluating the quality of sperm before it is used for artificial insemination (AI. This study was conducted to observe post thawing motility and abnormal morphology of spermatozoa Friesian Holstein (FH. The materials were used 10 straws of FH bulls in the form of 0.25 ml. A total of 10 straws then divided into two treatment groups of  thawing  in water at 37°C and water 8°C, respectively. An examination of the motility and morphology of spermatozoa abnormalities performed every two hours for two times. Calculating the percentage of sperm motility was done by calculating the percentage of spermatozoa moving forward in the field of view under a microscope with a magnification of 10x. the percentage of abnormal spermatozoa was  assessed by William's stain. Spermatozoa morphology was observed by using a microscope magnification of 100x. Abnormalities of spermatozoa were calculated from a total of 200 spermatozoa, either normal or abnormal. At the same thawing time, the motility of FH cattle sperms post thawing in water temperature 37°C had a higher preference than that of post thawing in water temperature 8°C, although it was not significantly different (P > 0.05.  Based on morphological aspects, frozen semen used in this study is within the tolerance limit for the total percentage of abnormal sperm morphology between 12% to 23% and normal morphology between 70% to 88%.

  7. Influence of selected factors on bovine spermatozoa cold shock resistance

    Directory of Open Access Journals (Sweden)

    Luděk Stádník

    2015-01-01

    Full Text Available The objectives of this study were to determine the effects of sire, extender, and addition of Low Density Lipoprotein (LDL to extenders used on the percentage rate of spermatozoa survival after cold shock. Two groups of extenders were compared: without LDL addition (control variants and LDL enriched (experimental variants. Three extenders were used: AndroMed®, Bioxcell®, and Triladyl®. Experimental variants included 4–8% LDL addition into the AndroMed® and Bioxcell® extenders, and 6–10% LDL addition into the Triladyl® extender. In total, 12 samples of fresh semen were collected from 4 bulls during a period of 8 weeks. Bovine spermatozoa cold shock resistance (1 ± 1 °C, 10 min was evaluated by the percentage rate of live sperm using eosin-nigrosine staining immediately and after heat incubation (37 ± 1 °C, 120 min. The results showed the effect of sire as important and individual differences between selected sires in their sperm resistance against cold shock were confirmed. AndroMed® and Bioxcell® were found to be providing better protection of bull semen to cold shock compared to Triladyl® due to lower decline of live sperm proportion. Our results detected a positive effect of LDL addition on sperm resistance against cold shock, especially on lower decrease of live sperm percentage rate after 120 min of the heat test (P < 0.05. Further studies are needed to assess the optimal concentration of LDL in various kinds of extenders as well to state ideal time and temperature conditions for ensuring LDL reaction with sperm.

  8. Lipsome-mediated uptake of exogenous DNA by Sahiwal cattle spermatozoa

    Directory of Open Access Journals (Sweden)

    Thomas V. Babu

    Full Text Available Aim: To investigate the influence of lipofection treatment and exogenous DNA uptake on the quality of sahiwal cattle spermatozoa. Materials and Methods: Semen collected from sahiwal bulls (n=7 were evaluated separately for color, volume, mass activity, concentration, motility and viability using standard procedures. Pooled sperm samples from selected bulls (n=3 were transfected with a model gene construct enhanced green fluorescent protein (p-EGFP via lipofection method and confirmed the genome integration by PCR technique. Furthermore the effect of transfection on spermatozoa was assessed based on apotosis, viability and motility. Results: In the current investigation sahiwal bulls were selected based on their breeding records and better semen characteristics. Although the transfected sperm samples failed to show florescence under fluorescence microscope, PCR studies confirmed the successful uptake of the p-EGFP gene in to the host sperm cell genome. Moreover transfected samples showed a significant reduction in the viability and motility without causing any DNA damage induced apoptosis as demonstrated by DNA Ladder assay. [Vet World 2012; 5(10.000: 621-627

  9. Quantification of Y-bearing spermatozoa of beef cattle throught ciscontinuous Percoll gradient centrifugation using fluorescent in situ hybridization

    Directory of Open Access Journals (Sweden)

    Ngernpradub, A.

    2007-09-01

    Full Text Available An appropriated procedure for proportion of Y-bearing spermatozoa is needed, especially to monitor sperm separation techniques. In this study was aimed to determine the Y-bearing spermatozoa proportion in bull semen through pre and post 6 layers discontinuous Percoll centrifugation and by using fluorescent in situ hybriddization (FISH technique with digoxigenin (Dig-labeled bovine Y-specific probe, which prepared by polymerase chain reaction (PCR using BC1.2 primers. This results fould that the metaphase male lymphocyte was clearly shown green-yellow fluorescence spot on the short arm of Y-chromosome, whereas an interphase male lymphocyte showed this signal within nuclei, in conversely, both interphase and metaphase female lymphocytes were not shown this signal. An average interphase male lymphocyte obviously showed the signal 96.8%. The proportion of Y-bearing spermatozoa in semen pre and post discontinuous Percoll centrifugation were 48.78% and 41.68%, respectively (P>0.05. These results indicated that FISH protocol with Dig-labeled bovine Y-specific probe could be used to mornitor the proportion of Y-bearing spermatozoa through discontinuous Percoll centrifugation in bull semen which after sperm separation, the proportion of Y-chromosome trend to be not significantly reduced.

  10. Different extenders in the cryopreservation of bovine epididymal spermatozoa.

    Science.gov (United States)

    Papa, Patrícia M; Papa, Frederico O; Oliveira, Letícia A; Guasti, Priscilla N; Castilho, Caliê; Giometti, Ines Cristina

    2015-10-01

    The objective of this study was to evaluate the effects of two different egg yolk extenders incubated with or without Sperm Talp on the motility and plasma membrane integrity of cryopreserved bovine epididymal spermatozoa after freezing. Twenty-five testicles with epididymides from mature bulls were collected at the abattoir. Epididymal sperm recovery was performed by retrograde flushing using a skim milk-extender (Botu-Semen™). After recovery, sperm were incubated either without or with Sperm Talp and then submitted to centrifugation. For the freezing process, half of the testes were processed with Tris egg yolk extender, and half were processed with Botu-Bov™ egg yolk extender. Samples incubated in Sperm Talp exhibited better results than epididymal spermatozoa that were incubated without Sperm Talp (psperm from the bovine epididymides if the sperm were previously incubated with Sperm Talp. The extenders examined in this work did not differ in their effect on plasma membrane integrity after freezing. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Effects of severe chronic scrotal Dermatophilus congolensis (kirchi) infection on semen characteristics in Zebu/Friesian crossbred bulls and effect of long-acting terramycin chemotherapy.

    Science.gov (United States)

    Sekoni, V O

    1993-07-01

    The semen characteristics of 12 Zebu/Friesian crossbred bulls, aged 2 to 3 years, were studied during a 21-month period. At the 12th month of the study, the commencement of the rainy season, the bulls were infected naturally with Dermatophilus congolensis . Lesions were scattered over the body and limbs, but were particularly pronounced on the scrotum. Monthly treatments with injection of terramycin were begun as soon as lesions were detected and continued until the end of the study. The lesions worsened and became pronounced particularly on the scrotum of all the bulls. Scrotal scab formation caused by infection became prominent at the 14th month of the study. Until that period, the bulls had normal semen characteristics. From the 15th month until the end of the study, there was progressive deterioration of semen characteristics in all the bulls; this was manifested by some or all of the following effects: decreased volume, increased percentage of dead spermatozoa, increased percentage of total sperm morphological abnormalities, decreased percentage of progressively motile spermatozoa, oligospermia and terminal azoospermia. Therefore, severe chronic scrotal dermatophilosis may be a significant cause of infertility or sterility in bulls.

  12. Breed and other effects on reproductive traits and breeding soundness categorization in young beef bulls in Florida.

    Science.gov (United States)

    Chenoweth, P J; Chase, C C; Thatcher, M J; Wilcox, C J; Larsen, R E

    1996-11-01

    Yearling, grass-fed, beef bulls at the USDA Subtropical Agricultural Research Station, Brooksville, Florida, were assessed for physical and semen traits in January, April, July and October of 1991 (Trial 1) and 1992 (Trial 2). Bulls were given a breeding soundness evaluation (BSE) using revised semen and scrotal circumference (SC) criteria. In Trial 1, the bulls consisted of Angus (n = 15), Brahman (n = 14), Hereford (n = 15) and Senepol (n = 14). In Trial 2, the breeds were Angus (n = 15), Brahman (n = 16), Romosinuano (n = 13) and Nellore x Brahman (n = 9). Trial bulls generally showed delayed growth compared with grain-fed bulls in temperate environments. Breed influenced semen traits (percentage sperm motility, normal spermatozoa and those with primary abnormalities) in both trials. Temperate Bos taurus breeds (Angus, Hereford) were generally superior to Bos indicus breeds (Brahman, Nellore x Brahman). Tropically-adapted Bos taurus breeds (Senepol, Romosinuano) were intermediate for those traits tested. In general, tropically-adapted Bos taurus breeds were more similar in reproductive development to temperate Bos taurus than to Bos indicus breeds. Breed by test period interactions occurred and were mainly influenced by delayed sexual maturity of Bos indicus bulls. Qualitative semen traits increased with bull age, particularly from 12 to 18 mo. Scrotal circumference development was slower in the Bos indicus breeds. Bulls of satisfactory BSE status at 18.1 to 22 mo of age were 73.9% in Trial 1 and 58.5% in Trial 2. Brahman bulls had the least satisfactory BSE scores in both years (Trial 1, 44.4%; Trial 2, 22.2%). Most bulls failed to achieve satisfactory BSE status due to a small SC relative to age (Trial 1, 66%; Trial 2, 72%). The most efficacious use of the BSE was > or = 15 mo in Bos taurus bulls and > 18 mo for Bos indicus bulls. Although the BSE has proven to be useful for the assessment of young, pasture-raised bulls in semi-tropical environments, use of SC

  13. RELATIONSHIP BETWEEN LEVEL OF COPPER IN BOVINE SEMINAL PLASMA AND SPERMATOZOA MOTILITY

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    Zuzana Kňažická

    2013-02-01

    Full Text Available The aim of this study was to evaluate relationship between copper (Cu concentration of bovine seminal plasma and spermatozoa motility. Semen samples were collected from 13 breeding bulls. The motility analysis was carried out using the Computer Assisted Sperm Analysis (CASA system. The mean value for the percentage of motile spermatozoa (MOT was 92.46±3.99% and the progressive motility of the spermatozoa (PROG as 90.23±4.02%. The seminal plasma Cu concentrations were analyzed by UV/VIS spectrophotometry. The total Cu concentration of the seminal plasma was 4.28±1.47 μM/L. The correlation analysis revealed a strong negative correlation between MOT and seminal plasma Cu concentration (rp=-0.781; P<0.01 as well as between PROG and Cu content in the seminal plasma (rp=-0.726; P<0.01. The data obtained from this study clearly indicated that concentration of copper in seminal plasma negatively affects the spermatozoa motility parameters and subsequently might cause reproductive alteration in male sexual functions.

  14. Effect of ejaculate, bull, and a double swim-up sperm processing method on sperm sex ratio.

    Science.gov (United States)

    Madrid-Bury, Ninoska; Fernández, Raúl; Jiménez, Adela; Pérez-Garnelo, Sonia; Moreira, Pedro Nuno; Pintado, Belén; de la Fuente, Julio; Gutiérrez-Adán, Alfonso

    2003-08-01

    Offspring gender preselection has applications of considerable economic, health and ecological interest. In this study we analysed modifications of the percentages of spermatozoa bearing Y and X chromosomes when semen samples are submitted to a double swim-up technique as a possible method for producing embryos of known sex with in vitro fertilisation protocols. As an initial experiment to provide accurate evaluation of the method we determined the possible incidence of natural deviations in the primary sex ratio between bulls or ejaculates, analysing the percentage of Y-chromosome DNA bearing spermatozoa (%Y-CDBS) with a polymerase chain reaction (PCR) amplification of X- and Y-specific fragments. Ejaculates were tested by direct semiquantitative PCR sexing and by sexing blastocysts produced in vitro with these spermatozoa. Bulls and ejaculates did not have any effect on the %Y-CDBS or on the sex ratio of embryos produced in vitro using these ejaculates. However, our double swim-up sperm preparation method produced differences in %Y-CDBS in some of the sperm fractions, suggesting that there are intrinsic differences in capacitation of X- and Y-bearing spermatozoa that might be used to produce embryos of the desired sex with in vitro fertilisation.

  15. Testicular Cell Indices and Peripheral Blood Testosterone Concentrations in Relation to Age and Semen Quality in Crossbred (Holstein Friesian×Tharparkar Bulls

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    S. K. Rajak

    2014-11-01

    Full Text Available Present study analyzed the changes in peripheral blood testosterone concentrations and testicular cytogram in relation to age and semen quality in crossbred males. Three different age groups of crossbred males viz. bull calves (6 months, n = 5, young bulls (15 months, n = 5 and adult bulls (4 to 6 years, n = 8 were utilized for the study. Testicular fine needle aspiration cytology technique was used to quantify testicular cytology and their indices. Peripheral blood testosterone concentrations were measured using enzyme-linked immunosorbent assay method. Semen samples collected from adult bulls were microscopically evaluated for quality parameters. Mean peripheral blood testosterone concentrations in bull calves, young bulls and adult bulls were 2.28±0.09 ng/mL, 1.42±0.22 ng/mL and 5.66±1.08 ng/mL respectively, and that in adult bulls were significantly different (p<0.01 from young bulls and bull calves. There was no significant difference between the proportion of different testicular cells in bull calves and young bulls. Between young and adult bulls, significant differences (p<0.01 were observed in the proportion of spermatocytes, spermatozoa, and sperm: Sertoli cell ratio. The proportions of Sertoli cells showed a significant difference (p<0.01 between the three age groups. The number of primary spermatocytes had a positive correlation with peripheral blood testosterone concentrations in bull calves (r = 0.719, p<0.01. Number of Sertoli cells per 100 germ cells was negatively correlated with blood testosterone concentration in young bulls (r = −0.713, p<0.01. Among different semen parameters in adult bulls, ejaculate volume (r = 0.790, p<0.05 had positive relationship, and sperm motility had significant negative correlation (r = −0.711, p<0.05 with testosterone concentrations. The number of Sertoli cells and Sertoli cell index had a positive correlation with various semen quality parameters (p<0.001. Results of the present study

  16. Danish holsteins favor bull offspring

    DEFF Research Database (Denmark)

    Græsbøll, Kaare; Kirkeby, Carsten; Nielsen, Søren Saxmose

    2015-01-01

    In a previous study from 2014 it was found that US Holstein cows that gave birth to heifer calves produced more milk than cows having bull calves. We wanted to assess whether this is also true for Danish cattle. Data from 578 Danish Holstein herds were analysed with a mixed effect model and contr......In a previous study from 2014 it was found that US Holstein cows that gave birth to heifer calves produced more milk than cows having bull calves. We wanted to assess whether this is also true for Danish cattle. Data from 578 Danish Holstein herds were analysed with a mixed effect model...... and contrary to the findings in the US, we found that cows produced higher volumes of milk if they had a bull calf compared to a heifer calf. We found a significantly higher milk production of 0.28% in the first lactation period for cows giving birth to a bull calf, compared to a heifer calf. This difference...... was even higher when cows gave birth to another bull calf, so having two bull calves resulted in a difference of 0.52% in milk production compared to any other combination of sex of the offspring. Furthermore, we found that farmer assisted calvings were associated with a higher milk yield. Cows...

  17. In-depth proteomic analysis of carp (Cyprinus carpio L) spermatozoa.

    Science.gov (United States)

    Dietrich, Mariola A; Arnold, Georg J; Fröhlich, Thomas; Ciereszko, Andrzej

    2014-12-01

    Using a combination of protein fractionation by one-dimensional gel electrophoresis and high performance liquid chromatography-electrospray ionization tandem mass spectrometry, we identified 348 proteins in carp spermatozoa, most of which were for the first time identified in fish. Dynein, tubulin, HSP90, HSP70, HSP60, adenosylhomocysteinase, NKEF-B, brain type creatine kinase, mitochondrial ATP synthase, and valosin containing enzyme represent high abundance proteins in carp spermatozoa. These proteins are functionally related to sperm motility and energy production as well as the protection of sperm against oxidative injury and stress. Moreover, carp spermatozoa are equipped with functionally diverse proteins involved in signal transduction, transcription, translation, protein turnover and transport. About 15% of proteins from carp spermatozoa identified here were also detected in seminal plasma which may be a result of leakage from spermatozoa into seminal plasma, adsorption of seminal plasma proteins on spermatozoa surface, and expression in both spermatozoa and cells secreting seminal plasma proteins. The availability of a catalog of carp sperm proteins provides substantial advances for an understanding of sperm function and for future development of molecular diagnostic tests of carp sperm quality, the evaluation of which is currently limited to certain parameters such as sperm count, morphology and motility or viability. The mass spectrometry data are available at ProteomeXchange with the dataset identifier PXD000877 (DOI: http://dx.doi.org/10.6019/PXD000877). Copyright © 2014 Elsevier Inc. All rights reserved.

  18. Influence of mitochondrial membrane potential of spermatozoa on in vitro fertilisation outcome.

    Science.gov (United States)

    Marchetti, P; Ballot, C; Jouy, N; Thomas, P; Marchetti, C

    2012-04-01

    To determine whether the outcome of in vitro fertilisation (IVF) is influenced by the percentage of spermatozoa with functional mitochondria, a total of 91 random couples undergoing IVF were included. Mitochondrial function was determined by flow cytometry and expressed as percentage of spermatozoa. Conventional sperm parameters were studied by light microscopy. Reproductive outcome parameters were fertilisation rate, embryo quality and clinical pregnancy. It was found that the fertilisation rate was correlated with the percentage of spermatozoa (r = 0.24, P = 0.01) as well as with the percentage of highly motile spermatozoa. However, we did not find any relationship between the percentage of spermatozoa and embryo quality. Nevertheless, no patient who exhibited less than 64% of spermatozoa achieved pregnancy. It is concluded that determination of Δψ(m) provides accurate information to guide physicians to identify male patients for whom IVF will be unlikely to result in pregnancy. Therefore, we suggest that the percentage of spermatozoa may contribute to identify the most appropriate treatment for an individual patient. © 2011 Blackwell Verlag GmbH.

  19. Simultaneous evaluation of plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential in bovine spermatozoa by flow cytometry.

    Science.gov (United States)

    Kanno, Chihiro; Kang, Sung-Sik; Kitade, Yasuyuki; Yanagawa, Yojiro; Takahashi, Yoshiyuki; Nagano, Masashi

    2016-08-01

    The present study aimed to develop an objective evaluation procedure to estimate the plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential of bull spermatozoa simultaneously by flow cytometry. Firstly, we used frozen-thawed semen mixed with 0, 25, 50, 75 or 100% dead spermatozoa. Semen was stained using three staining solutions: SYBR-14, propidium iodide (PI), and phycoerythrin-conjugated peanut agglutinin (PE-PNA), for the evaluation of plasma membrane integrity and acrosomal integrity. Then, characteristics evaluated by flow cytometry and by fluorescence microscopy were compared. Characteristics of spermatozoa (viability and acrosomal integrity) evaluated by flow cytometry and by fluorescence microscopy were found to be similar. Secondly, we attempted to evaluate the plasma membrane integrity, acrosomal integrity, and also mitochondrial membrane potential of spermatozoa by flow cytometry using conventional staining with three dyes (SYBR-14, PI, and PE-PNA) combined with MitoTracker Deep Red (MTDR) staining (quadruple staining). The spermatozoon characteristics evaluated by flow cytometry using quadruple staining were then compared with those of staining using SYBR-14, PI, and PE-PNA and staining using SYBR-14 and MTDR. There were no significant differences in all characteristics (viability, acrosomal integrity, and mitochondrial membrane potential) evaluated by quadruple staining and the other procedures. In conclusion, quadruple staining using SYBR-14, PI, PE-PNA, and MTDR for flow cytometry can be used to evaluate the plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential of bovine spermatozoa simultaneously.

  20. Soft Computing for Human Spermatozoa Morphology

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    Jia Lu

    2005-06-01

    Full Text Available A fuzzy data fusion approach was used for human sperm morphology recognition analysis using 3-CCD camera image data. Fuzzy operators were used to create the relationship between abnormal sperm morphology and normal sperm aberrations after digitizing the images. The approaches have focused on frequencies of sperm with either abnormal morphology in semen analysis samples. In order to establish whether various shapes of membership functions, the authors would classify the morphology according to their head, tail, and neck shapes into symmetrical, asymmetrical, irregular and amorphous categories based on the fuzzy region. The images results demonstrated how these findings facilitated approaches on the 100 semen samples. The average probability of morphology recognition analysis was equal to 95% and the average probability of unknown parameter was equal to 4.5%. The fuzzy fusion morphology provided a unified and consistent framework to express different shapes of human spermatozoa.

  1. Hormonal induction of spermatozoa from amphibians with Rana temporaria and Bufo bufo as anuran models.

    Science.gov (United States)

    Uteshev, V K; Shishova, N V; Kaurova, S A; Browne, R K; Gakhova, E N

    2012-01-01

    The use of hormonally induced spermatozoa expressed in urine (HISu) is a valuable component of reproduction technologies for amphibians. Five protocols for sampling HISu from the European common frog (Rana temporaria) were compared: (1) pituitary extracts, (2) 0.12 µg g⁻¹ luteinising hormone-releasing hormone analogue (LHRHa), (3) 1.20 µg g⁻¹ LHRHa, (4) 11.7 IU g⁻¹ human chorionic gonadotrophin (hCG) and (5) 23.4 IU g⁻¹ hCG (g⁻¹ = per gram bodyweight). From 1 to 24h after administration we assessed the number and concentration of spermatozoa in spermic urine and in holding water, and in urine the percentage of motile spermatozoa and their progressive motility. The protocol using 1.20 µg g⁻¹ LHRHa gave the highest total sperm numbers (650 × 10⁶) and the highest percentage (40%) of samples with sperm concentrations above 200 × 10⁶ mL⁻¹. The percentage motility and progressive motility was similar from all protocols. Considerable amounts of spermatozoa were expressed by R. temporaria into their holding water. We tested hormonal priming and spermiation in the common toad (Bufo bufo) using 0.13 µg g⁻¹ LHRHa administered 24h before a final spermiating dose of 12.8 IU g⁻¹ hCG. No spermatozoa were expressed in holding water. Priming resulted in 35% more spermatozoa than without; however, there were no differences in sperm concentrations. Primed B. bufo produced spermatozoa with significantly higher percentage motility, but not progressive motility, membrane integrity, or abnormal spermatozoa than unprimed males.

  2. Detection of spermatozoa following consensual sexual intercourse.

    Science.gov (United States)

    Astrup, Birgitte Schmidt; Thomsen, Jørgen Lange; Lauritsen, Jens; Ravn, Pernille

    2012-09-10

    In cases of sexual assault, the finding of semen can provide crucial evidence. The presence of spermatozoa serves as proof of a sexual act and may give the identity of the alleged perpetrator through DNA-profiling. In most western countries, there are guidelines for standardized examinations of sexual assault victims. For an objective evaluation of the findings, substantial knowledge of aspects regarding consensual sexual intercourse is crucial. The aim of this study was to examine detection frequencies and genital sampling sites of spermatozoa following consensual sexual intercourse. In a prospective setting, 60 women underwent forensic examination following consensual sexual intercourse. Specimens were obtained from the external genitalia, the posterior fornix and the cervical canal, and examined using the Papanicolau stain and standard light microscopy. We found that 88% of possible cases were positive for spermatozoa. The posterior fornix was significantly better than the other sites for detection of spermatozoa and the number of spermatozoa decreased significantly over time. In a large sub-group of women who reported that no intra-vaginal ejaculation had taken place during their latest intercourse, a significant number (14%) had spermatozoa in the vagina. Spermatozoa were best recovered from the posterior fornix, but spermatozoa were also present on swabs taken from the external genitalia. Detection of spermatozoa is thus possible in cases where a speculum examination is denied. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  3. Effects of female bovine plasma collected at different days of the estrous cycle on epididymal spermatozoa motility.

    Science.gov (United States)

    Nait Mouloud, M; Ouennoughi, F; Yaiche, L; Kaidi, R; Iguer-Ouada, M

    2017-03-15

    The aim of this study was to assess the effects of female bovine plasma collected at different days of the reproductive cycle on epididymal spermatozoa motility and to test hypothesis that the subpopulations pattern of motile spermatozoa is affected by this treatment. Blood plasma samples were collected from five Holstein Friesian cows at different stages of the estrous cycle (days 0, 5, 10, 12 and 18), one pregnant cow and one adult bull and were diluted 1:9 (V/V) with normal saline. Female charcoal-treated plasma, Bull plasma and saline were used as controls. Semen samples were obtained from cauda epididymidis through retrograde flushing and diluted in saline to approximately 60 × 106 sperm/ml. The extended semen was diluted 1:2 (V/V) with tested media and motility was evaluated at 15 min and then every hour for 6 h using a computer-assisted semen analysis. Multivariate clustering procedure was applied to identify and quantify specific subpopulations within the semen samples. The statistical analysis clustered all the motile spermatozoa into three separate subpopulations with defined patterns of movement: Subpopulation 1 poorly motile and non-progressive spermatozoa (39.3%), subpopulation 2 including the fastest and the most vigorous spermatozoa (46.4%) and subpopulation 3 represented by slow, non-vigorous but linear spermatozoa (14.3%). Initially, sperm samples supplemented with female, male or female charcoal-treated plasma stimulated equally total motility and spermatozoa belonging to subpopulation 2 regardless of the estrous cycle stage. After 1-h incubation, the motility of these both categories of spermatozoa (total motile and those assigned to subpopulation 2) is enhanced and maintained more in day 12, 18 and pregnant cow plasma than in female plasma from earlier stage of the estrous cycle (day 0, 5 and 10), male plasma and female-charcoal treated plasma. In conclusion, the overall results showed that female plasma stimulated significantly sperm

  4. EFFECT OF ANTIBIOTICS IN EXTENDER ON FERTILITY OF LIQUID BUFFALO BULL SEMEN

    Directory of Open Access Journals (Sweden)

    S. AKHTER, M. SAJJAD, S. M. H. ANDRABI1, N. ULLAH1 AND M. QAYYUM

    2007-01-01

    Full Text Available This study was carried out to determine if a new antibiotic combination comprising of gentamycin, tylosin and linco-spectin (GTLS in extender is suitable for improvement in fertility of liquid buffalo bull semen through artificial insemination (AI. Two consecutive ejaculates per week (4 weeks were collected from three Nili-Ravi buffalo bulls of known fertility by using artificial vagina. The pooled ejaculates were split-sampled and diluted with skimmed milk extender (37oC; 10x106 motile spermatozoa/ml containing either SP (streptomycin 1000 μg/ml and penicillin 1000 iu/ml or GTLS (gentamycin 500 μg/ml, tylosin 100 μg/ml, lincomycin 300 μg/ml, and spectinomycin 600 μg/ml. Liquid semen was stored at 5°C for seven days. Fertility, based on 90-days first service pregnancy rate, was determined under field conditions. The fertility rates for SP-based vs. GTLS-containing liquid semen of buffalo bull were 58.55 and 60.00%, respectively, the difference was non significant. The fertility rates also did not differ (P>0.05 due to antibiotics at different days of storage of liquid semen at 5oC. In conclusion, GTLS, in skimmed milk extender compared to SP, did not significantly improve the fertility of chilled buffalo bull semen.

  5. Effect of glutamine supplementation and replacement of tris-egg yolk based extender with defatted cow milk on spermatozoa quality after equilibration and thawing

    Directory of Open Access Journals (Sweden)

    Vasundhara Dawra

    2015-08-01

    Full Text Available Aim: The present study was designed to evaluate the effect of supplementation of glutamine and replacement of Tris-egg yolk (TE based buffer with defatted cow milk on the spermatozoa quality after equilibration and thawing. Materials and Methods: Semen was collected from five Bhadawari bulls biweekly, and a total of 30 ejaculates were taken. The semen ejaculates were pooled and divided into three equal parts. The pooled semen was diluted by TE based extender (control, TE + glutamine (8 mM (T1 and 50% TE + 50% deffated cow milk + glutamine (8 mM (T2. At two stages viz. after equilibration and after 12 h of cryopreservation (thawed samples, progressive motility, percent live, and percent acrosomal damage of the spermatozoa was assessed. Results: Supplementation of glutamine improved (p<0.05 the spermatozoa quality with respect to the progressive motility, percent live and acrosomal damage both post-equilibration and post-thaw. T2 improved (p<0.05 the spermatozoa quality as compared to control, however; it was less (p<0.05 effective as compared T1 both post-equilibration and post-thaw. Conclusion: From the results of present study it can be concluded that glutamine supplementation was effective in maintaining post-equilibration and post-thaw spermatozoa quality whereas defatted cow milk was not as effective as TE based buffer in the extender in improving the spermatozoa quality.

  6. Altered chromatin condensation of heat-stressed spermatozoa perturbs the dynamics of DNA methylation reprogramming in the paternal genome after in vitro fertilisation in cattle.

    Science.gov (United States)

    Rahman, Mohammad Bozlur; Kamal, Md Mostofa; Rijsselaere, Tom; Vandaele, Leen; Shamsuddin, Mohammed; Van Soom, Ann

    2014-10-01

    Shortly after penetration of the oocyte, sperm DNA is actively demethylated, which is required for totipotent zygotic development. Aberrant DNA methylation is thought to be associated with altered chromatin condensation of spermatozoa. The objectives of this study were to investigate the dynamics of DNA methylation reprogramming in the paternal pronucleus and subsequent fertilisation potential of heat-stressed bull spermatozoa having altered chromatin condensation. Hence, bovine zygotes (n=1239) were collected at three different time points (12, 18 and 24h post insemination, hpi), and stained with an antibody against 5-methylcytosine. Fluorescence intensities of paternal and maternal pronuclei were measured by ImageJ. DNA methylation patterns in paternal pronuclei derived from heat-stressed spermatozoa did not differ between time points (P>0.05), whereas control zygotes clearly showed demethylation and de novo methylation at 18 and 24hpi, respectively. Moreover, heat-stressed spermatozoa showed a highly reduced (Pfertilisation rate compared with non-heat-stressed or normal control spermatozoa (53.7% vs 70.2% or 81.5%, respectively). Our data show that the normal pattern of active DNA demethylation followed by de novo methylation in the paternal pronucleus is perturbed when oocytes are fertilised with heat-stressed spermatozoa, which may be responsible for decreased fertilisation potential.

  7. Bulling among yearling feedlot steers.

    Science.gov (United States)

    Pierson, R E; Jensen, R; Braddy, P M; Horton, D P; Christie, R M

    1976-09-01

    In a survey to determine the cause of illness and deaths among yearling feedlot cattle, bulling was found to be one of the major problems. During the years 1971-1974, 54,913 (2.88%) steers became bullers and represented an annual loss of around +325,000. Some of the causes of bulling were found to be hormones, either as implants or in the feed. In 1974, from 1,988 necropsies, it was determined that 83 steers died from riding injuries.

  8. Freezability of water buffalo spermatozoa is improved with the addition of catalase in cryodiluent.

    Science.gov (United States)

    Ali, L; Hassan Andrabi, S M; Ahmed, H; Hussain Shah, A A

    Catalase enzyme is usually distributed in mammalian seminal plasma, where it decomposes hydrogen peroxide into water and oxygen and enhances sperm survivability. To evaluate the effect of catalase (0, 100, 200 or 300 IU/ml) added in tris-citric acid (TCA) based extender on motion characteristics, viability and DNA integrity of bubaline spermatozoa at post dilution (PD) and post thawing (PT) stages of cryopreservation. Collection of semen was done in four Nili-Ravi bulls with an artificial vagina (42 degree C). Qualified semen samples from each bull were further subdivided into four aliquots for dilution with the experimental TCA extender containing either 0.0 (T1), 100 IU (T2), 200 IU (T3) or 300 IU (T4) catalase (activity12660 U/mg). At PT, mean computer progressive motility, average path velocity, straight line velocity, curvilinear velocity, visual motility and DNA integrity were higher (P catalase fortified treatment groups as compared with control. Regarding plasma membrane integrity and supra-vital plasma membrane integrity, at PT the mean values were higher (P catalase at a concentration of 300IU/ml in TCA cryodiluent improved the freezability of water buffalo spermatozoa.

  9. Uncle Sam vs. John Bull.

    Science.gov (United States)

    Mellini, Peter

    1990-01-01

    Compares John Bull and Uncle Sam as iconographic symbols, respectively personifying male images of the British and United States national characters. Recounts their origins, evolutions, and representative values, and includes cartoons depicting the evolution. Describes female counterparts: Britannia and Columbia/Liberty. (CH)

  10. Identification of putative fertility markers in seminal plasma of crossbred bulls through differential proteomics.

    Science.gov (United States)

    Aslam, M K Muhammad; Kumaresan, A; Sharma, Vinay K; Tajmul, Md; Chhillar, Shivani; Chakravarty, A K; Manimaran, A; Mohanty, T K; Srinivasan, A; Yadav, Savita

    2014-12-01

    Sub-fertility is a major problem in crossbred bulls leading to disintegration of breeding systems and huge economic loss. Identification of some potential biomarkers to determine the latent fertility of bulls accurately has long been the interest of researchers. In this study, we analyzed the proteome of seminal plasma (SP) from bulls with varying fertility to identify the fertility-associated proteins. The proteomic profile of high- and low-fertile bulls was compared by two-dimensional difference gel electrophoresis and differentially expressed proteins were identified through matrix-assisted laser desorption/ionization-time of flight/mass spectrometry. Out of the 18 differentially expressed proteins (P bulls and 9 were overexpressed in SP of low-fertile bulls. The differential expressions ranged from 1.5- to 5.5-fold between the two groups, where protection of telomeres-1 protein (POT1) was highly overexpressed (2.9-fold) in high-fertile group and prostaglandin E2 receptor EP3 (PTGER3) was highly abundant (5.5-fold) in low-fertile group. The protein interaction network was elucidated using STRING software tool, and the functional bioinformatics analysis was done using Blast2Go software. Most of the differentially expressed proteins were found to be involved in cellular processes and biological regulation with binding and catalytic function. It is inferred that the expression of certain proteins in the SP varied with bull fertility, and concurrent appraisal of their expression along with other fertility assays may help in determining bull fertility. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. Computer assisted semen analysis for quantification of motion characteristics of bull sperm during cryopreservation cycle

    Directory of Open Access Journals (Sweden)

    M. N. Sundararaman

    Full Text Available Aim: The study was undertaken to quantify and to analyze the changes in the motion characteristics of bull spermatozoa during various stages of cryopreservation cycle. Materials and Methods: Using computer assisted semen analysis (CASA technique, 26 ejaculates, collected from two Jersey bulls were analyzed for motility, head behaviour and swimming pattern of spermatozoa on dilution, pre-freeze and post-thaw stages of cryopreservation. French straw technique was employed for deep-freezing of semen using liquid nitrogen. Results: Equilibration of diluted semen at 5 C has significantly (P< 0.01 reduced sperm motility, progressive motility, path velocity, and progressive velocity. Beat cross frequency was also affected significantly (P<0.05 by equilibration. Freezing and thawing processes drastically affected all the motility, velocity and head behaviour characteristics (P< 0.01. Conclusion: CASA facilitate objective evaluation sperm motion characteristics. Adoption of CASA technique has the potential for improvements in evaluation of semen thereby the quality of frozen semen for fertility can be enhanced. [Vet World 2012; 5(12.000: 723-726

  12. Conventional and fluorescent based semen quality assessment in Karan Fries bulls

    Directory of Open Access Journals (Sweden)

    A. Panmei

    2015-10-01

    Full Text Available Aim: The present study was carried out on semen ejaculates of 15 Karan Fries (KF bulls maintained at Artificial Breeding Research Centre, National Dairy Research Institute, Karnal, India with an objective to evaluate the relationship between the conventional and fluorescent based semen quality analysis of the bulls. Materials and Methods: A total of 96 ejaculates were collected from 15 KF (Holstein Friesian [HF] crossbred bulls. Semen were evaluated for color, volume, mass activity (MA and percentage of individual motility (IM, sperm concentration, percent live spermatozoa, hypo-osmotic swelling test and acrosome integrity, chromatin integrity, sperm viability, and membrane integrity. Data were analyzed using SPSS software package for descriptive analysis. The correlation between rankings of sires based on conventional and fluorescent semen parameters were calculated by Spearman’s rank correlation coefficient. Results: The average ejaculates volume (ml, sperm concentration (106/ml, MA, IM (%, live (%, morphological abnormalities (%, host (%, acrosome integrity (%, chromomycin A3 (CMA3 (%, SYBR-PI (%, and fluorescent isothiocyanate-peanut agglutinin (FITC-PNA (% were 4.57±0.36, 1162.98±97.93, 2.95±0.09, 60.8±1.22, 71.41±2.10, 9.31±1.15, 65.5±1.81, 86.6±1.59, 3.53±0.43, 65.39±2.23 and 74.47±2.53, respectively. Rank correlations were found to be significant for SYBR-PI and FITC-PNA with most of the parameters evaluated by conventional methods. Overall, among conventional criteria, IM revealed ranking of bulls almost similar to that of fluorescent criteria. Conclusion: Overview of our results indicated that, among conventional criteria, MA and IM revealed ranking of bulls almost similar to that of fluorescent criteria.

  13. Protective effect of crocetin on bovine spermatozoa against oxidative stress during in vitro fertilization.

    Science.gov (United States)

    Sapanidou, V; Taitzoglou, I; Tsakmakidis, I; Kourtzelis, I; Fletouris, D; Theodoridis, A; Lavrentiadou, S; Tsantarliotou, M

    2016-11-01

    Oxidative stress is one of the major factors that contribute to poor semen quality and low rates of in vitro fertilization. Crocetin, a main constituent of saffron (Crocus sativus L.) possesses potent antioxidant activity, by scavenging reactive oxygen species (ROS) and/or enhancing the activity of intracellular antioxidant enzymes. The aim of this study was to investigate, for the first time, the effect of crocetin on the quality characteristics of bull spermatozoa and fertilization rate. For this reason, frozen/thawed bovine spermatozoa were incubated with crocetin (1, 2.5, and 5 μm), for 120 or 240 min, in the presence of a negative control, and evaluated in terms of motility, viability, acrosomal status, DNA fragmentation index, intracellular ROS, and lipid peroxidation. In order to evaluate the impact of crocetin on cleavage and blastocyst rate, the compound was added in the IVF medium at the previously identified optimal concentration (2.5 μm). The results indicate that incubation of spermatozoa with 2.5 μm of crocetin resulted in a statistically significant lower production of superoxide anion and hydrogen peroxide, lower lipid peroxidation, and in better maintenance of motility parameters, viability, and acrosomal integrity, with a very small number of cells with DNA fragmentation, compared to the other groups (p fertilization medium also resulted in a significant increase in acrosome-reacted spermatozoa and blastocyst production, compared to the control group (p fertilizing ability, directly and/or indirectly, by regulating ROS concentration and lipid peroxidation. © 2016 American Society of Andrology and European Academy of Andrology.

  14. Effects of GnRH vaccination in wild and captive African Elephant bulls (Loxodonta africana on reproductive organs and semen quality.

    Directory of Open Access Journals (Sweden)

    Imke Lueders

    Full Text Available Although the African elephant (Loxodonta africana is classified as endangered by the International Union for Conservation of Nature (IUCN, in some isolated habitats in southern Africa, contraception is of major interest due to local overpopulation. GnRH vaccination has been promoted as a non-invasive contraceptive measure for population management of overabundant wildlife. We tested the efficacy of this treatment for fertility control in elephant bulls.In total, 17 male African elephants that were treated with a GnRH vaccine were examined in two groups. In the prospective study group 1 (n = 11 bulls, ages: 8-36 years, semen quality, the testes, seminal vesicles, ampullae and prostate, which were all measured by means of transrectal ultrasound, and faecal androgen metabolite concentrations were monitored over a three-year period. Each bull in the prospective study received 5 ml of Improvac® (1000 μg GnRH conjugate intramuscularly after the first examination, followed by a booster six weeks later and thereafter every 5-7 months. In a retrospective study group (group 2, n = 6, ages: 19-33 years, one examination was performed on bulls which had been treated with GnRH vaccine for 5-11 years.In all bulls of group 1, testicular and accessory sex gland sizes decreased significantly after the third vaccination. In six males examined prior to vaccination and again after more than five vaccinations, the testis size was reduced by 57.5%. Mean testicular height and length decreased from 13.3 ± 2.6 cm x 15.2 ± 2.8 cm at the beginning to 7.6 ± 2.1 cm x 10.2 ± 1.8 cm at the end of the study. Post pubertal bulls (>9 years, n = 6 examined prior to vaccination produced ejaculates with viable spermatozoa (volume: 8-175 ml, sperm concentration: 410-4000x106/ml, total motility: 0-90%, while after 5-8 injections, only 50% of these bulls produced ejaculates with a small number of immotile spermatozoa. The ejaculates of group 2 bulls (vaccinated >8 times were

  15. Estimation of the potential fertility based upon non-return rates of bulls: using polyacrylamide gel instead of cervical mucus in the sperm penetration test.

    Science.gov (United States)

    Taş, M; Bacinoglu, S; Cirit, U; Ozgümüş, S; Kaşgöz, H; Pabuccuoğlu, S

    2007-10-15

    In the present study, we aimed to develop a polyacrylamide gel that could be used instead of bovine cervical mucus in the cervical mucus penetration test (CMPT) to obtain coherent and replicable results in bulls. The frozen semen samples of six Holstein bulls, which were divided into two fertility groups as low and high according to their non-return rate (NRR), were used. In this study, the modified CMPT (mCMPT) was carried out within 0.25 mL transparent plastic straws with an inner diameter 1.7 mm. The penetration ability of spermatozoa to bovine cervical mucus and to polyacrylamide gels swollen with two different solutions [NaCl (G1) and PBS (G2)] was compared. For the penetration test, the straws filled with cervical mucus and both gels were dipped into thawed semen samples and incubated at 37 degrees C for 15 min. After the incubation, straws were frozen in liquid nitrogen vapour and stored at -20 degrees C. On the evaluation day, the frozen straws were cut at 1.5-1.75 cm (penetration distance range=PDR1), 3.25-3.5 cm (PDR2) and 5.0-5.25 cm (PDR3), beginning from open-end of the straws. The separated frozen parts were then immediately transferred onto special counting slides by pushing with a mandrel and left to thaw. Thawed samples were covered with cover glass and penetrated spermatozoa in these parts were counted. The relation between the results and fertility of bulls was determined. In the tests performed using mucus, the number of spermatozoa determined in the high fertility group was found to be higher at PDR3 (p<0.0001) compared to the low fertility group, while in G1 spermatozoa number was significantly higher at PDR1 and PDR3 (p<0.0001). However, in G2 medium, no significant difference was observed between either of the fertility groups with respect to spermatozoa number determined at all distance ranges. In the study, we have determined that the gel swollen with NaCl produces better results and this gel can be used instead of bovine cervical mucus

  16. Sperm in poor quality semen from bulls during heat stress have a lower affinity for binding hydrogen-3 heparin

    Energy Technology Data Exchange (ETDEWEB)

    Ax, R.L.; Gilbert, G.R.; Shook, G.E.

    1987-01-01

    Binding assays with (/sup 3/H) heparin were performed using spermatozoa collected prior to, during, and following summer heat stress to dairy bulls. Ejaculates collected in August 1983 after a period of ambient temperatures exceeding 29.4/sup 0/C exhibited a high frequency of abnormal sperm, and motility was reduced in some samples. Sperm in samples collected during heat stress possessed dissociation constants for binding (/sup 3/H) heparin ranging from 134.5 to 163.2 nmol. In contrast, sperm in semen collected prior to and after heat stress had significantly lower dissociation constants (higher affinity) for (/sup 3/H)heparin, 12.9 to 56.4 nmol. The number of binding sites for (/sup 3/H) heparin on sperm did not change among collection periods. It was concluded that the binding affinity for (/sup 3/H) heparin may reflect membrane integrity of bull sperm.

  17. Effect of semen preparation on casa motility results in cryopreserved bull spermatozoa.

    Science.gov (United States)

    Contri, Alberto; Valorz, Claudio; Faustini, Massimo; Wegher, Laura; Carluccio, Augusto

    2010-08-01

    Computer-assisted sperm analyzers (CASA) have become the standard tool for evaluating sperm motility and kinetic patterns because they provide objective data for thousands of sperm tracks. However, these devices are not ready-to-use and standardization of analytical practices is a fundamental requirement. In this study, we evaluated the effects of some settings, such as frame rate and frames per field, chamber and time of analysis, and samples preparations, including thawing temperature, sperm sample concentration, and media used for dilution, on the kinetic results of bovine frozen-thawed semen using a CASA. In Experiment 1, the frame rate (30-60 frame/s) significantly affected motility parameters, whereas the number of frames per field (30 or 45) did not seem to affect sperm kinetics. In Experiment 2, the thawing protocol affects sperm motility and kinetic parameters. Sperm sample concentration significantly limited the opportunity to perform the analysis and the kinetic results. A concentration of 100 and 50 x 10(6) sperm/mL limited the device's ability to perform the analysis or gave wrong results, whereas 5, 10, 20, and 30 x 10(6) sperm/mL concentrations allowed the analysis to be performed, but with different results (Experiment 3). The medium used for the dilution of the sample, which is fundamental for a correct sperm head detection, affects sperm motility results (Experiment 4). In this study, Makler and Leja chambers were used to perform the semen analysis with CASA devices. The chamber used significantly affected motility results (Experiment 5). The time between chamber loading and analysis affected sperm velocities, regardless of chamber used. Based on results recorded in this study, we propose that the CASA evaluation of motility of bovine frozen-thawed semen using Hamilton-Thorne IVOS 12.3 should be performed using a frame rate of 60 frame/s and 30 frames per field. Semen should be diluted at least at 20 x 10(6) sperm/mL using PBS. Furthermore, it is necessary to consider the type of chamber used and perform the analysis within 1 or 2 min, regardless of the chamber used. Copyright 2010 Elsevier Inc. All rights reserved.

  18. Cryopreservation of bull semen is associated with carbonylation of sperm proteins.

    Science.gov (United States)

    Mostek, Agnieszka; Dietrich, Mariola Aleksandra; Słowińska, Mariola; Ciereszko, Andrzej

    2017-04-01

    Artificial insemination with cryopreserved semen enables affordable, large-scale dissemination of gametes with superior genetics. However, cryopreservation can cause functional and structural damage to spermatozoa that is associated with reactive oxygen species (ROS) production, impairment of sperm motility and decreased fertilizing potential, but little attention has been paid to protein changes. The goal of this study was to investigate the oxidative modifications (measured as carbonylation level changes) of bull spermatozoa proteins triggered by the cryopreservation process. Flow cytometry and computer-assisted sperm analysis were used to evaluate changes in viability, ROS level and motility of spermatozoa. Western blotting, in conjunction with two-dimensional electrophoresis (2D-oxyblot) and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight spectrometry, was employed to identify and quantify the specifically carbonylated spermatozoa proteins. Cryopreservation decreased motility and viability but increased the number of ROS-positive cells. We identified 11 proteins (ropporin-1, outer dense fiber protein 2, glutathione S-transferase, triosephosphate isomerase, capping protein beta 3 isoform, actin-related protein M1, actin-related protein T2, NADH dehydrogenase, isocitrate dehydrogenase, cilia- and flagella-associated protein 161, phosphatidylethanolamine-binding protein 4) showing differences in protein carbonylation in response to cryopreservation. The identified proteins are associated with cytoskeleton and flagella organization, detoxification and energy metabolism. Moreover, almost all of the identified carbonylated proteins are involved in capacitation. Our results indicate for the first time that cryopreservation induces oxidation of selected sperm proteins via carbonylation. We suggest that carbonylation of sperm proteins could be a direct result of oxidative stress and potentially lead to disturbances of capacitation

  19. Non-genomic effects of vitamin D in human spermatozoa

    DEFF Research Database (Denmark)

    Blomberg Jensen, Martin; Dissing, Steen

    2012-01-01

    The spectrum for vitamin D (VD) mediated effects has expanded in recent years. Activated VD (1,25(OH)(2)D(3)) binds to the VD receptor (VDR) and mediates non-genomic effects through the alternative ligand binding-pocket (VDR-ap) or regulates gene transcription through the genomic binding...... specificity, and CYP24A1 expression is positively correlated with all semen variables and suggested as a marker for both semen quality and VD responsiveness. Moreover, spermatozoa are transcriptionally silent and are therefore a unique model to study non-genomic effects. 1,25(OH)(2)D(3) induced a rapid...

  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

    2017-06-01

    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 < .05) in the 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 < .05). Progressive motility tended to be increased in the 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. © 2017 Blackwell Verlag GmbH.

  1. Integrity of the plasma membrane, the acrosomal membrane, and the mitochondrial membrane potential of sperm in Nelore bulls from puberty to sexual maturity

    Directory of Open Access Journals (Sweden)

    L.S.L.S. Reis

    2016-06-01

    Full Text Available ABSTRACT This study evaluated the plasma membrane integrity, acrosomal membrane integrity, and mitochondrial membrane potential of Nelore bull sperm from early puberty to early sexual maturity and their associations with sperm motility and vigor, the mass motility of the spermatozoa (wave motion, scrotal circumference, and testosterone. Sixty Nelore bulls aged 18 to 19 months were divided into four lots (n=15 bulls/lot and evaluated over 280 days. Semen samples, collected every 56 days by electroejaculation, were evaluated soon after collection for motility, vigor and wave motion under an optical microscope. Sperm membrane integrity, acrosomal integrity, and mitochondrial activity were evaluated under a fluorescent microscope using probe association (FITC-PSA, PI, JC-1, H342. The sperm were classified into eight integrity categories depending on whether they exhibited intact or damaged membranes, an intact or damaged acrosomal membrane, and high or low mitochondrial potential. The results show that bulls have a low amount of sperm with intact membranes at puberty, and the sperm show low motility, vigor, and wave motion; however, in bulls at early sexual maturity, the integrity of the sperm membrane increased significantly. The rate of sperm membrane damage was negatively correlated with motility, vigor, wave motion, and testosterone in the bulls, and a positive correlation existed between sperm plasma membrane integrity and scrotal circumference. The integrity of the acrosomal membrane was not influenced by puberty. During puberty and into early sexual maturity, bulls show low sperm mitochondrial potential, but when bulls reached sexual maturity, high membrane integrity with high mitochondrial potential was evident.

  2. Entosis Acts as a Novel Way within Sertoli Cells to Eliminate Spermatozoa in Seminiferous Tubule

    Directory of Open Access Journals (Sweden)

    Nisar Ahmed

    2017-05-01

    Full Text Available The present study was designed to investigate the hypothesis that in vivo entosis is a novel pathway for eliminating spermatozoa in the seminiferous tubules (ST during hibernation of the Chinese soft-shelled turtle. Western blot analysis revealed that the expression of LAMP1 in the testis was significantly higher during hibernation than that during non-hibernation. Immunohistochemistry reaction showed that LAMP1-positive substance was distributed within the Sertoli cells of the testis. Further examination by transmission electron microscopy (TEM, many degraded spermatozoa being enwrapped within large entotic vacuoles in Sertoli cells. The nucleus and the flagellum of the spermatozoa were shown to be decomposed and digested inside entotic vacuoles within Sertoli cells. More than two spermatozoa heads were always observed in each internalized vacuoles. Deserving note is that, a number of different autophagosomes, including initial autophagic vesicles and degradative autophagic vesicles were found inside the entotic vacuoles of the Sertoli cells during hibernation. At the end of hibernation, entotic vacuoles and their autophagosomes disappeared, and numerous large lipid droplets (LDs appeared within the Sertoli cells. Adherens junctions were apparent between Sertoli cells and developing germ cells, which is the ultrastructural basis of entosis. Taken together, the results presented here show that in the turtle: (1 entosis with internal autophagosomes can take place within normal body cells during hibernation; (2 spermatozoa, as a highly differentiated cell can be internalized and degraded within Sertoli cell by entosis in vivo, which is in favor of the next reproductive cycle in the turtle.

  3. Interaction between basigin and monocarboxylate transporter 2 in the mouse testes and spermatozoa

    Directory of Open Access Journals (Sweden)

    Cheng Chen

    2016-01-01

    Full Text Available Basigin is a member of the immunoglobulin superfamily and plays various important roles in biological events including spermatogenesis. To examine the basigin molecular variants during spermatogenesis and sperm maturation in the mouse, immunoprecipitated basigin samples from testis and epididymal spermatozoa were analyzed by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS. The results demonstrated that basigin molecules from the testis and spermatozoa were separable into two major bands and that the differences in the molecular sizes were possibly because of an endoproteolytic cleavage. Since basigin is known to be a chaperone for the monocarboxylate transporter 1 (MCT1, the localization of basigin, MCT1 and MCT2 was examined during postnatal testicular development. Immunohistochemical studies showed different expression patterns of MCT1 and MCT2. MCT1 was localized on the surface of spermatogonia, spermatocytes, and spermatids. In contrast, MCT2 appeared on the principal piece of spermatozoa in the testis, where basigin was also observed. In mature epididymal spermatozoa, MCT2 was located on the midpiece, where basigin co-localized with MCT2 but not with MCT1. Furthermore, MCT2 was immunoprecipitated with basigin in mouse testes and sperm. These results suggest that basigin has a functional role as a binding partner with MCT2 in testicular and epididymal spermatozoa.

  4. bulls with unilateral gonadal hypoplasia and aplasia

    African Journals Online (AJOL)

    Heritability of reproduction disturbances in bulls of Swedish Red and White cattle (SRB\\. Nordt, Vet. Med. 2,943. KISER, T.E., MILVAE, R.A., HAFS, H.D., OXENDER, W.D.. & LOUIS, T.M., 1978. Comparison of testosterone and andronstenedione secretion in bulls given prostaglandin (F2s or luteinizing hormone. J. Anim. Sci.

  5. Bull trout recovery: Monitoring and evaluation guidance

    Science.gov (United States)

    U.S. Fish and Wildlife Service (USFWS)

    2008-01-01

    Bull trout (Salvelinus confluentus) is an imperiled species of char native to the Pacific Northwest. Combinations of habitat degradation (e.g., Fraley and Shepard 1989), barriers to migration (e.g., Rieman and McIntyre 1995), and the introduction of non-natives (e.g., Leary et al. 1993) have led to the decline of bull trout populations across their...

  6. ABNORMAL CHROMATIN CONDENSATION IN SPERMATOZOA AND DNA FRAGMENTATION IN SPERMATOZOA: IS THERE A CORRELATION?

    Directory of Open Access Journals (Sweden)

    E. E. Bragina

    2017-01-01

    Full Text Available Introduction. In the last decade, an understanding of the two-fold nature of genetic apparatus damage in spermatozoa has emerged: abnormal chromatin condensation (“immature” chromatin, ICH related to defective protaminization and leading to altered epigenetic regulation of the early embryogenesis, and disruption of DNA integrity, i. e. DNA fragmentation (DFS.Objective. Study of the correlation between abnormal chromatin condensation in spermatozoa and DFS.Materials and methods. The study included spermatozoa of 54 fertile males (1st group, control, 46 patients with primary infertility (2nd group, and 111 patients whose wives had a history of pregnancy abnormalities or failures of the assisted reproductive technology (ART, i. e. arrested embryonic development (3rd group. Presence of ICH was identified by quantitative electron microscopy, presence of DFS by TUNEL. Study of ICH and DFS in the same spermatozoon was conducted using correlation microscopy (TUNEL with subsequent ultrastructural analysis of the labeled cells.Results. The number of ICH spermatozoa significantly differed in the 3rd group from the control group (29.26 ± 13.49 vs. 22.43 ± 9.54; p = 0.006. The number of ICH spermatozoa in the 2nd group was higher than in the control group, but the difference wasn’t statistically significant (p = 0.061. A significant difference in the number of spermatozoa with residual cytoplasm on the head was observed between the fertile group and 2nd and 3rd groups (p = 0.0001 and p = 0.0006, respectively and on the neck (p = 0.0002 and p = 0.0003, respectively. Number of DFS spermatozoa in the second group significantly differed from the control (21.40 ± 11.88 vs. 13.70 ± 7.00, p = 0.03, but this difference wasn’t observed for the 3rd group. A very weak correlation between the number of DFS and ICH spermatozoa was observed in all three groups (r = 0.18, r = 0.33, and r = 0.01, respectively. Forty-six (46 spermatozoa were studied using

  7. Impact of silymarin enriched semen extender on bull sperm preservability

    Directory of Open Access Journals (Sweden)

    El-Sheshtawy RI

    2017-02-01

    Full Text Available Objective: To explore the effect of silymarin on bull spermatozoa during cooling and cryopreservation. Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk diluents, purified silymarin powder (obtained from the milk thistle silybum marianum, purchased from Unipharma, Al Obour city, Egypt, was soaked in Tris-citric acid-fructose diluent for 48 h at 10 ℃ making a stock solution (70 mg/mL, from this stock solution we obtained concentrations of 0.18 mg/mL, 0.36 mg/mL, 0.54 mg/mL, 0.72 mg/mL, 0.90 mg/mL in addition to the control (0.00 mg/mL reaching a final volume of 5 mL in each tube. Egg yolk was added to each tube to obtain silymarin enriched semen extender (SEE with 20% egg yolk, cooled slowly up to 5 曟 and equilibrated for 4 h. Semen was packed into 0.25 mL polyvinyl French straws (IMV, France. After equilibration periods, the straws were placed horizontally on a rack and frozen in a vapor 4 cm above liquid nitrogen for 10 min and were then dipped in liquid nitrogen. Extended semen was subjected to evaluation (motility, alive%, abnormality%, intact sperm membrane (HOST% and conception rate in both chilled and frozen semen. Results: Table 1 revealed that Sperm motility of the concentrations 2, 3 and 4 after 8 d of chilling were significantly (P<0.02 higher than control. Sperm motility of the concentration 2 (45.00%±2.89% after 9 d of chilling was higher than control and the other concentrations. Addition of SEE in concentration 1 and 2 gave post thawing sperm motility as high as the control (47.50±2.81 and 45.00±2.58, respectively while other concentration have lower effects on motility as compared to the control. Addition of silymarin improved post thawing alive% and was significantly higher (P<0.000 1 than the control. SEE decreased significantly (P<0.000 1 the % of post thawing abnormal sperm in concentration 3 and 4 (11.83±0.65 and 16.00±0.58, respectively. SEE improved significantly (P<0.018 the % of post

  8. Detrimental Effects of Non-Functional Spermatozoa on the Freezability of Functional Spermatozoa from Boar Ejaculate

    Science.gov (United States)

    Martinez-Alborcia, Maria J.; Valverde, Anthony; Parrilla, Inmaculada; Vazquez, Juan M.; Martinez, Emilio A.; Roca, Jordi

    2012-01-01

    In the present study, the impact of non-functional spermatozoa on the cryopreservation success of functional boar spermatozoa was evaluated. Fifteen sperm-rich ejaculate fractions collected from five fertile boars were frozen with different proportions of induced non-functional sperm (0 –native semen sample-, 25, 50 and 75% non-functional spermatozoa). After thawing, the recovery of motile and viable spermatozoa was assessed, and the functional of the spermatozoa was evaluated from plasma membrane fluidity and intracellular reactive oxygen species (ROS) generation upon exposure to capacitation conditions. In addition, the lipid peroxidation of the plasma membrane was assessed by the indirect measurement of malondialdehyde (MDA) generation. The normalized (with respect to a native semen sample) sperm motility (assessed by CASA) and viability (cytometrically assessed after staining with Hoechst 33342, propidium iodide and fluorescein-conjugated peanut agglutinin) decreased (psperm subpopulation before freezing showed the highest (pmembrane fluidity (assessed with Merocyanine 540). These findings indicate that non-functional spermatozoa in the semen samples before freezing negatively influence the freezability of functional spermatozoa. PMID:22567165

  9. Filamentous actin detected in rat spermatozoa.

    Science.gov (United States)

    Vogl, A W; Genereux, K; Pfeiffer, D C

    1993-01-01

    In this paper we report the positive staining of epididymal spermatozoa and testicular cells (late spermatids and spermatozoa) with fluorescent phallotoxins. Staining is most obvious with rhodamine phalloidin, but is also detectible with NBD-phallacidin. Specific fluorescence is emitted as a linear tract along the dorsal curvature of the head and as an inverted V-shaped structure in what appears to be the anterior aspect of the post-acrosomal region. We conclude that filamentous actin occurs in the heads of rat spermatozoa. Moreover, we speculate that this filamentous actin is concentrated in two regions of the perinuclear theca; in the subacrosomal space along the dorsal curvature of the nucleus, and in the post-acrosomal region in an area termed the ventral spur.

  10. Selection of High-Quality Spermatozoa May Be Promoted by Activated Vitamin D in the Woman

    DEFF Research Database (Denmark)

    Bøllehuus Hansen, Lasse; Rehfeld, Anders; de Neergaard, Rosanna

    2017-01-01

    Context: The vitamin D receptor (VDR) and enzymes involved in activation (CYP2R1, CYP27B1) and inactivation (CYP24A1) of vitamin D are expressed in ovary, testes, and spermatozoa. Objective: Determine responsiveness to 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] in spermatozoa from normal and infertile...... men, and identify the site of exposure and how 1,25(OH)2D3 influences sperm function. Design: Spermatozoa expressing VDR, CYP2R1, CYP27B1, and CYP24A1 were analyzed in normal and infertile men. 25-Hydroxyvitamin D (25-OHD), 24,25-dihydroxyvitamin D [24,25(OH)2D3], and 1,25(OH)2D3 were measured...... measured in fluids from 245 men and 13 women, while 74 oocytes and 17 semen donors were used for sperm-function tests. Main Outcome Measures: VDR and CYP24A1 expressions in spermatozoa, fluid concentrations of 25-OHD, 24,25(OH)2D3, and 1,25(OH)2D3, and 1,25(OH)2D3-induced effects on intracellular calcium...

  11. Impaired protamination and sperm DNA damage in a Nellore bull with high percentages of morphological sperm defects in comparison to normospermic bulls

    Directory of Open Access Journals (Sweden)

    J.T. Carreira

    2015-04-01

    Full Text Available The routine semen evaluation assessing sperm concentration, motility and morphology, does not identify subtle defects in sperm chromatin architecture. Bulls appear to have stable chromatin, with low levels of DNA fragmentation. However, the nature of fragmentation and its impact on fertility remain unclear and there are no detailed reports characterizing the DNA organization and damage in this species. The intensive genetic selection, the use of artificial insemination and in vitro embryo production associated to the cryopreservation process can contribute to the chromatin damage and highlights the importance of sperm DNA integrity for the success of these technologies. Frozen-thawed semen samples from three ejaculates from a Nellore bull showed high levels of morphological sperm abnormalities (55.8±5.1%, and were selected for complementary tests. Damage of acrosomal (76.9±8.9% and plasma membranes (75.7±9.3% as well as sperm DNA strand breaks (13.8±9.5% and protamination deficiency (3.7±0.6% were significantly higher compared to the values measured in the semen of five Nellore bulls with normospermia (24.3±3.3%; 24.5±6.1%; 0.6±0.5%; 0.4±0.6% for acrosome, plasma membrane, DNA breaks and protamine deficiency, respectively (P<0.05. Motility and percentage of spermatozoa with low mitochondrial potential showed no differences between groups. This study shows how routine semen analyses (in this case morphology may point to the length and complexity of sperm cell damage emphasizing the importance of sperm function testing.

  12. CURCUMIN IN MALE FERTILITY: EFFECTS ON SPERMATOZOA VITALITY AND OXIDATIVE BALANCE

    Directory of Open Access Journals (Sweden)

    Eva Tvrdá

    2015-02-01

    Full Text Available The aim of this study was to assess the dose- and time-dependent effects of curcumin on bovine spermatozoa during short-term (0h, 2h, 6h and long-term (12h, 24h in vitro culture periods. Semen samples were collected from 20 adult breeding bulls, and diluted in physiological saline solution containing 0.5% DMSO together with 0, 1, 5, 10, 50 and 100 μM/L of curcumin. Spermatozoa motion parameters were determined using the SpermVisionTM and CASA (Computer Assisted Semen Analyzer system. Cell viability was measured using the metabolic activity MTT assay, and the nitroblue-tetrazolium (NBT test was used to assess the intracellular superoxide formation. The CASA analysis revealed that concentrations of 50 μM/L and 10 μM/L of curcumin were able to significantly prevent the decrease of motility and progressive motility (P<0.001 in case of group B and P<0.01 in case of group C over all time periods of the in vitro incubation. At the same time, supplementation of concentrations ranging from 50 μM/L to 5 μM/L of curcumin led to a significant preservation of the cell viability in comparison to the control (P<0.001 in case of groups B and C; P<0.05 in case of group D. Concentrations in between 50 μM/L and 5 μM/L of curcumin demonstrated antioxidant properties, translated in a significant reduction of the intracellular superoxide production throughout the in vitro culture (P<0.001. The results indicate that the addition of curcumin, especially in concentrations between 50 μM and 10 μM to the culture medium could be beneficial for a complex enhancement of spermatozoa activity and protection against complications resulting from in vitro culture.

  13. Effects of the platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen (AI) with reduced sperm motility.

    Science.gov (United States)

    Lecewicz, M; Kordan, W; Majewska, A; Kamiński, S; Dziekońska, A; Mietelska, K

    2016-01-01

    The aim of the study was to determine the effects of platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen with reduced sperm motility used for artificial insemination. The aim of experiment 1 was to identify the optimal concentration of the phospholipid able to preserve sperm viability. Cryopreserved semen was treated with different PAF concentrations: 1×10(-5) M, 1×10(-6) M, 1×10(-7) M, 1×10(-8) M and 1×10(-9) M. The experiment demonstrated that PAF at concentration 1×10(-9) M increased most the sperm viability parameters (motility parameters, plasma membrane integrity and mitochondrial function) after 120 min of incubation of thawed semen at 37°C. Cryopreserved bull semen with reduced sperm motility (below 70%) was supplemented with PAF in a concentration of 1×10(-9) M. A statistically significant increase in sperm motility, percentage of linear motile spermatozoa and VSL value was observed after 120 min incubation of sperm with 1×10(-9) M PAF. Sperm supplementation with PAF also had positive effects on plasma membrane integrity and percentage of spermatozoa with preserved mitochondrial transmembrane potential, but the differences were not statistically significant. The results indicated positive effects of PAF supplementation at a concentration of 1×10(-9) M on the selected sperm quality parameters in cryopreserved bull semen with reduced motility.

  14. PERUBAHAN VIABILITAS DAN STRUKTUR SUBSELULER SPERMATOZOA DOMBA SETELAH PENGERINGBEKUAN

    Directory of Open Access Journals (Sweden)

    Takdir Saili

    2009-12-01

    Full Text Available Several methods i.e. cooling, freezing, and freeze-drying have been widely used to preserve spermatozoa with various degree of success. Freeze-drying appears to provide a method to preserve spermatozoa in a dry state without requiring liquid nitrogen for storing frozen spermatozoa. Freeze-drying procedures can have a detrimental effect on plasma membrane and acrosomal cap of the spermatozoa. In this experiment study, the viability and subcellular changes of freeze-dried ram spermatozoa were evaluated using staining method and scanning electron microscopy. The results revealed that all freeze-dried spermatozoa were dead following evaluation using eosin staining and Hoechst-propidium iodide staining methods. Morover, plasma membrane and acrosomal cap of freeze-dried ram spermatozoa was disrupted observed using scanning electron microscope.

  15. In-depth proteomic analysis of boar spermatozoa through shotgun and gel-based methods.

    Science.gov (United States)

    Feugang, Jean M; Liao, Shengfa F; Willard, Scott T; Ryan, Peter L

    2018-01-18

    Mature spermatozoa contain numerous epididymal and seminal plasma proteins, which full identification through high-throughput technologies may allow for a better understanding of the sperm biology. Therefore, we conducted a global proteomic analysis of boar spermatozoa through shotgun and gel-based methodologies. The total proteins were extracted from mature spermatozoa and subjecsted to proteome analyses. Functional analyses of gene ontology representations and pathway enrichments were conducted on the shotgun dataset, followed by immunology and gene expression validations. Shotgun and gel-based approaches allowed the detection of 2728 proteins and 2123 spots, respectively. Approximately 38% and 59% of total proteins were respectively fully and partially annotated, and 3% were unknown. Gene ontology analysis indicated high proportions of proteins associated with intracellular and cytoplasm localizations, protein and nucleic acid binding, hydrolase and transferase activities, and cellular, metabolic, and regulation of biological processes. Proteins associated with phosphorylation processes and mitochondrial membranes, nucleic acid binding, and phosphate and phosphorous metabolics represented 77% of the dataset. Pathways associated with oxidative phosphorylation, citrate cycle, and extra-cellular matrix-receptor interaction were significantly enriched. Protein complex, intracellular organelle, cytoskeletal parts, fertilization and reproduction, and gap junction pathway were significantly enriched within the top 116 highly abundant proteins. Nine randomly selected protein candidates were confirmed with gel-based identification, immunofluorescence detection, and mRNA expression. This study offers an in-depth proteomic mapping of mature boar spermatozoa that will enable comparative and discovery research for the improvement of male fertility.

  16. CHARACTERISTIC OF FROZEN-THAWED EPIDIDYMAL SPERMATOZOA AND REFRIGERATED STORAGE OF RAM SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    N.W.K. Karja

    2014-10-01

    Full Text Available Post-mortem spermatozoa recovery is an important technique for obtaining germplasm reservesfrom genetically valuable animals or endangered species. The purpose of this study was to studywhether ram spermatozoa within epididymides stored at 4º C for 24 and 48 h remain their motility andviability. The characteristic of ram epididymal spermatozoa after freezing and thawing was alsoobserved. Six pairs of ram testes with attached epididymides were used in this study. The motility ofcontrol spermatozoa was well maintained throughout the dilution procedure (83.3±1.1, 80±1.3, and80±1.3% for collection, Niwa and Sasaki freezing -1 extender (NSF-1 and NSF-2 groups respectively;but declined (P<0.05 after freezing and thawing (38.3±3.1%. Motile and viable spermatozoa could berecovered from epididymides up to 48 h of storage, although their quality declined significantly(P<0.05 as post-mortem storage time increased (motility: 83±1.2, 67±3.0, and 46±5.1; viability:84.2±2.4, 73±2.8, and 66.6±2.6 % for control, 24 h and 48 h group respectively. These data indicatethat ram epididymides could be stored at 4º C for 48 h when epididymal spermatozoa cannot beimmmediately collected and cryopreserved. These storage conditions might be possible to use forepididymal sperm recovery in wild ruminants.

  17. Mycoplasma genitalium attaches to human spermatozoa

    DEFF Research Database (Denmark)

    Svenstrup, Helle Friis; Fedder, Jens; Abraham-Peskir, Joanna

    2003-01-01

    BACKGROUND: Mycoplasma genitalium causes urogenital diseases in men and women and is presumed to be sexually transmitted. We wanted to investigate whether spermatozoa could serve as vectors for M.genitalium in order to cause upper genital diseases in women. METHODS: By use of Nomarski light...... sperm. This ability may be important in the process of causing female genital diseases and infertility....

  18. Time trends, environmental factors and genetic basis of semen traits collected in Holstein bulls under commercial conditions.

    Science.gov (United States)

    Karoui, Sofiene; Díaz, Clara; Serrano, Magdalena; Cue, Roger; Celorrio, Idoia; Carabaño, María J

    2011-03-01

    The fact that results of artificial insemination (AI) are declining in highly selected dairy cattle populations has added a renewed interest to the evaluation of male fertility. Data from 42,348 ejaculates collected from 1990 to 2007 on 502 Holstein bulls were analysed in a Bayesian framework to provide estimates of the evolution of semen traits routinely collected in AI centres throughout the last decades of intense selection for production traits and estimate genetic parameters. The traits under consideration were volume (VOL), concentration (CONC), number of spermatozoa per ejaculate (NESPZ), mass motility score (MM), individual motility (IM), and post-thawing motility (PTM). The environmental factors studied were year-season and week of collection, which account for changes in environmental and technical conditions along time, age at collection, ejaculate order, time from previous collection (TPC) and time between collection and freezing (TCF) (only for PTM). Bull's inbreeding coefficient (Fi), bull's permanent environmental and additive genetic effects were also considered. The use of reduced models was evaluated using the Bayes factor. For all the systematic effects tested, strong or very strong evidence in favour of including the effect in the model was obtained, except for Fi for motility traits and TCF for PTM. No systematic time trends for environment or bull effects were observed, except for PTM, which showed an increasing environmental trend, associated with improvements in freezing-thawing protocols. Heritability estimates were moderate (0.16-0.22), except for IM, which presented a low value (0.07). Genetic correlations among motilities and between motilities and CONC were large and positive [0.38-0.87], VOL showed a negative correlation with CONC (-0.13) but with ample HPD 95%. The magnitude of heritabilities would allow an efficient selection if required and grants the use of these traits as indicators of the sperm viability component of bulls

  19. Copper-Induced Spermatozoa Head Malformation Is Related to Oxidative Damage to Testes in CD-1 Mice.

    Science.gov (United States)

    Zhang, Zhuo-Wei; Zhi-GangTan; Qiao, Na; Kang, Zhen-Long; Chen, Zhi-Ling; Hu, Lian-Mei; Yang, Zeng-Ming; Li, Ying

    2016-10-01

    The molecular mechanism for copper toxicity on spermatozoa quality in mice is not well understood. In a 4-week experiment, we challenged 24, 6-week-old male CD-1 mice with twice-a-week intraperitoneal copper chloride injections and evaluated spermatozoa quality, copper levels in the testes, serum testosterone, the expression of key antioxidant glutathione peroxidase 5 (GPx5), and the regulated androgen receptor (AR) in the mice testes. We compared these outcomes for four groups of six mice given doses of 0, 1.25, 2.5, 5.0 mg/kg weight copper chloride twice a week for 4 weeks. The mice demonstrated a copper increase spermatozoa head malformation in a dose-response manner. However, we observed no changes in spermatozoa viability and acrosome integrity in the ratio of mouse body weight to testes weight or in the histomorphology of the testes as the average copper level increased. Results of our RT-PCR assays, immunohistochemical tests, ELISA, and histochemistry analyses indicated that testis GPx5 expression was increased, AR expression in the testes was decreased, serum testosterone was decreased, and the activity of 3β-hydroxysteroid dehydrogenase was decreased as the copper dose increased. In conclusion, these data show that sublethal exposure to copper induces spermatozoa head malformation and influences both mRNA and protein levels of GPx5 and AR which is related to copper resides in the testes.

  20. The roles of protein disulphide isomerase family A, member 3 (ERp57) and surface thiol/disulphide exchange in human spermatozoa-zona pellucida binding.

    Science.gov (United States)

    Wong, Chi-Wai; Lam, Kevin K W; Lee, Cheuk-Lun; Yeung, William S B; Zhao, Wei E; Ho, Pak-Chung; Ou, Jian-Ping; Chiu, Philip C N

    2017-04-01

    Are multimeric sperm plasma membrane protein complexes, ERp57 and sperm surface thiol content involved in human spermatozoa-zona pellucida (ZP) interaction? ERp57 is a component of a multimeric spermatozoa-ZP receptor complex involved in regulation of human spermatozoa-ZP binding via up-regulation of sperm surface thiol content. A spermatozoon acquires its fertilization capacity within the female reproductive tract by capacitation. Spermatozoa-ZP receptor is suggested to be a composite structure that is assembled into a functional complex during capacitation. Sperm surface thiol content is elevated during capacitation. ERp57 is a protein disulphide isomerase that modulates the thiol-disulphide status of proteins. The binding ability and components of protein complexes in extracted membrane protein fractions of spermatozoa were studied. The roles of capacitation, thiol-disulphide reagent treatments and ERp57 on sperm functions and sperm surface thiol content were assessed. Spermatozoa were obtained from semen samples from normozoospermic men. Human oocytes were obtained from an assisted reproduction programme. Blue native polyacrylamide gel electrophoresis, western ligand blotting and mass spectrometry were used to identify the components of solubilized ZP/ZP3-binding complexes. The localization and expression of sperm surface thiol and ERp57 were studied by immunostaining and sperm surface protein biotinylation followed by western blotting. Sperm functions were assessed by standard assays. Several ZP-binding complexes were isolated from the cell membrane of capacitated spermatozoa. ERp57 was a component of one of these complexes. Capacitation significantly increased the sperm surface thiol content, acrosomal thiol distribution and ERp57 expression on sperm surface. Sperm surface thiol and ERp57 immunoreactivity were localized to the acrosomal region of spermatozoa, a region responsible for ZP-binding. Up-regulation of the surface thiol content or ERp57 surface

  1. In vitro exposure of bull sperm cells to DMSA-coated maghemite nanoparticles does not affect cell functionality or structure.

    Science.gov (United States)

    Caldeira, Denise Ferreira; Paulini, Fernanda; Silva, Renata Carvalho; Azevedo, Ricardo Bentes de; Lucci, Carolina Madeira

    2017-06-28

    Magnetic nanoparticles can be used in different areas of biology. It is therefore important to know the effects of such nanomaterials on germline cells as they may traverse the blood-testis barrier. This work aimed to evaluate the response of bull sperm after exposure to a magnetic fluid containing DMSA-coated maghemite nanoparticles (MNP-DMSA) in order to determine nanotoxicity. Bull sperm was incubated with MNP-DMSA at final concentrations of 0.06, 0.03 or 0.015 mg Fe/mL. Sperm kinetics, plasma membrane integrity and acrosome reaction were evaluated over a 4 h incubation period. The sperm cells were also evaluated by transmission electron microscopy. Exposure of bull sperm to MNP-DMSA did not affect sperm kinetics or integrity. Neither ultrastructural damage of sperm cells nor uptake of nanoparticles by the spermatozoa was observed. In conclusion, MNP-DMSA does not affect sperm function or structure under the conditions tested.

  2. Sexual behavior and its relationship with semen quality parameters in Sahiwal breeding bulls

    Directory of Open Access Journals (Sweden)

    Shushant Singh

    2015-06-01

    Full Text Available Aim: The study was conducted at Artificial Breeding Research Centre, NDRI, Karnal, to determine the sexual behavior and its relationship with semen quality parameters in Sahiwal breeding bulls. Materials and Methods: A total of 63 ejaculates were collected from six adult Sahiwal bulls (age ~47 mo and bwt ~466 kg, to study the relationship of sexual behavior and semen quality. The degree of association between different variables was estimated by Pearson’s correlation coefficient method. Results: The results depicted that, sexual aggressiveness showed significantly high positive correlation with libido score (LS and sexual behavior score (SBS. Reaction time (RT and total time taken in mounts (TTTM had a significant negative correlation with LS and SBS. Penile erection score and penile protrusion score (PPS both had a significant positive correlation with ejaculatory thrust score, mating ability score, and SBS. Results of correlation among seminal attributes and with sexual behavior depicted that ejaculate volume had positive significant correlation with initial progressive motility (IPM, sperm concentration (SCON, head abnormality, total abnormality, hypo-osmotic swelling test (HOST, acrosomal integrity (AI whereas, mass activity had positive significant correlation with IPM, SCON, non-eosinophilic spermatozoa count (NESC, HOST, AI, RT and TTTM and IPM had positive significant correlation with SCON, NESC, HOST, AI, and TTTM, whereas and HOST had positive significant correlation with AI. Among seminal attributes, SCON had a positive significant correlation with PPS where as head abnormalities had a positive significant correlation with RT and TTTM. Conclusion: It can be concluded that the relationship of sexual behavior and semen quality parameters are reflecting that the sexual behavior of individual bulls is important to harvest good quality and quantity of semen as desired type of sexual preparation can be provided.

  3. Canine epididymal spermatozoa: A hidden treasure with great potential.

    Science.gov (United States)

    Luvoni, G C; Morselli, M G

    2017-04-01

    The hidden treasure represented by epididymal spermatozoa has great potential in the current reproductive technologies in dogs. In case of azoospermia or when a donor male accidentally dies or undergoes orchiectomy, the retrieval of epididymal spermatozoa opens new possibilities to generate progeny. Spermatozoa can be collected by different techniques from ex vivo or in vivo testicles and can be cryopreserved for a future use. Freeze tolerance of canine epididymal spermatozoa seems lower than that of ejaculated spermatozoa; however, puppies were born after artificial insemination with frozen epididymal semen, other than with fresh and chilled. Even though several aspects need to be further investigated, advances have been recently made in the use of epididymal spermatozoa in assisted reproduction in dogs. © 2016 Blackwell Verlag GmbH.

  4. Phosphopeptide analysis of rodent epididymal spermatozoa.

    Science.gov (United States)

    Baker, Mark A; Hetherington, Louise; Weinberg, Anita; Velkov, Tony

    2014-12-30

    Spermatozoa are quite unique amongst cell types. Although produced in the testis, both nuclear gene transcription and translation are switched off once the pre-cursor round cell begins to elongate and differentiate into what is morphologically recognized as a spermatozoon. However, the spermatozoon is very immature, having no ability for motility or egg recognition. Both of these events occur once the spermatozoa transit a secondary organ known as the epididymis. During the ~12 day passage that it takes for a sperm cell to pass through the epididymis, post-translational modifications of existing proteins play a pivotal role in the maturation of the cell. One major facet of such is protein phosphorylation. In order to characterize phosphorylation events taking place during sperm maturation, both pure sperm cell populations and pre-fractionation of phosphopeptides must be established. Using back flushing techniques, a method for the isolation of pure spermatozoa of high quality and yield from the distal or caudal epididymides is outlined. The steps for solubilization, digestion, and pre-fractionation of sperm phosphopeptides through TiO2 affinity chromatography are explained. Once isolated, phosphopeptides can be injected into MS to identify both protein phosphorylation events on specific amino acid residues and quantify the levels of phosphorylation taking place during the sperm maturation processes.

  5. Bull Trout Spawning Surveys: Kootenai National Wildlife Refuge

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Bull trout are listed as threatened under the Endangered Species Act and Myrtle Creek was designated as critical habitat for bull trout this year. Myrtle Creek flows...

  6. Vesicular transfer of membrane components to bovine epididymal spermatozoa.

    Science.gov (United States)

    Schwarz, A; Wennemuth, G; Post, H; Brandenburger, T; Aumüller, G; Wilhelm, B

    2013-09-01

    Epididymosomes (apocrine secreted epididymal vesicles) are assumed to play a crucial role in sperm maturation. Our aim has been to analyze the fusogenic properties of bovine epididymosomes and their involvement in the transfer of membrane components (lipids, proteins, plasma membrane Ca(2+)-ATPase 4 [PMCA4]) into bovine sperm. The fusogenic properties of epididymosomes with spermatozoa were investigated in vitro by using octadecyl rhodamine-B (R18)-labeled epididymosomes. Spermatozoa isolated from the epididymal caput showed a higher fusion rate than those taken from the cauda. The fusion rate was dependent on pH and time. Furthermore, the lipid and protein content in spermatozoa changed during epididymal transit and after in vitro fusion with epididymosomes. Following the in vitro fusion of caput spermatozoa with epididymosomes, the cholesterol/total phospholipid ratio of the sperm plasma membrane decreased. The effect was comparable with the cholesterol/total phospholipid ratio of native cauda spermatozoa. Co-incubation experiments of spermatozoa with biotinylated epididymosomes additionally revealed that proteins were transferred from epididymosomes to sperm. To examine the potential transfer of epididymis-derived PMCA4 to spermatozoa, immunofluorescence analysis and Ca(2+)-ATPase activity assays were performed. In caput spermatozoa, the PMCA4 fluorescence signal was slightly raised and Ca(2+)-ATPase activity increased after in vitro fusion. Thus, our experiments indicate significant changes in the lipid and protein composition of epididymal sperm following interaction with epididymosomes. Moreover, our results substantiate the presumption that PMCA4 is transferred to spermatozoa via epididymosomes.

  7. Profiling of sperm proteins and association of sperm PDC-109 with bull fertility.

    Science.gov (United States)

    Somashekar, Lakshminarayana; Selvaraju, Sellappan; Parthipan, Sivashanmugam; Ravindra, Janivara Parameswaraiah

    2015-01-01

    The composition of sperm proteins influences the fertilizing ability of sperm and hence the present study was conducted (i) to profile sperm proteins expression patterns in bulls of differing fertility index and (ii) to identify and relate the abundant sperm proteins with bull fertility. The semen samples were collected from Holstein-Friesian bulls (n = 12) varying in conception rate (CR) (high/low). The frozen semen straws (three ejaculates, from each bull) were used to study (a) sperm kinetic parameters, (b) plasmalemma integrity, (c) mitochondrial membrane potential, and (d) chromatin distribution. Three bulls were randomly selected from each group (n = 3) and the neat sperm pellets were subjected to percoll purification, followed by protein isolation using 0.1% Triton X100. The sperm kinetic parameters, plasmalemma integrity, mitochondrial membrane potential, and the chromatin distribution did not differ significantly between groups. The number of acidic (pI; 3.1-5.6, 37%) and basic (pI; 7.9-10.0, 27%) proteins and their pattern of expression varied significantly (p sperm protein spots in 2D-gel electrophoresis (2DE) were identified as seminal plasma protein PDC-109 (i.e., protein with N-terminus aspartic acid, D and carboxy terminus cystine, having 109 amino acids) and its isoform and spermadhesin-1 (SPADH1). The western blot analysis confirmed the presence of PDC-109 isoform proteins at 15.4 kDa (pI 5.3 and 5.5). The seminal plasma protein PDC-109 was abundant in the low fertile when compared to the high fertile group (p sperm proteins may influence sperm fertility and sperm PDC-109 levels above a certain threshold affects bull fertility.

  8. Spermatogonial stem cells in the bull

    NARCIS (Netherlands)

    Aponte, P.M

    2009-01-01

    In the testis a complex process, called spermatogenesis, generates millions of spermatozoa per day. At the start of this process there are spermatogonial stem cells (SSCs) that have the ability to divide either into new stem cells (self-renewal) or daughter cells committed to develop into

  9. DNA fragmentation in chicken spermatozoa during cryopreservation.

    Science.gov (United States)

    Gliozzi, T M; Zaniboni, L; Cerolini, S

    2011-06-01

    Semen cryopreservation is fundamental both for the practice of artificial insemination, and for the conservation of genetic resources in cryobanks; nevertheless, there is still not an efficient standard freezing procedure assuring a steady and suitable level of fertility in fowl, and consequently there is no systematic use of frozen semen in the poultry industry. This study examined changes in motility (CASA), cell membrane integrity (Ethidium Bromide (EtBr) exclusion procedure and stress test) and DNA fragmentation (neutral comet assay) in fowl spermatozoa before, during and after cryopreservation and storage at -196 °C. An optimized comet assay for chicken semen was studied and applied to the analyses. Semen collected from 18 Mericanel della Brianza (local Italian breed) male chicken breeders was frozen in pellets and thawed in a water bath at 60 °C. Measurements were performed on fresh semen soon after dilution, after equilibration with 6% dimethylacetamide at 4 °C (processed semen) and after thawing. Sperm DNA damage occurred during cryopreservation of chicken semen and the proportion of spermatozoa with damaged DNA significantly increased from 6.2% in fresh and 6.4% in processed semen to 19.8% in frozen-thawed semen. The proportion of DNA in the comet tail of damaged spermatozoa was also significantly affected by cryopreservation, with an increase found from fresh (26.3%) to frozen-thawed (30.9%) sperm, whereas processed semen (30.1%) didn't show significant differences. The proportion of total membrane damaged spermatozoa (EtBr exclusion procedure) did not increase by 4 °C equilibration time, and greatly and significantly increased by cryopreservation; the values recorded in fresh, processed and frozen semen were 2.9, 5.6, and 66.7% respectively. As regards the proportion of damaged cells in the stress test, all values differed significantly (7.1% fresh semen, 11.7% processed semen, 63.7% frozen semen). Total motility was not affected by equilibration (52

  10. Optimizing age of bull at first use in relation to fertility of Murrah breeding bulls

    OpenAIRE

    Mir, M. A.; Chakravarty, A. K.; Gupta, A.K.; B. C. Naha; V. JAMUNA; Patil, C. S.; Singh, A. P.

    2015-01-01

    Aim: The aim of the present investigation was to optimize the age at first use (AAFU) of semen of Murrah breeding bulls, which will help in early selection of bulls under progeny testing program for improving the reproductive performance in the herd. Materials and Methods: The data on AAFU, conception rate based on first A.I. (CRFAI), overall conception rate (OCR), and birth weight (B.WT) of 57 Murrah bulls during 1993-2014 at NDRI center pertaining to 14 sets of Network Project on Buffalo Im...

  11. Leptin and leptin receptor are detectable in equine spermatozoa but are not involved in in vitro fertilisation.

    Science.gov (United States)

    Lange-Consiglio, Anna; Corradetti, Bruna; Perrini, Claudia; Bizzaro, Davide; Cremonesi, Fausto

    2016-04-01

    In human and swine, leptin (OB) has been identified in seminal plasma and leptin receptors (OB-R) on the cell surface of spermatozoa, indicating that spermatozoa are a target for OB. This hormone has also been detected in follicular fluid (FF) in women and mares, although its role requires further study. The aims of this study were to investigate the immunolocalisation and the expression of OB and OB-R in equine spermatozoa and to evaluate the involvement of OB in equine in vitro fertilisation (IVF). Since progesterone (P) and OB are both found in FF, the individual and combined effects of these two hormones were studied in equine IVF and compared with the results obtained from the use of FF for in vitro sperm preparation. For the first time, we were able to identify OB and OB-R mRNA and their corresponding proteins in equine spermatozoa. When spermatozoa were treated with OB, there was a decrease in the three motility parameters VSL, STR and LIN, commonly associated with hyperactivation, whilst the acrosome reaction rate increased (Phorse, OB is involved in capacitation and hyperactivation but not in sperm penetration.

  12. Scrotal circumference in young beef bulls: Relationships to growth ...

    African Journals Online (AJOL)

    dairy cattle, but natural mating accounts for most of the pregnancies achieved each ... Grou'th test data from Phase C (National Beef Cattle. Performance and Progenv Testing Scheme) from 129. Simmentaler bulls. 97 Santa Gertrudis bulls and 100. Hereford bulls were obtained from the different Cattle. Breeders' Societies of ...

  13. High-fat diet reprograms the epigenome of rat spermatozoa and transgenerationally affects metabolism of the offspring

    DEFF Research Database (Denmark)

    de Castro Barbosa, Thais; Ingerslev, Lars R; Alm, Petter S

    2016-01-01

    OBJECTIVES: Chronic and high consumption of fat constitutes an environmental stress that leads to metabolic diseases. We hypothesized that high-fat diet (HFD) transgenerationally remodels the epigenome of spermatozoa and metabolism of the offspring. METHODS: F0-male rats fed either HFD or chow diet...... for 12 weeks were mated with chow-fed dams to generate F1 and F2 offspring. Motile spermatozoa were isolated from F0 and F1 breeders to determine DNA methylation and small non-coding RNA (sncRNA) expression pattern by deep sequencing. RESULTS: Newborn offspring of HFD-fed fathers had reduced body weight...... and pancreatic beta-cell mass. Adult female, but not male, offspring of HFD-fed fathers were glucose intolerant and resistant to HFD-induced weight gain. This phenotype was perpetuated in the F2 progeny, indicating transgenerational epigenetic inheritance. The epigenome of spermatozoa from HFD-fed F0 and their F...

  14. Detection automatique de bulles HI en expansion

    Science.gov (United States)

    Daigle, Anik

    Plusieurs cavités en expansion observées dans le gaz d'hydrogène neutre galactique ont été associées à la présence d'étoiles massives présentant d'intenses vents stellaires ou à des explosions de supernovae. Ces bulles, qui sont remplies de gaz très chaud ( T = 10 5 -10 7 K), tracent donc à la fois les étoiles massives et la phase chaude et ionisée du gaz interstellaire. La morphologie de ces bulles est instable et généralement perturbée par la multitude de structures et de phénomènes existant dans le milieu interstellaire. Leur détection par inspection visuelle est donc difficile et subjective. L'objet de cette thèse est le développement d'une technique de détection automatique et objective de bulles en expansion dans le gaz d'hydrogène neutre de la Voie lactée. Les bulles à progéniteurs stellaires présentent toutes des vitesses d'expansion comprises entre 6 et 19 km s -1 . Une telle caractéristique peut généralement être reconnue dans les spectres en vitesse des cubes de données à 21 cm de l'hydrogène neutre. Notre technique de détection a donc été fondée sur la reconnaissance automatique de la signature d'une expansion à [6, 19] km s -1 dans les spectres en vitesse. La caractérisation et la généralisation de la signature dynamique propre à une bulle en expansion a été confiée à des réseaux de neurones artificiels. Des validations subséquentes, dont certaines liées à la morphologie, ont ensuite été appliquées. La technique a été testée sur onze bulles connues, dont dix ont pu être détectées. La technique a aussi été employée pour la réalisation d'un relevé dans la région du bras de Persée. La distribution spatiale des objets détectés s'est révélée suivre la distribution des étoiles du disque de la Voie lactée. De plus, la distribution selon le rayon et selon la vitesse d'expansion des bulles détectées de rayons plus petits que 10 pc correspondent aux distributions dérivées du mod

  15. Structured decision making for conservation of bull trout (Salvelinus confluentus) in Long Creek, Klamath River Basin, south-central Oregon

    Science.gov (United States)

    Benjamin, Joseph R.; McDonnell, Kevin; Dunham, Jason B.; Brignon, William R.; Peterson, James T.

    2017-06-21

    With the decline of bull trout (Salvelinus confluentus), managers face multiple, and sometimes contradictory, management alternatives for species recovery. Moreover, effective decision-making involves all stakeholders influenced by the decisions (such as Tribal, State, Federal, private, and non-governmental organizations) because they represent diverse objectives, jurisdictions, policy mandates, and opinions of the best management strategy. The process of structured decision making is explicitly designed to address these elements of the decision making process. Here we report on an application of structured decision making to a population of bull trout believed threatened by high densities of nonnative brook trout (S. fontinalis) and habitat fragmentation in Long Creek, a tributary to the Sycan River in the Klamath River Basin, south-central Oregon. This involved engaging stakeholders to identify (1) their fundamental objectives for the conservation of bull trout, (2) feasible management alternatives to achieve their objectives, and (3) biological information and assumptions to incorporate in a decision model. Model simulations suggested an overarching theme among the top decision alternatives, which was a need to simultaneously control brook trout and ensure that the migratory tactic of bull trout can be expressed. More specifically, the optimal management decision, based on the estimated adult abundance at year 10, was to combine the eradication of brook trout from Long Creek with improvement of downstream conditions (for example, connectivity or habitat conditions). Other top decisions included these actions independently, as well as electrofishing removal of brook trout. In contrast, translocating bull trout to a different stream or installing a barrier to prevent upstream spread of brook trout had minimal or negative effects on the bull trout population. Moreover, sensitivity analyses suggested that these actions were consistently identified as optimal across

  16. bulls with unilateral gonadal hypoplasia and aplasia

    African Journals Online (AJOL)

    maintenance of fertility of bulls with gonadal hypoplasia as observed by other ... that the unilaterally affected animals have normal fertility, while the bilaterally affected animals have reduced fertility or total infertility. Gonadal hypoplasia has been reported in numerous .... Hereditary forms of sterility in Swedish cattle breeds.

  17. Bull Riding Injuries In Central Queensland

    Directory of Open Access Journals (Sweden)

    Ryan James Livingston

    2012-07-01

    Full Text Available Background Bull riding is an increasingly popular and growing professional sport in Australia. This is the first national study that investigates bull riding-related injuries. Method A six-year retrospective study of patients admitted to Rockhampton Base Hospital with acute injuries sustained whilst bull riding. Patients were identified from the Rockhampton Hospital international coding system and surgical audit excel databases. Supporting information was found from patient chart review. Results Thirty-eight patients were admitted during the study. Injuries increased from 2008. The most common injuries were to limbs (52%, chest (15% and brain (10%. Life-threatening injuries were all caused by a direct kick or trampling by the bull; 5% of patients needed air transfer to Brisbane, and 10% to Rockhampton for their acute care. The only complication was infection of open wounds. The average hospital stay was 2.2 (range= 1-5, SD= 1.1 days and 64% of patients required operative intervention. Conclusion Patients that had been kicked or trampled should be identified as having potentially life-threatening injuries, and transferred for review at an appropriate facility. Due to the high risk of infection all contaminated wounds should be washed out formally and receive antibiotics. Protective equipment should be encouraged among riders.

  18. Primary mandibular hemangiosarcoma in a bull

    Science.gov (United States)

    Poulsen, Keith P.; McSloy, Alexandra C.; Perrier, Melanie; Prichard, Michael A.; Steinberg, Howard; Semrad, Susan D.

    2008-01-01

    Disseminated pulmonary and subcutaneous-muscular hemangiosarcoma at the left hemimandible was diagnosed postmortem in a 2-year-old Jersey bull that presented with a 7-day history of facial swelling from suspected traumatic injury. Hemangiosarcoma is uncommon in cattle and has never been reported to affect the bones of the skull. PMID:19043489

  19. Development of bull trout sampling protocols

    Science.gov (United States)

    R. F. Thurow; J. T. Peterson; J. W. Guzevich

    2001-01-01

    This report describes results of research conducted in Washington in 2000 through Interagency Agreement #134100H002 between the U.S. Fish and Wildlife Service (USFWS) and the U.S. Forest Service Rocky Mountain Research Station (RMRS). The purpose of this agreement is to develop a bull trout (Salvelinus confluentus) sampling protocol by integrating...

  20. Optimization of IVF pregnancy outcomes with donor spermatozoa.

    Science.gov (United States)

    Wang, Jeff G; Douglas, Nataki C; Prosser, Robert; Kort, Daniel; Choi, Janet M; Sauer, Mark V

    2009-03-01

    To identify risk factors for suboptimal IVF outcomes using insemination with donor spermatozoa and to define a lower threshold that may signal a conversion to fertilization by ICSI rather than insemination. Retrospective, age-matched, case-control study of women undergoing non-donor oocyte IVF cycles using either freshly ejaculated (N=138) or cryopreserved donor spermatozoa (N=69). Associations between method of fertilization, semen sample parameters, and pregnancy rates were analyzed. In vitro fertilization of oocytes with donor spermatozoa by insemination results in equivalent fertilization and pregnancy rates compared to those of freshly ejaculated spermatozoa from men with normal semen analyses when the post-processing motility is greater than or equal to 88%. IVF by insemination with donor spermatozoa when the post-processing motility is less than 88% is associated with a 5-fold reduction in pregnancy rates when compared to those of donor spermatozoa above this motility threshold. When the post-processing donor spermatozoa motility is low, fertilization by ICSI is associated with significantly higher pregnancy rates compared to those of insemination. While ICSI does not need to be categorically instituted when using donor spermatozoa in IVF, patients should be counseled that conversion from insemination to ICSI may be recommended based on low post-processing motility.

  1. The usefulness and significance of assessing rapidly progressive spermatozoa

    Science.gov (United States)

    Björndahl, Lars

    2010-01-01

    It is possible and clinically relevant to distinguish between slow and rapid progressive spermatozoa in basic semen analysis. This is discussed in light of the different purposes of semen analysis for the subfertile couple and the male patient. The two groups of progressive spermatozoa should be distinguished to help ensure that pertinent information available in the semen sample is not neglected. PMID:20111079

  2. DNA fragmentation in spermatozoa: a historical review.

    Science.gov (United States)

    Rex, A S; Aagaard, J; Fedder, J

    2017-07-01

    Sperm DNA Fragmentation has been extensively studied for more than a decade. In the 1940s the uniqueness of the spermatozoa protein complex which stabilizes the DNA was discovered. In the fifties and sixties, the association between unstable chromatin structure and subfertility was investigated. In the seventies, the impact of induced DNA damage was investigated. In the 1980s the concept of sperm DNA fragmentation as related to infertility was introduced as well as the first DNA fragmentation test: the Sperm Chromatin Structure Assay (SCSA). The terminal deoxynucleotidyl transferase nick end labelling (TUNEL) test followed by others was introduced in the nineties. The association between DNA fragmentation in spermatozoa and pregnancy loss has been extensively investigated spurring the need for a therapeutic tool for these patients. This gave rise to an increased interest in the aetiology of DNA damage. The present decade continues within this research area. Some of the more novel methods recently submerging are sorting of cells with increased DNA fragmentation and hyaluronic acid (HA) binding techniques. The clinical value of these tests remains to be elucidated. In spite of half a century of research within the area, this analysis is not routinely implemented into the fertility clinics. The underlying causes are multiple. The abundance of methods has impeded the need for a clinical significant threshold. One of the most promising methods was commercialized in 2005 and has been reserved for larger licensed laboratories. Myriads of reviews and meta-analyses on studies using different assays for analysis of DNA fragmentation, different clinical Artificial Reproductive Treatments (ART), different definitions of successful ART outcome and small patient cohorts have been published. Although the area of DNA fragmentation in spermatozoa is highly relevant in the fertility clinics, the need for further studies focusing on standardization of the methods and clinical

  3. Obesity and Bariatric Surgery Drive Epigenetic Variation of Spermatozoa in Humans.

    Science.gov (United States)

    Donkin, Ida; Versteyhe, Soetkin; Ingerslev, Lars R; Qian, Kui; Mechta, Mie; Nordkap, Loa; Mortensen, Brynjulf; Appel, Emil Vincent R; Jørgensen, Niels; Kristiansen, Viggo B; Hansen, Torben; Workman, Christopher T; Zierath, Juleen R; Barrès, Romain

    2016-02-09

    Obesity is a heritable disorder, with children of obese fathers at higher risk of developing obesity. Environmental factors epigenetically influence somatic tissues, but the contribution of these factors to the establishment of epigenetic patterns in human gametes is unknown. Here, we hypothesized that weight loss remodels the epigenetic signature of spermatozoa in human obesity. Comprehensive profiling of the epigenome of sperm from lean and obese men showed similar histone positioning, but small non-coding RNA expression and DNA methylation patterns were markedly different. In a separate cohort of morbidly obese men, surgery-induced weight loss was associated with a dramatic remodeling of sperm DNA methylation, notably at genetic locations implicated in the central control of appetite. Our data provide evidence that the epigenome of human spermatozoa dynamically changes under environmental pressure and offers insight into how obesity may propagate metabolic dysfunction to the next generation. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. The impact of lead and cadmium on selected motility, prooxidant and antioxidant parameters of bovine seminal plasma and spermatozoa.

    Science.gov (United States)

    Tvrdá, Eva; Kňažická, Zuzana; Lukáčová, Jana; Schneidgenová, Monika; Goc, Zofia; Greń, Agnieszka; Szabó, Csaba; Massányi, Peter; Lukáč, Norbert

    2013-01-01

    The aim of this study was to investigate the effects of lead (Pb) and cadmium (Cd) content on basic motility characteristics (motility-MOT, progressive motility-PROG) as well as selected markers of the prooxidant-antioxidant balance (catalase-CAT, glutathione-GSH, malondialdehyde-MDA) in bovine seminal plasma and spermatozoa. Twenty five semen samples were collected from breeding bulls and used in the study. Motility analysis was carried out using the Computer Assisted Sperm Analysis (CASA) system. The samples were centrifuged, fractions of seminal plasma and spermatozoa were separated, lysates were prepared from the sperm cell fractions. Pb and Cd concentrations were determined by the voltametric method (ASV), antioxidants and MDA were analyzed by UV/Vis spectrophotometry. The analysis showed that the average concentration of Pb in the seminal plasma was 0.23 ± 0.02 μg/mL, while its amount in the sperm cells was significantly higher (0.41 ± 0.07 μg/mL; P 0.05). The correlation analysis revealed that both heavy metals were significantly negatively correlated with MOT and PROG (P male fertility.

  5. In vitro production of Sudanese camel (Camelus dromedarius) embryos from epididymal spermatozoa and follicular oocytes of slaughtered animals.

    Science.gov (United States)

    Abdelkhalek, A E; Gabr, Sh A; Khalil, W A; Shamiah, Sh M; Pan, L; Qin, G; Farouk, M H

    2017-03-28

    Application of assisted reproductive technology in camelidea, such as artificial insemination (AI) and embryo transfer, has been slow in comparison to that for other livestock species. In Egypt, there are few attempts to establish in vitro maturation (IVM) and fertilization (IVF) techniques in dromedary camel. The present study was carried out to produce Sudanese camel embryos using in vitro matured oocytes and epididymal spermatozoa. Dromedary camel ovaries were collected from abattoirs and then, the oocytes were aspirated from all the visible follicles on the ovarian surface (~2-8 mm in a diameter). Meanwhile, Fetal Dromedary Camel Serum (FDCS) was obtained from camel fetuses after slaughtering. Thereafter, only Cumulus Oocyte Complexes (COCs) were matured in vitro in the Tissue Culture Medium (TCM-199) complemented with 10% FDCS. Spermatozoa required for in vitro fertilization were collected from testes (epididymal cauda) of the slaughtered camel bulls. The results clearly showed that the maturation rate of oocytes at metaphase II was about 59.5% while the fertilization rate was around 70.4%. Intriguingly, the embryo rates determined were 13.1%, in 2-cell; 0.0%, in 4-cell; 34.7%, in 8-16% cell; 39.1%, in morula and 13.1% in a blastocyst stage. This study represented a successful in vitro production of Sudanese dromedary camel embryos from epididymal sperm cells and in vitro matured oocytes recovered from slaughtered camels.

  6. The effect of paternal bull on milk fat composition of dairy cows of different breeds

    Directory of Open Access Journals (Sweden)

    Katarína Kirchnerová

    2015-05-01

    Full Text Available Intake of milk fat in human nutrition is important because of unsaturated and especially essential fatty acids (FAs, linoleic and α-linolenic acid, and conjugated linoleic acid (CLA, which is found only in meat and milk of ruminants. The objective of our study was to investigate the effect of paternal bulls on fatty acids composition in milk fat of dairy cows of different breeds. The milk samples were taken in total from 299 dairy cows from 11 dairy farms. In experiment Holstein (H, n = 105, Red Holstein (R, n = 120 and Pinzgau (P, n = 74 breeds originated from different bulls were used. Individual milk samples were analyzed for fatty acids in milk fat using gas chromatography (apparatus GC Varian 3800, Techtron, USA, using FID detector in capillary column Omegawax 530; 30 m. In the chromatography records there were identified 54 fatty acids inclusive of particular isomers. Their relative proportions were expressed in percent's (%. Among the studied breeds, the highest content of conjugated linoleic acid (CLA - 0.67%, essential FAs (EFA - 2.98%, monounsaturated FAs (MUFA - 25.84% and the lowest atherogenic index (AI - 3.10 was at breed P. Within this breed there was high variability and daughters of bull COS1 achieved significant above-average values of CLA content 1.07%, EFA 3.71%, MUFA 29.93% and under breed average AI = 2.40. The group of daughters of NOB3 was significant lower in CLA, 0.50% as compared with an average of P breed. . From the breed H bull MTY2 showed significantly higher value of 0.62% CLA, EFA 3.42%, 34.29% MUFA and lower value of AI, 1.9 as compared to H breed average. Statistically significantly lower levels of CLA 0.29% and 21.46% MUFA and higher AI 3.72 in milk fat of his daughters, bull STY3 may be considered as potential worser of these properties. At the breed R bull MOR506 showed in compar to the breed average significantly higher value of the EFA 3.80% and also the higher content of CLA 0.50% and MUFA 25

  7. Dysregulation of long noncoding RNAs in mouse testes and spermatozoa after exposure to cadmium.

    Science.gov (United States)

    Gao, Fengxin; Zhang, Peng; Zhang, Hongyan; Zhang, Yunhui; Zhang, Yunwen; Hao, Qingyun; Zhang, Xiaoning

    2017-02-26

    There is increasing evidence that cadmium (Cd) exposure can cause male subfertility and even complete infertility in mammals. Long noncoding (lnc) RNAs are critical for spermatogenesis, and their dysregulation might lead to male infertility. However, whether they are involved in Cd-induced subfertility is unknown. Here we found that intraperitoneal exposure to Cd in mice led to male subfertility indicated by reductions in testicular sperm production and motility, and by abnormal morphology. Testicular and sperm RNAs were used to investigate lncRNA expression profiles by strand-specific RNA sequencing at the transcriptome level to help determine any RNA-related mechanisms in Cd-induced subfertility. The Cd-treated testes and spermatozoa exhibited aberrant expression profiles for lncRNAs and mRNAs. Of the lncRNAs, there were 139 with upregulated expression and 174 with downregulated expression in testes; in contrast, 685 were upregulated and 375 were downregulated in spermatozoa. For mRNA expression, 214 were upregulated and 226 were downregulated in testes; 272 were upregulated and 111 were downregulated in spermatozoa. Gene ontology and pathway analyses showed that the functions of differentially expressed lncRNA targets and mRNAs were closely linked with many processes involved in spermatogenesis. Additionally, many newly identified lncRNAs showed inducible expression, suggesting that they might be good candidate markers for Cd-induced male reproductive toxicity. This study provides a preliminary database for further exploring lncRNA-related mechnisms in male infertility induced by Cd. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Assessment of bovine spermatozoa viability using different cooling protocols prior to cryopreservation

    Directory of Open Access Journals (Sweden)

    Paulo B.D. Gonçalves

    2010-11-01

    Full Text Available The aim of our study was to evaluate the effect of different cooling rates on the post-thawing quality of bovine spermatozoa. Ejaculated semen from a 24-month-old Jersey bull was collected using an artificial vagina and diluted in a commercial extender to evaluate spermatozoan concentration and motility subjectively before cooling and freezing and after thawing. Straws were allocated to four cooling curves: rapid (RD, semi-rapid (SRD, semi-slow (SSLW and slow (SLW. The temperature was decreased from 25ºC to 4ºC in 10, 50, 110 and 135 min, which represents a cooling rate of 2.06, 0.40, 0.18 and 0.15ºC/min, respectively. Then straws were frozen and stored at -196ºC. After thawing, one aliquot of each straw was used for evaluation. Spermatozoan integrity and mitochondrial function were evaluated using a combination of fluorescent probes containing 100 mg/mL FITC-PSA, 0.5 μg/mL PI and 153 μM JC-1. At the end of cooling, spermatozoan motility did not differ among RD (63.3%, SRD (66.7%, SSLW (66.7% and SLW (80.0%. However, normal spermatozoan morphology was lower in SRD (84.8% compared to RD (91.7%, SSLW (91.7% and SLW (90.3% (P<0.05. In thawed semen, spermatozoan motility and normal morphology did not differ among RD (40.0%; 88.8%, SRD (43.3%; 82.5%, SSLW (40.0%; 87.2% and SLW (36.7%; 88.0%. The percentage of damaged spermatozoa, including plasma and acrosome membrane damage and low mitochondrial potential, was higher in RD compared to the others (P<0.05. In conclusion, a rapid cooling curve is detrimental to the spermatozoa and affects the post-thaw spermatozoan integrity of bovine frozen semen.

  9. Optimizing age of bull at first use in relation to fertility of Murrah breeding bulls

    Directory of Open Access Journals (Sweden)

    M. A. Mir

    2015-04-01

    Full Text Available Aim: The aim of the present investigation was to optimize the age at first use (AAFU of semen of Murrah breeding bulls, which will help in early selection of bulls under progeny testing program for improving the reproductive performance in the herd. Materials and Methods: The data on AAFU, conception rate based on first A.I. (CRFAI, overall conception rate (OCR, and birth weight (B.WT of 57 Murrah bulls during 1993-2014 at NDRI center pertaining to 14 sets of Network Project on Buffalo Improvement at ICAR-National Dairy Research Institute, Karnal, Haryana, India were adjusted for significant environmental influences and subsequently analyzed. Simple and multiple regression models were used for prediction of CRFAI and OCR of Murrah breeding bulls. Comparative evaluation of three developed models (I-III showed that Model III, having AAFU and B.WT, fulfill the accuracy of model as revealed by high coefficient of determination, low mean sum of squares due to error, low conceptual predictive value, and low Bayesian information criterion. Results: The results revealed that the average predicted CRFAI was highest (39.95% at 4.5 years of age at first A.I/use. Similarly, average predicted OCR was highest (41.05% at 4.5 years of age at first A.I/use of Murrah bulls. Conclusion: In organized herd under progeny testing program, Murrah bulls should be used at young age, i.e. prior to 3.5 years, which is expected to result in 5.08% better CRFAI and 1.63% better OCR in comparison to Murrah bulls used after 4.5 years of age.

  10. Optimizing age of bull at first use in relation to fertility of Murrah breeding bulls.

    Science.gov (United States)

    Mir, M A; Chakravarty, A K; Gupta, A K; Naha, B C; Jamuna, V; Patil, C S; Singh, A P

    2015-04-01

    The aim of the present investigation was to optimize the age at first use (AAFU) of semen of Murrah breeding bulls, which will help in early selection of bulls under progeny testing program for improving the reproductive performance in the herd. The data on AAFU, conception rate based on first A.I. (CRFAI), overall conception rate (OCR), and birth weight (B.WT) of 57 Murrah bulls during 1993-2014 at NDRI center pertaining to 14 sets of Network Project on Buffalo Improvement at ICAR-National Dairy Research Institute, Karnal, Haryana, India were adjusted for significant environmental influences and subsequently analyzed. Simple and multiple regression models were used for prediction of CRFAI and OCR of Murrah breeding bulls. Comparative evaluation of three developed models (I-III) showed that Model III, having AAFU and B.WT, fulfill the accuracy of model as revealed by high coefficient of determination, low mean sum of squares due to error, low conceptual predictive value, and low Bayesian information criterion. The results revealed that the average predicted CRFAI was highest (39.95%) at 4.5 years of age at first A.I/use. Similarly, average predicted OCR was highest (41.05%) at 4.5 years of age at first A.I/use of Murrah bulls. In organized herd under progeny testing program, Murrah bulls should be used at young age, i.e. prior to 3.5 years, which is expected to result in 5.08% better CRFAI and 1.63% better OCR in comparison to Murrah bulls used after 4.5 years of age.

  11. 78 FR 40079 - Special Local Regulations; Red Bull Flugtag Miami, Biscayne Bay; Miami, FL

    Science.gov (United States)

    2013-07-03

    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Red Bull Flugtag Miami... Park, in Miami, Florida, during the Red Bull Flugtag event. The Red Bull Flugtag is scheduled to take... United States during the Red Bull Flugtag. C. Discussion of Proposed Rule On September 21, 2013, Red Bull...

  12. 77 FR 60302 - Special Local Regulations; Red Bull Flugtag Miami, Biscayne Bay; Miami, FL

    Science.gov (United States)

    2012-10-03

    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Red Bull Flugtag Miami... during the Red Bull Flugtag. The Red Bull Flugtag is scheduled to take place on Saturday, November 3... United States during the Red Bull Flugtag. C. Discussion of the Final Rule On November 3, 2012, Red Bull...

  13. DNA integrity of fresh and frozen canine epididymal spermatozoa.

    Science.gov (United States)

    Varesi, Sara; Vernocchi, Valentina; Morselli, Maria Giorgia; Luvoni, Gaia Cecilia

    2014-12-01

    The aims of this study were to evaluate: (1) the effect of cryopreservation on DNA fragmentation of canine epididymal spermatozoa, and (2) the potential protective effect of melatonin on post-thaw sperm quality (motility, morphology, acrosomal and DNA integrity). Epididymal spermatozoa were collected after orchiectomy of ten dogs. Sperm samples were frozen in the presence or absence of melatonin (1mM). DNA fragmentation index (percentage of spermatozoa with fragmented DNA) was similar in fresh samples (3.3±3.6) and samples frozen with (4.2±3.8) or without (3.6±3.7) melatonin. Sperm motility was significantly (psperm motility. Proportions of spermatozoa with normal morphology were similar in fresh and frozen samples, irrespective of the presence of melatonin in the extender. Acrosome integrity was significantly decreased (pepididymal spermatozoa was not affected by the freezing procedure, and the presence of melatonin did not preserve motility and acrosome integrity which were adversely affected by cryopreservation. The evaluation of DNA status of thawed gametes is particularly relevant for epididymal spermatozoa since these spermatozoa are usually stored and used in assisted reproductive techniques. Copyright © 2014 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  14. Organic meat quality of dual purpose young bulls supplemented with pea (Pisum sativum L.) or soybean.

    Science.gov (United States)

    Corazzin, Mirco; Piasentier, Edi; Saccà, Elena; Bazzoli, Ilario; Bovolenta, Stefano

    2018-02-01

    One of the main constraints established by organic legislation that limits the development of the rearing of young bulls is the ban on the use of genetically modified organisms (GMO). Most of the worldwide cultivated soybean is GMO, therefore the use of alternative protein sources should be evaluated. In this study, the effect of dietary substitution of soybean with pea (Pisum sativum L.) on carcass characteristics and meat quality of dual purpose young bulls reared following the organic method was investigated. Twenty-four young bulls of Rendena breed were randomly assigned to two diet treatments differing in protein supplement (soybean (SB) or field pea (FP)). Carcass characteristics and meat chemical composition, colour, cooking loss and Warner-Bratzler shear force did not differ between groups. Regarding meat fatty acid composition, SB showed higher concentrations of C18:0 and C18:1 t and lower C16:1n-9c, C14:0, C17:1n-9c and C18:1n-9c than FP. In descriptive sensory analysis, trained judges were not able to differentiate meats from SB and FP, which also had similar overall liking expressed by consumers. The results of this study indicate that FP can replace SB in the diet of dual purpose young bulls with only a minor influence on fatty acid composition and no effect on carcass characteristics and meat quality. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  15. Morphological and acrosomal changes of canine spermatozoa during epididymal transit

    Science.gov (United States)

    2013-01-01

    Background During epididymal transit, functional and structural modifications leading to full maturation enable male gametes to reach, recognize and fertilize the oocytes. In dogs, little is known on the modifications of spermatozoa during the passage in the epididymis. The aim of this study was to describe the motility, morphology and acrosomal patterns of canine spermatozoa retrieved from the epididymis caput, corpus and cauda. Results After the dilution required for the collection of epididymal content, sperm motility was significantly higher (P orchiectomy for medical purposes. Further investigations should be also focused on the potential use of spermatozoa recovered from other epididymal regions. PMID:23442223

  16. Spermatozoa protein alterations in infertile men with bilateral varicocele

    Directory of Open Access Journals (Sweden)

    Ashok Agarwal

    2016-01-01

    Full Text Available Among infertile men, a diagnosis of unilateral varicocele is made in 90% of varicocele cases and bilateral in the remaining varicocele cases. However, there are reports of under-diagnosis of bilateral varicocele among infertile men and that its prevalence is greater than 10%. In this prospective study, we aimed to examine the differentially expressed proteins (DEP extracted from spermatozoa cells of patients with bilateral varicocele and fertile donors. Subjects consisted of 17 men diagnosed with bilateral varicocele and 10 proven fertile men as healthy controls. Using the LTQ-orbitrap elite hybrid mass spectrometry system, proteomic analysis was done on pooled samples from 3 patients with bilateral varicocele and 5 fertile men. From these samples, 73 DEP were identified of which 58 proteins were differentially expressed, with 7 proteins unique to the bilateral varicocele group and 8 proteins to the fertile control group. Majority of the DEPs were observed to be associated with metabolic processes, stress responses, oxidoreductase activity, enzyme regulation, and immune system processes. Seven DEP were involved in sperm function such as capacitation, motility, and sperm-zona binding. Proteins TEKT3 and TCP11 were validated by Western blot analysis and may serve as potential biomarkers for bilateral varicocele. In this study, we have demonstrated for the first time the presence of DEP and identified proteins with distinct reproductive functions which are altered in infertile men with bilateral varicocele. Functional proteomic profiling provides insight into the mechanistic implications of bilateral varicocele-associated male infertility.

  17. Proteomics: a subcellular look at spermatozoa

    Directory of Open Access Journals (Sweden)

    Yadav Satya P

    2011-03-01

    Full Text Available Abstract Background Male-factor infertility presents a vexing problem for many reproductively active couples. Many studies have focused on abnormal sperm parameters. Recent advances in proteomic techniques, especially in mass spectrometry, have aided in the study of sperm and more specifically, sperm proteins. The aim of this study was to review the current literature on the various proteomic techniques, and their usefulness in diagnosing sperm dysfunction and potential applications in the clinical setting. Methods Review of PubMed database. Key words: spermatozoa, proteomics, protein, proteome, 2D-PAGE, mass spectrometry. Results Recently employed proteomic methods, such as two-dimensional polyacrylamide gel electrophoresis, mass spectrometry, and differential in gel electrophoresis, have identified numerous sperm-specific proteins. They also have provided a further understanding of protein function involved in sperm processes and for the differentiation between normal and abnormal states. In addition, studies on the sperm proteome have demonstrated the importance of post-translational modifications, and their ability to bring about physiological changes in sperm function. No longer do researchers believe that in order for them to elucidate the biochemical functions of genes, mere knowledge of the human genome sequence is sufficient. Moreover, a greater understanding of the physiological function of every protein in the tissue-specific proteome is essential in order to unravel the biological display of the human genome. Conclusion Recent advances in proteomic techniques have provided insight into sperm function and dysfunction. Several multidimensional separation techniques can be utilized to identify and characterize spermatozoa. Future developments in bioinformatics can further assist researchers in understanding the vast amount of data collected in proteomic studies. Moreover, such advances in proteomics may help to decipher metabolites

  18. Proteomics: a subcellular look at spermatozoa.

    Science.gov (United States)

    du Plessis, Stefan S; Kashou, Anthony H; Benjamin, David J; Yadav, Satya P; Agarwal, Ashok

    2011-03-22

    Male-factor infertility presents a vexing problem for many reproductively active couples. Many studies have focused on abnormal sperm parameters. Recent advances in proteomic techniques, especially in mass spectrometry, have aided in the study of sperm and more specifically, sperm proteins. The aim of this study was to review the current literature on the various proteomic techniques, and their usefulness in diagnosing sperm dysfunction and potential applications in the clinical setting. Review of PubMed database. Key words: spermatozoa, proteomics, protein, proteome, 2D-PAGE, mass spectrometry. Recently employed proteomic methods, such as two-dimensional polyacrylamide gel electrophoresis, mass spectrometry, and differential in gel electrophoresis, have identified numerous sperm-specific proteins. They also have provided a further understanding of protein function involved in sperm processes and for the differentiation between normal and abnormal states. In addition, studies on the sperm proteome have demonstrated the importance of post-translational modifications, and their ability to bring about physiological changes in sperm function. No longer do researchers believe that in order for them to elucidate the biochemical functions of genes, mere knowledge of the human genome sequence is sufficient. Moreover, a greater understanding of the physiological function of every protein in the tissue-specific proteome is essential in order to unravel the biological display of the human genome. Recent advances in proteomic techniques have provided insight into sperm function and dysfunction. Several multidimensional separation techniques can be utilized to identify and characterize spermatozoa. Future developments in bioinformatics can further assist researchers in understanding the vast amount of data collected in proteomic studies. Moreover, such advances in proteomics may help to decipher metabolites which can act as biomarkers in the detection of sperm impairments and

  19. Polymorphism in TNP-1 gene of Murrah buffalo bulls

    African Journals Online (AJOL)

    hope&shola

    medium supplemented with 10% egg albumin. The spermatozoa were centrifuged at 300 rpm for 10 min; the pellet was layered with. 3 to 4 drops of Ham's Fl0-medium in Eppendorf tubes and incubated at 37°C in an atmosphere of 5% CO2. Subsequent to incubation, 5 µl spermatozoa were spread onto glass slide and ...

  20. Bovine epididymal spermatozoa: Resistance to cryopreservation and binding ability to oviductal cells.

    Science.gov (United States)

    Cunha, A T M; Carvalho, J O; Kussano, N R; Martins, C F; Mourão, G B; Dode, M A N

    2016-12-01

    In this study we examined quality, longevity and ability of epididymal sperm (EP) to bind to oviduct explants (OE) after cooling and cryopreservation. Ejaculated (EJ) and EP sperm from seven bulls were evaluated before, during and after cryopreservation for total (TM), progressive motility (PM), sperm morphology, plasma membrane integrity (PMI) and acrosome integrity (ACI). For longevity, cryopreserved EP, EJ and a third group of cells in which EP spermatozoa were incubated with seminal plasm (SP) for 10 min after thawing (EPP group), were compared, and the groups were analyzed at 0, 3, 6, and 24 h for all parameters. Sperm from each group were co-incubated with OE for 30 min, 6 h, and 24 h for binding evaluation. Data were analyzed by the generalized linear models SAS 9.1 (P sperm. However, a reduction in motility occurred in the EJ sperm after cooling, while in EP group such reduction occurred only after cryopreservation. At 6 h of incubation EP and EPP had higher PMI and ACI than EJ (P  0.05) for all groups either at 30 min or 24 h. We conclude that EP are more resistant to cooling than EJ, and can bind to OE similarly to EJ. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Effect of lactose, skim milk and Tris diluents on frozen buffalo spermatozoa

    Directory of Open Access Journals (Sweden)

    Ali Rastegarnia

    2007-08-01

    Full Text Available The composition of the extender in which semen is diluted before freezing plays a major role in successful cryopreservation of spermatozoa. This study was carried out to identify the suitable buffer for cryopreservation of buffalo semen. Sixteen split pooled ejaculates from two buffalo bulls possessing more than 70% visual sperm motility, were diluted at 370c either in lactose, skim milk or Tris extenders. The diluted semen was cooled to 40c within 2 hours, equilibrated at 40c for 4-6 hours following the addition of glycerol, filled in 0.5 ml French straws and frozen in a programmable cell freezer before plunging into liquid nitrogen. Semen was thawed at 370c for 30 seconds after 48 hours of storage inside liquid nitrogen. Post thaw visual sperm motility, plasma membrane integrity and acrosome morphology of each semen sample were assessed by warm plate microscopy at 370c, hypo-osmotic swelling test (HOST and giemsa staining, respectively. Analysis of variance revelated that percentage of post thaw visual sperm motility (Mean± standard deviation tended to be higher in Tris (50±3.6 than skim milk (44.5±2.5 and lower in lactose (24.4±10.5 extenders (P

  2. Effect of various levels of catalase antioxidant in semen extenders on lipid peroxidation and semen quality after the freeze-thawing bull semen

    Directory of Open Access Journals (Sweden)

    Reza Asadpour

    2011-11-01

    Full Text Available The objective of this study was to evaluate effect of different concentrations of catalase in two extenders on motility, viability and lipid peroxidation bull spermatozoa during semen freezing process. Thirty ejaculates collected from ten Holstein bulls were pooled and evaluated at 37 °C. Pool ejaculated was split into two main experimental groups, 1 and 2. In experiment 1, specimen was diluted to a final concentration of 30 × 106 spermatozoa with citrate-egg yolk and in experiment 2; specimen was diluted with tris-egg yolk extender to the same concentration. In both experiments diluted semen was divided into three aliquots, including a control and two test groups. Each aliquot was rediluted with an equal volume of extender either without (control or with one of the antioxidants contained one of the following antioxidants: catalase (CAT; 100 IU mL-1 catalase (CAT; 200 IU mL-1 and control group. No significant differences were observed in sperm viability and motility following addition of catalase enzyme at concentration of 100 IU mL-1 and 200 IU mL-1 to citrate-egg yolk extender. But the highest sperm viability was achieved by addition of 100 IU mL-1 and 200 IU mL-1 catalase to tris-egg yolk semen extender compared with the control group (P < 0.05. Malondialdehyde levels did not change with addition of catalase in both extenders compared with the control group. The obtained results provide a new approach to the cryopreservation of bull semen, and could positively contribute to intensive cattle production.

  3. Debunking the Effects of Taurine in Red Bull Energy Drink

    OpenAIRE

    Kim, Woojae

    2003-01-01

    Red Bull is a carbonated beverage that initially gained wide popularity in the U.S. during the late nineties. Taking root amongst college campuses, it appeared throughout underground clubs and eventually entered mainstream pop-culture. The manufactures claim that drinking Red Bull enhances physical endurance, concentration and reaction speed (1,6). The main ingredients of Red Bull include sugar, taurine, glucuronolactone and caffeine. It is hypothesized that the combinatorial influences o...

  4. Lactate dehydrogenase activity of rat epididymis and spermatozoa: Effect of constant light

    Directory of Open Access Journals (Sweden)

    RH Ponce

    2009-12-01

    Full Text Available During its passage through the epididymis, the gamete undergoes a process of “maturation” leading to the acquisition of its fertilizing ability. The epididymis displays regional variations in the morphology and metabolic properties of its epithelium which are relevant for the progressive development of mature sperm characteristics. The epididymis has spontaneous peristaltic contractions and receives sympathetic innervation that is modulated by melatonin, a hormone synthesized and released by the pineal gland. Constant lighting disrupts melatonin synthesis and secretion. We have studied the effect of constant light on lactate dehydrogenase (LDH; EC 1.1.1.27 and its isozyme C4 activities and protein content in whole epididymis, epididymal tissue and in spermatozoa from caput and cauda segments. Animals were exposed from birth to an illumination schedule of 14 h light: 10 h dark (group L:D. At 60 days of age one group of animals was submitted to constant light over 50 days (group L:L. In order to test the fertilizing ability, the rats of each group were mated with soliciting estrous females. The percentage of pregnancies in females mated with males maintained in L:L was remarkably lower than those in females mated with males maintained in the L:D photoperiod (44% and 88% respectively. Constant light increased protein concentration and LDH activity in caput as well as in cauda of total epididymis. On the contrary, in epididymal tissue, the protein content decreased in both epididymal sections compared with controls. When enzymatic activity was expressed in Units per spermatozoa, constant light induced a significant reduction of total LDH and LDHC4 in caput and cauda spermatozoa while LDH activity of epididymal tissue was not affected. In spite of the decrease in LDH per sperm cell when rats were exposed to constant light, in total epididymis (epididymis tissue plus sperm cells content and in spermatozoa, values of enzyme activities expressed per

  5. MOLECULAR ANALYSIS OF HUMAN SPERMATOZOA: POTENTIAL FOR INFERTILITY RESEARCH

    Science.gov (United States)

    Gordon Research Conference: Mammalian Gametogenesis and Embryogenesis New London, CT, July 1-6, 2000Molecular Analysis of Human Spermatozoa: Potential for Infertility ResearchDavid Miller 1, David Dix2, Robert Reid 3, Stephen A Krawetz 3 1Reproductive ...

  6. D-penicillamine and granulosa cells can effectively extend the fertile life span of bovine frozen-thawed spermatozoa in vitro: effect on fertilization and polyspermy.

    Science.gov (United States)

    Pavlok, A

    2000-03-15

    The effect of D-penicillamine on the fertile life span of frozen-thawed bull spermatozoa was studied in Experiment 1. After thawing, the washed spermatozoa were incubated for 8 h in fertilization medium with 1 mg/mL polyvinyl alcohol (PVA) and 0.5 mg/mL D-penicillamine. The addition of cumulus-free oocytes together with 10% of bovine serum (BOS) + 4 IU/mL heparin to the 8-h incubated spermatozoa resulted in high fertilization and polyspermy rates (97/103, 94.2% and 49/97, 50.5%, respectively). When BOS was substituted with 3 mg/mL of BSA, the fertilization and polyspermy rates decreased to 33/91 (36.3%) and 3/33 (9.1%), respectively. The total absence of fertilization was observed after substitution of proteins with PVA. The 8-h sperm incubation in fertilization medium without D-penicillamine and following fertilization in medium + different capacitation supplements resulted in the total absence of or a very low fertilization rate. In Experiment 2, the first set of cumulus-oocyte complexes (COC) and cumulus-free oocytes were fertilized for 8 h with nonincubated spermatozoa. The second set of COC and cumulus-free oocytes were fertilized in the same wells with spermatozoa after removal of the first set. The fertilization rate for the first set of COC and cumulus-free oocytes was 65/67 (97%) and 73/73 (100%), respectively, with 21/65 (32.3%) and 32/73 (43.8%) polyspermy, respectively. In the second set, the high penetration rate (67/73, 91.8%) was observed only for COC, while that for cumulus-free oocytes (19/76, 25%) was significantly lower (P polyspermy. In medium + BSA + 10 or 100 IU/mL heparin, the fertilization rate of COC was 64/72 (88.9%) and 70/79 (88.6%), respectively, with polyspermy at 2/64 (3.1%) and 7/70 (10%), respectively. In medium + BOS + 10 or 100 IU/mL heparin, the fertilization rate was 79/82 (96.3%) and 79/80 (98.7%), respectively, with a significantly (P polyspermy rate (34/79, 43% and 30/79, 38%, respectively). The vitality and capacitation

  7. Artificial neural networks for the definition of kinetic subpopulations in electroejaculated and epididymal spermatozoa in the domestic cat.

    Science.gov (United States)

    Contri, Alberto; Zambelli, Daniele; Faustini, Massimo; Cunto, Marco; Gloria, Alessia; Carluccio, Augusto

    2012-09-01

    This study was designed for the identification of different sperm kinetic subpopulations in feline semen using artificial neural networks (ANNs) and for the evaluation of the effect of ejaculation on motility patterns of these subpopulations. Seven tomcats presented for routine orchiectomy were electroejaculated, and after 5 days, orchiectomized and epididymal tail sperms were collected. Sperm motility characteristics were evaluated using a computer-assisted sperm analyzer that provided individual kinetic characteristics of each spermatozoon. A total of 23 400 spermatozoa for electroejaculated and 9200 for epididymal tail samples were evaluated using a multivariate approach, comprising principal component analysis and ANN classification. The multivariate approach allowed the identification and characterization of three different and well-defined sperm subpopulations. There were significant differences before (epididymal tail spermatozoa) and after (electroejaculated sperm) ejaculation in sperm kinetic subpopulation characteristics. In both epididymal and ejaculated samples, the majority of subpopulation was characterized by high velocity and progressiveness; however, the electroejaculated samples showed significantly higher values, suggesting that the microenvironment of the epididymal tail could affect the sperm motility or, alternatively, seminal plasma could increase the kinetic characteristics of the spermatozoa, indicating that only after ejaculation, the spermatozoa express their motility potential. Nevertheless, further studies are required to clarify the functional significance of each kinetic subpopulation.

  8. Cryopreservation of spermatozoa of black marlin, Makaira indica (Teleostei: Istiophoridae).

    Science.gov (United States)

    van der Straten, K M; Leung, L K-P; Rossini, R; Johnston, S D

    2006-01-01

    As a first step towards the development of a method for the cryopreservation of black marlin spermatozoa, this study investigated the effect of dimethylsulfoxide (DMSO) concentration and pellet size on post-thaw spermatozoal motility. Spermatozoa were recovered from the spermatic duct of testes retrieved post-mortem from four adult black marlin caught in the Coral Sea spawning grounds of Australia. Undiluted spermatozoa were stored on ice for 4 to 10 hours during transport to shore, then evaluated for motility after activation in seawater (1:10 v:v). Spermatozoa were prepared for cryopreservation in pellets by extension (1:3 v:v) in a defined fish Ringer's solution to give two final DMSO concentrations of 2.5% or 5.0%. Diluted spermatozoa were frozen directly on a dry ice block in pellet sizes of either 0.25 ml or 0.50 ml. Frozen pellets were thawed in a water bath at 40 degrees C for 60 seconds and assessed for post-thaw motility following activation in seawater. Spermatozoa recovered within 50 minutes of death and chilled on ice for 4 to 10 hours showed a mean (+/- SEM) motility immediately following activation of 91.6 +/- 7.9%. 50% of the spermatozoa remained motile for approximately 4 to 5 minutes. Following cryopreservation, mean motility declined significantly across all cryoprotectant and pellet size combinations (P < 0.001) but spermatozoa frozen in 2.5% DMSO showed higher motility than those frozen in 5.0% DMSO (P = 0.014). Pellet size had no effect on post-thaw motility (P = 0.179).

  9. Papaverine-sensitive phosphodiesterase activity is measured in bovine spermatozoa.

    Science.gov (United States)

    Bergeron, A; Hébert, A; Guillemette, C; Laroche, A; Poulin, M-P; Aragon, J P; Leclerc, P; Sullivan, R; Blondin, P; Vigneault, C; Richard, F J

    2017-01-01

    Cyclic adenosine monophosphate (cAMP) plays a crucial role as a signaling molecule for capacitation, motility, and acrosome reaction in mammalian spermatozoa. It is well-known that cAMP degradation by phosphodiesterase (PDE) enzyme has a major impact on sperm functions. This study was undertaken to characterize cAMP-PDE activity in bovine spermatozoa. Total cAMP-PDE activity in cauda epididymal and ejaculated spermatozoa was 543.2 ± 49.5 and 1252.6 ± 86.5 fmoles/min/10(6) spermatozoa, respectively. Using different family-specific PDE inhibitors, we showed that in cauda epididymal and ejaculated spermatozoa, the major cAMP-PDE activity was papaverine-sensitive (44.5% and 57.5%, respectively, at 400 nm, papaverine is a specific inhibitor of the PDE10 family). These data are supporting the functional presence of PDE10 in bovine spermatozoa and were further confirmed by western blot to be PDE10A. Using immunocytochemistry, we showed immunoreactive signal for PDE10A present on the post-acrosomal region of the head and on the flagella of ejaculated spermatozoa. Using papaverine, we showed that it promotes tyrosine phosphorylation of sperm proteins, phosphorylation of Erk1 and Erk2, and Ca(2+) release from Ca(2+) store. These results suggest that PDE10 is functionally present in bovine spermatozoa and is affecting different molecular events involved in capacitation, most probably by cAMP local regulation. © 2016 American Society of Andrology and European Academy of Andrology.

  10. The effect of mature elephant bull introductions on ranging patterns of resident bulls: Phinda Private Game Reserve, South Africa

    Directory of Open Access Journals (Sweden)

    Heleen Druce

    2006-12-01

    Full Text Available Increasing popularity of wildlife viewing has resulted in a rapid increase in small, enclosed reserves in South Africa. The African elephant is one of the many species that has been reintroduced into these reserves for eco-tourism. These elephant populations were established as young (smaller that 10 years old orphans from prior Kruger National Park culling operations. Consequently, this abnormal sex and age structure of these introduced populations has influenced their behavioural and spatial ecology. In Pilanesberg National Park, this abnormal behaviour was corrected by introducing older bulls and culling certain problem elephants. In July 2003, three older bulls (29–41 years old were introduced into Phinda Private Game Reserve, KwaZulu-Natal, South Africa in order to normalise the bull age structure. These introduced bulls were monitored intensively after release, as was the resident bull population, both before and after introduction of the older bulls. The introduced bulls settled into restricted ranges separate from the family groups. All the resident bulls decreased their home ranges at first, with most increasing their home ranges a year later. The resident bulls’ change in ranging patterns was due more to ecological factors than to the influence of the mature bull introduction. This study indicates that the introduction of older male elephants into small populations does not pose major risks or animal welfare concerns.

  11. REAC technology as optimizer of stallion spermatozoa liquid storage.

    Science.gov (United States)

    Berlinguer, Fiammetta; Pasciu, Valeria; Succu, Sara; Cossu, Ignazio; Caggiu, Sabrina; Addis, Daniela; Castagna, Alessandro; Fontani, Vania; Rinaldi, Salvatore; Passino, Eraldo Sanna

    2017-02-08

    REAC technology (acronym for Radio Electric Asymmetric Conveyor) is a technology platform for neuro and bio modulation. It has already proven to optimize the ions fluxes at the molecular level and the molecular mechanisms driving cellular asymmetry and polarization. This study was designed to verify whether this technology could extend spermatozoa life-span during liquid storage, while preserving their functions, DNA integrity and oxidative status. At 0, 24, 48, and 72 h. of storage at 4 °C, a battery of analyses was performed to assess spermatozoa viability, motility parameters, acrosome status, and DNA integrity during REAC treatment. Spermatozoa oxidative status was assessed by determining lipid peroxidation, the activity of superoxide dismutase (SOD), and the total antioxidant capacity. During liquid storage REAC treated spermatozoa, while not showing an increased viability nor motility compared to untreated ones, had a higher acrosome (p > 0.001) and DNA integrity (p > 0.01). Moreover, the analysis of the oxidative status indicated that the mean activity of the intracellular superoxide dismutase (SOD) was significantly higher in REAC treated spermatozoa compared to untreated controls (p  0.05). The REAC efficacy on spermatozoa oxidative status was also evidenced by the higher trolox equivalent antioxidant capacity (TEAC) found in both the cellular extract (p management of male infertility.

  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. Calving distributions of individual bulls in multiple-sire pastures.

    Science.gov (United States)

    Abell, Kaitlynn M; Theurer, Miles E; Larson, Robert L; White, Brad J; Hardin, David K; Randle, Richard F; Cushman, Robert A

    2017-04-15

    The objective of this project was to quantify patterns in the calving rate of sires in multiple-sire pastures over seven years at a large-scale cow-calf operation. Data consisted of reproductive and genomic records from multiple-sire breeding pastures (n = 33) at the United States Meat Animal Research Center (USMARC) from 2007 to 2013. Calving intervals were analyzed in 21-day periods. A ranking system for each bull was developed based on the calving rate per pasture over the breeding season, with Rank 1 = the bull with greatest calving rate, Rank 3 = the bull with the least calving rate, and Rank 2 = all other bulls. A total of 179 bulls and 3703 calves were successfully genotyped over seven years. A uniform distribution described the expected percentage of calves sired per rank within pasture. Rank 1 bulls sired 113% greater calves than the expected pasture-average, Rank 2 bulls sired 6% less than expected, and Rank 3 bulls sired 81% less than expected. A rank by calving interval interaction effect was identified (P Rank 1 bull in calving interval 1 produced a greater average percent of the total calf crop over the entire season, compared to a Rank 2 and Rank 3 bull. The calving rate for individual sires is not homogeneous and there is a large difference between bulls siring the greatest and least number of calves. More research is needed to determine how rank changes over multiple breeding years and its association with dominance, libido, and fertility. Copyright © 2017. Published by Elsevier Inc.

  14. A minimum number of motile spermatozoa are required for successful fertilisation through artificial intrauterine insemination with husband's spermatozoa.

    Science.gov (United States)

    Cao, S; Zhao, C; Zhang, J; Wu, X; Zhou, L; Guo, X; Shen, R; Ling, X

    2014-06-01

    Intrauterine insemination (IUI) is an effective, noninvasive, relatively simple and cheap method of infertility treatment. Many factors that affect IUI outcomes have been studied. However, there is no consensus about the optimal number of motile spermatozoa inseminated (NMSI) required for a reasonable chance of pregnancy after IUI. In this retrospective study, we aimed to assess the relationship between NMSI and the pregnancy rate after IUI with husband's spermatozoa. Couples who had either primary or secondary infertility for more than one year were recruited from the Department of Reproduction, Nanjing Maternity and Child Health Hospital, China, between January 2007 and December 2010. Overall, 1153 IUI cycles with husband's spermatozoa were performed in 645 women after ovarian stimulation. Factors that have previously been associated with a successful fertilisation after IUI were assessed. A total pregnancy rate of 13.88% was obtained. The pregnancy rate was only 4.05% if less than 2 × 10(6) motile spermatozoa were used, but this rose to 14.55% when more than 2 × 10(6) motile spermatozoa were inseminated. We therefore conclude that IUI can be performed when the NMSI exceeds 2 × 10(6) . With this recommendation, IUI with husband's spermatozoa can be used to treat many more infertile couples. © 2013 Blackwell Verlag GmbH.

  15. Effect of Selected Factors on Qualitative and Quantitative Semen Parameters of Czech Fleckvieh Bulls

    Directory of Open Access Journals (Sweden)

    Michaela Paldusová

    2016-01-01

    Full Text Available The aim of the study was to estimate genetic characteristics for important qualitative and quantitative parameters of Czech Fleckvieh bulls’ ejaculate and to define factors which influence these parameters. a total number of 2,929 entries about samples from 163 bulls of a selected artificial insemination centre were used for calculation. the analysed ejaculate characteristics were: volume of ejaculate, sperm concentration, total sperm count, native sperm activity and activity after thawing. In all evaluated semen characteristics, the age of individual, coefficient of inbreeding, interval between colection and year of colection were statistically significant (P < 0.05. In the case of season of colection, statistically significance (P < 0.01 was proved in all monitored semen parameters, except concetration of spermatozoa where statistically significant difference was not found (P > 0.05. Estimated coefficients of heritability correspond to the values standardly specified for reproductive parameters (h2 = 0.003 – 0.14, except for the volume of ejaculate (h2 = 0.31. the highest value of genetic correlation was recorded between the volume of ejaculate and the total sperm count (rG = 0.9, on the contrary, the lowest value was between the volume of ejaculate and the sperm concentration (rG = 0.32.

  16. Cellulitis in a Red Kandhari Bull : A Case Report

    Directory of Open Access Journals (Sweden)

    M M Pathan

    2012-06-01

    Full Text Available A case of cellulitis caused by mixed infection of Staphylococcus spp. and Corynebacterium spp in a Red Kandhari bull leading to death of animal was autopsied at the department. It is being a case of cellulitis in a Red Kandhari bull and placed on record. [Vet. World 2012; 5(3.000: 183-184

  17. A watershed-scale monitoring protocol for bull trout

    Science.gov (United States)

    Dan Isaak; Bruce Rieman; Dona Horan

    2009-01-01

    Bull trout is a threatened species native to the Pacific Northwest that has been selected as Management Indicator Species on several national forests. Scientifically defensible procedures for monitoring bull trout populations are necessary that can be applied to the extensive and remote lands managed by the U.S. Forest Service. Distributional monitoring focuses...

  18. Multiple Congenital Defects In A Bunaji Bull | Ibrahim | Nigerian ...

    African Journals Online (AJOL)

    A congenitally malformed two- and-half year-old bull was acquired from Zaria abattoir for clinical and laboratory investigations. Clinical observations revealed that the bull had kyphoscoliosis and notomelia, with the two extra hind limbs attached to the right lumbosacral aspect, and a common hypoplastic pelvic bone for the ...

  19. Obesity and Bariatric Surgery Drive Epigenetic Variation of Spermatozoa in Humans

    DEFF Research Database (Denmark)

    Donkin, Ida; Versteyhe, Soetkin; Ingerslev, Lars R.

    2016-01-01

    Obesity is a heritable disorder, with children of obese fathers at higher risk of developing obesity. Environmental factors epigenetically influence somatic tissues, but the contribution of these factors to the establishment of epigenetic patterns in human gametes is unknown. Here, we hypothesized...... and offers insight into how obesity may propagate metabolic dysfunction to the next generation....... that weight loss remodels the epigenetic signature of spermatozoa in human obesity. Comprehensive profiling of the epigenome of sperm from lean and obese men showed similar histone positioning, but small non-coding RNA expression and DNA methylation patterns were markedly different. In a separate cohort...

  20. Effect of natural neosporosis on bull sperm quality.

    Science.gov (United States)

    Bahrami, Somayeh; Hamidinejat, Hossein; Fatemi-Tabatabaei, Seyed Reza; Sardarifar, Saeed

    2018-01-01

    Neospora is one of the protozoans that can infect the male and female's reproduction system. Despite the existence of N. caninum in the genitalia, its effect on sperm characteristics was not studied yet. Therefore, the aim of this study was to investigate the influence of natural neosporosis on the sperm parameters of bulls. Using 30 bulls with neosporosis diagnosed by modified agglutination test and enzyme-linked immunosorbent assay (ELISA) and 15 healthy bulls, some sperm parameters such as sperm concentration, viability, motility, and morphology were studied and compared. Also, the activity of super oxide dismutase (SOD), glutathione peroxidase (GPX), and malondialdehyde (MDA) level as the biomarker of lipid peroxidation was investigated. Results showed that sperm concentration, viability, and motility were significantly lower in bulls with neosporosis in the present study. There were no significant differences in activities of SOD and MDA level but GPX activity was significantly increased in infected bulls.

  1. A new reciprocal translocation in a subfertile bull

    Directory of Open Access Journals (Sweden)

    Darré Roland

    2000-11-01

    Full Text Available Abstract Three bulls of the Montbéliarde breed that exhibited fertility rates lower than 30% following more than 400 artificial inseminations were examined. Semen quality (sperm motility and morphology from these bulls was normal. Fertilizing ability estimated from in vitro embryo production results was studied for two of them. In vitro production rate was very low for one bull (A and normal for the other (B. Cytogenetic analyses were carried out on the three bulls using chromosome banding techniques. These analyses revealed a reciprocal translocation (12;17(q22;q14 in bull B. Based on family analyses, the hypothesis of a de novo origin of this rearrangement is proposed.

  2. Status of Oregon's Bull Trout.

    Energy Technology Data Exchange (ETDEWEB)

    Buchanan, David V.; Hanson, Mary L.; Hooton, Robert M.

    1997-10-01

    Limited historical references indicate that bull trout Salvelinus confluentus in Oregon were once widely spread throughout at least 12 basins in the Klamath River and Columbia River systems. No bull trout have been observed in Oregon's coastal systems. A total of 69 bull trout populations in 12 basins are currently identified in Oregon. A comparison of the 1991 bull trout status (Ratliff and Howell 1992) to the revised 1996 status found that 7 populations were newly discovered and 1 population showed a positive or upgraded status while 22 populations showed a negative or downgraded status. The general downgrading of 32% of Oregon's bull trout populations appears largely due to increased survey efforts and increased survey accuracy rather than reduced numbers or distribution. However, three populations in the upper Klamath Basin, two in the Walla Walla Basin, and one in the Willamette Basin showed decreases in estimated population abundance or distribution.

  3. Testicular Histomorphometric Evaluation of Zebu Bull Breeds

    Directory of Open Access Journals (Sweden)

    Paulo Antônio Terrabuio Andreussi

    2014-12-01

    Full Text Available The objective of this study was to evaluate the quantitative histology and testicular biometrics in zebu bulls of different breeds. Testicular fragments of Nelore (n=10, Polled Nelore (n=6, Gir (n=5, Guzerat (n=5 and Tabapuã bulls (n=5 were used. The fragments were perfusion-fixed in Karnovsky solution, embedded in glycol methacrylate and stained with toluidine blue-1% sodium borate. The Nelore animals had a higher tubular volumetric proportion (85.2% and greater height of the seminiferous epithelium (73.2 µm than the Gir, Guzerat and Tabapuã breeds. The Nelore animals also had a higher volumetric proportion of Leydig cells (5.2% than the Guzerat and Tabapuã breeds. There was no significant difference for any of these parameters between the Nelore and Polled Nelore breeds. The gonadosomatic index, seminiferous tubule diameter, cross-sectional area of the seminiferous tubule and tubule length (total length and length per gram of testicular parenchyma did not vary among the breeds studied. The morphometric parameters evaluated suggested that the genetic selection applied to the Nelore and Polled Nelore breeds improved the efficiency of spermatogenesis in these breeders.

  4. Hericium caput-medusae (Bull.:Fr.) Pers. polysaccharide enhance innate immune response, immune-related genes expression and disease resistance against Aeromonas hydrophila in grass carp (Ctenopharyngodon idella).

    Science.gov (United States)

    Gou, Changlong; Wang, Jiazhen; Wang, Yuqiong; Dong, Wenlong; Shan, Xiaofeng; Lou, Yujie; Gao, Yunhang

    2018-01-01

    The objective was to add 0, 400, 800 or 1200 mg/kg of Hericium caput-medusae polysaccharide (HCMP) to the basal diet of grass carp (Ctenopharyngodon idella) and determine effects on humoral innate immunity, expression of immune-related genes and disease resistance. Adding HCMP enhanced (P < 0.05) bactericidal activity at 1, 2 and 3 weeks and also lysozyme activity, complement C3, and SOD activity at 2 and 3 weeks. Supplementing 800 or 1200 mg/kg of HCMP for 2 or 3 weeks increased (P < 0.05) serum concentrations of total protein, albumin and globulin. Two immune-related genes (IL-1β and TNF-α) were up-regulated (P < 0.05) in HCMP supplemented groups given 800 or 1200 mg/kg HCMP after 2 and 3 weeks of feeding. Expression of anti-inflammatory cytokine IL-10 was down-regulated (P < 0.05) after receiving 800 or 1200 mg/kg HCMP for 2 or 3 weeks. Fish fed 800 mg/kg HCMP had maximal disease resistance against Aeromonas hydrophila (65.4%). In conclusion, HCMP enhanced immune response and expression of immune-related genes and increased disease resistance against Aeromonas hydrophila in grass carp, with greatest effects in fish given 800 mg/kg HCMP for 3 weeks. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. In‑depth mapping of human testicular and epididymal proteins and their functional association with spermatozoa.

    Science.gov (United States)

    Liu, Xuexia; Liu, Fujun

    2015-07-01

    The mammalian testis and epididymis are responsible for spermatozoa production and maturation, which contributes to male fertility. Predominantly expressed proteins in the testis and epididymis were suggested to be involved in the key functions or pathways in spermatogenesis and sperm maturation. To further investigate these proteins and their associations with sperm, large protein profiles of human testis and epididymis were mapped. Predominantly‑expressed testicular (173) and epididymal (244) secreted proteins were further screened and functionally characterized. Differential expression levels of solute carrier family 2 (facilitated glucose transporter), member 3, solute carrier family 25 (carnitine/acylcarnitine translocase), member 20, WAP‑type four‑disulfide core domain protein 8 and prostate and testis expressed 1 were validated using western blot and immunohistochemical analyses. The results may provide novel insight into the understanding of testicular and epididymal physiology and function, and facilitate sperm maturation research.

  6. In-depth mapping of human testicular and epididymal proteins and their functional association with spermatozoa

    Science.gov (United States)

    LIU, XUEXIA; LIU, FUJUN

    2015-01-01

    The mammalian testis and epididymis are responsible for spermatozoa production and maturation, which contributes to male fertility. Predominantly expressed proteins in the testis and epididymis were suggested to be involved in the key functions or pathways in spermatogenesis and sperm maturation. To further investigate these proteins and their associations with sperm, large protein profiles of human testis and epididymis were mapped. Predominantly-expressed testicular (173) and epididymal (244) secreted proteins were further screened and functionally characterized. Differential expression levels of solute carrier family 2 (facilitated glucose transporter), member 3, solute carrier family 25 (carnitine/acylcarnitine translocase), member 20, WAP-type four-disulfide core domain protein 8 and prostate and testis expressed 1 were validated using western blot and immunohistochemical analyses. The results may provide novel insight into the understanding of testicular and epididymal physiology and function, and facilitate sperm maturation research. PMID:25760095

  7. Effect of cholesterol supplementation on cryosurvival of goat spermatozoa

    Directory of Open Access Journals (Sweden)

    Sunita Behera

    2015-12-01

    Full Text Available Aim: Sperm membrane cholesterol influences cryodamage during cryopreservation. The present study was carried out to evaluate the effect of varying cholesterol levels in Tris based extenders on the freezability of sexually healthy Malabari buck semen. Materials and Methods: A total of 48 ejaculates from two adults healthy sexually healthy Malabari bucks were utilized for the study. The collected and pooled ejaculates were divided into four groups with Group I serving as Control - I, Group II and III were treated with 1 mg and 2 mg of cholesterol-loaded-cyclodextrin (CLC/120 × 106 spermatozoa, respectively, and Group IV treated with 1 mg methyl-β-cyclodextrin (MβCD served as Control - II. Manual freezing was carried out to cryopreserve the treated and control spermatozoa. Results: Treatment of semen samples with CLC resulted in improved maintenance of sperm motility at pre-freeze and post-thaw stages of cryopreservation without affecting hypo-osmotic swelling response. Treatment of semen with 1 mg of CLC/120 × 106 spermatozoa was observed to be better than treatment with 2 mg of CLC/120 × 106 spermatozoa. In general, MβCD treatment was found to result in significantly lower sperm characteristics than those of Control - I and CLC treatment at pre-feeze and post-thaw stages and when incubated up to 4 h. Conclusion: Cholesterol treatment of sexually healthy Malabari buck semen was found to hold promise for improving cryopreser-vability of spermatozoa.

  8. Oxidative phosphorylation versus glycolysis: what fuel do spermatozoa use?

    Science.gov (United States)

    du Plessis, Stefan S; Agarwal, Ashok; Mohanty, Gayatri; van der Linde, Michelle

    2015-01-01

    Spermatozoa are highly specialized cells. Adenosine triphosphate (ATP), which provides the energy for supporting the key functions of the spermatozoa, is formed by 2 metabolic pathways, namely glycolysis and oxidative phosphorylation (OXPHOS). It is produced in the mitochondria through OXPHOS as well as in the head and principal piece of the flagellum through glycolysis. However, there is a great discrepancy as to which method of ATP production is primarily utilized by the spermatozoa for successful fertilization. Mitochondrial respiration is considered to be a more efficient metabolic process for ATP synthesis in comparison to glycolysis. However, studies have shown that the diffusion potential of ATP from the mitochondria to the distal end of the flagellum is not sufficient to support sperm motility, suggesting that glycolysis in the tail region is the preferred pathway for energy production. It is suggested by many investigators that although glycolysis forms the major source of ATP along the flagellum, energy required for sperm motility is mainly produced during mitochondrial respiration. Nevertheless, some studies have shown that when glycolysis is inhibited, proper functioning and motility of spermatozoa remains intact although it is unclear whether such motility can be sustained for prolonged periods of time, or is sufficiently vigorous to achieve optimal fertilization. The purpose of this article is to provide an overview of mammalian sperm energy metabolism and identify the preferred metabolic pathway for ATP generation which forms the basis of energy production in human spermatozoa during fertilization.

  9. Quality of canine spermatozoa retrieved by percutaneous epididymal sperm aspiration.

    Science.gov (United States)

    Varesi, S; Vernocchi, V; Faustini, M; Luvoni, G C

    2013-02-01

    To investigate the feasibility of percutaneous epididymal sperm aspiration in dogs and whether it might provide a population of epididymal spermatozoa similar to the population that can be obtained by processing isolated epididymis caudae. Concentration and total sperm number, motility, morphology and acrosomal integrity of spermatozoa retrieved by percutaneous epididymal sperm aspiration, in vitro aspiration and mincing of the cauda of the epididymis were compared. Percutaneous epididymal sperm aspiration is a feasible procedure to retrieve a population of spermatozoa in dogs. Quality is similar to that of spermatozoa collected in vitro, although a wide variation amongst animals was observed. In case of ejaculation failure due to pathological conditions in dogs, the collection of spermatozoa from the cauda of the epididymis could be an option for providing gametes for assisted reproductive technologies. Percutaneous epididymal sperm aspiration can be used in dogs with compromised reproductive performance, in which orchiectomy cannot be performed for medical or owner reasons. Further studies aimed to investigate whether the percutaneous epididymal sperm aspiration technique might be feasible for repeated semen collection and to accurately evaluate side effects are required. © 2013 British Small Animal Veterinary Association.

  10. 77 FR 55139 - Safety Zone; Chicago Red Bull Flugtag, Lake Michigan, Chicago, IL

    Science.gov (United States)

    2012-09-07

    ... SECURITY Coast Guard 33 CFR Part 165 RIN 1625-AA00 Safety Zone; Chicago Red Bull Flugtag, Lake Michigan... restrict vessels from a portion of Lake Michigan for the Red Bull Flugtag event. This temporary safety zone... September 8, 2012 Red Bull North America will sponsor their Red Bull Flugtag event on the waters of Lake...

  11. 78 FR 57061 - Special Local Regulation; Red Bull Flugtag Miami, Biscayne Bay; Miami, FL

    Science.gov (United States)

    2013-09-17

    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulation; Red Bull Flugtag Miami..., during the Red Bull Flugtag. The Red Bull Flugtag is scheduled to take place on September 21, 2013. The... published a Notice of Proposed Rulemaking (NPRM) entitled USCG-2013-0180 Special Local Regulation; Red Bull...

  12. Changes in the Antioxidant Capacity and Iron-Binding Properties of Bovine Spermatozoa Following In Vitro Incubation with Ferrous or Ferric Iron

    Directory of Open Access Journals (Sweden)

    Eva Tvrdá

    2015-10-01

    Full Text Available The aim of this study was to assess the impact of ferrous (Fe2+ or ferric (Fe3+ iron on the antioxidant capacity and the ability to bind iron of bovine spermatozoa at specific time intervals (0h, 2h, 8h, 16h and 24h during an in vitro culture. 35 semen samples were collected from 7 adult breeding bulls and diluted in physiological saline solution supplemented with different concentrations (0, 1, 5, 10, 50, 100, 200, 500, 1000 μmol/L of FeCl2 or FeCl3. Spermatozoa motility was assessed using the SpermVisionTM CASA (Computer aided sperm analysis system. The ferric reducing ability of plasma (FRAP assay was applied to study the antioxidant capacity of the samples, while the ability of the sample to bind excess iron was determined using the Total iron-binding capacity (TIBC test. Both ferrous and ferric iron exhibited a dose- and time-dependent impact on the spermatozoa motility. Concentrations ≥50 µmol/L FeCl2 and ≥100 µmol/L FeCl3 led to a significant decrease of spermatozoa motion (P<0.001, while concentrations below 10 µmol/L FeCl2 and 50 µmol/L FeCl3 proved to preserve the parameter (P<0.001. The FRAP assay revealed that both ferrous as well as ferric iron had a similar effect on the FRAP marker of the samples: high concentrations led to a dramatic and significant (P<0.001 increase of the parameter, followed by a notable decrease of the reducing ability in the subsequent time periods, whose intensity was dependent upon the time, oxidation state of iron, as well as the time of analysis. Furthermore, supplementation of FeCl2 and FeCl3 had an impact on the capacity of the sperm culture to bind free iron, reflected in a significant decrease of the parameter (P<0.001 early on (Time 2h in case of high doses of both oxidative states of this biometal. In a direct comparison, ferrous iron has been shown to be more toxic than ferric iron.  Results from this in vitro study show that high concentrations of both forms of iron are toxic, while

  13. Morphological and acrosomal changes of canine spermatozoa during epididymal transit.

    Science.gov (United States)

    Varesi, Sara; Vernocchi, Valentina; Faustini, Massimo; Luvoni, Gaia Cecilia

    2013-02-26

    During epididymal transit, functional and structural modifications leading to full maturation enable male gametes to reach, recognize and fertilize the oocytes. In dogs, little is known on the modifications of spermatozoa during the passage in the epididymis. The aim of this study was to describe the motility, morphology and acrosomal patterns of canine spermatozoa retrieved from the epididymis caput, corpus and cauda. After the dilution required for the collection of epididymal content, sperm motility was significantly higher (Pepididymal regions. A significantly increased prevalence of tail defects, mainly represented by single bent tails, was observed in the corpus compared to caput (Psperm with cytoplasmic droplets decreased from the proximal to the distal region of the epididymis. Particularly, proximal cytoplasmic droplets were more frequently found in spermatozoa collected from the caput epididymis than in the corpus (Pepididymal regions.

  14. Characteristics of donkey spermatozoa along the length of the epididymis.

    Science.gov (United States)

    Contri, A; Gloria, A; Robbe, D; De Amicis, I; Carluccio, A

    2012-01-01

    In mammals, the epididymis has numerous interrelated functions including absorptive and secretory activity that affect luminal environment and cell membrane, and the maturation and storage of sperm. Spermatozoa acquire their motility and fertilizing ability during their passage through the epididymis and the motility of epididymal spermatozoa should be a balance between the maturation of flagellum and the inhibition of the flagellar machinery. In this study maturational change in sperm characteristics were evaluated in the epididymis of donkey. Spermatozoa collected from four portions of the epididymis (head, cranial corpus, caudal corpus, tail) were compared before and after ejaculation for viability, mitochondrial activity, kinetic parameters, and morphology. A significant increase in the mitochondrial activity along the epididymis was reported, suggesting a possible involvement in the motion mechanism. This should be corroborated by the significant correlation between mitochondrial activity and the total and progressive motility and the increase in velocities of spermatozoa recorded by computer-assisted sperm analysis. The percentage of most of the abnormal spermatozoa were similar in all tracts, with a great variability between jackasses. Only the bent midpiece percentage decreased significantly along epididymis. A significant increase in the percentage of distal cytoplasmic droplets (DCD), and a simultaneous decrease in the proximal cytoplasmic droplets (PCD), was found. The DCD fell down after ejaculation suggesting the late loss of the cytoplasmic residual (DCD) in the donkey, as hypothesized in the stallion. Because the prevalence of PCD were similar in both tail epididymal and ejaculated spermatozoa, a defect of the maturative process in the PCD sperm should be speculated. Copyright © 2012 Elsevier Inc. All rights reserved.

  15. 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.

  16. EFFECT OF DIFFERENT DILUENTS ON ROOSTER SPERMATOZOA APOPTOSIS

    Directory of Open Access Journals (Sweden)

    Lenka Kuželová

    2013-02-01

    Full Text Available The aim of this study was to observe the apoptosis of rooster spermatozoa and to compare the effect of commercial insemination diluent and saline solution on rooster spermatozoa apoptosis. Semen samples were collected once a week from roosters lines Lohmann Light (n=30. The one heterospermic sample was diluted with saline solution at a ratio of 1:100 and the second was diluted with commercial insemination diluent (Avian diluents; IMV Technologies, France in the same ratio 1:100 at room temperature. The percentage of apoptotic spermatozoa was estimated by fluorescent staining (Yo-Pro-1 0.5, 1 and 2 hours after semen collection and analyzed by fluorescent microscopy. There were no significant differences in sperms’ membranes quality after half an hour and one hour between diluted semen in both heterospermic samples. Significantly higher rate (P<0.05 of Yo-Pro-1 – positive cells was observed in sample diluted in commercial diluents (22.03% vs. 35.91% of apoptotic spermatozoa after two hours. We concluded that rooster semen could be diluted by saline solution which is cheaper and more accessible for practical The aim of this study was to observe the apoptosis of rooster spermatozoa and to compare the effect of commercial insemination diluent and saline solution on rooster spermatozoa apoptosis. Semen samples were collected once a week from roosters lines Lohmann Light (n=30. The one heterospermic sample was diluted with saline solution at a ratio of 1:100 and the second was diluted with commercial insemination diluent (Avian diluents; IMV Technologies, France in the same ratio 1:100 at room temperature. The percentage of apoptotic spermatozoa was estimated by fluorescent staining (Yo-Pro-1 0.5, 1 and 2 hours after semen collection and analyzed by fluorescent microscopy. There were no significant differences in sperms’ membranes quality after half an hour and one hour between diluted semen in both heterospermic samples. Significantly higher

  17. Bull Trout Distribution, Pacific Northwest (updated March, 2006)

    Data.gov (United States)

    Pacific States Marine Fisheries Commission — This dataset is a record of fish distribution and activity for BULL TROUT contained in the StreamNet database. This feature class was created based on linear event...

  18. Bull Moose Tube Company - Clean Water Act Public Notice

    Science.gov (United States)

    The EPA is providing notice of a proposed Administrative Penalty Assessment against the Bull Moose Tube Company, a business located at 1819 Clarkson Road, Chesterfield, MO, 63017, for alleged violations at the facility located at 406 East Industrial Drive,

  19. Protective features of resveratrol on human spermatozoa cryopreservation may be mediated through 5' AMP-activated protein kinase activation.

    Science.gov (United States)

    Shabani Nashtaei, M; Amidi, F; Sedighi Gilani, M A; Aleyasin, A; Bakhshalizadeh, Sh; Naji, M; Nekoonam, S

    2017-03-01

    Biochemical and physical modifications during the freeze-thaw process adversely influence the restoration of energy-dependent sperm functions required for fertilization. Resveratrol, a phytoalexin, has been introduced to activate 5' AMP-activated protein kinase which is a cell energy sensor and a cell metabolism regulator. The cryoprotection of resveratrol on sperm cryoinjury via activation of AMP-activated protein kinase also remains to be elucidated. Our aim, thus, was to investigate: (i) the presence and intracellular localization of AMP-activated protein kinase protein; (ii) whether resveratrol may exert a protective effect on certain functional properties of fresh and post-thaw human spermatozoa through modulation of AMP-activated protein kinase. Spermatozoa from normozoospermic men were incubated with or without different concentrations of Compound C as an AMP-activated protein kinase inhibitor or resveratrol as an AMP-activated protein kinase activator for different lengths of time and were then cryopreserved. AMP-activated protein kinase is expressed essentially in the entire flagellum and the post-equatorial region. Viability of fresh spermatozoa was not significantly affected by the presence of Compound C or resveratrol. However, although Compound C caused a potent inhibition of spermatozoa motility parameters, resveratrol did not induce negative effect, except a significant reduction in motility at 25 μm for 1 h. Furthermore, resveratrol significantly increased AMP-activated protein kinase phosphorylation and mitochondrial membrane potential and decreased reactive oxygen species and apoptosis-like changes in frozen-thawed spermatozoa. Nevertheless, it was not able to compensate decreased sperm viability and motility parameters following cryopreservation. In contrast, Compound C showed opposite effects to resveratrol on AMP-activated protein kinase phosphorylation, reactive oxygen species, apoptosis-like changes, mitochondrial membrane potential, and

  20. 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...

  1. Static magnetic field reduced exogenous oligonucleotide uptake by spermatozoa using magnetic nanoparticle gene delivery system

    Energy Technology Data Exchange (ETDEWEB)

    Katebi, Samira; Esmaeili, Abolghasem, E-mail: aesmaeili@sci.ui.ac.ir; Ghaedi, Kamran

    2016-03-15

    Spermatozoa could introduce exogenous oligonucleotides of interest to the oocyte. The most important reason of low efficiency of sperm mediated gene transfer (SMGT) is low uptake of exogenous DNA by spermatozoa. The aim of this study was to evaluate the effects of static magnetic field on exogenous oligonucleotide uptake of spermatozoa using magnetofection method. Magnetic nanoparticles (MNPs) associated with the labeled oligonucleotides were used to increase the efficiency of exogenous oligonucleotide uptake by rooster spermatozoa. We used high-field/high-gradient magnet (NdFeB) to enhance and accelerate exogenous DNA sedimentation at the spermatozoa surface. Flow cytometry analysis was performed to measure viability and percentage of exogenous oligonucleotide uptake by sperm. Flow cytometry analysis showed a significant increase in exogenous oligonucleotide uptake by rooster spermatozoa (P<0.001) when spermatozoa were incubated in exogenous oligonucleotide solution and MNPs. However, by applying static magnetic field during magnetofection method, a significant decrease in exogenous oligonucleotide uptake was observed (P<0.05). Findings of this study showed that MNPs were effective to increase exogenous oligonucleotide uptake by rooster spermatozoa; however unlike others studies, static magnetic field, was not only ineffective to enhance exogenous oligonucleotide uptake by rooster spermatozoa but also led to reduction in efficiency of magnetic nanoparticles in gene transfer. - Highlights: • Core/shell type Iron oxide nanoparticles were used as a novel and efficient method. • This method increases exogenous DNA uptake by rooster spermatozoa. • Static magnetic field decreased DNA uptake by rooster spermatozoa.

  2. Cryopreservation of very low numbers of spermatozoa from male patients undergoing infertility treatment using agarose capsules.

    Science.gov (United States)

    Hatakeyama, Shota; Tokuoka, Susumu; Abe, Hiroyuki; Araki, Yasuyuki; Araki, Yasuhisa

    2017-07-01

    This study tried to cryopreserve low numbers of spermatozoa from men undergoing infertility treatments by inserting into agarose capsules. The capsules were transferred into a drop of cryoprotectant solution and injected 3-4 motile spermatozoa that were selected by the swim-up method by conventional intracytoplasmic sperm injection. These capsules were put on a Cryotop(®) and frozen in liquid nitrogen vapor, and then submerged into liquid nitrogen and subsequently thawed and recovered. The motile spermatozoa in the capsules were counted. Eventually, we cryopreserved 2142 motile spermatozoa in 702 agarose capsules from 26 male patients and 1356 (63%) spermatozoa maintained their motility after thawing. The spermatozoa motility rates after thawing (MRAT) ranged from 20.0% (5/25) to 95.1% (58/61) among patients. The median MRAT was 68.3% (interquartile range 46.1-75.7). The total number of motile spermatozoa collected by swim-up method strongly correlated with MRAT (r = 0.746). It was possible to cryopreserve spermatozoa from male patients undergoing infertility treatment using agarose capsules. However, there were wide differences in MRAT among patients. It seems the spermatozoa from semen where there were many motile spermatozoa may have higher freezing resistance. Further studies using this method in cryptozoospermic semen, testicular and epididymal spermatozoa are required.

  3. CHARACTERISTICS AND FERTILITY OF SUMATRAN TIGER SPERMATOZOA CRYOPRESERVED WITH DIFFERENT SUGARS.

    Science.gov (United States)

    Wayan Kurniani Karja, Ni; Fahrudin, Mokhamad; Setiadi, Mohamad Agus; Tumbelaka, Ligaya Ita; Sudarwati, Retno; Hastuti, Yohana Tri; Mulia, Bongot Huas; Widianti, Ardyta; Sultan, Keni; Terazono, Tsukasa; Namula, Zhao; Taniguchi, Masayasu; Tanihara, Fuminori; Takemoto, Tatsuya; Kikuchi, Kazuhiro; Sato, Yoko; Otoi, Takeshige

    Cryopreservation of semen is one of the most important methods for the preservation of endangered tigers. This study evaluated the effects of sugar supplementation on the cryosurvival of spermatozoa from Sumatran tigers (Panthera tigris sumatrae). The post-thaw characteristics and fertility of spermatozoa cryopreserved with different sugars (glucose, lactose, and trehalose) were evaluated using heterologous in-vitro fertilisation with cat oocytes. All parameters of post-thaw spermatozoa significantly decreased as compared with those of fresh spermatozoa. The index of sperm motility for semen cryopreserved with lactose was significantly higher than that for semen cryopreserved with trehalose. The percentage of total fertilisation for tiger spermatozoa cryopreserved with trehalose was significantly lower than that for control cat spermatozoa. Our findings indicated that supplementation with lactose or glycerol as the main sugar in the egg yolk extender resulted in a better motility and fertility potential for post-thawed spermatozoa.

  4. Ultrastructural studies on the epididymal spermatozoa in the rhesus ...

    Indian Academy of Sciences (India)

    tribpo

    Abstract. Ultrastructural studies on the spermatozoa in different regions of the epididymis of the rhesus monkey have shown that the process of sperm maturation is associated with the caudad migration of the cytoplastmic droplet, a reduction in the volume of the cytoplasmic droplet, and an obvious wrinkling of the plasma ...

  5. The Role of Oviductal Cells in Activating Stallion Spermatozoa

    NARCIS (Netherlands)

    Leemans, Bart; Gadella, Bart M.|info:eu-repo/dai/nl/115389873; Stout, Tom A.E.|info:eu-repo/dai/nl/304828831; De Schauwer, Catharina; Nelis, Hilde; Hoogewijs, Maarten; Van Soom, Ann

    2016-01-01

    Conventional in vitro fertilization is poorly successful with equine gametes. Although stallion spermatozoa bind to the zona pellucida in vitro, they fail to acrosome react and cannot therefore penetrate into the perivitelline space. Failed sperm penetration most likely relates to the absence in in

  6. Spermatozoa morphology and characteristics of male wistar rats ...

    African Journals Online (AJOL)

    This study was aimed at determining the effects of the ethanolic extract of the whole fruit of Lagenaria breviflora Robert on male fertility by evaluating some andrological parameters of the Wistar rat such as morphology of spermatozoa, sperm count, motility, liveability and volume of the semen. Histopathology of the testis was ...

  7. The effect of sperm morphology and testicular spermatozoa ...

    African Journals Online (AJOL)

    Objective. To determine the correlation between sperm morphology groups (strict criteria) and testicular spermatozoa, and day 2 and 3 embryo quality in intracytoplasmic sperm injection (ICSI) and in vitro fertilisation (IVF) cases. Methods. A retrospective study was done of 2 402 IVF and ICSI-fertilised embryos classified as ...

  8. Elimination of apoptotic boar spermatozoa using magnetic activated cell sorting

    Directory of Open Access Journals (Sweden)

    Janko Mrkun

    2014-01-01

    Full Text Available One of the features of apoptosis is the externalization of phosphatidylserine which could be used to remove apoptotic cells from semen preparations. Magnetic-activated cell sorting using annexin V-conjugated microbeads which bind to phosphatidylserine could be used to enhance semen quality. Twelve boar semen samples after 3 days of liquid storage at 16­­–17 °C were subjected to magnetic-activated cell sorting. Bound and unbound fractions and control samples were subjected to flow cytometry following the staining of spermatozoa with Annexin V conjugated with Alexa Fluor 488 and propidium iodide. Four subpopulations were obtained: live, early apoptotic live, late apoptotic, early necrotic dead and late necrotic dead. The frequency of early apoptotic and late necrotic spermatozoa was significantly higher (P P P P < 0.05 lower proportion of morphologically normal spermatozoa was observed in both fractions compared to control (67.2 ± 17.0%. Boar spermatozoa were separated by the above method for the first time, however, the results showed this method to be inappropriate for boar semen separation under the tested conditions.

  9. Acetylcarnitine hydrolase activity in bovine caudal epididymal spermatozoa

    Energy Technology Data Exchange (ETDEWEB)

    Bruns, K.; Foster, R.A.; Casillas, E.R.

    1986-05-01

    Recently, the authors identified mM concentrations of acetylcarnitine in epidiymal fluids and have investigated the metabolism of acetylcarnitine by bovine and hamster caudal epididymal spermatozoa. (1-/sup 14/C)acetyl-L-carnitine is oxidized to /sup 14/CO/sub 2/ by washed, intact hamster and bovine sperm at maximal rates of 8.4 and 15.2 nmol/hr/10/sup 7/ cells respectively. Conversely, the carnitine moiety of acetyl-L-(/sup 3/H-methyl)carnitine is not accumulated by sperm under similar conditions. Hydrolysis of (/sup 3/H)acetyl-L-carnitine and competition of uptake of (/sup 3/H)acetate by unlabeled acetate was demonstrated in incubations of intact cells of both species. The amount of (/sup 3/H)acetate accumulated in the incubation medium is time-dependent and also depends on the concentration of unlabeled acetate. A partial solubilization of acetylcarnitine hydrolase activity from washed, intact bovine caudal epididymal spermatozoa in buffer or 0.01% Triton X-100 is observed. There is an enrichment of acetylcarnitine hydrolase activity in purified plasma membranes from bovine caudal epididymal spermatozoa when compared to the activity present in broken cell preparations or other cellular fractions. The results suggest that acetylcarnitine is a substrate for spermatozoa as they traverse the epididymis.

  10. Coitus Interruptus: Are there spermatozoa in the pre-ejaculate ...

    African Journals Online (AJOL)

    Methods: A literature search was performed using PubMed, and Google Scholar search engines. Literature reviewed included reviews, and original articles that evaluated the presence of spermatozoa in the pre-ejaculatory fluid. Articles reporting about coitus interruptus as a method of contraception were also reviewed.

  11. Cryosurvival of goat spermatozoa in tris-egg yolk extender ...

    African Journals Online (AJOL)

    The effect of melatonin supplementation in semen extenders on cryosurvival of spermatozoa obtained from West African Dwarf (WAD) goat bucks was studied. Tris-egg yolk extenders supplemented with different levels of melatonin (0, 2, 4, 6 and 8 mM) were diluted with semen samples. The diluted semen samples were ...

  12. Breeding soundness evaluations of Senepol bulls in the US Virgin Islands.

    Science.gov (United States)

    Godfrey, R W; Dodson, R E

    2005-02-01

    The breeding soundness evaluation (BSE) was used to evaluate Senepol (Bos taurus) bulls (n = 495) on St. Croix over a 7-year period. Young, unproven bulls (10-26 months of age) and breeding bulls (16 months to 8.5 years) were tested prior to sale or use in breeding. Inbreeding coefficients were determined for a subset of bulls (n = 290). The percentage of bulls passing the BSE increased (P 20 months. The proportion of all bulls that failed the BSE and received an Unsatisfactory rating for scrotal circumference or sperm motility decreased (P 90 to Senepol bulls raised under tropical conditions had a low probability of passing the BSE at young ages, but the passing rate increased with age. Older Senepol bulls were more likely to fail the BSE due to abnormal sperm morphology than due to inadequate testicular size or sperm motility. To prevent unnecessary culling, a BSE should not be performed on Senepol bulls <16 months old.

  13. Effect of oleic-linoleic acid and ?-sitosterol to freezing extender of bulls and stallions semen

    Directory of Open Access Journals (Sweden)

    Érika Saltiva Cruz Bender

    2015-06-01

    Full Text Available Addition of polyunsaturated fatty acids and/or cholesterol to a freezing diluent can modify the sperm plasma membrane composition, influencing its behavior during cryopreservation, thus, favoring seminal cryoresistance. The present study aimed to evaluate the effects of the addition of oleic-linoleic acid, (OLA; ?-sitosterol (?-sit, a plant analog of cholesterol; and OLA + ?-sit in combination to a freezing diluent, on the cryopreservation bull and stallion semen. The following variables were analyzed: motility/vigor, plasma and acrosomal membrane integrity (by Trypan Blue/Giemsa staining, mitochondrial activity (by DAB staining, and lipid peroxidation (by a TBARS assays. The lipids were added according to experimental treatments: C – control group, A1 and A2 – OLA at concentrations of 37 ?M and 74 ?M, B1 and B2 – ?-sit at concentrations of 1 ?g mL-1 and 2 ?g mL-1; AB1 and AB2 – OLA 37 ?M + ?-sit 1 ?g mL-1 and OLA 74 ?M + ?-sit 2 ?g mL-1, respectively. The study was divided into three experiments; in Experiment 1, the concentrations of the groups A1, B1, and AB1 were evaluated, whereas in Experiment 2 the concentrations of the groups A2, B2, and AB2 were analyzed, both experiments were performed with bull semen. We conducted Experiment 3 using equine semen with the addition of lipids at all of the concentrations described. Data were subjected to analysis of variance, using the GLM procedure of SAS, with treatment means compared by Duncan test considering 5% significance. These variables differed significantly after thawing the semen post-collection. However, there was no significant difference between treatments when variables were compared within the same time point, except for Experiment 2, where there was a decrease in motility and vigor decrease post-thaw in the groups following ?-sit addition (C – 51.0 ± 13.7%/2.9 ± 0.4; B2 – 35.8 ± 15.8%/2.3 ± 0.6; AB2 – 38.5 ± 16.6%/2.5 ± 0.5, respectively; p < 0.05. In conclusion, the

  14. Composition and significance of detergent resistant membranes in mouse spermatozoa.

    Science.gov (United States)

    Nixon, Brett; Bielanowicz, Amanda; McLaughlin, Eileen A; Tanphaichitr, Nongnuj; Ensslin, Michael A; Aitken, R John

    2009-01-01

    Mammalian spermatozoa acquire the ability to fertilize an oocyte as they ascend the female reproductive tract. This process is characterized by a complex cascade of biophysical and biochemical changes collectively know as "capacitation." The attainment of a capacitated state is accompanied by a dramatic reorganization of the surface architecture to render spermatozoa competent to recognize the oocyte and initiate fertilization. Emerging evidence indicates that this process is facilitated by molecular chaperone-mediated assembly of a multimeric receptor complex on the sperm surface. However, the mechanisms responsible for gathering key recognition molecules within this putative complex have yet to be defined. In this study, we provide the first evidence that chaperones partition into detergent resistant membrane fractions (DRMs) within capacitated mouse spermatozoa and co-localize in membrane microdomains enriched with the lipid raft marker, G(M1) ganglioside. During capacitation, these microdomains coalesce within the apical region of the sperm head, a location compatible with a role in sperm-zona pellucida interaction. Significantly, DRMs isolated from spermatozoa possessed the ability to selectively bind to the zona pellucida of unfertilized, but not fertilized, mouse oocytes. A comprehensive proteomic analysis of the DRM fractions identified a total of 100 proteins, a number of which have previously been implicated in sperm-oocyte interaction. Collectively, these data provide compelling evidence that mouse spermatozoa possess membrane microdomains that provide a platform for the assembly of key recognition molecules on the sperm surface and thus present an important mechanistic insight into the fundamental cell biological process of sperm-oocyte interaction. (c) 2008 Wiley-Liss, Inc.

  15. Effect of cooling (4°C) and cryopreservation on cytoskeleton actin and protein tyrosine phosphorylation in buffalo spermatozoa.

    Science.gov (United States)

    Naresh, Sai

    2016-02-01

    Semen cryopreservation is broadly utilized as a part of the bovine reproducing industry, a large portion of the spermatozoa does not survive and the majority of those that do survive experience various molecular and physiological changes that influence their fertilizing capacity. The main aim of this study is to determine the effect of cooling (4 °C) and cryopreservation on cytoskeleton actin, tyrosine phosphorylation and quality of buffalo spermatozoa, and to determine the similarity between in vitro capacitation and cryopreservation induced capacitation like changes. To achieve this, Western blot was used to examine the changes in actin expression and protein tyrosine phosphorylation, whereas changes in actin polymerization, localization of actin and protein tyrosine phosphorylation during capacitation and cryopreservation were evaluated by indirect immunofluorescence technique. Localization studies revealed that the actin localized to flagella and acrosome membrane regions and following, capacitation it migrated towards the acrosome region of sperm. Time dependent increase in actin polymerization and protein tyrosine phosphorylation was observed during in vitro capacitation. The cooling phase (4 °C) and cryopreservation processes resulted in the loss/damage of cytoskeleton actin. In addition, we performed the actin polymerization and protein tyrosine phosphorylation in cooled and cryopreserved buffalo spermatozoa. Interestingly, cooling and cryopreservation induces actin polymerization and protein tyrosine phosphorylation, which were similar to in vitro capacitation (cryo-capacitation). These changes showed 1.3 folds reduction in the sperm quality parameters which includes motility, viability and plasma membrane integrity. Furthermore, our findings indicate that cooling and cryopreservation damages the cytoskeleton actin and also induces capacitation like changes such as protein tyrosine phosphorylation and actin polymerization. This could be one of the

  16. Effects of supplementation of iodixanol to semen extender on quality and fertilization ability of frozen-thawed Thai native bull sperm.

    Science.gov (United States)

    Chuawongboon, Phirawit; Sirisathien, Saksiri; Pongpeng, Jatuporn; Sakhong, Denpong; Nagai, Takashi; Vongpralub, Thevin

    2017-09-01

    This study investigates the effects of iodixanol supplementation in varied concentrations to Tris egg yolk (TEY) extender on the quality and fertilization ability of frozen-thawed sperm of Thai native bulls. Each ejaculate was divided into four different groups, as follows: sperm were treated with TEY extender (control group) and TEY extender supplemented with three different concentrations of iodixanol (1.25%, 2.50% and 5.00%). Semen straws were frozen in liquid nitrogen vapor. After thawing, sperm motility characteristics, viability, plasma membrane integrity and acrosome integrity were determined. Also, frozen-thawed spermatozoa from all groups were used for in vitro fertilization and artificial insemination (AI) in natural estrus Thai native cows. The results showed that the post-thaw quality of the 2.50% iodixanol group was superior to the other iodixanol groups (P fertilization ability and pregnancy rate after AI (P > 0.05). It can be concluded that the supplementation of 2.50% iodixanol extender significantly improves the progressive motility, viability, plasma membrane integrity and acrosome integrity of cryopreserved semen from Thai native bulls, but it has no beneficial effect on in vitro fertilization ability and pregnancy rate after AI. © 2017 Japanese Society of Animal Science.

  17. Adrenal involvement in the biostimulatory effect of bulls

    Directory of Open Access Journals (Sweden)

    Berardinelli James G

    2007-08-01

    Full Text Available Abstract Background The objective was to evaluate if cortisol concentrations are associated with the resumption of luteal activity in postpartum, primiparous cows exposed to bulls. The hypotheses were that 1 interval from start of exposure to resumption of luteal activity; 2 proportions of cows that resumed luteal function during the exposure period; and 3 cortisol concentrations do not differ among cows exposed or not exposed to bulls (Exp. 1, and cows continuously exposed to bull or steer urine (Exp. 2. Methods In Exp. 1, 28 anovular cows were exposed (BE; n = 13 or not exposed (NE; n = 15 to bulls for 30 d at 58 d after calving. In Exp. 2, 38 anovular cows were fitted with a controlled urine delivery device at 45 d after calving and exposed continuously (24 h/d to bull (BUE; n = 19 or steer (SUE; n = 19 urine. Length of exposure was ~64 d. Blood samples were collected from each cow on D 0 and every 3 d throughout exposure periods in both experiments and assayed for progesterone. Cortisol was assayed in samples collected on D 0, 8, 16, and 24 in Exp. 1; and, D 0, 19, 38, and 57 in Exp. 2. Results In Exp. 1, interval from the start of exposure to resumption of luteal activity was shorter (P Conclusion We conclude that the physical presence of bulls stimulates resumption of luteal activity and is coincident with increased cortisol concentrations, and hypothesize a possible association between adrenal activation and the biostimulatory effect of bulls.

  18. Spawning and rearing behavior of bull trout in a headwaterlake ecosystem

    Science.gov (United States)

    Lora B. Tennant,; Gresswell, Bob; Guy, Christopher S.; Michael H. Meeuwig,

    2015-01-01

    Numerous life histories have been documented for bull trout Salvelinus confluentus. Lacustrine-adfluvial bull trout populations that occupy small, headwater lake ecosystems and migrate short distances to natal tributaries to spawn are likely common; however, much of the research on potamodromous bull trout has focused on describing the spawning and rearing characteristics of bull trout populations that occupy large rivers and lakes and make long distance spawning migrations to natal headwater streams. This study describes the spawning and rearing characteristics of lacustrine-adfluvial bull trout in the Quartz Lake drainage, Glacier National Park, USA, a small headwater lake ecosystem. Many spawning and rearing characteristics of bull trout in the Quartz Lake drainage are similar to potamodromous bull trout that migrate long distances. For example, subadult bull trout distribution was positively associated with slow-water habitat unit types and maximum wetted width, and negatively associated with increased stream gradient. Bull trout spawning also occurred when water temperatures were between 5 and 9 °C, and redds were generally located in stream segments with low stream gradient and abundant gravel and cobble substrates. However, this study also elucidated characteristics of bull trout biology that are not well documented in the literature, but may be relatively widespread and have important implications regarding general characteristics of bull trout ecology, use of available habitat by bull trout, and persistence of lacustrine-adfluvial bull trout in small headwater lake ecosystems.

  19. Evaluating bull fertility based on non-return method

    Directory of Open Access Journals (Sweden)

    Prka Igor

    2012-01-01

    Full Text Available In order to evaluate the results of reproductive cows and heifers, different parameters of fertility are used, such as the service period, insemination index, intercalving time and others, and of the breeding bulls the values obtained through non-return. An ejaculate is taken up for further processing by veterinary centres only provided it meets the prescribed quality parameters. Rating semen parameters includes a macroscopic (volume, colour, consistency, smell and pH and a microscopic evaluation (mobility, density, percentage of live sperm and abnormal and damaged sperm. In addition to sperm quality and the fertility of the female animal, the results of the non-return method are also influenced by a number of exogenous causes (season, age, race, insemination techniques that have no small impact on the end result of insemination - pregnancy. In order to obtain more objective results of the fertility of bulls the following tasks were undertaken, namely: 1. to calculate with the non-return method the fertility of bulls in over 10,000 cows inseminated for the first time during a period of 6 years; and 2. to analyze the impact of semen quality, season, age of cow and bull, and the bull breed on the results of fertility.

  20. Proteomic comparison of detergent-extracted sperm proteins from bulls with different fertility indexes

    National Research Council Canada - National Science Library

    D'Amours, Olivier; Frenette, Gilles; Fortier, Marlene; Leclerc, Pierre; Sullivan, Robert

    2010-01-01

    .... Frozen semen from 23 Holstein bulls with documented fertility was used. According to their ‘fertility solution’ (SOL), as calculated by the Canadian dairy network, bulls were divided into four groups...

  1. Proteomic comparison of detergent-extracted sperm proteins from bulls with different fertility indexes

    National Research Council Canada - National Science Library

    Olivier D'Amours; Gilles Frenette; Marlène Fortier; Pierre Leclerc; Robert Sullivan

    2010-01-01

    .... Frozen semen from 23 Holstein bulls with documented fertility was used. According to their ‘fertility solution’ (SOL), as calculated by the Canadian dairy network, bulls were divided into four groups...

  2. Cytological detection of spermatozoa: comparison of three staining methods.

    Science.gov (United States)

    Allery, J P; Telmon, N; Mieusset, R; Blanc, A; Rougé, D

    2001-03-01

    Sperm detection can be an important factor in confirming sexual assault in cases of rape. This paper compares three of the most commonly used staining methods cited in the scientific literature: Christmas tree. hematoxylin-eosin, and alkaline fuchsin. The population studied was composed of 174 consenting women seen at the Male Infertility Center in Toulouse. France. The date of their last sexual intercourse was accurately known. Alkaline fuchsin did not seem effective in detecting spermatozoa in vaginal samples. Compared with hematoxylin-eosin, Christmas tree stain appeared to be the most useful test in the first 72 h. Two external factors were associated with decreased detection of spermatozoa: time since in tercourse and sperm volume.

  3. Conventional slow freezing cryopreserves mouflon spermatozoa better than vitrification.

    Science.gov (United States)

    Pradiee, J; Esteso, M C; Castaño, C; Toledano-Díaz, A; Lopez-Sebastián, A; Guerra, R; Santiago-Moreno, J

    2017-04-01

    This work examines the effectiveness of a TCG (Tris, citric acid, glucose, 6% egg yolk and 5% glycerol) and a TEST (TES, Tris, glucose, 6% egg yolk and 5% glycerol) sperm extender in the freezing of mouflon spermatozoa at slow cooling rates, using different pre-freezing equilibration times (2-3 hr). It also examines the tolerance of mouflon spermatozoa to different concentrations of cryoprotectants (5, 10, 20% glycerol; 5%, 10%, 20% dimethyl sulfoxide; 6% polyvinylpyrrolidone) and/or sucrose (100, 300, 500 mm). The highest quality (p freezing rates (60-85°C min -1 ), independent of the concentration of cryoprotectants. The lowest sucrose concentration (100 mm) provided the highest (p freezing. © 2016 Blackwell Verlag GmbH.

  4. Prostasome-like vesicles stimulate acrosome reaction of pig spermatozoa

    Directory of Open Access Journals (Sweden)

    Marcianò Vito

    2008-01-01

    Full Text Available Abstract Background The presence of small membranous particles characterizes the male genital fluids of different mammalian species. The influence of semen vesicles, denominated prostasomes, on sperm functional properties has been well documented in humans, but their biological activity is scarcely known in other species. The present work investigated prostasome-like vesicles in pig semen for their ability to interact with spermatozoa and to affect acrosome reaction. Methods Prostasome-like vesicles have been isolated from pig seminal plasma by high-speed centrifugation and Sephadex G-200 gel chromatography. Morphology of purified vesicles has been checked by scanning electron microscopy while their protein pattern has been investigated by SDS-PAGE. Then prostasome- like vesicles have been incubated with pig spermatozoa and their ability to interact with sperm has been tested by the aminopeptidase assay. In addition, the efficiency of vesicles to influence the acrosome reaction has been investigated by assessing the sperm acrosomal status by the PI/FITC-PNA (propidium iodide/fluorescein isothiocyanate-labeled peanut agglutinin stainings. Results Purified vesicles revealed a complex protein pattern with the occurrence of bands in the high, medium and low molecular weight range. However, the two major bands were observed at ~90 kDa and ~60 kDa. A vesicle-mediated transfer of aminopeptidase to sperm cells has been also detected. Furthermore, a significant increase of acrosome reaction extent has been revealed in spermatozoa incubated with prostasome-like vesicles in comparison to control sperm. Conclusion This is the first report demonstrating that pig prostasome-like vesicles are able, in vitro, to interact with spermatozoa and to stimulate the acrosome reaction. These findings lead to hypothesize a transfer of molecules from vesicles to sperm membrane, thus sensitizing male gametes to undergo the acrosome reaction

  5. Bull'S eye maculopathy in a patient taking sertraline.

    Science.gov (United States)

    Mason, John O; Patel, Shyam A

    2015-01-01

    To report a very rare case of bilateral Bull's eye maculopathy caused by sertraline. Clinical case report and literature review. A 14-year-old girl. A 14-year-old girl with no significant medical history developed bilateral Bull's eye maculopathy after taking sertraline for 1 year. Diagnostic work-up, which consisted of electrooculography, electroretinography, and genetic testing, yielded no abnormal results. Three-year follow-up examination after discontinuation of sertraline showed no improvement in visual acuity (20/200 bilaterally) or retinal pigment epithelium. This is the second published case, and according to the National Registry of Drug-Induced Ocular Side Effects, this is only the fifth reported case of presumed sertraline maculopathy. Although Bull's eye maculopathy related to sertraline is rare, physicians and patients need to be aware of the possible toxicity.

  6. Rhéologie de suspensions de bulles

    OpenAIRE

    Morini, Romain; Tocquer, Laurent; Chateau, Xavier; Ovarlez, Guillaume; Pitois, Olivier

    2015-01-01

    Un dispositif original a été développé pour fabriquer des suspensions monodisperses de bulles (diamètre 700 ¼m) dans un fluide newtonien très visqueux (viscosité dynamique 100 Pa.s). La viscosité de ces suspensions a été mesurée dans les régimes dilué et faiblement concentré (fraction volumique en bulles comprises entre 2% et 15%) dans une géométrie Couette. La viscosité des suspensions dépend de la viscosité du fluide suspendant, de la fraction volumique en bulles et d'un nombre sans ...

  7. Effects of epidural lidocaine anesthesia on bulls during electroejaculation.

    Science.gov (United States)

    Falk, A J; Waldner, C L; Cotter, B S; Gudmundson, J; Barth, A D

    2001-02-01

    Two experiments were conducted to determine whether caudal epidural lidocaine anesthesia reduces a stress response to electroejaculation. In the 1st experiment, changes in cortisol and progesterone concentrations in serial blood samples were used to assess the stress response to restraint (control), transrectal massage, caudal epidural injection of saline, electroejaculation after caudal epidural injection of lidocaine, and electroejaculation without epidural lidocaine. In the 2nd experiment, behavioral responses were subjectively scored in bulls that were electroejaculated with or without caudal epidural lidocaine anesthesia. Cortisol and progesterone concentrations were significantly elevated after electroejaculation, whether or not bulls received caudal epidural anesthesia. Elevations in cortisol and progesterone were lower and fewer bulls vocalized during electroejaculation when given caudal epidural anesthesia; however, the differences were not significant.

  8. Oviduct binding and elevated environmental ph induce protein tyrosine phosphorylation in stallion spermatozoa.

    Science.gov (United States)

    Leemans, Bart; Gadella, Bart M; Sostaric, Edita; Nelis, Hilde; Stout, Tom A E; Hoogewijs, Maarten; Van Soom, Ann

    2014-07-01

    Sperm-oviduct binding is an essential step in the capacitation process preparing the sperm for fertilization in several mammalian species. In many species, capacitation can be induced in vitro by exposing spermatozoa to bicarbonate, Ca(2+), and albumin; however, these conditions are insufficient in the horse. We hypothesized that binding to the oviduct epithelium is an essential requirement for the induction of capacitation in stallion spermatozoa. Sperm-oviduct binding was established by coincubating equine oviduct explants for 2 h with stallion spermatozoa (2 × 10(6) spermatozoa/ml), during which it transpired that the highest density (per mm(2)) of oviduct-bound spermatozoa was achieved under noncapacitating conditions. In subsequent experiments, sperm-oviduct incubations were performed for 6 h under noncapacitating versus capacitating conditions. The oviduct-bound spermatozoa showed a time-dependent protein tyrosine phosphorylation response, which was not observed in unbound spermatozoa or spermatozoa incubated in oviduct explant conditioned medium. Both oviduct-bound and unbound sperm remained motile with intact plasma membrane and acrosome. Since protein tyrosine phosphorylation can be induced in equine spermatozoa by media with high pH, the intracellular pH (pHi) of oviduct explant cells and bound spermatozoa was monitored fluorometrically after staining with BCECF-AM dye. The epithelial secretory cells contained large, alkaline vesicles. Moreover, oviduct-bound spermatozoa showed a gradual increase in pHi, presumably due to an alkaline local microenvironment created by the secretory epithelial cells, given that unbound spermatozoa did not show pHi changes. Thus, sperm-oviduct interaction appears to facilitate equine sperm capacitation by creating an alkaline local environment that triggers intracellular protein tyrosine phosphorylation in bound sperm. © 2014 by the Society for the Study of Reproduction, Inc.

  9. Storage and Release of Spermatozoa from the Pre-Uterine Tube Reservoir

    Science.gov (United States)

    Freeman, Sarah L.; England, Gary C.W.

    2013-01-01

    In mammals, after coitus a small number of spermatozoa enter the uterine tube and following attachment to uterine tube epithelium are arrested in a non-capacitated state until peri-ovulatory signalling induces their detachment. Whilst awaiting release low numbers of spermatozoa continually detach from the epithelium and the uterine tube reservoir risks depletion. There is evidence of attachment of spermatozoa to uterine epithelium in several species which might form a potential pre-uterine tube reservoir. In this study we demonstrate that: (1) dog spermatozoa attach to uterine epithelium and maintain flagellar activity, (2) in non-capacitating conditions spermatozoa progressively detach with a variety of motility characteristics, (3) attachment is not influenced by epithelial changes occurring around ovulation, (4) attachment to uterine epithelium slows capacitation, (5) capacitated spermatozoa have reduced ability to attach to uterine epithelium, (6) under capacitating conditions increased numbers of spermatozoa detach and exhibit transitional and hyperactive motility which differ to those seen in non-capacitating conditions, (7) detachment of spermatozoa and motility changes can be induced by post-ovulation but not pre-ovulation uterine tube flush fluid and by components of follicular fluid and solubilised zona pellucida, (8) prolonged culture does not change the nature of the progressive detachment seen in non-capacitating conditions nor the potential for increased detachment in capacitating conditions. We postulate that in some species binding of spermatozoa to uterine epithelium is an important component of the transport of spermatozoa. Before ovulation low numbers of spermatozoa continually detach, including those which are non-capacitated with fast forward progressive motility allowing the re-population of the uterine tube, whilst around the time of ovulation, signalling from as-yet unknown factors associated with follicular fluid, oocytes and uterine tube

  10. Causes and consequences of oxidative stress in spermatozoa.

    Science.gov (United States)

    Aitken, Robert John; Gibb, Zamira; Baker, Mark A; Drevet, Joel; Gharagozloo, Parviz

    2016-01-01

    Spermatozoa are highly vulnerable to oxidative attack because they lack significant antioxidant protection due to the limited volume and restricted distribution of cytoplasmic space in which to house an appropriate armoury of defensive enzymes. In particular, sperm membrane lipids are susceptible to oxidative stress because they abound in significant amounts of polyunsaturated fatty acids. Susceptibility to oxidative attack is further exacerbated by the fact that these cells actively generate reactive oxygen species (ROS) in order to drive the increase in tyrosine phosphorylation associated with sperm capacitation. However, this positive role for ROS is reversed when spermatozoa are stressed. Under these conditions, they default to an intrinsic apoptotic pathway characterised by mitochondrial ROS generation, loss of mitochondrial membrane potential, caspase activation, phosphatidylserine exposure and oxidative DNA damage. In responding to oxidative stress, spermatozoa only possess the first enzyme in the base excision repair pathway, 8-oxoguanine DNA glycosylase. This enzyme catalyses the formation of abasic sites, thereby destabilising the DNA backbone and generating strand breaks. Because oxidative damage to sperm DNA is associated with both miscarriage and developmental abnormalities in the offspring, strategies for the amelioration of such stress, including the development of effective antioxidant formulations, are becoming increasingly urgent.

  11. Activity of Chinchilla laniger spermatozoa collected by electroejaculation and cryopreserved.

    Science.gov (United States)

    Ponce, A A; Carrascosa, R E; Aires, V A; Fiol de Cuneo, M; Ruiz, R D; Ponzio, M F; Lacuara, J L

    1998-12-01

    Because reproductive studies and the application of assisted reproductive techniques are relevant issues for an endangered species such as Chinchila laniger, the availability of a source of viable spermatozoa becomes of utmost importance. In this paper, we evaluate several functional parameters (motility, viability, response to hypoosmotic swelling test and acrosomal integrity) of fresh or frozen-thawed spermatozoa. Electro-ejaculation trials (50-cyc/sec sinusoidal wave was applied for 5 of every 10 sec) were successful in all unanesthetized animals. After volume (108.3 +/- 12.0 microL, n = 15) and concentration (421.8 +/- 34.4 x 10(6) cells/mL, n = 15) measurements, the above mentioned parameters were determined. In frozen-thawed semen samples sperm motility, viability, hypoosmotic swelling test and acrosomal integrity were significantly lower than in fresh semen samples. The results clearly indicated that electro-ejaculation is a useful method for evaluating spermatozoa for genetic analysis or for used in Al in this species. In addition, the cryopreservation procedure in this study preserved adequate levels of functional sperm activity.

  12. Physical and morphological characteristics of Kankrej bull semen

    Directory of Open Access Journals (Sweden)

    Bharatkumar R. Patel

    Full Text Available Aim: Present investigation was carried out to study the physical characteristics of Kankrej bulls semen by evaluation of various semen parameters from neat semen and at various stages of semen preservation. Materials and Methods: A total of 60 ejaculates, 10 each from 6 mature Kankrej bulls, once in a week for 10 weeks, were collected and analyzed for various semen attributes. Result: The mean values for different seminal attributes were: ejaculate volume 4.84 ± 0.01 ml, pH 6.88 ± 0.01, mass motility 3.72 ± 0.02, sperm concentration 1253.83 ± 14.68 million / ml, individual motility 86.15 ± 0.30 per cent, live sperm count 90.58 ± 0.20 per cent, abnormal sperm count 4.24 ± 0.03 per cent and acrosomal integrity 81.17 ± 0.11 per cent. The colour of the Kankrej bull semen under the investigation was creamy white. Mean values of ejaculate volume, sperm concentration, live sperm count and acrosomal integrity of semen differed significantly (P < 0.05 among the bulls under investigation. The ejaculate volume was positively correlated with mass motility (+ 0.392 and sperm concentration (+ 0.385 and inversely proportional to the mass motility whereas mass motility positively correlated with volume (+0.392, individual sperm motility (+0.329 and live sperm count (+0.527. Conclusion: It can be concluded that the volume, pH, mass motility and sperm concentration of Kankrej bull semen were well comparable with other breeds of Indian cattle, however higher individual motility, live sperm count, acrosomal integrity and lower abnormal sperm count were recorded in the Kankrej bull semen. [Vet World 2013; 6(7.000: 405-408

  13. Numerical Chromosomal Abnormalities in Rat Epididymal Spermatozoa Following Chronic Cyclophosphamide Exposure1

    National Research Council Canada - National Science Library

    Tara S. Barton; Andrew J. Wyrobek; Francesca S. Hill; Bernard Robaire; Barbara F. Hales

    2003-01-01

    .... The goal of the present study was to determine the effect of chronic cyclophosphamide treatment during spermatogenesis on the frequency of numerical chromosomal anomalies in epididymal spermatozoa...

  14. Time budgets of finishing bulls housed in an uninsulated barn or at pasture

    Directory of Open Access Journals (Sweden)

    Leena Tuomisto

    2015-10-01

    Full Text Available This study aimed at comparing the behaviour of finishing bulls raised in an uninsulated barn (UB and at pasture (PAS. In experiment 1, dairy bulls were housed in an uninsulated barn (two groups of five bulls, 32 m2/pen or at pasture (groups of four and five bulls, 5000 m2/paddock. In experiment 2, Hereford bulls were housed in an uninsulated barn (three groups of four or five bulls, 32 m2/pen or at pasture (three groups of five bulls, 5000 m2/paddock. There were no differences in drinking, social licking, butting, other social behaviour, self-licking or idling between the UB and PAS bulls. The UB bulls spent more time in lying, ruminating, oral explorative and manipulative behaviour and rubbing and less time foraging and walking than the PAS bulls. The UB bulls performed more social licking and oral manipulation of objects and less mounting than the PAS bulls. These differences resulted most probably from the different feeding regimes and different space allowances.

  15. Reversible postural tachycardia syndrome due to inadvertent overuse of Red Bull.

    Science.gov (United States)

    Terlizzi, Rossana; Rocchi, Camilla; Serra, Maria; Solieri, Laura; Cortelli, Pietro

    2008-08-01

    Postural tachycardia syndrome associated with a vasovagal reaction was recorded in a young volleyball player after an excess intake of Red Bull as a refreshing energy drink. Considering the widespread use of Red Bull among young people who are often unaware of the drink's drug content, this case report suggest Red Bull be considered a possible cause of orthostatic intolerance.

  16. Whole-genome sequencing of 234 bulls facilitates mapping of monogenic and complex traits in cattle

    DEFF Research Database (Denmark)

    Daetwyler, Hans D; Capitan, Aurélien; Pausch, Hubert

    2014-01-01

    The 1000 bull genomes project supports the goal of accelerating the rates of genetic gain in domestic cattle while at the same time considering animal health and welfare by providing the annotated sequence variants and genotypes of key ancestor bulls. In the first phase of the 1000 bull genomes p...

  17. 77 FR 47334 - Safety Zone; Red Bull Flugtag, Delaware River; Camden, NJ

    Science.gov (United States)

    2012-08-08

    ... SECURITY Coast Guard 33 CFR Part 165 RIN 1625-AA00 Safety Zone; Red Bull Flugtag, Delaware River; Camden... to establish a safety zone for the ``Red Bull Flugtag Camden'', a marine event to be held on... Red Bull is sponsoring a Flugtag event along the Camden Riverfront. During this event participants...

  18. 75 FR 17106 - Safety Zone; Red Bull Air Race, Detroit River, Detroit, MI

    Science.gov (United States)

    2010-04-05

    ... SECURITY Coast Guard 33 CFR Part 165 RIN 1625-AA00 Safety Zone; Red Bull Air Race, Detroit River, Detroit... vessels from portions of the Detroit River during the Red Bull Air Race. This temporary safety zone is... in conjunction with the Red Bull Air Race. The safety zone will be in effect from 9 a.m. June 3, 2010...

  19. 75 FR 54026 - Safety Zone; Red Bull Flugtag, Delaware River, Camden, NJ

    Science.gov (United States)

    2010-09-03

    ... [Docket No. USCG-2010-0728] RIN 1625-AA00 Safety Zone; Red Bull Flugtag, Delaware River, Camden, NJ AGENCY... of the Delaware River during the Red Bull Flugtag event. The safety zone is necessary to protect... vessels and vessels from any debris in the water as a result from the event. Basis and Purpose Red Bull...

  20. 78 FR 38577 - Special Local Regulations; Red Bull Flugtag National Harbor Event, Potomac River; National Harbor...

    Science.gov (United States)

    2013-06-27

    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Red Bull Flugtag National...; Red Bull Flugtag National Harbor Event, Potomac River; National Harbor Access Channel, MD'' in the... safety of life on navigable waters of the United States during the Red Bull Flugtag National Harbor event...

  1. 75 FR 30708 - Safety Zone; Red Bull Air Race, Detroit River, Detroit, MI

    Science.gov (United States)

    2010-06-02

    ... SECURITY Coast Guard 33 CFR Part 165 RIN 1625-AA00 Safety Zone; Red Bull Air Race, Detroit River, Detroit... from portions of the Detroit River during the Red Bull Air Race. This temporary safety zone is... proposed rulemaking (NPRM) entitled Safety Zone; Red Bull Air Race, Detroit River, Detroit, MI in the...

  2. 78 FR 18274 - Special Local Regulations; Red Bull Flugtag National Harbor Event, Potomac River; National Harbor...

    Science.gov (United States)

    2013-03-26

    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Red Bull Flugtag National... during the ``Red Bull Flugtag National Harbor event,'' to be held on the waters of the Potomac River on... of National Harbor, Maryland, is sponsoring the Red Bull Flugtag National Harbor event, a competition...

  3. 77 FR 14965 - Special Local Regulations; Red Bull Candola, New River, Fort Lauderdale, FL

    Science.gov (United States)

    2012-03-14

    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Red Bull Candola, New River... east of the South Andrews Avenue Bascule Bridge in Fort Lauderdale, Florida for the Red Bull Candola... this rule because the Coast Guard did not receive necessary information about the Red Bull Candola...

  4. On-farm welfare and estimated daily carcass gain of slaughtered bulls

    NARCIS (Netherlands)

    Herva, T.; Virtala, A.M.; Huuskonen, A.; Saatkamp, H.W.; Peltoniemi, O.

    2009-01-01

    Welfare of growing bulls was evaluated using on-farm scoring modified by well-described test theory methods. Production parameters of the bulls were collected at slaughter. A positive relationship was observed between on-farm welfare, using the full A-Index score, and daily carcass gain of bulls.

  5. Bull trout distributions related to temperature regimes in four central Idaho streams

    Science.gov (United States)

    Susan B. Adams; Theodore C. Bjornn

    1997-01-01

    bull trout Salvelinus confluentus distributions and water temperature regimes were studied in four streams in the Weiser River basin, Idaho, in 1992 and 1993. bull trout occurred at elevations ranging from 1,472 m to 2,182 m and at densities up to 9.5 fish per 100 m2. Bull trout were sympatric with rainbow trout

  6. The bull as a source of trichomonosis and lumpy skin disease: An African perspective

    NARCIS (Netherlands)

    Irons, P.C.

    2007-01-01

    The use of bulls in breeding herds is regarded as a practical, labour-efficient way of producing a calf crop. However, the risks of a poor crop due to unsatisfactory performance of bulls are significant. The causes of sub- or infertility of the bull are classified under the headings Impotentia

  7. SCREENING OF BREEDING BULLS OF DIFFERENT BREEDS THROUGH KARYOTYPING

    Directory of Open Access Journals (Sweden)

    I. Ahmad, K. Javed1 and A. Sattar

    2004-10-01

    Full Text Available A study of chromosomal analysis for 200 breeding bulls of different breeds of cattle (Jersey, Holstein Friesian, Sahiwal and Cross-bred and Nili-Ravi buffalo, maintained at Semen Production Unit, Qadirabad and Livestock Experiment Station, Bhunikey (Pattoki was carried out. Micromethod was adopted for leukocyte culture and chromosomes were trapped at metaphase stage. The diploid number of chromosomes in all breeds of cattle was found to be 60 (58 autosomes and 2 sex chromosomes: XY, while this number in Nili-Ravi buffalo bulls was 50 (48 autosomes and 2 sex chromosomes: XY. All the autosomes and sex chromosomes in males of both species were found normal.

  8. 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

  9. Características seminais de touros Curraleiros e Holandeses submetidos à insulação escrotal Seminal characteristics of Curraleiro and Holstein bulls submitted to scrotal insulation

    Directory of Open Access Journals (Sweden)

    Tomaz Gelson Pezzini

    2006-05-01

    after insulation, returned to pre-insulation values sooner than that of Holstein bulls (51 days. Sperm vigor had a lower decrease after insulation in Curraleiro (15.2% than in Holstein bulls (47.8%. The total number of spermatozoa per ejaculate has dropped similarly in both groups after scrotal insulation (67.9 and 79.4% of drop for Curraleiro and Holstein bulls, respectively, between 16 and 23 days. The percentage of abnormal spermatozoa reached maximum values between 16 and 30 days after insulation and was greater in Holstein bulls (41.3±5.9% 30 days after the insult. Curraleiro are more tolerant to the effects of scrotal insulation on seminal characteristics than Holstein bulls.

  10. Subspecies and diet affect the expression of genes involved in lipid metabolism and chemical composition of muscle in beef cattle.

    Science.gov (United States)

    Teixeira, Priscilla D; Oliveira, Dalton M; Chizzotti, Mario L; Chalfun-Junior, Antonio; Coelho, Tamara C; Gionbelli, MateusP; Paiva, Luciano V; Carvalho, José Roberto R; Ladeira, Marcio M

    2017-11-01

    Fourteen Nellore and 14 Angus young bulls with BW of 381±11.8kg were randomly assigned into 2 feeding groups (whole shelled corn without forage (WSC) or corn silage and ground corn (GC)) to evaluate chemical composition and expression of genes involved in lipid metabolism in the longissimus thoracis (LT). We hypothesized that bulls fed the WSC diet have greater amounts of intramuscular fat and Angus have higher expression levels of PPAR and SREBF. Meat from Angus bulls had greater ether extract compared to Nellore (Pbeef of bulls fed this diet due to the reduced expression of SREBF1. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. The g.-165 T>C Rather than Methylation Is Associated with Semen Motility in Chinese Holstein Bulls by Regulating the Transcriptional Activity of the HIBADH Gene.

    Directory of Open Access Journals (Sweden)

    Shuai Zhang

    Full Text Available The 3-hydroxyisobutyrate dehydrogenase (HIBADH is regarded as a human sperm-motility marker. However, the molecular mechanisms involved in the regulation of expression of the HIBADH gene in bulls remain largely unknown. HIBADH was detected in the testis, epididymis, and sperm via reverse transcription polymerase chain reaction and Western blot analysis. It is also expressed in the seminiferous epithelium, spermatids, and the entire epididymis, as detected by immunohistochemistry. Furthermore, HIBADH was expressed in the neck-piece and mid-piece of bull spermatids, as shown in the immunofluorescence assay. Using serially truncated bovine HIBADH promoters and luciferase constructs, we discovered an 878 bp (-703 bp to +175 bp fragment that constitutes the core promoter region. One SNP g.-165 T>C of HIBADH was identified and genotyped in 307 Chinese Holstein bulls. Correlation analysis revealed that bulls with the TT genotype had higher initial sperm motility than those with the CC genotype (P C rather than methylation in the 5'-flanking region could affect the bovine sperm motility through the regulation of HIBADH gene transcriptional activity.

  12. Cyclic nucleotide phosphodiesterases in human spermatozoa and seminal fluid: Presence of an active PDE10A in human spermatozoa.

    Science.gov (United States)

    Maréchal, Loïze; Guillemette, Christine; Goupil, Serge; Blondin, Patrick; Leclerc, Pierre; Richard, François J

    2017-02-01

    Cyclic adenosine monophosphate (cAMP) plays a crucial role as a signaling molecule for sperm functions such as capacitation, motility and acrosome reaction. It is well known that cAMP degradation by phosphodiesterase (PDE) enzyme has a major impact on sperm functions. The present study was undertaken to characterize cAMP-PDE activity in human semen. cAMP-PDE activity was measured in human sperm and seminal plasma using family specific PDE inhibitors. Three sperm fractionation methods were applied to assess cAMP-PDE activity in spermatozoa. Western blots were used to validate the presence of specific family in sperm and seminal plasma. Using three sperm fractionation methods, we demonstrated that in human sperm, the major cAMP-PDE activity is papaverine-sensitive and thus ascribed to PDE10. In seminal plasma, total cAMP-PDE activity was 1.14±0.39fmol of cAMP hydrolyzed per minute per μg of protein. Using specific inhibitors, we showed that the major cAMP-PDE activity found in human seminal plasma is ascribed to PDE4 and PDE11. Western blot analysis, immunoprecipitation with a specific monoclonal antibody, and mass spectrometry confirmed the presence of PDE10 in human spermatozoa. This study provides the first demonstration of the presence of functional PDE10 in human spermatozoa and functional PDE4 and PDE11 in human seminal plasma. Since the contribution of cyclic nucleotides in several sperm functions is well known, the finding that PDE10 is an active enzyme in human spermatozoa is novel and may lead to new insight into fertility. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Species-specific and differential expression of BSP-5 and other BSP variants in normozoospermic and asthenozoospermic buffalo (Bubalus bubalis) and cattle (Bos taurus) seminal plasma.

    Science.gov (United States)

    Divyashree, B C; Roy, Sudhir C

    2018-01-15

    Binder of sperm-5 (BSP-5) is one of the fertility-associated proteins of cattle seminal plasma. Binding of sperm to the oviductal epithelium is mediated by BSP group of proteins. However, it is not clear, whether this protein is also involved in sperm motility. In the present study, attempts were made to characterize BSP-5 protein in both normozoospermic (NS) and asthenozoospermic (AS) Murrah buffalo (n = 18; Bubalus bubalis), Holstein Friesian (n = 8, Bos taurus) and Jersey cattle (n = 8; Bos taurus) bull seminal plasma and also study its expression pattern in these species. 1-D Western blot demonstrated three major BSP-5 immunoreactive protein bands (24.2 kDa, 20.5 kDa, and 12.3 kDa) in buffalo seminal plasma. Of these, the intensities of 24.2 and 20.5 kDa protein bands reduced significantly (P ≤ 0.05) in seminal plasma of AS group compared to that of NS group. On the contrary, the expression of 12.3 kDa protein band did not vary significantly between the groups. In Holstein Friesian seminal plasma, at least six BSP-5 immunoreactive protein bands (25.1, 23.6, 19.5, 13.8, 13.1 and 12.3 kDa) could be detected. Of these, the intensities of 23.6, 13.8/13.1 and 12.3 kDa protein bands decreased (P = 0.058, 0.111, 0.053) in AS group bulls compared to NS bulls. Holstein Friesian bull seminal plasma demonstrated a BSP-5 immunoreactive duplex protein band of 13.8/13.1 kDa, which was not evident in buffalo seminal plasma. In 2-D Western blot, a train of five BSP-5 immunoreactive duplex protein spots (Mr 21.0-27.6 kDa, pI of ∼3.9-5.1) was detected. Mass spectrometry of one of the representative duplex spot confirmed that these were BSP-5 and BSP-3 proteins, respectively. Indirect immunofluorescence studies showed that BSP-5 is primarily localized to the mid-piece/mitochondrial region of buffalo spermatozoa. To conclude, the findings of the present study could establish the significance and association of BSP-5 proteins in sperm motility and how their

  14. Enhanced early-life nutrition of Holstein bulls increases sperm production potential without decreasing postpubertal semen quality.

    Science.gov (United States)

    Dance, Alysha; Thundathil, Jacob; Blondin, Patrick; Kastelic, John

    2016-08-01

    Enhanced early-life nutrition (∼130% of required energy and protein) increased testes size and weight (∼20-25%) and reduced age at puberty (∼1 month) in beef and dairy bulls, compared with those fed 70% of dietary requirements. The objective was to determine effects of early-life (2-31 weeks) nutritional modulation on feed costs, predicted number of harvestable sperm and doses of semen, and semen quality. Calves (∼1 week old) were randomly allocated into three groups that were fed 4, 6, or 8 L/day of milk (low [n = 8], medium [n = 9], and high groups [n = 9], respectively) from ages 2 to 8 weeks. Thereafter, they were weaned, transitioned onto barley silage-based diets, to receive ∼70, 100, or 130% of recommended amounts of energy and protein (feed costs were ∼CDN$280 more per bull to feed high versus low diets from 2 to 31 weeks). After 31 weeks, all bulls were fed a medium diet. Semen was collected, by electroejaculation, from 51 to 73 weeks, extended, chilled, and cryopreserved. Bulls fed high nutrition were numerically younger (P = 0.45) at sexual maturity (sperm with ≥30% progressive motility, ≥70% morphologically normal, and ≤20% abnormal heads), first acceptable post-chill sperm motility (>50%; P = 0.66) and first acceptable post-thaw motility (>25% progressive; P = 0.25) than bulls in the low-nutrition group. Semen from three bulls per group was used for in vitro fertilization (total of 1249 bovine oocytes); there were no significant differences among groups in fertilization percentage (mean ± SEM of 68.0 ± 8.7, 77.1 ± 3.5, and 68.7 ± 4.5% for low, medium, and high, respectively) or blastocyst yield (31.5 ± 5.6, 41.4 ± 4.9, and 33.7 ± 4.6%). On the basis of analysis of 2D gels of sperm proteins, 380 spots were identified on the fused master gel, but no spots were differentially expressed across groups. Overall, there were no significant differences in semen quality or sperm function among bulls fed

  15. The ability of feline spermatozoa in different epididymal regions to undergo capacitation and acrosome reaction.

    Science.gov (United States)

    Kunkitti, Panisara; Bergqvist, Ann-Sofi; Sjunnesson, Ylva; Axnér, Eva

    2015-10-01

    The sperm maturation process that occurs in the epididymis is a necessary process for spermatozoa to acquire motility and the ability to undergo capacitation, which is an important key for fertilization. The aim of this study was to evaluate the ability of feline spermatozoa from different regions of the epididymis to undergo capacitation and acrosome reaction. Experiment I: epididymal spermatozoa from caput, corpus and cauda regions were placed in phosphate buffered saline (control medium) and in vitro fertilization medium (capacitating conditions). Sperm motility, motility patterns, plasma membrane integrity and tyrosine phosphorylation were evaluated at time 0 and 60min after incubation. Experiment II: spermatozoa were treated with 2μM of calcium ionophore (A23187) to induce the acrosome reaction and acrosome reaction was evaluated. The results showed a significant effect of region with a higher percentage of tyrosine phosphorylation in spermatozoa from the cauda than in the caput or corpus regions (P=0.0061; P=0.0088). Spermatozoa from corpus and cauda showed higher values in the majority of the measured motility parameters than spermatozoa from the caput (Pepididymal regions can undergo the acrosome reaction in vitro in response to induction by calcium ionophore with no difference between regions (P>0.05). Spermatozoa from all epididymal regions were able to undergo capacitation. Higher percentage of tyrosine phosphorylation in spermatozoa from the cauda reflect that they more easily underwent capacitation compared to spermatozoa from caput and corpus which required more time of incubation for capacitation. In conclusion feline epididymal spermatozoa from all regions can undergo capacitation and acrosome reaction in vitro and do not require incubation under capacitating conditions. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Feed efficiency and carcass and meat quality characteristics of bulls ...

    African Journals Online (AJOL)

    The inclusion of 8% SBP significantly improved panel ratings for tenderness, juiciness, beef flavour intensity and general acceptance, as well as number of chews before swallowing and the Warner Bratzler Shear value. Proximate analysis of the meat did not demonstrate differences between meat from the bulls fed C and ...

  17. Bulls, Goats, and Pedagogy: Engaging Students in Overseas Development Aid

    Science.gov (United States)

    Miles, William F. S.

    2009-01-01

    This article illustrates the profound learning that occurs--for students and instructor alike--when a class on third-world development attempts to undertake foreign aid. With undergraduate, graduate, and departmental money, I purchased bulls and carts for farmers, and goats for widows, in two West African villages. Such experiential learning…

  18. binding globulin gene of Bubalus bubalis bulls in Egypt

    Indian Academy of Sciences (India)

    Lillian

    This study was carried out on 67 adult buffalo bulls of the species Bubalus bubalis from different farms in. Damietta and .... result of the development and functional differentiation of the testis (Sharma et al. 1984 .... Gulia S., Sarkar M., Kumar V., Meyer H. H. D. and Prakash B. S. 2010 Divergent development of testosterone.

  19. Fusarium graminearum in a Papilloma Virus Infected Friesian Bull in ...

    African Journals Online (AJOL)

    Fusarium graminearum in a Papilloma Virus Infected Friesian Bull in Vom, Nigeria: Case Report. IO Fagbamila, CA Meseko, JA Adedeji, SS Ngulukun, Y Akalusi, JS Dalis, BO Akanbi, NJ Zwandor, J Okpara, PI Ankeli, OO Asala, L Taama, M Muhammad ...

  20. Bull. Chem. Soc. Ethiop. 1998, 12(1)

    African Journals Online (AJOL)

    Bull. Chem. Soc. Ethiop. 1998, 12(1), 69-77. ISSN 1011-3294. Printed in Ethiopia @ 1998 Chemical Society of Ethiopia ... The electrochemical cell and the reference electrodes used were the same as those described in ref. 6. RE ... potential. Further details are available in ref. 9. For comparison the capacitance values of.

  1. Bulls and bears: the stock market and clinical pathology research.

    Science.gov (United States)

    Khong, T Y

    2009-09-01

    To analyse the level of funded research in clinical pathology in a recent bear and bull market to act as a predictor for future funding during the current global financial crisis. The level of funding for research published in three clinical pathology journals in 2005 and 2008 to coincide with the bear market of March 2000 to October 2002 and with the subsequent bull market to October 2007 was determined using a Medline query. Other parameters examined were the type of article, affiliation of the first author and the pathology subspecialty. Approximately 30% of publications were funded and did not differ between the 2 years studied. Original research papers were more likely to be funded than case reports or reviews. Research from university departments of pathology was more likely to be funded than from hospital pathology departments but there were more publications from hospital pathology departments. The proportion of research in the different subspecialties that was funded did not differ significantly between each other and between 2005 and 2008. Based on data from the previous bear market, which was the longest and deepest of the post 1950 era, and the subsequent bull market, which led to the all-time high in the Dow Jones Industrial Index, funding for clinical pathology research does not seem to be affected by bull or bear markets.

  2. Cytotoxic triterpenoids from the mushroom Clavulina cinerea (Bull) J ...

    African Journals Online (AJOL)

    C. cinerea (Bull) J. Schröt (Lyophyllaceae) is among the many edible mushrooms in Kenya and is also traditionally regarded as a complementary medicine for chronically-ill people. The use of these mushrooms in the East African prompted this investigation in which the phytochemistry and potential anti-cancer activity was ...

  3. Record Litter Size for the Bull Shark, Carcharhinus leucas (Muller ...

    African Journals Online (AJOL)

    On the morning of 25 September 2013, a large female bull shark, Carcharhinus leucas, was landed in Port Victoria, Seychelles. It had been caught on an anchored long line set the previous evening, within 100 m of the main fishing quay. The female exhibited an unusually large girth for this heavy-set species. The shark ...

  4. Effective population size and genetic conservation criteria for bull trout

    Science.gov (United States)

    Bruce E. Rieman; F. W. Allendorf

    2001-01-01

    Effective population size (Ne) is an important concept in the management of threatened species like bull trout Salvelinus confluentus. General guidelines suggest that effective population sizes of 50 or 500 are essential to minimize inbreeding effects or maintain adaptive genetic variation, respectively....

  5. Welfare aspects of theriogenology: investigating alternatives to electroejaculation of bulls.

    Science.gov (United States)

    Palmer, Colin W

    2005-08-01

    Evaluation of the breeding soundness of bulls is an important management tool. Electroejaculation has been a reliable method of obtaining a semen sample for the purpose of evaluating breeding soundness, but is considered by some to be inhumane on the grounds that it is painful. This paper provides a review of studies conducted to find ways to both measure, as well as lessen, pain associated with electroejaculation, and to explore alternatives to electroejaculation in bulls. Changes in heart rate, serum cortisol, serum progesterone, relative aversion, and degrees of vocalization, struggling and lying down have been used to assess the pain associated with electroejaculation. Transrectal massage and artificial vaginas, and oxytocin and cloprostenol have been investigated as alternatives to, and facilitators of electroejaculation, respectively. Epidural, intravenous and topical anesthetics have been used to ameliorate the pain associated with electroejaculation. Serum progesterone and degrees of vocalization are useful for measuring the pain associated with electroejaculation in bulls. Transrectal massage and artificial vaginas are not as efficacious as electroejaculation for obtaining a semen sample and drugs used to facilitate or decrease pain associated with electroejaculation have not been efficacious enough to warrant use. Transrectal massage of the ampullae may be of some use as an alternative to electroejaculation in docile bulls and may be also be used to decrease the duration of subsequent electroejaculation. Pain associated with electroejaculation may be influenced by operator technique; therefore, operators of electroejaculator equipment must strive to apply electrical stimulation as gently as possible.

  6. The Missing Manuscript of Dr. Jose Delgado's Radio Controlled Bulls.

    Science.gov (United States)

    Marzullo, Timothy C

    2017-01-01

    Neuroscience systems level courses teach: 1) the role of neuroanatomical structures of the brain for perception, movement, and cognition; 2) methods to manipulate and study the brain including lesions, electrophysiological recordings, microstimulation, optogenetics, and pharmacology; 3) proper interpretation of behavioral data to deduce brain circuit operation; and 4) the similarities, differences, and ethics of animal models and their relation to human physiology. These four topics come together quite dramatically in Dr. Jose Delgado's 1960s famous experiments on the neural correlates of aggression in which he stopped bulls in mid-charge by electrically stimulating basal ganglia and thalamic structures. Technical documentation on these experiments is famously difficult to find. Here I translate and discuss a Spanish language article written by Dr. Delgado in 1981 for an encyclopedia on bull fighting published in Madrid. Here Dr. Delgado appears to give the most complete explanation of his experiments on microstimulation of bovine brains. Dr. Delgado's motivations, methods, and his interpretation of the bull experiments are summarized, as well as some accompanying information from his 1970 English language book: "Physical Control of the Mind." This review of Dr. Delgado's written work on the bull experiments can provide a resource to educators and students who desire to learn more about and interpret the attention-calling experiments that Dr. Delgado did on a ranch in Andalucía over 50 years ago.

  7. Bull's Mental Skills Questionnaire validation in an Afrikaans ...

    African Journals Online (AJOL)

    The purpose of this research was to validate the Bull's Mental Skills Questionnaire in an Afrikaans speaking population. An Afrikaans version of the scale was developed through a rigorous translation and back translation procedure. The scale was administered to a convenience sample of 674 first to third year, Human ...

  8. 75 FR 25794 - Regulated Navigation Area: Red Bull Air Race World Championship, Upper New York Bay, Lower Hudson...

    Science.gov (United States)

    2010-05-10

    ... SECURITY Coast Guard 33 CFR Part 165 RIN 1625-AA08 Regulated Navigation Area: Red Bull Air Race World... State Park, New Jersey and Ellis Island, New Jersey and New York for the Red Bull Air Race World... Register. Basis and Purpose Red Bull Air Race GmbH is sponsoring the Red Bull Air Race World Championship...

  9. Demographic characteristics of an adfluvial bull trout population in Lake Pend Oreille, Idaho

    Science.gov (United States)

    McCubbins, Jonathan L; Hansen, Michael J.; DosSantos, Joseph M; Dux, Andrew M

    2016-01-01

    Introductions of nonnative species, habitat loss, and stream fragmentation have caused the Bull Trout Salvelinus confluentus to decline throughout much of its native distribution. Consequently, in June 1998, the Bull Trout was listed under the U.S. Endangered Species Act as threatened. The Bull Trout has existed in Lake Pend Oreille and its surrounding tributaries since the last ice age, and the lake once supported a world-renowned Bull Trout fishery. To quantify the current status of the Bull Trout population in Lake Pend Oreille, Idaho, we compared the mean age, growth, maturity, and abundance with reports in a study conducted one decade earlier. Abundance was estimated by mark–recapture for Bull Trout caught in trap nets and gill nets set in Lake Pend Oreille during ongoing suppression netting of Lake Trout S. namaycushin 2007–2008. Bull Trout sampled in 2006–2008 were used to estimate age structure, survival, growth, and maturity. Estimated Bull Trout abundance was similar to that estimated one decade earlier in Lake Pend Oreille. Bull Trout residing in Lake Pend Oreille between 2006 and 2008 were between ages 4 and 14 years; their growth was fastest between ages 1 and 2 and slowed thereafter. Male and female Bull Trout matured at a similar age, but females grew faster than males, thereby maturing at a larger size. Our findings suggest that management has effectively addressed current threats to increase the likelihood of long-term persistence of the Bull Trout population in Lake Pend Oreille.

  10. Multiscale hydrogeomorphic influences on bull trout (Salvelinus confluentus) spawning habitat

    Science.gov (United States)

    Bean, Jared R; Wilcox, Andrew C.; Woessner, William W.; Muhlfeld, Clint C.

    2015-01-01

    We investigated multiscale hydrogeomorphic influences on the distribution and abundance of bull trout (Salvelinus confluentus) spawning in snowmelt-dominated streams of the upper Flathead River basin, northwestern Montana. Within our study reaches, bull trout tended to spawn in the finest available gravel substrates. Analysis of the mobility of these substrates, based on one-dimensional hydraulic modeling and calculation of dimensionless shear stresses, indicated that bed materials in spawning reaches would be mobilized at moderate (i.e., 2-year recurrence interval) high-flow conditions, although the asynchronous timing of the fall–winter egg incubation period and typical late spring – early summer snowmelt high flows in our study area may limit susceptibility to redd scour under current hydrologic regimes. Redd occurrence also tended to be associated with concave-up bedforms (pool tailouts) with downwelling intragravel flows. Streambed temperatures tracked stream water diurnal temperature cycles to a depth of at least 25 cm, averaging 6.1–8.1 °C in different study reaches during the spawning period. Ground water provided thermal moderation of stream water for several high-density spawning reaches. Bull trout redds were more frequent in unconfined alluvial valley reaches (8.5 versus 5.0 redds·km−1 in confined valley reaches), which were strongly influenced by hyporheic and groundwater – stream water exchange. A considerable proportion of redds were patchily distributed in confined valley reaches, however, emphasizing the influence of local physical conditions in supporting bull trout spawning habitat. Moreover, narrowing or “bounding” of these alluvial valley segments did not appear to be important. Our results suggest that geomorphic, thermal, and hydrological factors influence bull trout spawning occurrence at multiple spatial scales.

  11. 'ZP domain' of human zona pellucida glycoprotein-1 binds to human spermatozoa and induces acrosomal exocytosis.

    Science.gov (United States)

    Ganguly, Anasua; Bansal, Pankaj; Gupta, Tripti; Gupta, Satish K

    2010-09-11

    The human egg coat, zona pellucida (ZP), is composed of four glycoproteins designated as zona pellucida glycoprotein-1 (ZP1), -2 (ZP2), -3 (ZP3) and -4 (ZP4) respectively. The zona proteins possess the archetypal 'ZP domain', a signature domain comprised of approximately 260 amino acid (aa) residues. In the present manuscript, attempts have been made to delineate the functional significance of the 'ZP domain' module of human ZP1, corresponding to 273-551 aa fragment of human ZP1. Baculovirus-expressed, nickel-nitrilotriacetic acid affinity chromatography purified 'ZP domain' of human ZP1 was employed to assess its capability to bind and subsequently induce acrosomal exocytosis in capacitated human spermatozoa using tetramethyl rhodamine isothiocyanate conjugated Pisum sativum Agglutinin in absence or presence of various pharmacological inhibitors. Binding characteristics of ZP1 'ZP domain' were assessed employing fluorescein isothiocyanate (FITC) labelled recombinant protein. SDS-PAGE and immunoblot characterization of the purified recombinant protein (both from cell lysate as well as culture supernatant) revealed a doublet ranging from ~35-40 kDa. FITC- labelled 'ZP domain' of ZP1 binds primarily to the acrosomal cap of the capacitated human spermatozoa. A dose dependent increase in acrosomal exocytosis was observed when capacitated sperm were incubated with recombinant 'ZP domain' of human ZP1. The acrosome reaction mediated by recombinant protein was independent of Gi protein-coupled receptor pathway, required extra cellular calcium and involved both T- and L-type voltage operated calcium channels. Results described in the present study suggest that the 'ZP domain' module of human ZP1 has functional activity and may have a role during fertilization in humans.

  12. Complete staining of human spermatozoa and immature germ cells combined with phase contrast microscopy

    DEFF Research Database (Denmark)

    Michael, A Y; Drejer, J O; Bagger, P V

    1987-01-01

    A method combining Janus green B and Thymol blue stains the anterior part of the head, the nuclear membrane, middle piece, and tail of spermatozoa light green and the nucleus deep purple. The method provides excellent stained preparations for the evaluation of sperm morphology by phase contrast...... microscopy. It produces significantly less abnormal spermatozoa compared with the Papanicolaou stain....

  13. Cisplatin-DNA adduct formation in rat spermatozoa and its effect on fetal development

    NARCIS (Netherlands)

    Hooser, S.T.; Dijk-Knijnenburg, C.M. van; Waalkens-Berendsen, I.D.H.; Smits-van Prooije, A.E.; Snoeij, N.J.; Baan, R.A.; Fichtinger-Schepman, M.J.

    2000-01-01

    Exposure of males to some genotoxic chemicals causes DNA damage in spermatozoa resulting in embryotoxicity and developmental defects in their offspring. This study demonstrates that cisplatin-DNA adducts could be measured in spermatozoa following treatment with the antineoplastic drug, cisplatin.

  14. Cysteine protects rabbit spermatozoa against reactive oxygen species-induced damages

    Science.gov (United States)

    Fan, Xiaoteng; Pan, Yang; Lv, Shan; Pan, Chuanying; Lei, Anmin

    2017-01-01

    The process of cryopreservation results in over-production of reactive oxygen species, which is extremely detrimental to spermatozoa. The aim of this study was to investigate whether addition of cysteine to freezing extender would facilitate the cryosurvival of rabbit spermatozoa, and if so, how cysteine protects spermatozoa from cryodamages. Freshly ejaculated semen was diluted with Tris-citrate-glucose extender supplemented with different concentrations of cysteine. The motility, intact acrosomes, membrane integrity, mitochondrial potentials, 8-hydroxyguanosine level and sperm-zona pellucida binding capacity were examined. Furthermore, glutathione peroxidase (GPx) activity, glutathione content (GSH), and level of reactive oxygen species (ROS) and hydrogen peroxide of spermatozoa were analyzed. The values of motility, intact acrosomes, membrane integrity, mitochondrial potentials and sperm-zona pellucida binding capacity of the frozen-thawed spermatozoa in the treatment of cysteine were significantly higher than those of the control. Addition of cysteine to extenders improved the GPx activity and GSH content of spermatozoa, while lowered the ROS, DNA oxidative alterations and lipid peroxidation level, which makes spermatozoa avoid ROS to attack DNA, the plasma membrane and mitochondria. In conclusion, cysteine protects spermatozoa against ROS-induced damages during cryopreservation and post-thaw incubation. Addition of cysteine is recommended to facilitate the improvement of semen preservation for the rabbit breeding industry. PMID:28700739

  15. In vitro phagocytosis of boar spermatozoa by neutrophils from peripheral blood of sows

    NARCIS (Netherlands)

    Matthijs, A.; Harkema, W.; Engel, B.; Woelders, H.

    2000-01-01

    A considerable number of spermatozoa are used in each sow in routine artificial insemination. However, within a few hours after insemination, many spermatozoa are phagocytosed by polymorphonuclear leucocytes. Some aspects of sperm transport in the female genital tract in the sow have been thoroughly

  16. Artificial insemination with seminal plasma improves the reproductive performance of frozen-thawed boar epididymal spermatozoa.

    Science.gov (United States)

    Okazaki, Tetsuji; Akiyoshi, Teiichi; Kan, Masakazu; Mori, Manabu; Teshima, Hisanori; Shimada, Masayuki

    2012-01-01

    Frozen-thawed epididymal spermatozoa have good fertilization capability in vitro; however, their artificial insemination conception rate is less than half of that of frozen-thawed ejaculated spermatozoa. Because the addition of seminal plasma to the thawing solution enhances the in vivo fertilizing ability of frozen-thawed ejaculated spermatozoa, we hypothesized that the reproductive performance of frozen-thawed epididymal spermatozoa could also be improved by the inclusion of seminal plasma. When frozenthawed epididymal spermatozoa were incubated for up to 6 hours, the motility of the sperm significantly decreased in a time-dependent manner. The acrosomal membrane was damaged in the majority of frozen-thawed epididymal spermatozoa. The addition of seminal plasma to the thawing solution significantly decreased the percentage of sperm with abnormal acrosomes and increased their total motility in a dose-dependent manner. Furthermore, the addition of seminal plasma reduced the abundance of a 15-kDa tyrosinephosphorylated protein in frozen-thawed sperm, and the maximum effect was observed at 15% (vol/vol) seminal plasma. When cryopreserved epididymal spermatozoa from 3 different boars were thawed with a 15% (vol/vol) seminal plasma-containing solution, the conception rate and mean litter size obtained by artificial insemination were significantly increased as compared with those in the control without seminal plasma. From these results, we concluded that the addition of seminal plasma to the thawing solution is a key step in obtaining an optimal number of piglets by artificial insemination using frozen-thawed boar epididymal spermatozoa.

  17. Milk supplements in a glycerol free trehalose freezing extender enhanced cryosurvival of boar spermatozoa

    Directory of Open Access Journals (Sweden)

    Rukmali Athurupana

    2016-03-01

    Conclusion: 2% skim milk can be used as supplements for a glycerol-free trehalose and egg yolk-based extender to improve post-thaw survival of boar spermatozoa, whereas 2% coconut milk has an effect to protect boar spermatozoa from acrosome damage.

  18. Assessment of reproductive tissues of gilts born from magnetic nanoparticles-treated boar spermatozoa

    Science.gov (United States)

    Semen ejaculates contain a heterogeneous population of spermatozoa that may interfere with male fertility. Indeed, poor quality semen generally translates into low fertility rates that are attributed to higher proportions of damaged or abnormal spermatozoa in the ejaculates. It is likely that their ...

  19. Monitor and Protect Wigwam River Bull Trout for Koocanusa Reservoir; White River Bull Trout Enumeration Project Summary, Progress Report 2003.

    Energy Technology Data Exchange (ETDEWEB)

    Cope, R.

    2004-02-01

    This report summarizes the first year of a three-year bull trout (Salvelinus confluentus) enumeration project on the White River and is a co-operative initiative of the British Columbia Ministry of Water, Land, and Air Protection and Bonneville Power Administration. The White River has been identified as an important bull trout spawning tributary of the upper Kootenay River in southeastern British Columbia. The objective was to collect information on the returning adult spawning population to the White River through the use of a fish fence and traps, and to conduct redd surveys at the conclusion of spawning to provide an index of spawning escapement and distribution. The fence was installed on September 9th, 2003 and was operated continuously (i.e. no high-water or breaching events) until the fence was removed on October 9th, 2003. Estimation of the spawning population of White River bull trout was incomplete. This was due to a larger and more protracted out-migration than expected. As a result, the bull trout spawning population of the White River was estimated to be somewhere above 899 fish. In comparison, this represents approximately one third the population estimate of the 2003 Wigwam River bull trout spawning population. Based on redd index data, the number of bull trout per redd was over twice that of the Wigwam River or Skookumchuck Creek. This was expected as the index sites on the Wigwam River and Skookumchuck Creek cover the majority of the spawning area. This is not true on the White River. From previous redd counts, it is known that there are approximately twice as many redds in Blackfoot Creek as there are in the index site. Additionally, given the large size of the White River watershed and in particular, the large number of tributaries, there is a high likelihood that important bull trout spawning areas remain unidentified. Both floy tag and radio-telemetry data for the White River bull trout have identified extensive life history migrations

  20. Morphology and Ultrastructure of the Amazon River Dolphin (Inia geoffrensis) Spermatozoa.

    Science.gov (United States)

    Amaral, Rodrigo S; Da Silva, Vera M F; Valdez Domingos, Fabíola X; Martin, Anthony R

    2017-08-01

    The spermatozoa from seven adult Amazon river dolphins (Inia geoffrensis, CETACEA: INIIDAE) were analyzed by light and electron microscopy. The spermatozoa showed an elongated ellipsoid shaped head and a long tail with a well distinguishable midpiece. The head spermatozoa have a smooth surface like other odontocetes examined, with the exception of the Delphinidae family. The mean dimensions of the spermatozoa were within the range already reported for other cetaceans. The spermatozoa midpiece, as in other cetaceans, showed a random pattern of mitochondria, different from that described for other mammals. Further studies of sperm morphology of a wider spectrum of cetacean families could help to better understand the reproductive biology of these animals and the intergeneric and intrageneric relationships among them, as well as, among other mammals. Anat Rec, 300:1519-1523, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  1. EXTENDING THE VIABILITY OF SPERMATOZOA AND EGGS OF THE SEA URCHIN LYTECHINUS VARIEGATUS.

    Science.gov (United States)

    Malgarin, Jéssica; Resgalla, Charrid

    2015-01-01

    The storage of spermatozoa and eggs of the sea urchin Lytecninus variegatus can meet the demand of different human activities. To develop a protocol easy to reproduce for spermatozoa cryopreservation and cooling of the eggs of the sea urchin. Different formulations of artificial sea water were tested for their effectiveness in the freezing of sea urchin spermatozoa and storage of the eggs. Protocol for freezing of spermatozoa in liquid nitrogen presented the positive results when the cryoprotectant solution was diluted in artificial seawater free of calcium and magnesium. For the conservation of the eggs by cooling, the calcium-free artificial sea water, the calcium- and magnesium-free sea water, and the low-sodium water proved more efficient in preserving the integrity of the eggs. The results showed success in the freezing protocol of spermatozoa and cooling of the eggs mainly in artificial calcium- and magnesium-free sea water.

  2. Phagocytosis of spermatozoa by epithelial cells in the vagina of the lizard Hemidactylus mabouia (Reptilia, Squamata).

    Science.gov (United States)

    Nogueira, Katiane de Oliveira Pinto Coelho; Araújo, Vinícius Albano; Rodrigues Sartori, Sirlene Souza; Neves, Clóvis Andrade

    2011-07-01

    Hemidactylus mabouia is an Africa oviparous lizard that is now distributed on other continents and has been introduced to Brazil. In the majority of reptiles, the females have the ability to store spermatozoa in specialized regions of the genital tract. Considering that in H. mabouia the storage of spermatozoa is restricted to the region of the uterine tube, in this study we utilized optical and transmission electron microscopy to investigate the processes related to the large number of spermatozoa in the vagina. Although it was possible to visualize spermatozoa in the vagina, an ultrastructural analysis of the region revealed that significant phagocytosis occurs, which is mediated by the epithelial cells. Such a process indicates that the anterior portion of the vagina is related to the elimination of supernumerary or deficient spermatozoa and not storage. Copyright © 2010 Elsevier Ltd. All rights reserved.

  3. Paternity after vasectomy with two previous semen analyses without spermatozoa

    OpenAIRE

    Lucon, Marcos; Lucon, Antonio Marmo; Pasqualoto, Fabio Firmbach; Srougi, Miguel

    2007-01-01

    CONTEXT: The risk of paternity after vasectomy is rare but still exists. Overall failure to achieve sterility after vasectomy occurs in 0.2 to 5.3% of patients due to technical failure or recanalization. The objective of this report was to describe a rare but notable case of proven paternity in which the semen analyses had not given evidence of spermatozoa. CASE REPORT: A 44-year-old vasectomized man whose semen analyses had shown azoospermia became a father four years after sterilization. Bl...

  4. Evaluation of the damage in fish spermatozoa cryopreservation

    Science.gov (United States)

    Li, Jun; Liu, Qinghua; Zhang, Shicui

    2006-12-01

    Cryodamages occur during sperm cryopreservation. Cryopreservation of fish sperm usually results in marked decrease in sperm quality, such as swelling or disruption of the plasma membrane, mitochondrial dysfunction, diminished sperm motility, impaired velocity, shorter motility period, denaturation, and release of some enzymes from spermatozoa. In this paper, damages in morphology, physiology, biochemistry and metabolism, and genetic integrity of fish semen after cryopreservation are discussed. New approaches in assessment of fish thawed sperm quality such as computer assisted sperm analysis, flow cytometic analysis combined with fluorescent probes and single cell gel electrophoresis are also briefly reviewed.

  5. PLCz functional haplotypes modulating promoter transcriptional activity are associated with semen quality traits in Chinese Holstein bulls.

    Directory of Open Access Journals (Sweden)

    Qing Pan

    Full Text Available The sperm-specific phospholipase C zeta (PLCz is a candidate sperm-borne oocyte-activating factor that triggers a characteristic series of physiological stimuli via cytoplasmic Ca(2+ oscillations during fertilization. The molecular mechanisms involved in the regulation of PLCz gene expression remain largely unknown. To explore the genetic variations in the 5'-flanking region of the PLCz gene and their common haplotypes in Chinese Holstein bulls, as well as to determine whether these variations affect bovine semen quality traits and transcriptional activity, DNA samples were collected from Chinese Holstein bulls and sequenced for the identification of genetic variants in the 5'-flanking region of PLCz. Two genetic variants were identified, and their haplotypic profiles were constructed. The two novel genetic variations (g. -456 G>A and g. +65 T>C were genotyped in 424 normal Chinese Holstein bulls. Bioinformatics analysis revealed that both loci are in transcription factor binding sites of the core promoter region. The association studies revealed that the two genetic variations and their haplotype combinations significantly affected semen quality traits. Using serially truncated constructs of the bovine PLCz promoters and the luciferase reporter, we found that a 726 bp (-641 nt to +112 nt fragment constitutes the core promoter region. Furthermore, four haplotypes, H1H1 (GTGT, H2H2 (GCGC, H3H3 (ATAT, and H4H4 (ACAC, were significantly associated with semen quality traits and successfully transfected into MLTC-1 cell lines. The luciferase reporter assay showed that the different haplotypes exhibited distinct promoter activities. Maximal promoter activity was demonstrated by the H2H2 haplotypes, as compared with the other haplotypes. To the best of our knowledge, this study is the first report on genetic variants and their respective haplotypes in the 5'-flanking region of PLCz gene that can influence the semen quality of Chinese Holstein bulls as

  6. Fibroblast growth factor 2 (FGF2) is present in human spermatozoa and is related with sperm motility. The use of recombinant FGF2 to improve motile sperm recovery.

    Science.gov (United States)

    Garbarino Azúa, D J; Saucedo, L; Giordana, S; Magri, M L; Buffone, M G; Neuspiller, F; Vazquez-Levin, M H; Marín-Briggiler, C I

    2017-09-01

    Fibroblast growth factors (FGFs) and their receptors (FGFRs) regulate several functions of somatic cells. In a previous work, we reported FGFR expression in human spermatozoa and their involvement in motility. This study aimed to evaluate the presence and localization of fibroblast growth factor 2 (FGF2) in human spermatozoa, to determine the relationship of FGF2 levels with conventional semen parameters and to assess the effect of recombinant FGF2 (rFGF2) on sperm recovery in a selection procedure. Western immunoblotting analysis using an antibody against FGF2 revealed an 18-kDa band in sperm protein extracts. The protein was immunolocalized in the sperm flagellum and acrosomal region, as well as in all germ cells. Sperm FGF2 levels, assessed by flow cytometry, showed a positive (p recoveries, and increased (p recovery in selection techniques. © 2017 American Society of Andrology and European Academy of Andrology.

  7. Seminal plasma aids the survival and cervical transit of epididymal ram spermatozoa.

    Science.gov (United States)

    Rickard, J P; Pini, T; Soleilhavoup, C; Cognie, J; Bathgate, R; Lynch, G W; Evans, G; Maxwell, W M C; Druart, X; de Graaf, S P

    2014-11-01

    Seminal plasma purportedly plays a critical role in reproduction, but epididymal spermatozoa are capable of fertilisation following deposition in the uterus, calling into question the biological requirement of this substance. Through a combination of direct observation of spermatozoa in utero using probe-based Confocal Laser Endomicroscopy, in vivo assessment of sperm fertility and in vitro analysis of various sperm functional parameters, this study investigated the role of seminal plasma in spermatozoa transit through the cervix of the ewe. Following deposition in the cervical os, epididymal spermatozoa previously exposed to seminal plasma displayed an enhanced ability to traverse the cervix as evidenced by both significantly higher pregnancy rates and numbers of spermatozoa observed at the utero-tubal junction when compared with epididymal spermatozoa not previously exposed to seminal plasma. The beneficial effect of seminal plasma on sperm transport was clearly localised to transit through the cervix as pregnancy rates of spermatozoa deposited directly into the uterus were unaffected by exposure to seminal plasma. This phenomenon was not explained by changes to sperm motion characteristics, as seminal plasma had no effect on the motility, kinematic parameters or mitochondrial membrane potential of spermatozoa. Rather, in vitro testing revealed that seminal plasma improved the ability of epididymal spermatozoa to penetrate cervical mucus recovered from ewes in oestrus. These results demonstrate that the survival and transport of ram spermatozoa through the cervix of the ewe is not linked to their motility or velocity but rather the presence of some cervical penetration trait conferred by exposure to seminal plasma. © 2014 Society for Reproduction and Fertility.

  8. Temporary Restoration of Bull Trout Passage at Albeni Falls Dam

    Energy Technology Data Exchange (ETDEWEB)

    Paluch, Mark; Scholz, Allan; McLellan, Holly [Eastern Washington University Department of Biology; Olson, Jason [Kalispel Tribe of Indians Natural Resources Department

    2009-07-13

    This study was designed to monitor movements of bull trout that were provided passage above Albeni Falls Dam, Pend Oreille River. Electrofishing and angling were used to collect bull trout below the dam. Tissue samples were collected from each bull trout and sent to the U. S. Fish and Wildlife Service Abernathy Fish Technology Center Conservation Genetics Lab, Washington. The DNA extracted from tissue samples were compared to a catalog of bull trout population DNA from the Priest River drainage, Lake Pend Oreille tributaries, and the Clark Fork drainage to determine the most probable tributary of origin. A combined acoustic radio or radio tag was implanted in each fish prior to being transported and released above the dam. Bull trout relocated above the dam were able to volitionally migrate into their natal tributary, drop back downstream, or migrate upstream to the next dam. A combination of stationary radio receiving stations and tracking via aircraft, boat, and vehicle were used to monitor the movement of tagged fish to determine if the spawning tributary it selected matched the tributary assigned from the genetic analysis. Seven bull trout were captured during electrofishing surveys in 2008. Of these seven, four were tagged and relocated above the dam. Two were tagged and left below the dam as part of a study monitoring movements below the dam. One was immature and too small at the time of capture to implant a tracking tag. All four fish released above the dam passed by stationary receivers stations leading into Lake Pend Oreille and no fish dropped back below the dam. One of the radio tags was recovered in the tributary corresponding with the results of the genetic test. Another fish was located in the vicinity of its assigned tributary, which was impassable due to low water discharge at its mouth. Two fish have not been located since entering the lake. Of these fish, one was immature and not expected to enter its natal tributary in the fall of 2008. The other

  9. Ultrasound Imaging of Testes and Epididymides of Normal and Infertile Breeding Bulls

    Directory of Open Access Journals (Sweden)

    Khalid Mahmood Ali, Nazir Ahmad*, Nafees Akhtar, Shujait Ali, Maqbool Ahmad and Muhammad Younis1

    2011-10-01

    Full Text Available Echotexture of testes and epididymides from 10 slaughtered male buffaloes was studied. Diameter of testis and mediastinum testis was measured by ultrasound and compared with respective values taken by calipers. Testes and epididymides of another 10 fertile and 10 infertile breeding bulls were examined in vivo through manual palpation and ultrasound imaging. Semen quality of these bulls was also monitored. There were significant (P<0.01 positive correlations between ultrasound and calipers values of all parameters. The testicular parenchyma of fertile bulls was uniformly homogeneous and moderately echogenic. Epididymal tail was more heterogeneous and less echogenic, while epididymal head was homogeneous and less echogenic, than the testicular parenchyma. The epididymal body appeared as hypoechoic structure with echogenic margin. Among 10 infertile bulls, nine had poor semen quality, while one bull failed to give any ejaculate. On ultrasonography, six bulls showed abnormalities in their scrotal echotexture. Among these, one had an abundance of hyperechoic areas scattered in the testicular parenchyma, some of these showed acoustic shadowing, showing testicular degenerations with mineralization. The second bull showed many anechoic areas in the testes and epididymal head, demarcated from the rest of the organ by well defined margins. In the third bull, three-fourth of the right testis showed hyperechoic areas, suspected of testicular degeneration with mineralization. The fourth bull had two anechoic areas in one testis assumed to represent dilated blood vessel. The fifth bull showed small hyperechoic areas within the testicular parenchyma. The sixth bull showed an anechoic area with distinct hyperechogenic margin below the testicular tunics. The remaining four bulls had normal echogenicity of testes and epididymides in spite of poor semen quality. In conclusion, diagnostic ultrasound may be included in breeding soundness examination of breeding

  10. Detection of copy number variations and their effects in Chinese bulls

    KAUST Repository

    Zhang, Liangzhi

    2014-06-17

    Background: Copy number variations (CNVs) are a main source of genomic structural variations underlying animal evolution and production traits. Here, with one pure-blooded Angus bull as reference, we describe a genome-wide analysis of CNVs based on comparative genomic hybridization arrays in 29 Chinese domesticated bulls and examined their effects on gene expression and cattle growth traits.Results: We identified 486 copy number variable regions (CNVRs), covering 2.45% of the bovine genome, in 24 taurine (Bos taurus), together with 161 ones in 2 yaks (Bos grunniens) and 163 ones in 3 buffaloes (Bubalus bubalis). Totally, we discovered 605 integrated CNVRs, with more " loss" events than both " gain" and " both" ones, and clearly clustered them into three cattle groups. Interestingly, we confirmed their uneven distributions across chromosomes, and the differences of mitochondrion DNA copy number (gain: taurine, loss: yak & buffalo). Furthermore, we confirmed approximately 41.8% (253/605) and 70.6% (427/605) CNVRs span cattle genes and quantitative trait loci (QTLs), respectively. Finally, we confirmed 6 CNVRs in 9 chosen ones by using quantitative PCR, and further demonstrated that CNVR22 had significantly negative effects on expression of PLA2G2D gene, and both CNVR22 and CNVR310 were associated with body measurements in Chinese cattle, suggesting their key effects on gene expression and cattle traits.Conclusions: The results advanced our understanding of CNV as an important genomic structural variation in taurine, yak and buffalo. This study provides a highly valuable resource for Chinese cattle\\'s evolution and breeding researches. 2014 Zhang et al.; licensee BioMed Central Ltd.

  11. Spermatozoa of the 'primitive type' in Scutigerella (Myriapoda, Symphyla).

    Science.gov (United States)

    Dallai, R; Afzelius, B A

    2000-02-01

    The myriapod class Symphyla is of interest in that insects generally are assumed to be derived from symphylan-like ancestors. Male Symphyla form spermatophores that are picked up by the female. Both euspematozoa and paraspermatozoa are formed. In spite of the mode of fertilization their euspermatozoa were found to be of a kind that is typical of aquatic animals, so called 'primitive spermatozoa;' these are characterized by a short sperm head with a bilayered acrosome, a midpiece containing a few unmodified mitochondria, and a 9 + 2 flagellum. Scutigerella are unique among terrestrial arthropods in having 'primitive spermatozoa'; and together with horseshoe crabs they are only the second case in Arthropoda. Two further sperm plesiomorphies not found in other myriapods or insects are (1) the presence of a cytoplasmic canal housing the proximal flagellum and (2) the existence of microtubular triplets in centrioles, one of which acts as a basal body. Symphyla and Diplopoda both have a striated structure in the center of the subacrosomal material. The paraspermatozoa lack acrosome and nucleus but have a prominent crystal, a single mitochondrion, and two membrane systems. The structure of Scutigerella euspermatozoa is consistent with Symphyla being close to the stem group of Myriapoda plus Insecta.

  12. Bioluminescent magnetic nanoparticles as potential imaging agents for mammalian spermatozoa.

    Science.gov (United States)

    Vasquez, Erick S; Feugang, Jean M; Willard, Scott T; Ryan, Peter L; Walters, Keisha B

    2016-03-17

    Nanoparticles have emerged as key materials for developing applications in nanomedicine, nanobiotechnology, bioimaging and theranostics. Existing bioimaging technologies include bioluminescent resonance energy transfer-conjugated quantum dots (BRET-QDs). Despite the current use of BRET-QDs for bioimaging, there are strong concerns about QD nanocomposites containing cadmium which exhibits potential cellular toxicity. In this study, bioluminescent composites comprised of magnetic nanoparticles and firefly luciferase (Photinus pyralis) are examined as potential light-emitting agents for imaging, detection, and tracking mammalian spermatozoa. Characterization was carried out using infrared spectroscopy, TEM and cryo-TEM imaging, and ζ-potential measurements to demonstrate the successful preparation of these nanocomposites. Binding interactions between the synthesized nanoparticles and spermatozoon were characterized using confocal and atomic/magnetic force microscopy. Bioluminescence imaging and UV-visible-NIR microscopy results showed light emission from sperm samples incubated with the firefly luciferase-modified nanoparticles. Therefore, these newly synthesized luciferase-modified magnetic nanoparticles show promise as substitutes for QD labeling, and can potentially also be used for in vivo manipulation and tracking, as well as MRI techniques. These preliminary data indicate that luciferase-magnetic nanoparticle composites can potentially be used for spermatozoa detection and imaging. Their magnetic properties add additional functionality to allow for manipulation, sorting, or tracking of cells using magnetic techniques.

  13. Prolonged incubation of processed human spermatozoa will increase DNA fragmentation.

    Science.gov (United States)

    Nabi, A; Khalili, M A; Halvaei, I; Roodbari, F

    2014-05-01

    One of the causes of failure in ART is sperm DNA fragmentation which may be associated with long period of spermatozoa incubation at 37 °C. The objective was to evaluate the rate of sperm DNA fragmentation using the sperm chromatin dispersion (SCD) test after swim-up at different time intervals prior to use. In this prospective study, 21 normozoospermic specimens were analysed. The samples were incubated at 37 °C after preparation by direct swim-up. DNA fragmentation was assessed at different time intervals (0, 1, 2 and 3 h) using SCD test. Spermatozoa with no DNA fragmentation showed large- or medium-sized halos, and sperm cells with DNA fragmentation showed either a small halo or no halo. The rates of normal morphology and progressive motility after sperm processing were 72.33 ± 2.53% and 90 ± 1.02%, respectively. The rate of sperm DNA fragmentation was significantly higher after 2 h (8.81 ± 0.93%, P = 0.004) and 3 h (10.76 ± 0.89%, P fragmentation. Therefore, sperm samples intended for ART procedures should be used within 2 h of incubation at 37 °C. © 2013 Blackwell Verlag GmbH.

  14. In vitro fertilization using frozen-thawed feline epididymal spermatozoa from corpus and cauda regions.

    Science.gov (United States)

    Kunkitti, Panisara; Axnér, Eva; Bergqvist, Ann-Sofi; Sjunnesson, Ylva

    2016-10-01

    Epididymal sperm preservation offers a potential for rescuing genetic material from endangered or valuable animals after injury or death. Spermatozoa from corpus, as well as from cauda, have the capability to be motile and to undergo capacitation and can thus potentially be preserved for assisted reproductive technologies. In the present study, feline frozen-thawed epididymal spermatozoa from corpus and cauda regions were investigated for their ability to fertilize homologous oocytes and further embryo development in vitro. Epididymal spermatozoa from corpus and cauda of seven cats were cryopreserved and used for IVF. Cumulus-oocyte complexes (n = 419) were obtained from female cats after routine spaying. Frozen-thawed corpus epididymal spermatozoa showed similar properties of acrosome integrity, membrane integrity, and chromatin integrity as frozen-thawed spermatozoa from cauda except corpus spermatozoa showed lower motility (P epididymal spermatozoa was confirmed by similar number of embryos developing to the two- and four-cell stages compared with sperm from cauda (32.03% vs. 33.33%). However, oocytes fertilized with corpus spermatozoa had lower potential to develop to the blastocyst stage (6.79%) and had lower cell numbers compared to oocytes fertilized with cauda spermatozoa (14.08%). In conclusion, spermatozoa from corpus epididymis had a similar capability to fertilize homologous oocytes in vitro as sperm from cauda but resulted in fewer embryos developing to the blastocyst stage compared to spermatozoa from the cauda. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Environmental Enrichment in Kennelled Pit Bull Terriers (Canis lupus familiaris)

    OpenAIRE

    Kiddie, Jenna; Bodymore, Anna; Alex, Dittrich

    2017-01-01

    Simple Summary Rescue shelters, although set-up with good intentions, may lead to poor welfare of the animals that they house, especially when the shelter organisation is under-resourced. This study therefore investigated the behavioural effects of cheap and locally accessible objects added to the cages of rescued Pit Bull Terrier type dogs in the Philippines to determine whether it is possible for shelter organisations with limited funds and staffing to improve the welfare of their dogs with...

  16. Inquisitorial bulls : Ad Abolendam (1184 and Vergentis in Senium (1199

    Directory of Open Access Journals (Sweden)

    Leandro Duarte Rust

    2012-06-01

    Full Text Available In this paper we present two important medieval documents, the decretals Ad Abolendam (1184 and Vergentis in Senium (1199, in a bilingual edition, Latin-Portuguese. Usually described as "the founding texts of Inquisition", these bulls record the political scene at the end of the 12th century. Unpublished in Portuguese, the translation comes to public with a brief introductory text and some notes for historical information.

  17. Nutrient intake, digestibility and rumen metabolites in bulls fed rice ...

    African Journals Online (AJOL)

    Nutrient intake, digestibility and rumen metabolites were determined in rumen - cannulated bulls fed rice straw or straw supplemented with urea, groundnut hay or cotton seed cake. Total dry matter intake (DMI) ranged from 7.55 Lo 8.29kg/d or 3.66 to 4.04% of liveweight and from 6.48 to 7. 21 kg/d for organic matter.

  18. The Papal Bulls on Papyrus: an Approach to their Conservation

    OpenAIRE

    Bello, Carmen; Borrell, Àngels

    2013-01-01

    There are few Papal bulls on Papyrus, it is a support difficult to find and to preserve, and that’s why we have to consider them like an exceptional historic and cultural heritage, especially in Catalonia where some important collections are preserved. Our work over the years has been studying, documenting and preserving these extraordinary documents and setting some precedents which will serve as a good model for their documentation and for any action on conservation or restoration in the fu...

  19. Receptor mediated agglutination of human spermatozoa by spermagglutinating factor isolated from Staphylococcus aureus.

    Science.gov (United States)

    Kaur, Siftjit; Prabha, Vijay; Sarwal, Abha

    2010-12-01

    We examined spermagglutinating factor isolated from Staphylococcus aureus for evidence of receptor mediated agglutination of human spermatozoa. Binding to spermatozoa by spermagglutinating factor isolated from S. aureus with a high degree of specificity indicates receptor-ligand interaction. To examine this interaction we isolated and purified the ligand and the receptor. To assess receptor mediated agglutination of spermatozoa further we blocked spermagglutination induced by spermagglutinating factor in the presence of receptor. Spermagglutinating factor induced spermagglutination was competitively inhibited by adding purified receptor, indicating that sperm agglutinating factor isolated from S. aureus attaches to specific receptors on human spermatozoa. The spermagglutinating factor receptor was a protein with a molecular weight of approximately 57 kDa. Spermagglutinating factor induced spermagglutination and at higher concentrations had a spermicidal effect, which was inhibited by introducing the receptor. As observed on scanning electron microscopy studies, incubating spermatozoa with spermagglutinating factor showed profound morphological alterations. However, spermatozoa with normal morphology were noted when incubated with spermagglutinating factor in the presence of receptor, indicating that morphological alterations may account for spermatozoa agglutination by spermagglutinating factor. Results suggest that spermagglutinating factor isolated from S. aureus may bind specifically to sperm surface receptor sites before causing spermagglutination. Copyright © 2010 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

  20. Cryopreservation of kangaroo spermatozoa using alternative approaches that reduce cytotoxic exposure to glycerol.

    Science.gov (United States)

    McClean, Rhett; Zee, Yeng Peng; Holt, William V; Johnston, Stephen D

    2008-12-01

    Alternative techniques for the cryopreservation of kangaroo spermatozoa that reduced or eliminated the need for glycerol were investigated including; (1) freezing spermatozoa with 20% glycerol in pre-packaged 0.25 mL Cassou straws to enable rapid dilution of the glycerol post-thaw, (2) investigating the efficacy of 20% (v/v) dimethyl sulphoxide (DMSO) and dimethylacetamide (DMA-10%, 15% and 20% v/v) as cryoprotectants and (3) vitrification of spermatozoa with or without cryoprotectant (20% v/v glycerol, 20% v/v DMSO and 20% v/v DMA). Immediate in-straw post-thaw dilution of 20% glycerol and cryopreservation of spermatozoa in 20% DMSO produced no significant improvement in post-thaw viability of kangaroo spermatozoa. Spermatozoa frozen in 20% DMA showed post-thaw motility and plasma membrane integrity of 12.7+/-1.9% and 22.7+/-5.4%, respectively, while kangaroo spermatozoa frozen by ultra-rapid freezing techniques showed no evidence of post-thaw viability. The use of 10-20% DMA represents a modest but significant improvement in the development of a sperm cryopreservation procedure for kangaroos.

  1. [The immobilization effect of zinc ions on human spermatozoa in vitro].

    Science.gov (United States)

    Zhu, W H

    1991-08-01

    Based on the literature on the suppression of fertility by copper an zinc, a study on the effectiveness of zinc solution in suppressing the mobility of spermatozoa was conducted. 30 samples of semen from men 22-42 years of age were collected. Zinc was diluted in 4 mg/ml, 6 mg/ml, and 8 mg/ml of salt water. 0.2 ml of semen was mixed with 0.5 ml of each concentration of zinc solution. In comparison, another test tube of 0.2 ml of semen was mixed with 0.5 ml of salt water. Test results showed that increasing concentrations of zinc solution and increasing time were associated with the increasing spermatozoa immobilization. With the exception of the 4 mg/ml group, which in 5 and 30 minutes showed no significant difference in immobilization effect with the comparison group, the other groups all showed significant differences in immobilizing spermatozoa for the same amount of time. When the zinc concentration was 6-8 mg/ml, at 120 minutes, the immobilization effect was 100%. When the concentration reached 20 mg.ml, spermatozoa were immobilized in 20 seconds. The effects of zinc solution was basically to immobilize instead of killing the spermatozoa. As a potential agent for contraception, zinc ions could not only immobilize spermatozoa, they could also suppress the penetrating enzyme in spermatozoa, which will achieve the effect of infertility. Zinc is a hopeful agent in vaginal contraception.

  2. THE IMPACT OF NONYLPHENOL (NP ON THE SPERMATOZOA MOTILITY IN VITRO

    Directory of Open Access Journals (Sweden)

    Jana Lukáčová

    2012-06-01

    Full Text Available Nonylphenol (NP is an environmental endocrine disruptor that has toxic, estrogenic and carcinogenic effects in fish, amphibians and mammals. NP can result in male reproductive dysfunction, altered testicular development, decreased male fertility and a decline of spermatozoa count. The target of this in vitro study was to determine the effect of NP on the spermatozoa motility. Specifically, we examined the dose- and time-dependent effect of nonylphenol (1, 10, 100 and 200 µg/mL dissolved either in 0.1% dimethyl sulfoxide (DMSO or 0.1% ethanol (ETOH on the motility of bovine spermatozoa during several time periods (0 h, 2 h, 4 h and 6 h. The spermatozoa motility was determined by CASA (Computer Assisted Semen Analyzer system using the Sperm VisionTM program. The results showed a decreased spermatozoa motility in all experimental groups with the addition of NP. Significant differences (P<0.001 and P<0.05 between the control group and all experimental groups were recorded. The lowest motility of bovine spermatozoa was found at doses > 100 µg/mL of NP in comparison with the control group. The obtained data indicate that the exposure to high doses of NP has the negative effect on spermatozoa motility.

  3. Minimization of apoptosis-like changes in cryopreserved buffalo bull sperm by supplementing extender with Bcl-2 protein

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    Jasmer Dalal

    2016-05-01

    Full Text Available Aim: This study was aimed at evaluating the anti-apoptotic effects of Bcl-2 protein in cryopreserved buffalo bull sperm. Materials and Methods: A total 10 ejaculates from two buffalo bulls (5 each were collected using artificial vagina method, and semen was evaluated using a standard protocol. Semen was extended by Tris egg yolk extender supplemented with Bcl-2 protein at 5, 10, and 15 μM. Semen was cryopreserved at ultra-low temperature using traditional vapor freezing method. Prefreeze and post-thaw semen samples were evaluated for percent motility, viability, hypo-osmotic swelling test (HOST reactive sperms; status of mitochondrial membrane activity and status of sperm phospholipase A1 and phospholipase A2 activity. Results: There were no significant effects of Bcl-2 protein supplementation on pre-freeze sperm quality. Percent motility and active mitochondria in post-thaw Bcl-2 supplemented and control groups were also similar. However, viable sperms were significantly (p<0.05 higher (74.29±4.23% in Bcl-2 supplemented group (5 μM as compared to control (51.6±5.77%. The proportion of HOST reactive sperms was also higher (63.1±6.73% in Bcl-2 supplemented (5 μM group as compared to control (50.7±6.98%. The sperm with low PLA activity (non-apoptotic was significantly (p<0.05 higher in all the supplemented doses of Bcl-2 protein, i.e., at 5 μM (73.42±5.79%, 10 μM (75.51±6.22%, and 15 μM (74.78±5.89% as compared to control (60.23±4.45%. We found that Bcl-2 protein supplementation at 5 μM dose improved the postthaw semen quality indicated by higher viability, HOST reactive sperms, and sperm with low PLA activity (non-apoptotic sperms. Conclusion: Bcl-2 protein supplementation exerts its protective effect on spermatozoa against apoptosis-like changes developed during cryopreservation.

  4. Discerning between Giant Kelp and Bull Kelp in AVIRIS Imagery

    Science.gov (United States)

    Thompson, T. J.; Kudela, R. M.; Bausell, J.

    2016-12-01

    Kelp forests are an important staple of sub-tidal coastal ecosystems in the subtropical and temperate zones. Two species of kelp form the backbone of the northeastern Pacific coast's kelp forests: bull kelp (Nereocystis Luetkeana) and giant kelp (Macrocystis Pyrifera). In recent years, these species have been declining at an alarming rate due to warmer water temperatures and the rising abundance of purple urchins, which feed on kelp holdfasts. Additionally, in situ measurements have indicated that this rapid decline is especially severe for bull kelp. However, because of assumptions about the spectral similarity of these two kinds of kelp, oceanographers do not have a good idea of their relative prevalence on a large scale. In this study, data from 2013 AVIRIS runs are used to investigate the spectral separability of bull kelp and giant kelp. Two machine-learning-based methods of distinguishing are presented, one involving spectral angle comparison and the other using linear unmixing techniques. Reference endmembers are selected from kelp beds near Manchester and Fort Ross in northern California and Santa Barbara in southern California. These are then verified using survey data from Reef Check. The two methods are then tested on pixels taken from kelp beds in the vicinity of Monterey and Carmel, CA. The results show some spectral separability of the two kinds of kelp, as well as potential for application in large-scale surveying operations.

  5. Morphological alterations in cryopreserved spermatozoa of scallop Argopecten purpuratus Alteraciones morfológicas en espermatozoides criopreservados de concha de abanico Argopecten purpuratus

    OpenAIRE

    Carlos Espinoza; Martha Valdivia; Enrique Dupré

    2010-01-01

    The present work identifies and quantifies the morphological alterations of scallop Argopecten purpuratus spermatozoa caused by long-term cryopreservation. Percentages of motility, fertilization and injured spermatozoa were quantified by optic microscopy and scanned electron microscopy. These parameters were evaluated in sperm without treatment (CTR), spermatozoa incubated in cryoprotective solution but not freezed (ICS) and freezed-thawed spermatozoa (FTS). Spermatozoa of ICS treatment remai...

  6. Morphology of spermatozoa in the cauda epididymidis before and after electroejaculation and a comparison with ejaculated spermatozoa in the domestic cat.

    Science.gov (United States)

    Axnér, E; Holst, B S; Linde-Forsberg, C

    1998-10-15

    When 2 ejaculates are collected by electroejaculation from the domestic cat within a period of 10 min the first ejaculate has a higher proportion of abnormal spermatozoa than the second. The reason for this difference is not known for the domestic cat, but in other species long-term epididymal storage results in a higher proportion of abnormal spermatozoa. The aims of this study were to determine the proportions of abnormal spermatozoa in the cauda epididymidis and to ascertain if electroejaculation affects this proportion. Therefore the proportions of spermatozoa in the cauda epididymidis with different morphological abnormalities were compared before and after ejaculation. In addition, the proportion of morphologically abnormal spermatozoa in the epididymis was compared with that in the ejaculate. Nine privately-owned domestic cats were anesthetized, and one testicle was surgically removed. An ejaculate was collected by electroejaculation, after which the remaining testicle was ectomized. There were no significant differences in the proportions of different sperm abnormalities between the cauda epididymidis removed before ejaculation and the one removed after ejaculation. A significantly (P = 0.009) higher proportion of spermatozoa with tail abnormalities was found in the ejaculates compared with the cauda epididymides (11.1 and 1.6%, respectively), while, as expected, there was a lower proportion of spermatozoa with distal droplets in the ejaculates than in the cauda epididymides (35.1 and 75.9%, respectively). This new information contributes to the understanding of the etiology of sperm defects in the domestic cat, and is of importance when evaluating a semen sample in this species.

  7. Thiol changes during epididymal maturation: a link to flagellar angulation in mouse spermatozoa?

    Science.gov (United States)

    Ijiri, T W; Vadnais, M L; Huang, A P; Lin, A M; Levin, L R; Buck, J; Gerton, G L

    2014-01-01

    Caput epididymal wild-type spermatozoa and cauda epididymal spermatozoa from mice null for the adenylyl cyclase Adcy10 gene are immotile unless stimulated by a membrane-permeant cyclic AMP analogue. Both types of spermatozoa exhibit flagellar angulation where the head folds back under these conditions. As sperm proteins undergo oxidation of sulfhydryl groups and the flagellum becomes more stable to external forces during epididymal transit, we hypothesized that ADCY10 is involved in a mechanism regulating flagellar stabilization. Although no differences were observed in global sulfhydryl status between caput and cauda epididymal spermatozoa from wild-type or Adcy10-null mice, two-dimensional fluorescence difference gel electrophoresis was performed to identify specific mouse sperm proteins containing sulfhydryl groups that became oxidized during epididymal maturation. A-kinase anchor protein 4, fatty acid-binding protein 9 (FABP9), glutathione S-transferase mu 5 and voltage-dependent anion channel 2 exhibited changes in thiol status between caput and cauda epididymal spermatozoa. The level and thiol status of each of these proteins were quantified in wild-type and Adcy10-null cauda epididymal spermatozoa. No differences in the abundance of any protein were observed; however, FABP9 in Adcy10-null cauda epididymal spermatozoa contained fewer disulfide bonds than wild-type sperm cells. In caput epididymal spermatozoa, FABP9 was detected in the cytoplasmic droplet, principal piece, midpiece, and non-acrosomal area of the head. However, in cauda epididymal spermatozoa, this protein localized to the perforatorium, post-acrosomal region and principal piece. Together, these results suggest that thiol changes during epididymal maturation have a role in the stabilization of the sperm flagellum. © 2013 American Society of Andrology and European Academy of Andrology.

  8. Epididymis response partly compensates for spermatozoa oxidative defects in snGPx4 and GPx5 double mutant mice.

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    Anaïs Noblanc

    Full Text Available We report here that spermatozoa of mice lacking both the sperm nucleus glutathione peroxidase 4 (snGPx4 and the epididymal glutathione peroxidase 5 (GPx5 activities display sperm nucleus structural abnormalities including delayed and defective nuclear compaction, nuclear instability and DNA damage. We show that to counteract the GPx activity losses, the epididymis of the double KO animals mounted an antioxydant response resulting in a strong increase in the global H(2O(2-scavenger activity especially in the cauda epididymis. Quantitative RT-PCR data show that together with the up-regulation of epididymal scavengers (of the thioredoxin/peroxiredoxin system as well as glutathione-S-transferases the epididymis of double mutant animals increased the expression of several disulfide isomerases in an attempt to recover normal disulfide-bridging activity. Despite these compensatory mechanisms cauda-stored spermatozoa of double mutant animals show high levels of DNA oxidation, increased fragmentation and greater susceptibility to nuclear decondensation. Nevertheless, the enzymatic epididymal salvage response is sufficient to maintain full fertility of double KO males whatever their age, crossed with young WT female mice.

  9. Epididymis Response Partly Compensates for Spermatozoa Oxidative Defects in snGPx4 and GPx5 Double Mutant Mice

    Science.gov (United States)

    Noblanc, Anaïs; Peltier, Manon; Damon-Soubeyrand, Christelle; Kerchkove, Nicolas; Chabory, Eléonore; Vernet, Patrick; Saez, Fabrice; Cadet, Rémi; Janny, Laurent; Pons-Rejraji, Hanae; Conrad, Marcus; Drevet, Joël R.; Kocer, Ayhan

    2012-01-01

    We report here that spermatozoa of mice lacking both the sperm nucleaus glutathione peroxidase 4 (snGPx4) and the epididymal glutathione peroxidase 5 (GPx5) activities display sperm nucleus structural abnormalities including delayed and defective nuclear compaction, nuclear instability and DNA damage. We show that to counteract the GPx activity losses, the epididymis of the double KO animals mounted an antioxydant response resulting in a strong increase in the global H2O2-scavenger activity especially in the cauda epididymis. Quantitative RT-PCR data show that together with the up-regulation of epididymal scavengers (of the thioredoxin/peroxiredoxin system as well as glutathione-S-transferases) the epididymis of double mutant animals increased the expression of several disulfide isomerases in an attempt to recover normal disulfide-bridging activity. Despite these compensatory mechanisms cauda-stored spermatozoa of double mutant animals show high levels of DNA oxidation, increased fragmentation and greater susceptibility to nuclear decondensation. Nevertheless, the enzymatic epididymal salvage response is sufficient to maintain full fertility of double KO males whatever their age, crossed with young WT female mice. PMID:22719900

  10. Pracovní využití teriérů typu bull

    OpenAIRE

    TÖRÖKOVÁ, Jacquelina

    2013-01-01

    This work deals with the history of born, origin, domestication of the dog and various types of bull terriers. Pointing at the crossing of the Bulldog with a black terrier. This work also deals with the different types of dogs Bull, their character, description. This work answers the question, ?Why the pit bull is not dangerous to people", evaluates conflict situations, and shows how conflicts can occur. It mentions the current differences and cons of each breed. It places an attempt to creat...

  11. Inconsistent identification of pit bull-type dogs by shelter staff.

    Science.gov (United States)

    Olson, K R; Levy, J K; Norby, B; Crandall, M M; Broadhurst, J E; Jacks, S; Barton, R C; Zimmerman, M S

    2015-11-01

    Shelter staff and veterinarians routinely make subjective dog breed identification based on appearance, but their accuracy regarding pit bull-type breeds is unknown. The purpose of this study was to measure agreement among shelter staff in assigning pit bull-type breed designations to shelter dogs and to compare breed assignments with DNA breed signatures. In this prospective cross-sectional study, four staff members at each of four different shelters recorded their suspected breed(s) for 30 dogs; there was a total of 16 breed assessors and 120 dogs. The terms American pit bull terrier, American Staffordshire terrier, Staffordshire bull terrier, pit bull, and their mixes were included in the study definition of 'pit bull-type breeds.' Using visual identification only, the median inter-observer agreements and kappa values in pair-wise comparisons of each of the staff breed assignments for pit bull-type breed vs. not pit bull-type breed ranged from 76% to 83% and from 0.44 to 0.52 (moderate agreement), respectively. Whole blood was submitted to a commercial DNA testing laboratory for breed identification. Whereas DNA breed signatures identified only 25 dogs (21%) as pit bull-type, shelter staff collectively identified 62 (52%) dogs as pit bull-type. Agreement between visual and DNA-based breed assignments varied among individuals, with sensitivity for pit bull-type identification ranging from 33% to 75% and specificity ranging from 52% to 100%. The median kappa value for inter-observer agreement with DNA results at each shelter ranged from 0.1 to 0.48 (poor to moderate). Lack of consistency among shelter staff indicated that visual identification of pit bull-type dogs was unreliable. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  12. Joint disorder; a contributory cause to reproductive failure in beef bulls?

    Directory of Open Access Journals (Sweden)

    Ekman Stina

    2007-11-01

    Full Text Available Abstract The lame sire, unsound for breeding, can cause substantial economic loss due to reduced pregnancies in the beef-producing herd. To test the hypothesis that joint disorder is a possible cause of infertility in beef sires, right and left hind limb bones from 34 beef sires were examined postmortem to identify lesions in the femorotibial, femoropatellar (stifle, tarsocrural, talocalcaneus, and proximal intertarsal (tarsal joints. The bulls were slaughtered during or after the breeding season due to poor fertility results. Aliquots of the cauda epididymal contents taken postmortem from 26 bulls were used for sperm morphology evaluation. As a control, hind limbs (but no semen samples from 11 beef bulls with good fertility results were included. Almost all infertile bulls (30/34 had lesions in at least one joint. Twenty-eight bulls (28/30, 93% had lesions in the stifle joint, and 24 (24/28, 86% of these were bilateral. Fourteen bulls (14/30, 47% had lesions in the tarsal joint, and 10 (10/14, 71% of these were bilateral. Four bulls (4/34, 12% had no lesions, three bulls (3/34, 9% had mild osteoarthritis (OA, 5 (5/34, 15% moderate OA, 17 (17/34, 50% severe OA and 5 (5/34, 15% deformed OA. Almost all OA lesions (97% were characterized as lesions secondary to osteochondrosis dissecans. All the bulls with satisfactory sperm morphology (n = 12/34 had joint lesions, with mostly severe or deformed bilateral lesions (83%. Consequently, the most likely cause of infertility in these 12 bulls was joint disease. Almost all control bulls (10/11 had OA lesions, but most of them were graded as mild (55% or moderate (36%. None of the control bulls had severe lesions or deformed OA. We suggest that joint lesions should be taken into consideration as a contributory cause of reproductive failure in beef sires without symptoms of lameness.

  13. Viability of postmortal epididymal mouse spermatozoa during long-term hypothermic storage and cryopreservation.

    Science.gov (United States)

    Shishova, N V; Gakhova, E N; Mel'nikova, E V

    2011-08-01

    The study examined the effect of long-term hypothermic (4°C) storage of mouse carcasses on motility, cell membrane damage, in vitro survival and capacitation of epididymal spermatozoa before and after cryopreservation. It was shown that the number of spermatozoa with rectilinear forward motion decreased with increasing storage time. There were no significant changes in the total sperm motility and integrity of their plasmalemma. Pronounced effects of hypothermia and long-term storage of the mouse carcasses on cryocapacitation of spermatozoa during cryoconservation were demonstrated.

  14. Impact of cigarette smoking on histone (H2B) to protamine ratio in human spermatozoa and its relation to sperm parameters.

    Science.gov (United States)

    Hamad, M F; Shelko, N; Kartarius, S; Montenarh, M; Hammadeh, M E

    2014-09-01

    Smoking is strongly associated with abnormalities in histone-to-protamine transition and with alteration of protamine expression in human spermatozoa. A proper protamine to histone ratio is, however, essential for sperm chromatin maturity and DNA integrity. Alterations in these sperm nuclear proteins were observed in infertile men. The present prospective study is aimed at evaluating the possible relationship among smoking, semen quality and the histone-to-protamine transition ratio in mature spermatozoa. Histone H2B and protamine 1 (P1) and 2 (P2) were quantified using acid-urea polyacrylamide gel electrophoresis in the spermatozoa of 35 smokers and 19 non-smokers. Levels of lipid peroxidation marker malondialdehyde (MDA) were measured in seminal plasma by thiobarbituric acid assay. Cotinine concentrations were determined in seminal plasma using an enzyme-linked immunosorbent assay. Histone H2B levels in smokers (292.27 ± 58.24 ng/10(6)) were significantly higher (p = 0.001) than that of non-smokers (109.1 ± 43.70 ng/10(6)), besides, a significant difference (p > 0.0001) was found for the P1 and P2 ratio between smokers (1.71 ± 0.071) and non-smokers (1.05 ± 0.033). The H2B/(H2B+P1 + P2) ratio (0.29 ± 0.71) of smokers were significantly higher (p = sperm count, motility (p = 0.018), vitality (p = 0.009) and membrane integrity (p = 0.0001) than non-smokers. These results reveal that patients who smoke possess a higher proportion of spermatozoa with an alteration of the histone to protamine ratio than patients who do not smoke, and suggest that cigarette smoking may inversely affect male fertility. © 2014 American Society of Andrology and European Academy of Andrology.

  15. Bull Trout Life History, Genetics, Habitat Needs, and Limiting Factors in Central and Northeast Oregon. Annual Report 1996.

    Energy Technology Data Exchange (ETDEWEB)

    Bellerud, Blane L.; Gunckel, Stephanie; Hemmingsen, Alan R.; Buchanan, David V.; Howell, Philip J.

    1997-10-01

    This study is part of a multi-year research project studying aspects of bull trout life history, ecology and genetics. This report covers the activities of the project in 1996. Results and analysis are presented in the following five areas: (1) analysis of the genetic structure of Oregon bull trout populations; (2) distribution and habitat use of bull trout and brook trout in streams containing both species; (3) bull trout spawning surveys; (4) summary and analysis of historical juvenile bull trout downstream migrant trap catches in the Grande Ronde basin; and (5) food habits and feeding behavior of bull trout alone and in sympatry with brook trout.

  16. THE PERFORMANCE OF JAVA AND ONGOLE CROSSBRED BULL UNDER INTENSIVE FEEDING MANAGEMENT

    Directory of Open Access Journals (Sweden)

    C.M.S. Lestari

    2014-10-01

    Full Text Available This study was set up to evaluate the performance of Java and Ongole Crossbred (OC bulls fedconcentrate and rice straw. A total of four Java bulls and four OC bulls were used in this experiment. Thebulls were fed concentrates (50% of the total dry matter feed requirement and rice straw (ad libitum.The concentrates were consisted of rice bran, beer waste product, copra meal, minerals, with crudeprotein (CP and total digestible nutrients (TDN contents of 15.32% and 73.09%, respectively. Theaverage daily gain (ADG, dry matter intake (DMI, protein and energy intake, and feed conversion ratio(FCR were observed. The results of this study showed that the ADG, DMI, CP and TDN intake, andFCR were not significantly different (p> 0.05. The ADG of Java and OC bulls were 0.58 kg and 0.78kg, respectively. The averages of DMI, CP and TDN intake were 6.59 kg (2.09% of BW, 0.81 kg and4.34 kg for Java bulls whereas for OC bulls were 6.42 kg (2.11% of BW, 0.78 kg, and 4.20 kg,respectively. The FCR of Java bulls was 11.49 and those of OC bulls was 9.21. It can be concluded thatJava and OC bulls raised intensively and fed concentrate and rice straw had the similar performance.

  17. Enhanced early-life nutrition promotes hormone production and reproductive development in Holstein bulls.

    Science.gov (United States)

    Dance, Alysha; Thundathil, Jacob; Wilde, Randy; Blondin, Patrick; Kastelic, John

    2015-02-01

    Holstein bull calves often reach artificial insemination centers in suboptimal body condition. Early-life nutrition is reported to increase reproductive performance in beef bulls. The objective was to determine whether early-life nutrition in Holstein bulls had effects similar to those reported in beef bulls. Twenty-six Holstein bull calves were randomly allocated into 3 groups at approximately 1 wk of age to receive a low-, medium-, or high-nutrition diet, based on levels of energy and protein, from 2 to 31 wk of age. Calves were on their respective diets until 31 wk of age, after which they were all fed a medium-nutrition diet. To evaluate secretion profiles and concentrations of blood hormones, a subset of bulls was subjected to intensive blood sampling every 4 wk from 11 to 31 wk of age. Testes of all bulls were measured once a month; once scrotal circumference reached 26cm, semen collection was attempted (by electroejaculation) every 2 wk to confirm puberty. Bulls were maintained until approximately 72 wk of age and then slaughtered at a local abattoir. Testes were recovered and weighed. Bulls fed the high-nutrition diet were younger at puberty (high=324.3 d, low=369.3 d) and had larger testes for the entire experimental period than bulls fed the low-nutrition diet. Bulls fed the high-nutrition diet also had an earlier and more substantial early rise in LH than those fed the low-nutrition diet and had increased concentrations of insulin-like growth factor-I (IGF-I) earlier than the bulls fed the low-nutrition diet. Furthermore, we detected a temporal association between increased IGF-I concentrations and an early LH rise in bulls fed the high-nutrition diet. Therefore, we inferred that IGF-I had a role in regulating the early gonadotropin rise (in particular, LH) and thus reproductive development of Holstein bulls. Overall, these results support our hypothesis that Holstein bull calves fed a high-nutrition diet reach puberty earlier and have larger testes than

  18. SEMEN CHARACTERISTICS OF SAHIWAL BULLS IN RELATION TO AGE AND SEASON

    Directory of Open Access Journals (Sweden)

    Ahmad, M., M.T. Asmat, N.U. Rehman and M.Z. Khan*

    2003-12-01

    Full Text Available This study was undertaken to investigate the effects of age and season on the semen quality of 21 Sahiwal bulls kept under the similar managemental conditions at the Semen Production Unit, Qadirabad, District Sahiwal. These bulls were divided into 3 age groups i.e. group-I (upto 3 years, group-II (3-5 years and group-III (above five years. Depending on the photo period months of the year were grouped into 4 seasons i.e. Winter (November, December and January, Spring (February, March and April, Summer (May, June and July and Autumn (August, September & October.The results indicated that ejaculatory volume, motility percentage and doses produced by bulls of Group-II were significantly higher( P<0.05 than that of bulls of Group I and III. However mass activity, sperm concentration, pH and post thaw motility differed non significantly among bulls of three age groups. Generally all bulls showed better performance during Spring than other seasons of the year. It may be concluded that semen collected from mature bulls (adult bulls during the Spring is of better quality than the semen of young and old bulls and also in Spring the quality of semen is better than the other seasons of the year.

  19. Effect of ribonucleic acid (RNA) isolation methods on putative reference genes messenger RNA abundance in human spermatozoa.

    Science.gov (United States)

    Barragán, M; Martínez, A; Llonch, S; Pujol, A; Vernaeve, V; Vassena, R

    2015-07-01

    Although the male gamete participates in a significant proportion of infertility cases, there are currently no proven molecular markers of sperm quality. The search for significant gene expression markers is partially hindered by the lack of a recognized set of reference genes (RGs) to normalize reverse transcription quantitative PCR (RT-qPCR) data across studies. The aim of this study is to define a set of RGs in assisted reproduction patients undergoing different sample collection and RNA isolation methods. Twenty-two normozoospermic men were included in the study. From each man, semen was either cryopreserved by slow freezing or analyzed fresh, and, for each, RNA was extracted with either phenol-free or phenol-based methods. In two cases, both methods were used to isolate RNA. Twenty putative RGs were analyzed and their mRNA abundance across samples was estimated by RT-qPCR. To determine the genes whose steady-state mRNA abundance remains unchanged, three different algorithms (geNorm, BestKeeper and NormFinder) were applied to the qPCR data. We found that RGs such as GAPDH or ACTB, useful in other biological contexts, cannot be used as reference for human spermatozoa. It is possible to compare gene expression from fresh and cryopreserved sperm samples using the same isolation method, while the mRNA abundance of expressed genes becomes different depending on the RNA isolation technique employed. In our conditions, the most appropriate RGs for RT-qPCR analysis were RPLP1, RPL13A, and RPLP2. Published discrepancies in gene expression studies in human spermatozoa may be due in part to inappropriate RGs selection, suggesting a possible different interpretation of PCR data in several reports, which were normalized using unstable RGs. © 2015 American Society of Andrology and European Academy of Andrology.

  20. Monitor and Protect Wigwam River Bull Trout for Koocanusa Reservoir : Summary of the Skookumchuck Creek Bull Trout Enumeration Project Final Report 2000-2002.

    Energy Technology Data Exchange (ETDEWEB)

    Baxter, Jeremy; Baxter, James S.

    2002-12-01

    This report summarizes the third and final year of a bull trout (Salvelinus confluentus) enumeration project on Skookumchuck Creek in southeastern British Columbia. The fence and traps were operated from September 6th to October 11th 2002 in order to enumerate post-spawning bull trout. During the study period a total of 309 bull trout were captured at the fence. In total, 16 fish of undetermined sex, 114 males and 179 females were processed at the fence. Length and weight data, as well as recapture information, were collected for these fish. An additional 41 bull trout were enumerated upstream of the fence by snorkeling prior to fence removal. Coupled with the fence count, the total bull trout enumerated during the project was 350 individuals. Several fish that were tagged in the lower Bull River were recaptured in 2002, as were repeat and alternate year spawners previously enumerated in past years at the fence. A total of 149 bull trout redds were enumerated on the ground in 2002, of which 143 were in the 3.0 km index section (river km 27.5-30.5) that has been surveyed over the past six years. The results of the three year project are summarized, and population characteristics are discussed.

  1. Heat Shock Protein member A2 forms a stable complex with angiotensin converting enzyme and protein disulfide isomerase A6 in human spermatozoa.

    Science.gov (United States)

    Bromfield, Elizabeth G; McLaughlin, Eileen A; Aitken, Robert John; Nixon, Brett

    2016-02-01

    Given the importance of the chaperone Heat Shock Protein A2 (HSPA2) in the regulation of male fertility, this study aimed to identify and characterize additional proteins that may rely on the activity of this chaperone in human spermatozoa. In view of the findings in this study we propose that angiotensin converting enzyme (ACE) and protein disulfide isomerase A6 (PDIA6) are novel interacting proteins of HSPA2 and that this multimeric complex may participate in key elements of the fertilization cascade. The molecular chaperone HSPA2 plays a pivotal role in the remodelling of the sperm surface during capacitation. Indeed, human spermatozoa that are deficient in HSPA2 protein expression lack the ability to recognize human oocytes, resulting in repeated IVF failure in a clinical setting. Moreover, our recent work has shown that defective HSPA2 function induced by oxidative stress leads to the aberrant surface expression of one of its interacting proteins, arylsulfatase A, and thus contributes to a loss of sperm-zona pellucida adhesion. Human spermatozoa were collected from fertile donors, capacitated and prepared for Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) analysis. Protein complexes resolved via BN-PAGE were excised and their constituents were identified using mass spectrometry. The interactions between ACE, PDIA6 and HSPA2 were then confirmed using immunoprecipitation and proximity ligation assays and the localization of these proteins was assessed in isolated spermatozoa and commercially available human testis tissue sections. Finally, pharmacological inhibition of ACE was performed to assess the role of ACE in human sperm capacitation. Herein we have identified ACE and PDIA6 as potential HSPA2-interacting proteins and shown that this assemblage resides in membrane raft microdomains located in the peri-acrosomal region of the sperm head. Additionally, the surface expression of PDIA6, but not ACE, was shown to be dynamically regulated during sperm

  2. Image cytometer method for automated assessment of human spermatozoa concentration

    DEFF Research Database (Denmark)

    Egeberg, D L; Kjaerulff, S; Hansen, C

    2013-01-01

    to investigator bias. Here we show that image cytometry can be used to accurately measure the sperm concentration of human semen samples with great ease and reproducibility. The impact of several factors (pipetting, mixing, round cell content, sperm concentration), which can influence the read-out as well...... as inter-operator and -cytometer variation on two different image cytometers (NC-3000 and SP-100) were evaluated. Furthermore, 725 semen samples were assessed both by manual assessment (WHO recommended method) and by image cytometry and tight correlations between the measured concentrations were shown....... Moreover, by evaluation of repeated measurements it appeared that image cytometry produced more consistent and accurate measurements than manual counting of human spermatozoa concentration. In conclusion, image cytometry provides an appealing substitute of manual counting by providing reliable, robust...

  3. Effect of coculturing spermatozoa with oviductal cells on the incidence of polyspermy in pig in vitro fertilization.

    Science.gov (United States)

    Dubuc, A; Sirard, M A

    1995-07-01

    It is known that oviductal cells play an important role in fertilization in vivo. We were interested in the effect of those cells on spermatozoa and their influence on the incidence of polyspermy in pig in vitro fertilization (IVF) when cocultured with spermatozoa. Oviductal cells are believed to select a highly fecund population of spermatozoa. By coculturing spermatozoa with oviductal cells it is possible to reduce the number of spermatozoa confronting the eggs at fertilization, reducing the incidence of polyspermy. In our study, spermatozoa were cocultured with oviductal cells for 30 min so that they could bind to the oviductal cells. Both bound and unbound spermatozoa were used for fertilization. The results show that the spermatozoa that were bound to the oviductal cells were capable of fertilizing the eggs, but the nonbound spermatozoa had a reduced penetration incidence. With the bound sperm, polyspermy incidence decreased and the two-pronuclei proportion increased. We also found that with time the spermatozoa released from the cells had better motility than those that did not bind. Therefore, it is our belief that oviductal cells can be used for porcine IVF resulting in a lower polyspermy incidence and higher pronuclei incidence.

  4. Growth and reproductive development from weaning through 20 months of age among breeds of bulls in subtropical Florida.

    Science.gov (United States)

    Chase, C C; Chenoweth, P J; Larsen, R E; Olson, T A; Hammond, A C; Menchaca, M A; Randel, R D

    1997-02-01

    To determine the effect of breed on growth and reproductive development, weaned bulls in each of 2 yr were managed as a single group for approximately a year. In Year 1, the study group consisted of 24 Angus, 24 Brahman, 20 Hereford and 14 Senepol bulls, while in Year 2, it contained 25 Angus, 17 Brahman. 13 Romosinuano and 9 Nellore x Brahman bulls. Body and testicular growth measurements were recorded at 6-wk intervals. At approximately 1 yr of age and quarterly thereafter (4 periods), bulls were evaluated for libido, pubertal status, and GnRH-induced LH and testosterone secretion. Significant breed-by-age interactions occurred for most growth measurements. Brahman bulls (Bos indicus ) were (P Senepol and Romosinuano bulls (Bos taurus ). Libido scores were lowest for Brahman and Nell ore x Brahman bulls (Bos indicus ). highest for Angus and Hereford bulls (temperate Bos taurus breeds) and intermediate for Senepol and Romosinuano bulls (tropical Bos taurus breeds; P Senepol, Romosinuano and Nellore x Brahman bulls (tropical breeds). In conclusion, reproductive development of Senepol and Romosinuano bulls (tropical Bos taurus breeds) was more similar to Angus and Hereford bulls (temperate Bos taurus breeds) than to Brahman and Nellore x Brahman bulls (Bos indicus ).

  5. KAJI BANDING KUALITAS SPERMATOZOA SAPI SIMMENTAL, LIMOUSIN, DAN FRIESIAN HOLSTEIN TERHADAP PROSES PEMBEKUAN

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    Komariah (Komariah

    2013-10-01

    Full Text Available Penelitian ini bertujuan untuk mempelajari dan membandingkan kualitas spermatozoa beku sapi Simmental, Limousin, dan Friesian Holstein (FH di Balai Inseminasi Buatan Lembang (BIB, Bandung, Jawa Barat. Jumlah sapi yang digunakan pada penelitian ini adalah 24 ekor sapi jantan yang terdiri atas 8 Simmental, 8 Limousin, dan 8 FH berumur 4 tahun dengan kisaran bobot badan 800-900 kg. Penelitian menggunakan data sekunder dan primer BIB Lembang bulan November sampai Desember 2010. Data sekunder yang diambil adalah data motilitas spermatozoa segar meliputi, before freezing (BF, post thawing motility (PTM, longivitas dan data primer yaitu nilai recovery rate. Hasil yang diperoleh dari penelitian adalah motilitas spermatozoa segar dan before freezing sapi Simmental lebih tinggi (P<0,05 dibandingkan sapi Limousin dan FH, sedangkan hasil PTM, longivitas, dan recovery rate tidak berbeda nyata pada ketiga bangsa tersebut. (Kata kunci: Freezability, Spermatozoa, Simmental, Limousin, Friesian Holstein

  6. Ultrastructure of spermatozoa of Solifuges (Arachnida, Solifugae): possible characters for their phylogeny?

    Science.gov (United States)

    Klann, A E; Bird, T; Peretti, A V; Gromov, A V; Alberti, G

    2009-04-01

    The ultrastructure of spermatozoa is a widely accepted source of characters for phylogenetic studies. In this study the fine structure of sperm cells of representatives of six different New and Old World families (Ammotrechidae, Daesiidae, Eremobatidae, Galeodidae, Karschiidae, Solpugidae) of solifuges (Arachnida, Solifugae) were investigated in order to reveal putative characters suitable for subsequent systematic and phylogenetic analyses. The spermatozoa of solifuges represent a relatively simple type of sperm cells. In general, their spermatozoa are roundish, oval shaped (Ammotrechidae, Daesiidae, Eremobatidae, Solpugidae) or plate-shaped (Karschiidae) with or without membrane protuberances and devoid of a flagellum. Only in Galeodidae, very conspicuous thin and elongated sperm cells occur. The spermatozoa either occur as single cells (Eremobatidae, Solpugidae) or in groups of loose knit cells (Ammotrechidae) or in highly ordered groups (Karschiidae). In contrast to the other families studied here, within the Galeodidae and in the genus Blossia (Daesiidae) sperm cells surrounded by a secretion sheath, clearly representing coenospermia, could be observed.

  7. Differences in the expression of microRNAs and their predicted gene targets between cauda epididymal and ejaculated boar sperm.

    Science.gov (United States)

    Chang, Yu; Dai, Ding-Hui; Li, Yuan; Zhang, Yan; Zhang, Ming; Zhou, Guang-Bin; Zeng, Chang-Jun

    2016-12-01

    Mammalian spermatozoa gradually mature and acquire fertility during the transition from the testis to the caput and cauda epididymis, after which they are stored at the tail of the epididymis and the ampulla of vas deferens. During ejaculation, mixing of spermatozoa with the secretions of accessory sex glands leads to their dilution and changes in their function. Although remarkable progress has been made toward the understanding of changes in spermatozoa biochemistry and function before and after ejaculation, it is unknown whether microRNAs (miRNAs) are involved in regulating the function of spermatozoa during the transition between the cauda epididymis and ejaculation. In this study, 48 miRNAs were selected for analysis on the basis of their potential involvement in spermatogenesis, sperm maturation, and quality parameters markers. The differential expression levels of these 48 miRNAs between the caudal epididymis and fresh ejaculates of boar spermatozoa were determined. We found that 15 miRNAs were significantly differentially expressed (eight downregulated and seven upregulated) between boar cauda epididymal and fresh spermatozoa. Five miRNAs hypothesized to be involved in sperm apoptosis were further tested to demonstrate their influence over the expression of their target mRNAs using quantitative reverse-transcription polymerase chain reaction. Together, our findings suggest that these differentially expressed miRNAs are associated with the functional regulation of spermatozoa between cauda epididymis and ejaculation. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. SEMEN PRODUCTION AND PRODUCTIVE LIFE OF SAHIWAL BULLS: RELATIONSHIP WITH GENETIC WORTH

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    M. S. KHAN, A. A. BHATTI, S. A. BHATTI1 AND M. ASHIQ2

    2007-01-01

    Full Text Available Objective of the present study was to document the semen producing ability, productive life and genetic ability for lactation milk yield of Sahiwal bulls used for artificial insemination (AI in Punjab and to find the impact of AI bulls on the improvement of Sahiwal cattle. Data from Semen Production Unit (SPU, Qadirabad, Sahiwal, Pakistan were used for this purpose. A repeatability animal model was used for estimation of breeding values for lactation milk yield. Productive life of a bull was calculated as a difference between culling age and the age at first ejaculation. Number of bulls brought to SPU varied from 9 to 102 for any year. Average number of doses of semen produced by any bull for a year varied from 724 to 5745. On the average, 238 bulls produced 17143 ± 1164 semen doses during their average stay of 5.4 ± 0.2 years. About 50% of the bulls stayed for less than four years at the SPU; with a maximum range of 14 years. Progeny tested bulls (n=90 produced 5000 and 10000 semen doses (Y in three and four years of stay (X, respectively (Y = 24.8 + 2.3635 X - 0.0112 X2. To produce 20,000 doses, it is predicted that bulls need to stay for six and a half years at the SPU. There was no association between breeding values for lactation milk yield estimated under a repeatability animal model (EBVs and number of semen doses produced (r = 0.17 and EBVs and number of daughters. Lack of genetic superiority of bulls used indicated that AI did not bring desired genetic improvement in Sahiwal cattle in the present situation. Modifications for judicious utilization of bulls are suggested along with improvements in data recording.

  9. Fatty acid content in epididymal fluid and spermatozoa during sperm maturation in dogs

    OpenAIRE

    Ramos Angrimani, Daniel S.; Nichi, Marcilio; Losano, Jo?o Diego A.; Lucio, Cristina F.; Lima Veiga, Gisele A.; Franco, M?rcia V. M. Junqueira; Vannucchi, Camila I.

    2017-01-01

    Abstract Background During sperm maturation, there is a reorganization of fatty acids from plasmatic membrane of the spermatozoa, which allows higher membrane integrity and acquisition of sperm motility. However, the fatty acid profile during sperm maturation remains unclear in dogs. Thus, the aim of this study was to identify the fatty acids from the epididymal spermatozoa and plasma during the sperm maturation, and observed changes in ...

  10. Fragmentasi DNA Spermatozoa: Penyebab, Deteksi, dan Implikasinya pada Infertilitas Laki-Laki

    Directory of Open Access Journals (Sweden)

    Silvia W. Lestari

    2015-12-01

    Full Text Available Prediksi fertilitas laki-laki dapat dilakukan dengan analisis semen. Analisis semen konvensionalmerupakan pemeriksaan sederhana dan tidak mahal, tetapi memiliki variabilitas yang tinggi.Integritas DNA spermatozoa penting untuk transmisi informasi genetik. Fragmentasi DNAspermatozoa sebagai akibat gangguan spermatogenesis, maturasi spermatozoa, stres oksidatifdan infeksi, dapat menyebabkan infertilitas laki-laki, gangguan perkembangan embrio dan abortusberulang. Hubungan fragmentasi DNA spermatozoa dengan luaran teknologi reproduksi berbantu(TRB mengarahkan fragmentasi DNA spermatozoa sebagai pemeriksaan infertilitas laki-laki. Dariberbagai metode fragmentasi DNA spermatozoa yang umum dilakukan, sperm chromatin dispersion(SCD merupakan metode pemeriksaan fragmentasi DNA spermatozoa yang sederhana, akuratdan tidak mahal, sehingga dapat dilaksanakan di laboratorium andrologi. Selain menghasilkandiagnosis yang lebih baik, pemeriksaan fragmentasi DNA spermatozoa juga menggambarkanprognosis infertilitas termasuk luaran program TRB. Kata kunci: infertilitas laki-laki, fragmentasi DNA spermatozoa, SCD   Sperm DNA Fragmentation: Etiology, Detection and Implicationto Male Infertility Abstract The prediction of male fertility is determined by semen analysis. The conventional semenanalysis is simple and inexpensive but prone to variability. The integrity of sperm DNA is essentialfor the transmission of genetic information. Fragmentation of sperm DNA as result of disruptionin spermatogenesis and sperm maturation, oxidative stress, and infection may lead to maleinfertility, abnormal embryonic development and recurrent abortion. The association betweensperm DNA fragmentation and diminished reproductive outcomes has led to the introduction ofsperm DNA fragmentation testing on the clinical assessment of male infertility. Of all the spermDNA fragmentation tests, sperm chromatin dispersion (SCD test is quite simple, accurate, andinexpensive to be conducted on

  11. IN VITRO EFFECTS OF COPPER ON THE MOTILITY AND VIABILITY OF SPERMATOZOA

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    Zuzana Kňažická

    2012-06-01

    Full Text Available Copper (Cu is an environmental risk factor which has various effects on the animal and human organism. The target of this study was to investigate the effects of Cu on motility and viability of spermatozoa in vitro. Specifically, we examined the dose- and time-dependent effect of copper (II chloride (CuCl2 on the survival of spermatozoa during different time periods (Time 0 h, 1 h, 2 h. The percentage of motile spermatozoa was determined after exposure to concentrations of 3.9; 7.8; 15.6; 31.2; 62.5; 125; 250; 500; 1000 µmol.dm-3 of CuCl2 using the Sperm VisionTM CASA (Computer Assisted Semen Analyzer system. The cell viability was measured by the MTT (metabolic activity assay. The initial spermatozoa motility showed slightly increased values at doses < 31.20 μmol.dm-3 of CuCl2 compared to the control group. In this time, the lowest spermatozoa motility was recorded significantly (P<0.001 in the group A using the highest dose of CuCl2 (1000 μmol.dm-3. After 1 h of cultivation we proved that the average motility values decreased proportionally to the increasing concentration of CuCl2. The low doses (< 7.80 μmol.dm-3 of CuCl2 increased the spermatozoa motility and concurrently of mitochondrial activity (Time 2 h. The obtained data confirm that Cu (in the form CuCl2 at high doses acts as a toxic element on the spermatozoa motility and it has a destructive effect on the mitochondrial complex, which is necessary for their life processes. The low concentrations (< 7.80 μmol.dm-3 of CuCl2 stimulated the mitochondrial activity of cells and maintained of spermatozoa motility during the short-term of cultivation.

  12. Reduced levels of intracellular calcium releasing in spermatozoa from asthenozoospermic patients

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    García Juan F

    2009-02-01

    Full Text Available Abstract Background Asthenozoospermia is one of the most common findings present in infertile males characterized by reduced or absent sperm motility, but its aetiology remains unknown in most cases. In addition, calcium is one of the most important ions regulating sperm motility. In this study we have investigated the progesterone-evoked intracellular calcium signal in ejaculated spermatozoa from men with normospermia or asthenozoospermia. Methods Human ejaculates were obtained from healthy volunteers and asthenospermic men by masturbation after 4–5 days of abstinence. For determination of cytosolic free calcium concentration, spermatozoa were loaded with the fluorescent ratiometric calcium indicator Fura-2. Results Treatment of spermatozoa from normospermic men with 20 micromolar progesterone plus 1 micromolar thapsigargin in a calcium free medium induced a typical transient increase in cytosolic free calcium concentration due to calcium release from internal stores. Similar results were obtained when spermatozoa were stimulated with progesterone alone. Subsequent addition of calcium to the external medium evoked a sustained elevation in cytosolic free calcium concentration indicative of capacitative calcium entry. However, when progesterone plus thapsigargin were administered to spermatozoa from patients with asthenozoospermia, calcium signal and subsequent calcium entry was much smaller compared to normospermic patients. As expected, pretreatment of normospermic spermatozoa with both the anti-progesterone receptor c262 antibody and with progesterone receptor antagonist RU-38486 decreased the calcium release induced by progesterone. Treatment of spermatozoa with cytochalasin D or jasplakinolide decreased the calcium entry evoked by depletion of internal calcium stores in normospermic patients, whereas these treatments proved to be ineffective at modifying the calcium entry in patients with asthenozoospermia. Conclusion Our results suggest

  13. Influence of the breed of bull (Bos taurus indicus vs. Bos taurus taurus) and the breed of cow (Bos taurus indicus, Bos taurus taurus and crossbred) on the resistance of bovine embryos to heat.

    Science.gov (United States)

    Eberhardt, Bruno G; Satrapa, Rafael A; Capinzaiki, Cláudia R L; Trinca, Luzia A; Barros, Ciro M

    2009-08-01

    In vitro studies have shown that Bos taurus indicus (B. t. indicus) embryos submitted to heat shock at early stages of development are better able to survive as compared to Bos taurus taurus embryos. Embryo genotype influences resistance to heat shock thus leading to the question as to whether embryos sired by thermo-tolerant breeds exhibit the same resistance to heat shock. In the present study the influence of both oocyte and semen, on the resistance to heat shock (HS) at early stages of in vitro development, was assessed in B. t. indicus [Nelore (N) breed], B. t. taurus [Holstein (H) and Angus (A) breeds] and crossbreds. In Experiment 1, Nelore and crossbred oocytes were collected from slaughterhouse ovaries and fertilized with spermatozoa from Nelore and Angus bulls. Presumptive embryos were collected and randomly assigned to control (39 degrees C) or HS at 12, 48 or 96 h post insemination (hpi; 41 degrees C for 12h) treatments. The cleavage rates and proportion of embryos developing to the blastocyst and hatched blastocyst stages were recorded on Days 2, 8 and 10, respectively. Heat shock treatment decreased development of both Nelore and crossbred embryos. There was a significant interaction between time (12, 48 or 96 hpi) and temperature for blastocyst rates, i.e., the embryos became more thermotolerant as development proceeded. In Experiment 2, oocytes from Nelore and Holstein cows were fertilized with semen from bulls of either Nelore or Angus breeds, and subjected to 12 h HS at 96 hpi. Heat shock at 96 hpi, decreased embryo development. Additionally, cowxtreatment and bullxtreatment interactions were significant for blastocyst rates, i.e., both breed of cow and breed of bull affected the decline in blastocyst rate caused by heat shock treatment. In conclusion, the present results indicate that Nelore embryos (indicus) are more resistant to heat shock than Holstein (taurus) at early stages of in vitro development, and that embryos become more thermo

  14. Evaluation of the function of fresh and frozen-thawed sex-sorted and non-sorted stallion spermatozoa using a heterologous oocyte binding assay.

    Science.gov (United States)

    Clulow, J R; Evans, G; Maxwell, W M C; Morris, L H A

    2010-01-01

    The aim of the present study was to evaluate the potential oocyte binding ability and functional integrity of fresh or frozen-thawed, sex-sorted or non-sorted stallion spermatozoa. In the absence of effective IVF procedures in the horse, a heterologous sperm-binding assay was used as an indicator of fertilising capacity to assess differences in the ability of stallion spermatozoa to bind to bovine oocytes. The functional integrity of four treatment groups was assessed: (1) fresh non-sorted spermatozoa; (2) fresh sex-sorted spermatozoa; (3) frozen-thawed non-sorted spermatozoa; and (4) frozen-thawed sex-sorted spermatozoa. Spermatozoa found in association with the zona pellucida of the bovine oocytes were deemed 'attached' or 'bound' depending on their characterisation as either acrosome intact or acrosome reacted, respectively. Significantly less frozen-thawed spermatozoa were found attached to the oocytes compared with fresh spermatozoa. No significant differences were identified between the number of attached sex-sorted and non-sorted frozen-thawed spermatozoa. However, significantly more sex-sorted than non-sorted fresh spermatozoa were found attached to the oocytes after 1 h coincubation, although after 3 h coincubation this difference was no longer apparent. In conclusion, sex-sorted fresh and frozen-thawed stallion spermatozoa are functionally capable of attaching and binding to bovine oocytes in vitro. Furthermore, fresh sex-sorted spermatozoa attach better than non-sorted spermatozoa, suggesting that they have a more advanced capacitation-like status.

  15. Spermatozoa as a transport system of large unilamellar lipid vesicles into the oocyte.

    Science.gov (United States)

    Geerts, N; McGrath, J; Stronk, J N; Vanderlick, T K; Huszar, G

    2014-04-01

    In addition to their role as man-made membranes, vesicles continue to be investigated as carriers for drug delivery. While most research focuses on their injectable properties, here a new delivery strategy is proposed. It is shown that spermatozoa can transport vesicles of variable composition. For human spermatozoa, the vesicles started to show binding after 20 mol% of the nonbinding vesicle backbone lipids were substituted with positive, negative, cerebroside or ganglioside lipids. Vesicle binding is a dynamic process with constant 'on' and 'off' binding. The physiological and motility attributes of the spermatozoa are not affected by the attached vesicles. Sperm swimming characteristics changed only marginally. Also, the activation status of the acrosomal membrane, tested with the fluorescent probe Pisum sativum agglutinin, was not affected by vesicle binding. Moreover, the hyaluronic acid-binding test showed that viable, fully developed spermatozoa will attach and remain bound to hyaluronic acid-coated slides regardless of vesicle binding. Therefore a new 'hybrid' delivery system was created with human spermatozoa, and tested with a mouse IVF system. Large unilamellar vesicles physisorbed to mouse spermatozoa can not only penetrate the mouse oocytes in these proof-of-principle experiments, but also deliver the cargo placed within the vesicles. Copyright © 2013 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  16. Receptor dependent immobilization of spermatozoa by sperm immobilization factor isolated from Escherichia coli: proof of evidence.

    Science.gov (United States)

    Kumar, Vijay; Prabha, Vijay; Kaur, Siftjit; Kaur, Kiranjeet; Singh, S K

    2011-08-01

    To examine the receptor-ligand interaction between E. coli and spermatozoa resulting in sperm immobilization. Sperm immobilization factor (SIF) was isolated and purified from filtrate of E. coli suspension. Receptor on human spermatozoa for SIF was made to isolated and purified by salt extraction, gel permeation and ion exchange chromatography. Receptor dependent immobilization of spermatozoa by SIF was confirmed by competitive inhibition by addition of the purified receptor and binding to sperm receptor by Fluorescin Isothiocynate (FITC) labelled SIF using fluorescence microscopy. Heat labile, non-dialyzable, sperm immobilization factor of ∼56 kDa was isolated and purified from E. coli. Using SIF as a tool, receptor of 113 kDa could be isolated and purified from human spermatozoa. Addition of purified receptor completely inhibited sperm immobilization and death induced by SIF suggesting receptor-dependent immobilization of spermatozoa. Further incubation of sperm with FITC labelled SIF resulted in staining of whole sperm. The study provides evidence of receptor-ligand interaction between E. coli and spermatozoa. © 2011 The Japanese Urological Association.

  17. Hypotonic resistance of boar spermatozoa: sperm subpopulations and relationship with epididymal maturation and fertility.

    Science.gov (United States)

    Druart, Xavier; Gatti, Jean-Luc; Huet, Sylvie; Dacheux, Jean-Louis; Humblot, Patrice

    2009-02-01

    Hypotonic resistance of boar spermatozoa was investigated by measuring the ratio of live/dead spermatozoa (SYBR-14/propidium iodide) by flow cytometry after hypotonic stress. The survival rate of ejaculated spermatozoa incubated in hypotonic solutions ranging from 3 to 330 mmol/kg followed a sigmoid curve that fitted a simple logistic model. The critical osmolality value (Osm(crit)) at which 50% of spermatozoa died was determined with this model. Hypotonic resistance of spermatozoa increased with temperature between 15 and 39 degrees C and decreased after hydrogen superoxide treatment, but was not modified during 8 days of preservation in Beltsville thawing solution. Hypotonic resistance markedly decreased during epididymal maturation and after ejaculation as Osm(crit) at 15 degrees C was 54.7+/-3.2, 68.5+/-10.6, 116.7+/-2.1 and 194.3+/-3.7 mmol/kg for the caput, corpus, cauda and ejaculated spermatozoa respectively. Hypo-osmotic stress of 100 mmol/kg revealed a sperm subpopulation exhibiting increased hypotonic resistance compared with the whole ejaculate (Osm(crit)=67.8+/-2.1 mmol/kg). Consistent differences were observed between lean and standard breeds (Pietrain versus Large White) and between boars within the same breed. According to data collected by artificial insemination centers during a large-scale field trial, hypotonic resistance of ejaculates was found to be positively correlated with in vivo fertility.

  18. Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

    Directory of Open Access Journals (Sweden)

    Quynh Thu Nguyen

    2016-11-01

    Full Text Available Mammalian spermatozoa utilize ATP as the energy source for key functions on the route to fertilization. ATP and its precursor nucleotides ADP and AMP are regularly investigated in sperm physiology studies, mostly by bioluminescence assays. Assay results vary widely, mainly due to different efficiencies in nucleotide extraction and prevention of their enzymatic degradation. Here, we describe a revised, validated protocol for efficient phosphatase inhibition and adenine nucleotide extraction resulting in consistently high ATP concentrations exceeding previously reported values for boar spermatozoa up to 20-fold. The revised assay is applicable for determining ATP concentrations and adenylate energy charge in extracts from fresh and frozen samples, thereby allowing simultaneous assessment of semen samples from long-term storage experiments. After validation, the assay was applied to liquid-preserved boar spermatozoa stored at 17°C and 5°C for 24 and 72 h. Cooling to 5°C, but not storage duration, reduced ATP concentration in spermatozoa (P<0.05, which was accompanied by the appearance of AMP and ADP in the preservation medium. ATP and energy charge were highly correlated to the proportion of membrane-intact spermatozoa, supporting the idea of nucleotides leaking through disrupted membranes in cold-shocked cells. The present assay allows highly standardized studies of energy metabolism in spermatozoa.

  19. Function and culture requirements of snow leopard (Panthera uncia) spermatozoa in vitro.

    Science.gov (United States)

    Roth, T L; Howard, J G; Donoghue, A M; Swanson, W F; Wildt, D E

    1994-08-01

    Electroejaculates from eight snow leopards were used to determine how the motility of spermatozoa was influenced by (i) type of media (Ham's F10, PBS, human tubal fluid or RPMI-1640); (ii) holding temperature (23 degrees C versus 37 degrees C); (iii) washing of spermatozoa and (iv) a sperm metabolic enhancer, pentoxifylline. The duration of sperm motility was assessed by evaluating samples in each treatment every hour for 6 h and a sperm motility index (a value combining percentage sperm motility and rate of forward progression) calculated. Spermatozoa from the Ham's F10, PBS and PBS plus pentoxifylline treatments were also co-incubated with zona-intact, domestic cat eggs that were fixed and evaluated for spermatozoa bound to the zona pellucida, penetrating the outer and inner layers of the zona pellucida and within the perivitelline space. During the 6 h co-incubation, the sperm motility index in PBS with pentoxifylline was greater (P 0.05) of holding temperature. Pentoxifylline supplementation enhanced (P snow leopard spermatozoa in the inner layer of the zona pellucida, and there were no spermatozoa in the perivitelline space.(ABSTRACT TRUNCATED AT 250 WORDS)

  20. Characterization of sulfhydryl proteins involved in the maintenance of flagellar straightness in hamster spermatozoa.

    Science.gov (United States)

    Cornwall, G A; Chang, T S

    1990-01-01

    Hamster caput epididymal spermatozoa exhibit a marked 90-180 degree bend when induced to acquire progressive motility in vitro (Cornwall et al, 1988). Flagellar bending is prevented by oxidizing sperm sulfhydryl (SH) groups with diamide. In the present study, the authors examined the SH proteins involved in sperm flagellar bending using two-dimensional polyacrylamide gel electrophoresis and monobromobimane, an SH-specific fluorescent dye. Proteins extracted from samples containing bent caput spermatozoa contained more reduced SH than the same proteins extracted from straight caput spermatozoa. To further characterize these sperm SH proteins, caudal epididymal spermatozoa that exhibited straight flagella were induced to undergo flagellar bending by treatment with the SH reductant dithiothreitol. Specific proteins of 85-94 kDa, 50-55 kDa and 35-40 kDa, which were rich in SH groups when extracted from samples containing bent caput spermatozoa, were also SH-rich in samples containing bent cauda spermatozoa. These studies suggest that oxidation of specific SH proteins may be important for maintaining sperm flagellar morphology.

  1. [The reproductive function in experimentally-produced monozygotic twin bulls].

    Science.gov (United States)

    Braun, J; Schams, D; Brem, G

    1990-06-01

    Reproductive functions were evaluated in 12 experimentally derived monozygotic cattle twins (6 pairs). In 4 twin pairs GnRH (20 micrograms Receptal i.m.) was administered in monthly intervals from 6 through 12 months of age. LH-secretion was determined by radioimmunoassay up to 6 hours after the GnRH-challenge in hourly collected peripheral blood samples. In 5 twin pairs (age 14 to 20 months) semen was collected 5 times and processed for cryoconservation. In fresh and in frozen-thawed semen samples sperm motility was determined by means of a computerized system. Twins showed a remarkable uniform response to the GnRH-challenge. One set of twins (HF-breed) had a significantly reduced response compared to the other pairs (DFV-breed). Both individuals of this particular pair had a low percentage of progressively motile spermatozoa. In pairs with physiological sperm motility, variance of this parameter within pairs and between pairs was not different. In contrast, twin uniformity concerning the average velocity of motile spermatozoa was high; differences between set of twins explained most of the total variance. According to this study, reproductive parameters in monozygotic cattle twins are strongly influenced by twin uniformity.

  2. Evaluation of bison (Bison bison) semen from Yellowstone National Park, Montana, USA, bulls for Brucella abortus shedding.

    Science.gov (United States)

    Frey, Rebecca K; Clarke, P Ryan; McCollum, Matt P; Nol, Pauline; Johnson, Kammy R; Thompson, Brent D; Ramsey, Jennifer M; Anderson, Neil J; Rhyan, Jack C

    2013-07-01

    To determine if bison (Bison bison) bulls from Yellowstone National Park (YNP), Montana, USA, shed an infective dose of Brucella abortus in semen, 50 YNP bulls were captured on public lands in Montana during the winter and early spring (April-May) of 2010 and 2011. The bulls were immobilized, and blood and semen samples were collected for serology and Brucella culture. Thirty-five bulls (70%) were antibody-positive, and B. abortus was cultured from semen in three (9%) of the 35 antibody-positive or suspect bulls, though not at concentrations considered an infective dose. Eight bulls (six antibody-positive, two negative) had palpable lesions of the testes, epididymides, or seminal vesicles consistent with B. abortus infection. Breeding soundness exams and semen analysis suggested that antibody-positive bulls were more likely to have nonviable ejaculate (8/35; 23%) than bulls without detectable antibody (2/15; 13%).

  3. Adaptive Management of Bull Trout Populations in the Lemhi Basin

    Science.gov (United States)

    Peterson, James T.; Tyre, Andrew J.; Converse, Sarah J.; Bogich, Tiffany L.; Miller, Damien; Post van der Burg, Max; Thomas, Carmen; Thompson, Ralph J.; Wood, Jeri; Brewer, Donna; Runge, Michael C.

    2011-01-01

    The bull trout Salvelinus confluentus, a stream-living salmonid distributed in drainages of the northwestern United States, is listed as threatened under the Endangered Species Act because of rangewide declines. One proposed recovery action is the reconnection of tributaries in the Lemhi Basin. Past water use policies in this core area disconnected headwater spawning sites from downstream habitat and have led to the loss of migratory life history forms. We developed an adaptive management framework to analyze which types of streams should be prioritized for reconnection under a proposed Habitat Conservation Plan. We developed a Stochastic Dynamic Program that identified optimal policies over time under four different assumptions about the nature of the migratory behavior and the effects of brook trout Salvelinus fontinalis on subpopulations of bull trout. In general, given the current state of the system and the uncertainties about the dynamics, the optimal policy would be to connect streams that are currently occupied by bull trout. We also estimated the value of information as the difference between absolute certainty about which of our four assumptions were correct, and a model averaged optimization assuming no knowledge. Overall there is little to be gained by learning about the dynamics of the system in its current state, although in other parts of the state space reducing uncertainties about the system would be very valuable. We also conducted a sensitivity analysis; the optimal decision at the current state does not change even when parameter values are changed up to 75% of the baseline values. Overall, the exercise demonstrates that it is possible to apply adaptive management principles to threatened and endangered species, but logistical and data availability constraints make detailed analyses difficult.

  4. Polymyxin B effects on motility parameters of cryopreserved bull semen

    Directory of Open Access Journals (Sweden)

    Mojtaba Rashedi

    2017-01-01

    Full Text Available Objective: To evaluate the effect of adding different values of polymyxin B (PMB to bull semen on various motility parameters of post-thawed semen such as total motility, progressive motility and velocity parameters using kinetic parameters of sperm by Computer Assisted Sperm Analysis. Methods: Gram negative bacteria release lipopolysaccharide, which induces the apoptotic pathway. Antibiotics are added to semen in order to prevent bacterial contaminations in bovine semen. These antibiotics kill the bacteria especially gram negative bacteria. Therefore, their endotoxins are released during bacteriolysis and bind to the head region and midpiece of sperm. PMB is a bactericidal antibiotic against multidrug resistant gram-negative bacteria and is able to neutralize the toxic effects of the released endotoxin. This study was performed on 3-year old Taleshi bulls. Results: The results showed both positive and negative significant effects of PMB on semen quality. Total motility and progressive motility were significantly increased (P<0.000 1 by 100 μg per mL of PMB (55.2% and 48.8% respectively against the control groups (43.5% and 37.7%, respectively. Moreover, they were significantly decreased (P<0.000 1 by 1 000 μg per mL of PMB (35.2% and 28.8% respectively against the control groups (43.5% and 37.7% respectively in above-mentioned parameters. In Computer Assisted Semen Analyzer, parameter VAP was significantly decreased (P<0.04 in 1 000 μg (69.6 μm/s against the control group (78.7 μm/s. Finally, using PMB in processing cryopreserved bull semen is advised, but before using it, the rate of endotoxins must be measured. Conclusions: We advise using PMB after measuring endotoxin concentration; In vitro, in vivo and in field fertilization, adding other sperm evaluation factors such as acrosomal integrity, DNA integrity, mitochondrial function to PMB treated semen.

  5. Michael Bull, Sound Moves. Ipod Culture and Urban Experience

    OpenAIRE

    Lisa McCormick

    2013-01-01

    Sound Moves, ouvrage non traduit de l'universitaire britannique Michael Bull, paru en 2007, a une place importante dans l'élaboration du champ de recherche sur l'écoute et les pratiques musicales appareillées en situation. Dans la continuité de son premier ouvrage (2000), ce deuxième opus poursuit l'étude de la « musicalisation » de notre quotidien au travers d’appareils mobiles, l'iPod en particulier. Dans la filiation d'un cadre théorique hérité des cultural studies et situé au carrefour de...

  6. Reduced fertilization after ICSI and abnormal phospholipase C zeta presence in spermatozoa from the wobbler mouse.

    Science.gov (United States)

    Heytens, Elke; Schmitt-John, Thomas; Moser, Jakob M; Jensen, Nanna Mandøe; Soleimani, Reza; Young, Claire; Coward, Kevin; Parrington, John; De Sutter, Petra

    2010-12-01

    Failed fertilization after intracytoplasmic sperm injection (ICSI) can be due to a reduced oocyte-activation capacity caused by reduced concentrations and abnormal localization of the oocyte-activation factor phospholipase C (PLC) zeta. Patients with this condition can be helped to conceive by artificial activation of oocytes after ICSI with calcium ionophore (assisted oocyte activation; AOA). However some concern still exists about this approach. Mouse models could help to identify potential oocyte-activation strategies and evaluate their safety. In this study, the fertilizing capacity of wobbler sperm cells was tested and the efficiency of AOA with two exposures to ionomycin to restore fertilization and embryo development was studied. The quality of the obtained blastocysts was assessed and embryo transfer was performed to evaluate post-implantation development. The presence of PLCzeta in the spermatozoa and testis of the wobbler mouse was evaluated by PLCzeta immunostaining and quantitative reverse-transcription polymerase chain reaction. Sperm cells from wobbler mice had reduced fertilizing capacity and abnormalities in PLCzeta localization, but not in its expression. Artificially activating the oocytes restored fertilization and embryo development. Therefore, the wobbler mouse can be a model for failed fertilization after ICSI to study PLCzeta dynamics and aid in optimization of the AOA method. Copyright © 2010 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  7. A comparison of the performance of Holstein and Friesian bulls in ...

    African Journals Online (AJOL)

    The performance of 92 Holstein-Friesian bulls, which were accepted for progeny testing ander the South African National Dairy Animal Performance and Progeny Testing Scheme during 1982, 1983 and 1984, was compared. Bulls which were locally bred for more than one generation were not entered under the Scheme.

  8. 75 FR 2269 - Endangered and Threatened Wildlife and Plants; Revised Designation of Critical Habitat for Bull...

    Science.gov (United States)

    2010-01-14

    ... and Puget Sound), and 215,870 hectares (ha) (533,426 acres (ac)) of reservoirs or lakes are being... to waters of western North America. Bull trout range throughout the Columbia River and Snake River...-history form of bull trout is unique to the Coastal-Puget Sound population (64 FR 58921; November 1, 1999...

  9. A comparison of the performance of Holstein and Friesian bulls in ...

    African Journals Online (AJOL)

    The performance of 92 Holstein-Friesian bulls, which were accepted for progeny testing ander the South African. National Dairy Animal Performance and Progeny Testing Scheme during 1982, 1983 and 1984, was compared. Bulls which were locally bred for more than one generation were not entered under the Scheme.

  10. Detection of Paratuberculosis in Breeding Bulls at Pakistani Semen Production Units: A Continuous Source of Threat

    National Research Council Canada - National Science Library

    Abbas, Muhammad; Munir, Muhammad; Khaliq, Syed Abdul; Haq, Muhammad Ikram Ul; Tanveer Khan, Muhammad; Qureshi, Zafar ul Ahsan

    2011-01-01

    .... Collectively, seroprevalence of 20.0% (47/235) in breeding bulls and 33.3% (6/18) in teaser bulls was observed, and thus it poses a potential threat of disease spread to a high number of heifers and cows through artificial...

  11. Evaluation of rice offal as energy source for fattening Bunaji bulls ...

    African Journals Online (AJOL)

    249kg and age range of 2-3 years to evaluate the feeding value of raw or parboiled rice offal as energy source for bull fattening. Four bulls per treatment were allotted to five dietary treatments in a 2x2 factorial arrangement with a common control.

  12. Genomic Prediction from Whole Genome Sequence in Livestock: The 1000 Bull Genomes Project

    DEFF Research Database (Denmark)

    Hayes, Benjamin J; MacLeod, Iona M; Daetwyler, Hans D

    Advantages of using whole genome sequence data to predict genomic estimated breeding values (GEBV) include better persistence of accuracy of GEBV across generations and more accurate GEBV across breeds. The 1000 Bull Genomes Project provides a database of whole genome sequenced key ancestor bulls...

  13. Use of NMR and NMR Prediction Software to Identify Components in Red Bull Energy Drinks

    Science.gov (United States)

    Simpson, Andre J.; Shirzadi, Azadeh; Burrow, Timothy E.; Dicks, Andrew P.; Lefebvre, Brent; Corrin, Tricia

    2009-01-01

    A laboratory experiment designed as part of an upper-level undergraduate analytical chemistry course is described. Students investigate two popular soft drinks (Red Bull Energy Drink and sugar-free Red Bull Energy Drink) by NMR spectroscopy. With assistance of modern NMR prediction software they identify and quantify major components in each…

  14. Physical, biotic, and sampling influences on diel habitat use by stream-dwelling bull trout

    Science.gov (United States)

    Nolan P. Banish; James T. Peterson; Russell F. Thurow

    2008-01-01

    We used daytime and nighttime underwater observation to assess microhabitat use by bull trout Salvelinus confluentus (N = 213) in streams of the intermountain western USA during the summers of 2001 and 2002. We recorded fish focal points and measured a set of habitat characteristics as well as habitat availability via line transects. Bull trout were...

  15. Positive effects of Red Bull® Energy Drink on driving performance during prolonged driving.

    Science.gov (United States)

    Mets, Monique A J; Ketzer, Sander; Blom, Camilla; van Gerven, Maartje H; van Willigenburg, Gitta M; Olivier, Berend; Verster, Joris C

    2011-04-01

    The purpose of this study was to examine if Red Bull® Energy Drink can counteract sleepiness and driving impairment during prolonged driving. Twenty-four healthy volunteers participated in this double-blind placebo-controlled crossover study. After 2 h of highway driving in the STISIM driving simulator, subjects had a 15-min break and consumed Red Bull® Energy Drink (250 ml) or placebo (Red Bull® Energy Drink without the functional ingredients: caffeine, taurine, glucuronolactone, B vitamins (niacin, pantothenic acid, B6, B12), and inositol) before driving for two additional hours. A third condition comprised 4 h of uninterrupted driving. Primary parameter was the standard deviation of lateral position (SDLP), i.e., the weaving of the car. Secondary parameters included SD speed, subjective driving quality, sleepiness, and mental effort to perform the test. No significant differences were observed during the first 2 h of driving. Red Bull® Energy Drink significantly improved driving relative to placebo: SDLP was significantly reduced during the 3rd (p Red Bull® Energy Drink significantly reduced the standard deviation of speed (p Red Bull® Energy Drink (p Red Bull® Energy Drink significantly improved each parameter. Red Bull® Energy Drink significantly improves driving performance and reduces driver sleepiness during prolonged highway driving.

  16. Bull Trout Population Assessment in the Columbia River Gorge : Annual Report 2000.

    Energy Technology Data Exchange (ETDEWEB)

    Byrne, Jim; McPeak, Ron

    2001-02-01

    We summarized existing knowledge regarding the known distribution of bull trout (Salvelinus confluentus) across four sub-basins in the Columbia River Gorge in Washington. The Wind River, Little White Salmon River, White Salmon River, and the Klickitat River sub-basins were analyzed. Cold water is essential to the survival, spawning, and rearing of bull trout. We analyzed existing temperature data, installed Onset temperature loggers in the areas of the four sub-basins where data was not available, and determined that mean daily water temperatures were <15 C and appropriate for spawning and rearing of bull trout. We snorkel surveyed more than 74 km (46.25 mi.) of rivers and streams in the four sub-basins (13.8 km at night and 60.2 km during the day) and found that night snorkeling was superior to day snorkeling for locating bull trout. Surveys incorporated the Draft Interim Protocol for Determining Bull Trout Presence (Peterson et al. In Press). However, due to access and safety issues, we were unable to randomly select sample sites nor use block nets as recommended. Additionally, we also implemented the Bull Trout/Dolly Varden sampling methodology described in Bonar et al. (1997). No bull trout were found in the Wind River, Little White Salmon, or White Salmon River sub-basins. We found bull trout in the West Fork Klickitat drainage of the Klickitat River Sub-basin. Bull trout averaged 6.7 fish/100m{sup 2} in Trappers Creek, 2.6 fish/100m{sup 2} on Clearwater Creek, and 0.4 fish/100m{sup 2} in Little Muddy Creek. Bull trout was the only species of salmonid encountered in Trappers Creek and dominated in Clearwater Creek. Little Muddy Creek was the only creek where bull trout and introduced brook trout occurred together. We found bull trout only at night and typically in low flow regimes. A single fish, believed to be a bull trout x brook trout hybrid, was observed in the Little Muddy Creek. Additional surveys are needed in the West Fork Klickitat and mainstem

  17. Monitor and Protect Wigwam River Bull Trout for Koocanusa Reservoir : Summary of the Skookumchuck Creek Bull Trout Enumeration Project, Annual Report 2001.

    Energy Technology Data Exchange (ETDEWEB)

    Baxter, James S.; Baxter, Jeremy

    2002-03-01

    This report summarizes the second year of a bull trout (Salvelinus confluentus) enumeration project on Skookumchuck Creek in southeastern British Columbia. An enumeration fence and traps were installed on the creek from September 6th to October 12th 2001 to enable the capture of post-spawning bull trout emigrating out of the watershed. During the study period, a total of 273 bull trout were sampled through the enumeration fence. Length and weight were determined for all bull trout captured. In total, 39 fish of undetermined sex, 61 males and 173 females were processed through the fence. An additional 19 bull trout were observed on a snorkel survey prior to the fence being removed on October 12th. Coupled with the fence count, the total bull trout enumerated during this project was 292 fish. Several other species of fish were captured at the enumeration fence including westslope cutthroat trout (Oncorhynchus clarki lewisi), Rocky Mountain whitefish (Prosopium williamsoni), and kokanee (O. nerka). A total of 143 bull trout redds were enumerated on the ground in two different locations (river km 27.5-30.5, and km 24.0-25.5) on October 3rd. The majority of redds (n=132) were observed in the 3.0 km index section (river km 27.5-30.5) that has been surveyed over the past five years. The additional 11 redds were observed in a 1.5 km section (river km 24.0-25.5). Summary plots of water temperature for Bradford Creek, Sandown Creek, Buhl Creek, and Skookumchuck Creek at three locations suggested that water temperatures were within the temperature range preferred by bull trout for spawning, egg incubation, and rearing.

  18. AKTIVITAS SPERMATOPROTECTIVE EKSTRAK DAUN JAMBU BIJI (Psidium guajava PADA JUMLAH SPERMATOZOA TIKUS PUTIH TERINDUKSI KADMIUM

    Directory of Open Access Journals (Sweden)

    W Christijanti

    2014-06-01

    Full Text Available Kadmium dapat bersifat meningkatkan aktivitas oksigen reaktif yang memicu munculnya radikal bebas. Zat aktif dalam daun jambu biji bersifat antioksidan yaitu suatu senyawa yang dapat berikatan atau menghambat terbentuknya radikal bebas. Penelitian ini bertujuan untuk mengkaji adanya hubungan  antara ekstrak daun jambu biji dengan jumlah spermatozoa tikus terinduksi kadmium. Penelitian eksperimental ini menggunakan 20 ekor tikus yang terinduksi kadmium melalui air minum dengan dosis 100ppm/ekor/hari. Kelompok kontrol mendapatkan ekstrak daun jambu biji 0 mg/ekor sedangkan kelompok I, II dan III berturut turut adalah 50 mg/ekor, 100 mg/ekor dan 150 mg/ekor. Pemberian ekstrak daun jambu biji selama 30 hari. Data jumlah spermatozoa dan kadar kadmium dianalisis dengan uji korelasi pada taraf uji 5%. Pada hari ke 31, testis diambil untuk diukur kadar kadmium dan jumlah spermatozoanya. Hasil penelitian menunjukkan adanya korelasi antara jumlah spermatozoa dan kadar kadmium dengan nilai sig 0,009 <0,05 dan berbanding terbalik sebesar  -0,567. Hal ini berarti semakin besar kadar kadium dalam testis maka semakin sedikit jumlah spermatozoa tikus dan semakin kecil kadar kadium dalam testis maka semakin banyak jumlah spermatozoa tikus. Simpulan dari penelitian ini adalah ekstrak daun jambu biji dapat berperan sebagai spermatoprotective tikus yang terpapar Cadmium. Cadmium can be used to increase the activity of reactive oxygen that triggers the emergence of free radicals. The active substances in guava leaves is an antioxidant compound that can bind or inhibit the formation of free radicals . This study aims to examine the relationship between guava leaf extract with the amount of rat spermatozoa induced by cadmium. This experimental study used 20 mice induced by cadmium through drinking water at  dose of 100ppm/mice/day. The control group received guava leaf extract of 0 mg/mice , while group I , II and III received 50 mg , 100 mg and 150 mg in 30 days

  19. IN VITRO ASSESSMENT OF IRON EFFECT ON THE SPERMATOZOA MOTILITY PARAMETERS

    Directory of Open Access Journals (Sweden)

    Zuzana Kňažická

    2012-08-01

    Full Text Available Iron (Fe is an essential element but on the other hand it could induce changes in reproductive system. The general objective of this in vitro study was at first to examine dose- and time-dependent effects of iron (ferrous sulphate heptahydrate - FeSO4.7H2O on the spermatozoa motility parameters, secondly expand the knowledge concerning direct action of this metal on the fertilization potential of the spermatozoa. The motility analysis was determined after exposure to concentrations of 3.9; 7.8; 15.6; 31.2; 62.5; 125; 250; 500; 1000 µmol.dm-3 of FeSO4.7H2O using the Sperm VisionTM CASA (Computer Assisted Semen Analyzer system during different time periods (Time 0 h, 2 h and 24 h. The highest percentage of motile spermatozoa was detected in the control group (95.41±1.32% (Time 0 h. After 2 h of cultivation with ferrous sulphate heptahydrate the motility spermatozoa significantly (P<0.001 increased at the concentrations ≤ 125 μmol.dm-3. The experimental administration at the doses ≥ 125 μmol.dm-3 FeSO4.7H2O inhibited the overall percentage of spermatozoa motility during Time 24 h. The identical spermatozoa motility was detected also for the percentage of progressive motile spermatozoa during all time periods. Detailed evaluation of spermatozoa distance average (DAP and velocity average (VAP path as well as amplitude of lateral head displacement (ALH revealed decrease in groups with concentrations ≥ 125 μmol.dm-3 FeSO4.7H2O in comparison with the control group during the long-term cultivation. Based on these results, we can conclude that the iron at the low concentrations maintains the spermatozoa motility parameters. This essential element has probably direct action on the fertilization potential of the spermatozoa, what could be used in assisted reproductive technologies.

  20. Effect of cryoprotectant solutes on water permeability of human spermatozoa.

    Science.gov (United States)

    Gilmore, J A; McGann, L E; Liu, J; Gao, D Y; Peter, A T; Kleinhans, F W; Critser, J K

    1995-11-01

    Osmotic permeability characteristics and the effects of cryoprotectants are important determinants of recovery and function of spermatozoa after cryopreservation. The primary purpose of this study was to determine the osmotic permeability parameters of human spermatozoa in the presence of cryoprotectants. A series of experiments was done to: 1) validate the use of an electronic particle counter for determining both static and kinetic changes in sperm cell volume; 2) determine the permeability of the cells to various cryoprotectants; and 3) test the hypothesis that human sperm water permeability is affected by the presence of cryoprotectant solutes. The isosmotic volume of human sperm was 28.2 +/- 0.2 microns3 (mean +/- SEM), 29.0 +/- 0.3 microns3, and 28.2 +/- 0.4 microns3 at 22, 11, and 0 degrees C, respectively, measured at 285 mOsm/kg via an electronic particle counter. The osmotically inactive fraction of human sperm was determined from Boyle van't Hoff (BVH) plots of samples exposed to four different osmolalities (900, 600, 285, and 145 mOsm/kg). Over this range, cells behaved as linear osmometers with osmotically inactive cell percentages at 22, 11, and 0 degrees C of 50 +/- 1%, 41 +/- 2%, and 52 +/- 3%, respectively. Permeability of human sperm to water was determined from the kinetics of volume change in a hyposmotic solution (145 mOsm/kg) at the three experimental temperatures. The hydraulic conductivity (Lp) was 1.84 +/- 0.06 microns.min-1.atm-1, 1.45 +/- 0.04 microns.min-1.atm-1, and 1.14 +/- 0.07 microns.min-1.atm-1 at 22, 11, and 0 degrees C, respectively, yielding an Arrhenius activation energy (Ea) of 3.48 kcal/mol. These biophysical characteristics of human spermatozoa are consistent with findings in previous reports, validating the use of an electronic particle counter for determining osmotic permeability parameters of human sperm. This validated system was then used to investigate the permeability of human sperm to four different cryoprotectant

  1. Extending the viability of emu spermatozoa during in vitro storage by manipulation of temperature and diluent potassium concentration.

    Science.gov (United States)

    Sood, S; Malecki, I A; Tawang, A; Martin, G B

    2012-01-01

    1. Survival of emu spermatozoa during in vitro storage is not affected by increasing the extracellular [K(+)] to the point where it does not adversely affect spermatozoa function. 2. In three experiments, the effects were studied of [K(+)] in a diluent in the range 12·5-80 mM/l on emu spermatozoa survival for up to 48 h at 5, 10 or 20°C. 3. At the end of the storage period, spermatozoa viability, motility, fertilising ability and morphology were measured. 4. In Experiment 1, spermatozoa viability and morphology were adversely affected after storage (P diluent containing 80 mM/l [K(+)] whereas spermatozoa motility decreased as [K(+)] increased from 12·5 to 80 mM/l. 5. In Experiment 2, during storage at 5°C, the spermatozoa viability was comparable among any of the diluents (standard or modified) but morphology was better (P diluents than in the standard E3 diluent. 6. In Experiment 3, after 48 h of storage in a diluent containing 40 mM/l of [K(+)], the spermatozoa functions were better preserved at 10°C than at 5 or 20°C. 7. It is concluded that a higher than physiological level of potassium can be used in a diluent without detrimental effect on emu spermatozoa survival during 48 h storage and that the best outcome was with storage at 10°C rather than 5 or 20°C.

  2. Hope for restoration of dead valuable bulls through cloning using donor somatic cells isolated from cryopreserved semen.

    Science.gov (United States)

    Selokar, Naresh L; Saini, Monika; Palta, Prabhat; Chauhan, Manmohan S; Manik, Radheysham; Singla, Suresh K

    2014-01-01

    Somatic cells were isolated from cryopreserved semen of 4 buffalo bulls, 3 of which had died over 10 years earlier, and were established in culture. The cells expressed cytokeratin-18, keratin and vimentin indicating that they were of epithelial origin. The cells were used as nuclear donors for hand-made cloning for producing buffalo embryos. The blastocyst rate and quality, as indicated by apoptotic index, were comparable among embryos produced using cells obtained from fresh or frozen-thawed semen or those obtained from conventional cell sources such as skin. Examination of the epigenetic status revealed that the global level of H3K27me3 but not that of H3K9/14ac and H4K5ac differed significantly (Pcloned embryos from different bulls. The relative mRNA abundance of HDAC1, DNMT1, P53 and CASPASE 3 but not that of DNMT3a differed in cells and in cloned embryos. Following transfer of 24 cloned embryos produced from fresh semen-derived cells to 12 recipients, one calf weighing 55 kg, which is now 6 months of age and is normal, was born through normal parturition. Following transfer of 20 embryos produced from frozen-thawed semen-derived cells to 10 recipients, 2 became pregnant, one of which aborted in the first trimester; the calf born was severely underweight (17 kg), and died 12 h after birth. The ability of cells derived from fresh and frozen-thawed semen to produce live offspring confirms the ability of these cells to be reprogrammed. Our findings pave the way for restoration of highly precious progeny-tested bulls, which has immense economic importance, and can also be used for restoration of endangered species.

  3. Hope for Restoration of Dead Valuable Bulls through Cloning Using Donor Somatic Cells Isolated from Cryopreserved Semen

    Science.gov (United States)

    Selokar, Naresh L.; Saini, Monika; Palta, Prabhat; Chauhan, Manmohan S.; Manik, Radheysham; Singla, Suresh K.

    2014-01-01

    Somatic cells were isolated from cryopreserved semen of 4 buffalo bulls, 3 of which had died over 10 years earlier, and were established in culture. The cells expressed cytokeratin-18, keratin and vimentin indicating that they were of epithelial origin. The cells were used as nuclear donors for hand-made cloning for producing buffalo embryos. The blastocyst rate and quality, as indicated by apoptotic index, were comparable among embryos produced using cells obtained from fresh or frozen-thawed semen or those obtained from conventional cell sources such as skin. Examination of the epigenetic status revealed that the global level of H3K27me3 but not that of H3K9/14ac and H4K5ac differed significantly (Pcloned embryos from different bulls. The relative mRNA abundance of HDAC1, DNMT1, P53 and CASPASE 3 but not that of DNMT3a differed in cells and in cloned embryos. Following transfer of 24 cloned embryos produced from fresh semen-derived cells to 12 recipients, one calf weighing 55 kg, which is now 6 months of age and is normal, was born through normal parturition. Following transfer of 20 embryos produced from frozen-thawed semen-derived cells to 10 recipients, 2 became pregnant, one of which aborted in the first trimester; the calf born was severely underweight (17 kg), and died 12 h after birth. The ability of cells derived from fresh and frozen-thawed semen to produce live offspring confirms the ability of these cells to be reprogrammed. Our findings pave the way for restoration of highly precious progeny-tested bulls, which has immense economic importance, and can also be used for restoration of endangered species. PMID:24614586

  4. Commercial communication of RED BULL on the Czech market

    OpenAIRE

    Gavrilova, Polina

    2017-01-01

    Cílem Bakalářské práce je definovat komerční komunikaci společnosti Red Bull GmbH na českém trhu. Teoretická část práce pomůže pochopit druhou, praktickou část. V práci bude čtenáři krok po kroku představena značka Red Bull a její skrytá identita ale také obraz v očích konzumenta. Umožněné to bude analýzou postavení společnosti na českém trhu, spolu s jejich jedinečným a originálním způsobem komunikace. Dále realizace případové studie Red Bullu se uskuteční skrz primárního výzkumu, který slou...

  5. Peningkatan Kualitas Spermatozoa Epididimis Kerbau Belang yang Dikriopreservasi dengan Beberapa Konsentrasi Sukrosa (THE QUALITY ENHANCEMENT OF EPIDIDYMAL SPERMATOZOA OF SPOTTED BUFFALO CRYOPRESERVING WITH VARIOUS SUCROSE CONCENTRATIONS

    Directory of Open Access Journals (Sweden)

    Muhammad Rizal

    2007-12-01

    Full Text Available Penelitian dilakukan untuk mengetahui kualitas spermatozoa epididimis kerbau belang selama proses pembekuan (kriopreservasi. Bahan pengencer yang digunakan adalah AndroMed® sebagai kontrol (AM serta kombinasi AndroMed® dengan 0,2% (S0,2 dan 0,4% sukrosa (S0,4. Hasil penelitian menunjukkan bahwa persentase motilitas spermatozoa epididimis kerbau belang setelah thawing dalam bahan pengencer AM (41%, nyata (P<0,05 lebih rendah dibandingkan dengan S0,2 (46% dan S0,4 (46%. Demikian pula halnya dengan persentase hidup spermatozoa setelah thawing dalam bahan pengencer AM, S0,2, dan S0,4 secara berturut-turut adalah 52,2; 59,8 dan 60,8% (P<0,05. Sementara itu, tidak terdapat perbedaan nyata (P>0,05 pada persentase membran plasma utuh (MPU setelah thawing dalam ketiga bahan pengencer. Persentase MPU setelah thawing perlakuan AM, S0,2, dan S0,4 adalah masing-masing 68; 68,8 dan 66,8%. Dapat disimpulkan bahwa penambahan sukrosa sebagai krioprotektan eksternal ke dalam bahan pengencer AndroMed® dapat meningkatkan kualitas spermatozoa epididimis kerbau belang setelah thawing.

  6. Sperm tail flexibility test: a simple test for selecting viable spermatozoa for intracytoplasmic sperm injection from semen samples without motile spermatozoa

    Directory of Open Access Journals (Sweden)

    Soares Jonathas Borges

    2003-01-01

    Full Text Available PURPOSE: The objective was to describe the results of the injection of immotile spermatozoa with flexible tails when only immotile spermatozoa are present in the semen sample. METHODS: A retrospective study was conducted to analyze the procedure results for 10 couples who participated in our intracytoplasmic sperm injection program. The sperm tail was considered flexible when it moved up and down independently of the head movement, and it was considered inflexible when the movement occurred together (tail plus head. The fertilization and pregnancy rate were analyzed. RESULTS: The normal fertilization rate (presence of 2 pronuclei was 30.3% (40/132, and the abnormal fertilization rate (presence of less than or more than 2 pronuclei was 6.81% (9/132. A total of 52 embryos were obtained with 9 transfer procedures performed (pregnancy rate: 11.12%. CONCLUSIONS: The sperm tail flexibility test (STFT is an easy and cost-effective way for selecting viable immotile spermatozoa and can be used as an alternative method for determining the viability of spermatozoa. This test seems to be a simple and risk-free method when compared to the swelling test.

  7. Effects of herd origin, AI stud and sire identification on genetic evaluation of Holstein Friesian bulls

    Directory of Open Access Journals (Sweden)

    Giovanni Bittante

    2010-01-01

    Full Text Available The purpose of this study was to estimate the effects of herd origin of bull, AI stud and sire identification number (ID  on official estimated breeding values (EBV for production traits of Holstein Friesian proven bulls. The data included 1,005  Italian Holstein-Friesian bulls, sons of 76 sires, born in 100 herds and progeny tested by 10 AI studs. Bulls were required  to have date of first proof between September 1992 and September 1997, to be born in a herd with at least one other  bull and to have sire and dam with official EBV when bull was selected for progeny testing. Records of sires with only one  son were also discarded. The dependent variable analyzed was the official genetic evaluation for a “quantity and quality  of milk” index (ILQ. The linear model to predict breeding values of bulls included the fixed class effects of herd origin of  bull, AI testing organization, birth year of bull, and estimated breeding values of sire and dam, both as linear covariates.  The R2of the model was 45% and a significant effect was found for genetic merit of sire (P   for herd origin of bull (P   nificant. The range of herd origin effect was 872 kg of ILQ. However, in this study, the causes of this result were not  clear; it may be due to numerous factors, one of which may be preferential treatment on dams of bulls. Analyses of resid-  uals on breeding value of proven bulls for ILQ showed a non significant effect of sire ID, after adjusting for parent aver-  age, herd origin effect and birth year effect. Although the presence of bias in genetic evaluation of dairy bulls is not evi-  dent, further research is recommended firstly to understand the reasons of the significant herd origin effect, secondly to  monitor and guarantee the greatest accuracy and reliability of genetic evaluation procedures. 

  8. Alternative strategies for the long-term preservation of spermatozoa.

    Science.gov (United States)

    Holt, W V

    1997-01-01

    A number of perceived future requirements for stored germplasm in agriculture, aquaculture, biotechnology and conservation are discussed in the present review. In the light of these broad demands, it is apparent that current approaches to gamete and embryo storage need considerable improvement, and that novel approaches to the technologies of germplasm preservation should be pursued if possible. The present article is presented in response to a request for novel future research ideas in this area. Early literature on desiccation, and later research into natural mechanisms of survival during desiccation, is considered in relation to the development of freeze-drying and vitrification methods. Developments in reproductive technologies themselves may mean that freeze-dried spermatozoa could realistically be used for direct intracytoplasmic microinjection of oocytes. Other radical methods that may achieve germplasm preservation are harvesting testicular cells in culture, cryopreserving immature or seasonally regressed testicular material, or a combination of both. Evidence from non-mammalian testicular culture systems, and recent success with the transmeiotic development of mouse spermatids in vitro, suggest that these approaches may eventually become feasible.

  9. Spermatozoa quality assessment: a combined holographic and Raman microscopy approach

    Science.gov (United States)

    De Angelis, Annalisa; Ferrara, Maria A.; Di Caprio, Giuseppe; Managò, Stefano; Sirleto, Luigi; Coppola, Giuseppe; De Luca, Anna Chiara

    2015-05-01

    Semen analysis is widely used as diagnostic tool for assessing male fertility, controlling and managing the animal reproduction. The most important parameters measured in a semen analysis are the morphology and biochemical alterations. For obtaining such information, non-invasive, label-free and non-destructive techniques have to be used. Digital Holography (DH) combined with Raman Spectroscopy (RS) could represent the perfect candidate for a rapid, non-destructive and high-sensitive morphological and biochemical sperm cell analysis. In this study, DH-RS combined approach is used for a complete analysis of single bovine spermatozoa. High-resolution images of bovine sperm have been obtained by DH microscopy from the reconstruction of a single acquired hologram, highlighting in some cases morphological alterations. Quantitative 3D reconstructions of sperm head, both normal and anomalous, have been studied and an unexpected structure of the post-acrosomal region of the head has been detected. Such anomalies have been also confirmed by Raman imaging analysis, suggesting the protein vibrations as associated Raman marker of the defect.

  10. The flagellar apparatus of spermatozoa in fish. Ultrastructure and evolution.

    Science.gov (United States)

    Mattei, X

    1988-01-01

    Chondrichthyes possess an evolved type of spermatozoa. Their flagellar apparatus is characterized by the presence of flagellar roots which form the axis of the midpiece, and the existence of one or two lateral elements associated with the axoneme. Osteichthyes, mainly teleosteans, show a great diversity of spermatic forms. The primitive spermatozoon with a 9 + 2 pattern flagellum is common. The primitive spermatozoon has evolved along different lines. The spermatic diversity which results from this is mainly evident in the structure of the flagellar apparatus. In the animal kingdom the primitive spermatozoon with a 9 + 2 pattern flagellum, present in primitive metazoa, is retained in phyla where external fertilization is maintained. The main evolutionary tendencies--elongation, aflagellarity or biflagellarity--are generally connected with the acquisition of internal fertilization. These evolutionary tendencies are found in teleosteans. It is not possible to link aflagellarity or biflagellarity of the gamete in certain fishes to this method of fertilization. Only the elongation of the spermatozoon is connected, in certain cases, with internal fertilization, but this cannot be taken as general.

  11. Expression

    Directory of Open Access Journals (Sweden)

    Wang-Xia Wang

    2014-02-01

    Full Text Available The miR-15/107 family comprises a group of 10 paralogous microRNAs (miRNAs, sharing a 5′ AGCAGC sequence. These miRNAs have overlapping targets. In order to characterize the expression of miR-15/107 family miRNAs, we employed customized TaqMan Low-Density micro-fluid PCR-array to investigate the expression of miR-15/107 family members, and other selected miRNAs, in 11 human tissues obtained at autopsy including the cerebral cortex, frontal cortex, primary visual cortex, thalamus, heart, lung, liver, kidney, spleen, stomach and skeletal muscle. miR-103, miR-195 and miR-497 were expressed at similar levels across various tissues, whereas miR-107 is enriched in brain samples. We also examined the expression patterns of evolutionarily conserved miR-15/107 miRNAs in three distinct primary rat brain cell preparations (enriched for cortical neurons, astrocytes and microglia, respectively. In primary cultures of rat brain cells, several members of the miR-15/107 family are enriched in neurons compared to other cell types in the central nervous system (CNS. In addition to mature miRNAs, we also examined the expression of precursors (pri-miRNAs. Our data suggested a generally poor correlation between the expression of mature miRNAs and their precursors. In summary, we provide a detailed study of the tissue and cell type-specific expression profile of this highly expressed and phylogenetically conserved family of miRNA genes.

  12. Characteristics of beef from intensively fed western Baggara bulls and heifers: quality attributes and chemical composition

    Directory of Open Access Journals (Sweden)

    I.M.A. Sharaf Eldin

    2013-06-01

    Full Text Available Fourteen samples of L. dorsi muscles were taken from western Baggara cattle, one sample from each of seven bulls and seven heifers randomly selected for slaughter at the end of an experimental feedlot feeding which lasted for 16 weeks at Kuku Research Station, Khartoum North, Sudan, to study sex effects on meat chemical composition and quality attributes. Moisture content of beef was higher in bulls meat than in heifers meat. Protein and ash content were significantly (P<0.001 higher in bulls meat, whereas fat content was significantly (P<0.001 higher in heifers meat than in bulls meat. Cooking loss of bulls meat was significantly (P<0.001 lower and water-holding capacity was also significantly (P<0.01 lower in the bulls meat than in heifers meat. Bull’s meat colour had low lightness (L and high redness (a and yellowness (b, as determined by Hunter Lab. Tristimulus colorimeter, as compared with heifers meat. Sensory panelist scores were higher for colour darkness and flavour intensity and lower for tenderness, juiciness and overall acceptability of bulls meat as compared with heifers meat.

  13. Associations between endotoxin-induced metabolic changes and temperament in Brahman bulls.

    Science.gov (United States)

    Burdick Sanchez, N C; Carroll, J A; Randel, R D; Vann, R C; Welsh, T H

    2014-02-01

    The influence of temperament on the alteration of metabolic parameters in response to a lipopolysaccharide(LPS) challenge was investigated. Brahman bulls were selected based on temperament score. Bulls (10 months; 211±5kg BW; n = 6, 8 and 7 for Calm, Intermediate and Temperamental groups, respectively) were fitted with indwelling jugular catheters to evaluate peripheral blood concentrations of glucose, blood urea nitrogen (BUN),non-esterified fatty acids (NEFA), insulin, epinephrine and cortisol before and after LPS administration (0.5 μg/kg BW LPS). Feed intake was also recorded. Intermediate bulls consumed more feed than the Temperamental bulls during the challenge (p = 0.046). Pre-LPS glucose (p = 0.401) and BUN (p = 0.222) did not differ among the temperament groups. However, pre-LPS insulin (p = 0.023) was lower, whereas pre-LPS NEFA (p metabolic responses of Brahman bulls following a provocative endotoxin challenge.Specifically, Temperamental bulls may preferentially utilize an alternate energy source (i.e. NEFA) to a greater degree than do bulls of Calm and Intermediate temperaments. The use of circulating NEFA from lipolysis may reduce the negative metabolic consequences of an immune response by allowing for a prompt answer to increasing energy demands required during immunological challenge, compared with the time required for glycogenolysis and gluconeogenesis.

  14. Diet overlap of top-level predators in recent sympatry: bull trout and nonnative lake trout

    Science.gov (United States)

    Guy, Christopher S.; McMahon, Thomas E.; Fredenberg, Wade A.; Smith, Clinton J.; Garfield, David W.; Cox, Benjamin S.

    2011-01-01

    The establishment of nonnative lake trout Salvelinus namaycush in lakes containing lacustrine–adfluvial bull trout Salvelinus confluentus often results in a precipitous decline in bull trout abundance. The exact mechanism for the decline is unknown, but one hypothesis is related to competitive exclusion for prey resources. We had the rare opportunity to study the diets of bull trout and nonnative lake trout in Swan Lake, Montana during a concomitant study. The presence of nonnative lake trout in Swan Lake is relatively recent and the population is experiencing rapid population growth. The objective of this study was to evaluate the diets of bull trout and lake trout during the early expansion of this nonnative predator. Diets were sampled from 142 bull trout and 327 lake trout during the autumn in 2007 and 2008. Bull trout and lake trout had similar diets, both consumed Mysis diluviana as the primary invertebrate, especially at juvenile stages, and kokanee Oncorhynchus nerka as the primary vertebrate prey, as adults. A diet shift from primarily M. diluviana to fish occurred at similar lengths for both species, 506 mm (476–545 mm, 95% CI) for bull trout and 495 mm (470–518 mm CI) for lake trout. These data indicate high diet overlap between these two morphologically similar top-level predators. Competitive exclusion may be a possible mechanism if the observed overlap remains similar at varying prey densities and availability.

  15. Monitor and Protect Wigwam River Bull Trout for Koocanusa Reservoir : Summary of the Skookumchuck Creek Bull Trout Enumeration Project, Annual Report 2000.

    Energy Technology Data Exchange (ETDEWEB)

    Baxter, James S.; Baxter, Jeremy

    2001-02-01

    An enumeration fence and traps were installed on Skookumchuck Creek from September 7 th to October 16 th to enable the capture of post-spawning bull trout emigrating out of the watershed. During the study period, a total of 252 bull trout were sampled through the enumeration fence. Length, weight, and sex were determined for all but one of the 252 bull trout captured. In total, one fish of undetermined sex, 63 males and 188 females were processed through the fence. A total of 67 bull trout were observed on a snorkel survey prior to the fence being removed on October 16 th . Coupled with the fence count, the total bull trout count during this project was 319 fish. Several other species of fish were captured at the enumeration fence including westslope cutthroat trout, Rocky Mountain whitefish, kokanee, sucker, and Eastern brook trout. Redds were observed during ground surveys in three different locations (river km 27.5- 28.5, km 29-30, and km 24-25). The largest concentration of redds were noted in the upper two sections which have served as the index sections over the past four years. A total of 197 bull trout redds were enumerated on the ground on October 4 th . The majority of redds (n=189) were observed in the 3.0 km index section (river km 27.5-30.5) that has been surveyed over the past four years. The additional 8 redds were observed in a 1.5 km section (river km 24.0-25.5). Summary plots of water temperature for Bradford Creek, Sandown Creek, Skookumchuck Creek at km 39.5, and Skookumchuck Creek at the fence site suggested that water temperatures were within the range preferred by bull trout for spawning, egg incubation, and rearing.

  16. Impact of a timed-release FSH treatment from 2 to 6 months of age in bulls II: Endocrinology, puberty attainment, and mature sperm production in Holstein bulls.

    Science.gov (United States)

    Harstine, B R; Cruppe, L H; Abreu, F M; Rodrigues, A D; DeJarnette, J M; Day, M L

    2018-01-01

    The use of genomic testing in the cattle industries has renewed an interest in hastening bull puberty. In prepubertal males, FSH facilitates Sertoli cell proliferation and testis maturation. The aim of this study was to determine the effect of prepubertal administration of a timed-release FSH (delivered in a hyaluronan solution) on hormone secretion, puberty attainment, and mature sperm production in Holstein bulls in an AI center. Bulls (n = 29) were randomly assigned to one of two treatment groups based on birth date and pedigree. Beginning at 62 days of age (Day 62), bulls were injected im every 3.5 days with either 30 mg FSH (Folltropin-V; NIH-FSH-P1 units) in a 2% hyaluronan solution (FSH-HA, n = 17) or saline (control, n = 12) until Day 170.5. Blood samples to assess FSH, activin A, and testosterone were collected prior to each treatment. Scrotal circumference (SC) and BW were measured monthly. Puberty assessment (ability to ejaculate 5 × 107 sperm, 10% motile) was initiated at Day 244. Average mature daily sperm production (3× wk collection, combined 2 ejaculates) was assessed from Day 571-627. In blood collected every 3.5 days, FSH concentrations within FSH-HA bulls were increased (P  0.1) in testosterone, BW, or SC. FSH-HA bulls attained puberty at a younger age than control bulls (278 ± 7.7 vs. 303 ± 9.1 days of age, P puberty. We propose this exogenous FSH delivered in hyaluronan initiates a positive feedback loop that includes an increase in activin A production observed on Day 86.5 and 107.5. However, differences in mature sperm production were not realized in this experiment. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Effect of procaine, pentoxifylline and trolox on capacitation and hyperactivation of stallion spermatozoa.

    Science.gov (United States)

    Ortgies, F; Klewitz, J; Görgens, A; Martinsson, G; Sieme, H

    2012-05-01

    Reasons for low in vitro fertilisation rates in the horse include the difficulties in inducing capacitation and/or hyperactivation of stallion spermatozoa. The aim of this study was to analyse the effect of noncapacitating and capacitating modified Whitten's (MW) and modified Tyrode's medium (MT) and treatment with procaine (5 mmol), pentoxifylline (3.5 mmol) and trolox (120 mmol) on motility (CASA), capacitation, acrosomal status, viability and mitochondrial membrane potential of stallion spermatozoa (n = 4). While there was no influence of MW and MT on sperm motility, a significant increase in the percentage of viable-capacitated spermatozoa was observed after incubation in capacitating MW (P Trolox had no detectable effect on either capacitation or hyperactivation. Procaine was the only agent that induced hyperactivation in terms of a reduced proportion of progressively motile spermatozoa, straight line velocity, straightness, linearity and beat-cross frequency and an increase in the amplitude of lateral head displacement (P < 0.05). The combination of capacitating Whitten's medium and procaine showed the best results for the induction of capacitation and hyperactivation in stallion spermatozoa; this was possible even after short-term incubation. © 2011 Blackwell Verlag GmbH.

  18. Bovine serum albumin: survival and osmolarity effect in bovine spermatozoa stored above freezing point.

    Science.gov (United States)

    Nang, C F; Osman, K; Budin, S B; Ismail, M I; Jaffar, F H F; Mohamad, S F S; Ibrahim, S F

    2012-05-01

    Liquid nitrogen preservation in remote farms is a limitation. The goal of this study was to determine optimum temperature above freezing point for bovine spermatozoa preservation using bovine serum albumin (BSA) as a supplementation. Pooled semen sample from three ejaculates was subjected to various BSA concentration (1, 4, 8 and 12 mg ml(-1)), before incubation in different above freezing point temperatures (4, 25 and 37 °C). Viability assessment was carried out against time from day 0 (fresh sample) until all spermatozoa become nonviable. Optimal condition for bovine spermatozoa storage was at 4 °C with 1 mg ml(-1) BSA for almost 7 days. BSA improved bovine spermatozoa viability declining rate to 44.28% at day 4 and 57.59% at day 7 compared to control, with 80.54% and 98.57% at day 4 and 7 respectively. Increase in BSA concentration did not improve sperm viability. Our results also confirmed that there was a strong negative correlation between media osmolarity and bovine spermatozoa survival rate with r = 0.885, P freezing point. © 2011 Blackwell Verlag GmbH.

  19. Effects of 1950 MHz W-CDMA-like signal on human spermatozoa.

    Science.gov (United States)

    Nakatani-Enomoto, Setsu; Okutsu, Miho; Suzuki, Satoshi; Suganuma, Ryota; Groiss, Stefan Jun; Kadowaki, Suguru; Enomoto, Hiroyuki; Fujimori, Keiya; Ugawa, Yoshikazu

    2016-09-01

    There are growing concerns about how electromagnetic waves (EMW) emitted from mobile phones affect human spermatozoa. Several experiments have suggested harmful effects of EMW on human sperm quality, motility, velocity, or the deoxyribonucleic acid (DNA) of spermatozoa. In this study, we analyzed the effects on human spermatozoa (sperm motility and kinetic variables) induced by 1 h of exposure to 1950 MHz Wideband Code Division Multiple Access (W-CDMA)-like EMW with specific absorption rates of either 2.0 or 6.0 W/kg, using a computer-assisted sperm analyzer system. We also measured the percentage of 8-hydroxy-2'-deoxyguanosine (8-OHdG) positive spermatozoa with flow cytometry to evaluate damage to DNA. No significant differences were observed between the EMW exposure and the sham exposure in sperm motility, kinetic variables, or 8-OHdG levels. We conclude that W-CDMA-like exposure for 1 h under temperature-controlled conditions has no detectable effect on normal human spermatozoa. Differences in exposure conditions, humidity, temperature control, baseline sperm characteristics, and age of donors may explain inconsistency of our results with several previous studies. Bioelectromagnetics. 37:373-381, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  20. EFFECTS OF BREED, SPERMATOZOA CONCENTRATION, AND STORAGE ON PROGRESSIVE MOTILITY OF EXTENDED BOAR SEMEN

    Directory of Open Access Journals (Sweden)

    Aleksandar Božić

    2011-12-01

    Full Text Available The classic technology of artificial insemination (AI often requires insemination doses to be kept for more than 24 hours, with a requirement that the degree of progressive motility at the moment of insemination not be below 65%. The aim of this paper was to determine the influence of breed, spermatozoa concentration, and storage time on the fertilization capacity of extended semen from native ejaculates of boars. The research included the following boar breeds: Duroc (n=34, Hampshire (n=30, Large White (n=42 and Swedish Landrace (n=32, from large pig farms in Vojvodina (Republic of Serbia. Two ejaculates were collected from each boar once monthly for 12 months (a total of 24 ejaculates per boar. There was statistically significant (p<0.01 influence of breed on the number of spermatozoa samples that maintained ≥ 65% progressive motility during 48 hours of storage in 1:4 dilution. There was also an influence of spermatozoa concentration on progressive motility. As spermatozoa concentration increased during storage, ≥ 65% progressive motility declined (P≤0.01 within 24 hours. The results show that it is necessary to determine the adequate dilution rate and storage time for each ejaculate, while taking into account spermatozoa concentration in the native semen.

  1. Proteomic Investigation of Ram Spermatozoa and the Proteins Conferred by Seminal Plasma.

    Science.gov (United States)

    Pini, Taylor; Leahy, Tamara; Soleilhavoup, Clement; Tsikis, Guillaume; Labas, Valerie; Combes-Soia, Lucie; Harichaux, Gregoire; Rickard, Jessica P; Druart, Xavier; de Graaf, Simon P

    2016-10-07

    Sperm proteomes have emerged for several species; however, the extent of species similarity is unknown. Sheep are an important agricultural species for which a comprehensive sperm proteome has not been produced. In addition, potential proteomic factors from seminal plasma that may contribute to improved fertility after cervical insemination are yet to be explored. Here we use liquid chromatography-tandem mass spectrometry to investigate the proteome of ejaculated ram spermatozoa, with quantitative comparison to epididymal spermatozoa. We also present a comparison to published proteomes of five other species. We identified 685 proteins in ejaculated ram spermatozoa, with the most abundant proteins involved in metabolic pathways. Only 5% of ram sperm proteins were not detected in other species, which suggest highly conserved structures and pathways. Of the proteins present in both epididymal and ejaculated ram spermatozoa, 7% were more abundant in ejaculated spermatozoa. Only two membrane-bound proteins were detected solely in ejaculated sperm lysates: liver enriched gene 1 (LEG1/C6orf58) and epidermal growth factor-like repeats and discoidin I-like domains 3 (EDIL3). This is the first evidence that despite its relatively complex proteomic composition, seminal plasma exposure leads to few novel proteins binding tightly to the ram sperm plasma membrane.

  2. A comparison between freezing methods for the cryopreservation of stallion spermatozoa.

    Science.gov (United States)

    Clulow, J R; Mansfield, L J; Morris, L H A; Evans, G; Maxwell, W M C

    2008-11-01

    The effects of sperm freezing concentration (40 x 10(6)mL(-1) vs. 400 x 10(6)mL(-1)), straw size (0.25 mL vs. 0.5 mL) and freezing method (liquid nitrogen vapour in a Styrofoam box vs. programmable freezing machine) were evaluated in a 2 x 2 x 2 factorial experimental design using 3 split ejaculates from each of 4 stallions. Immediately after thawing, the total motility and forward progressive motility of spermatozoa frozen at a concentration of 40 x 10(6)mL(-1) was higher than for spermatozoa frozen at 400 x 10(6)mL(-1). No significant differences were observed in the semen parameters assessed after cryopreservation in either 0.25 or 0.5 mL straws. However, the programmable freezer provided a more consistent and reliable freezing rate than liquid nitrogen vapour. We conclude that an effective protocol for the cryopreservation of stallion spermatozoa at low concentrations would include concentrations of 40 x 10(6)mL(-1) in 0.25 mL straws using a programmable freezer. This freezing protocol would be suitable for emerging sperm technologies such as sex-preselection of stallion spermatozoa as the sorting process yields only low numbers of spermatozoa in a small volume available for either immediate insemination or cryopreservation.

  3. In vitro fertilizing potential of urethral and epididymal spermatozoa collected from domestic cats (Felis catus).

    Science.gov (United States)

    Prochowska, S; Niżański, W

    2017-03-28

    The aim of this study was to provide a comparative analysis of in vitro fertilizing potential of frozen-thawed urethral and epididymal feline spermatozoa. Both types of semen were collected from 7 cats and cryopreserved in liquid nitrogen. To perform in vitro fertilization, both urethral and epididymal samples from the same individual were thawed and spermatozoa were co-incubated with in vitro matured cat oocytes. Obtained embryos were cultured in vitro for 7 days in a commercial medium. Cleavage rate, morula rate and blastocyst rate were calculated. Experiment was run in 10 replicates. The examined parameters showed no significant differences between urethral and epididymal spermatozoa (p>0.05). Cleavage rate and embryo's development were highly variable between replicates, even for the different sperm samples collected from one individual. There was no significant correlation between fertilizing capacity of two types of spermatozoa collected from the same male. In this study we confirmed that cryopreserved urethral spermatozoa have equally good fertilizing potential as epididymal ones, and both can be successfully used for in vitro fertilization in cats with the use of commercial medium.

  4. Effect of Red Bull energy drink on cardiovascular and renal function.

    Science.gov (United States)

    Ragsdale, Frances R; Gronli, Tyler D; Batool, Narjes; Haight, Nicole; Mehaffey, April; McMahon, Erin C; Nalli, Thomas W; Mannello, Carla M; Sell, Crystal J; McCann, Patrick J; Kastello, Gary M; Hooks, Tisha; Wilson, Ted

    2010-04-01

    Energy drink consumption has been anecdotally linked to the development of adverse cardiovascular effects in consumers, although clinical trials to support this link are lacking. The effects of Red Bull energy drink on cardiovascular and neurologic functions were examined in college-aged students enrolled at Winona State University. In a double-blind experiment where normal calorie and low calorie Red Bull were compared to normal and low calorie placebos, no changes in overall cardiovascular function nor blood glucose (mg/dL) were recorded in any participant (n = 68) throughout a 2-h test period. However, in the second experiment, nine male and twelve female participants subjected to a cold pressor test (CPT) before and after Red Bull consumption showed a significant increase in blood sugar levels pre- and post Red Bull consumption. There was a significant increase in diastolic blood pressure of the male volunteers immediately after submersion of the hand in the 5 degrees C water for the CPT. Under the influence of Red Bull, the increase in diastolic pressure for the male participants during the CPT was negated. There were no significant changes in the blood pressure of the female participants for the CPT with or without Red Bull. Finally, the CPT was used to evaluate pain threshold and pain tolerance before and after Red Bull consumption. Red Bull consumption was associated with a significant increase in pain tolerance in all participants. These findings suggest that Red Bull consumption ameliorates changes in blood pressure during stressful experiences and increases the participants' pain tolerance.

  5. Red Bull® energy drink increases consumption of higher concentrations of alcohol.

    Science.gov (United States)

    Roldán, Marta; Echeverry-Alzate, Victor; Bühler, Kora-Mareen; Sánchez-Diez, Israel J; Calleja-Conde, Javier; Olmos, Pedro; Boehm, Stephen L; Maldonado, Rafael; Rodríguez de Fonseca, Fernando; Santiago, Catalina; Gómez-Gallego, Felix; Giné, Elena; López-Moreno, Jose Antonio

    2017-09-22

    Mixing alcohol with caffeinated energy drinks is a common practice, especially among young people. In humans, the research on this issue has mainly focused on the use of the mass-marketed energy drinks themselves, whereas in animal models, it has focused on the individual effects of their active ingredients (i.e. caffeine). Here, we have characterized how Red Bull®, one of the most consumed caffeinated energy drink worldwide, modulates operant alcohol self-administration in Wistar rats. We found that animals readily and steadily responded for Red Bull (mean: 90 responses, 30 minutes and fixed-ratio 1), which was accompanied by locomotor stimulating effects (26 percent increase). The higher the concentration of alcohol (3-20 percent), the higher the consumption of alcohol (g/kg) and associated blood alcohol levels (91.76 percent) in the mixed Red Bull-alcohol group (60 percent increase). Blood caffeine levels in the Red Bull group were 4.69 μg/ml and 1.31 μg/ml in the Red Bull-alcohol group after the 30-minute session. Because Red Bull also contains 11 percent sucrose, we examined the time course of blood glucose as well as insulin and corticosterone. The correlation between intake of Red Bull and blood glucose levels was higher at 90 minutes than 5 minutes after its consumption, and there was no relationship with blood insulin or blood corticosterone levels. Red Bull did not alter extinction and reacquisition of responding for alcohol nor did it affect relapse-like drinking. Overall, our results suggest that Red Bull might be a vulnerability factor to develop alcoholism given that it intensifies the consumption of higher concentrations of alcohol. © 2017 Society for the Study of Addiction.

  6. Wigwam River Juvenile Bull Trout and Fish Habitat Monitoring Program : 2002 Data Report.

    Energy Technology Data Exchange (ETDEWEB)

    Cope, R.S. [Westslope Fisheries, Cranbrook, BC, Canada

    2003-03-01

    The Wigwam River bull trout (Salvelinus confluentus) and fish habitat monitoring program is a trans-boundary initiative implemented by the British Columbia Ministry of Water, Land, and Air Protection (MWLAP), in cooperation with Bonneville Power Administration (BPA). The Wigwam River is an important fisheries stream located in southeastern British Columbia that supports healthy populations of both bull trout and Westslope cutthroat trout (Figure 1). This river has been characterized as the single most important bull trout spawning stream in the Kootenay Region (Baxter and Westover 2000, Cope 1998). In addition, the Wigwam River supports some of the largest Westslope cutthroat trout (Oncorhynchus clarki lewisi) in the Kootenay Region. These fish are highly sought after by anglers (Westover 1999a, 1999b). Bull trout populations have declined in many areas of their range within Montana and throughout the northwest including British Columbia. Bull trout were blue listed as vulnerable in British Columbia by the B.C. Conservation Data Center (Cannings 1993) and although there are many healthy populations of bull trout in the East Kootenay they remain a species of special concern. Bull trout in the United States portion of the Columbia River were listed as threatened in 1998 under the Endangered Species Act by the U.S. Fish and Wildlife Service. The upper Kootenay River is within the Kootenai sub-basin of the Mountain Columbia Province, one of the eleven Eco-provinces that make up the Columbia River Basin. MWLAP applied for and received funding from BPA to assess and monitor the status of wild, native stocks of bull trout in tributaries to Lake Koocanusa (Libby Reservoir) and the upper Kootenay River. This task is one of many that were undertaken to ''Monitor and Protect Bull Trout for Koocanusa Reservoir'' (BPA Project Number 2000-04-00).

  7. Wigwam River Juvenile Bull Trout and Fish Habitat Monitoring Program : 2000 Data Report.

    Energy Technology Data Exchange (ETDEWEB)

    Cope, R.S.; Morris, K.J.

    2001-03-01

    The Wigwam River bull trout (Salvelinus confluentus) and fish habitat monitoring program is a trans-boundary initiative implemented by the British Columbia Ministry of Environment, Lands and Parks (MOE), in cooperation with Bonneville Power Administration (BPA). The Wigwam River is an important fisheries stream located in southeastern British Columbia that supports healthy populations of both bull trout and Westslope cutthroat trout (Figure 1.1). This river has been characterized as the single most important bull trout spawning stream in the Kootenay Region (Baxter and Westover 2000, Cope 1998). In addition, the Wigwam River supports some of the largest Westslope cutthroat trout (Oncorhynchus clarki lewisi) in the Kootenay Region. These fish are highly sought after by anglers (Westover 1999a, 1999b). Bull trout populations have declined in many areas of their range within Montana and throughout the northwest including British Columbia. Bull trout were blue listed as vulnerable in British Columbia by the B.C. Conservation Data Center (Cannings 1993) and although there are many healthy populations of bull trout in the East Kootenays they remain a species of special concern. Bull trout in the United States portion of the Columbia River were listed as threatened in 1998 under the Endangered Species Act by the U.S. Fish and Wildlife Service. The upper Kootenay River is within the Kootenai sub-basin of the Mountain Columbia Province, one of the eleven Eco-provinces that make up the Columbia River Basin. MOE applied for and received funding from BPA to assess and monitor the status of wild, native stocks of bull trout in tributaries to Lake Koocanusa (Libby Reservoir) and the upper Kootenay River. This task is one of many that was undertaken to ''Monitor and Protect Bull Trout for Koocanusa Reservoir'' (BPA Project Number 2000-04-00).

  8. Temporary Restoration of Bull Trout Passage at Albeni Falls Dam, 2008 Progress Report.

    Energy Technology Data Exchange (ETDEWEB)

    Bellgraph, Brian J. [Pacific Northwest National Laboratory

    2009-03-31

    The goal of this project is to provide temporary upstream passage of bull trout around Albeni Falls Dam on the Pend Oreille River, Idaho. Our specific objectives are to capture fish downstream of Albeni Falls Dam, tag them with combination acoustic and radio transmitters, release them upstream of Albeni Falls Dam, and determine if genetic information on tagged fish can be used to accurately establish where fish are located during the spawning season. In 2007, radio receiving stations were installed at several locations throughout the Pend Oreille River watershed to detect movements of adult bull trout; however, no bull trout were tagged during that year. In 2008, four bull trout were captured downstream of Albeni Falls Dam, implanted with transmitters, and released upstream of the dam at Priest River, Idaho. The most-likely natal tributaries of bull trout assigned using genetic analyses were Grouse Creek (N = 2); a tributary of the Pack River, Lightning Creek (N = 1); and Rattle Creek (N = 1), a tributary of Lightning Creek. All four bull trout migrated upstream from the release site in Priest River, Idaho, were detected at monitoring stations near Dover, Idaho, and were presumed to reside in Lake Pend Oreille from spring until fall 2008. The transmitter of one bull trout with a genetic assignment to Grouse Creek was found in Grouse Creek in October 2008; however, the fish was not found. The bull trout assigned to Rattle Creek was detected in the Clark Fork River downstream from Cabinet Gorge Dam (approximately 13 km from the mouth of Lightning Creek) in September but was not detected entering Lightning Creek. The remaining two bull trout were not detected in 2008 after detection at the Dover receiving stations. This report details the progress by work element in the 2008 statement of work, including data analyses of fish movements, and expands on the information reported in the quarterly Pisces status reports.

  9. Electroejaculation increased vocalization and plasma concentrations of cortisol and progesterone, but not substance P, in beef bulls.

    Science.gov (United States)

    Whitlock, B K; Coffman, E A; Coetzee, J F; Daniel, J A

    2012-09-01

    Electroejaculation is a reliable method of obtaining a semen sample for a bull breeding soundness examination, but is sometimes regarded as painful. Substance P is a neuropeptide involved in the integration of pain, stress, and anxiety. We hypothesized that substance P is a measure of pain in bulls following electroejaculation. The specific objective was to compare vocalization and plasma concentrations of cortisol, progesterone, and substance P immunoreactivity in bulls following electroejaculation. Nine Angus bulls (501.9 ± 14.3 kg) were used. Blood samples were collected at -60, -30, 0, 2, 10, 20, 30, 45, 60, 75, 90, 120 min relative to treatment. At Time 0, bulls were subject to electroejaculation, rectal probe insertion without electroejaculation, or no manipulation. Treatments were administered contemporaneously to three bulls. Treatments were repeated weekly until each bull had received each treatment in a 3 × 3 Latin square design. More bulls (P = 0.0147) in the electroejaculation group vocalized (5 of 9 bulls; 55.6%) when compared to controls (0 of 9 bulls; 0%). Mean plasma cortisol and progesterone concentration following electroejaculation in bulls were higher (P electroejaculation in bulls (77.2 ± 17.2 pg/mL) was not different (P = 0.6264) from probed (79.1 ± 17.2 pg/mL) or control bulls (93.4 ± 17.2 pg/mL). A significant increase in vocalization and plasma cortisol and progesterone concentrations in bulls following electroejaculation was likely owing to acute stress. However, the lack of a difference in plasma concentrations of substance P after electroejaculation was interpreted as a lack of pain associated with nociception. Copyright © 2012 Elsevier Inc. All rights reserved.

  10. Proteins associated with critical sperm functions and sperm head shape are differentially expressed in morphologically abnormal bovine sperm induced by scrotal insulation

    NARCIS (Netherlands)

    Saadi, H.A.S.; Riemsdijk, van E.L.C.; Dance, A.L.; Rajamanickam, G.D.; Kastelic, J.P.; Thundathil, J.C.

    2013-01-01

    The objective was to investigate expression patterns of proteins in pyriform sperm, a common morphological abnormality in bull sperm. Ejaculates were collected from sexually mature Holstein bulls (n = 3) twice weekly for 10 weeks (pre-thermal insult samples). Testicular temperature was elevated in

  11. Infertility in the dromedary bull: a review of causes, relations and implications.

    Science.gov (United States)

    Al-Qarawi, A A

    2005-06-01

    Research into infertility in the dromedary bull, as reported during the last two decades, is reviewed with emphasis on causes and effects. Reproductive activity of such animals is naturally limited by a breeding season, though with enough encouragement some may mate with oestrous females out of season but a full fertilization potential can in no way be expected. It is essential that any female presented to a bull is capable of reproducing. The presentation of a subfertile or infertile female due to infection or physiological abnormality will adversely affect the female's ability to conceive and, therefore, the apparent fertility rates of the bull she was put to. The average number of successful services a bull could be expected to perform is two per day. Dromedary bulls with large testes have larger sperm outputs and can cope with more than two females per day providing that they are given adequate periods of rest, 1-2 days every 10 days or so, in conjunction with appropriate nutrition throughout the season. Anabolic steroids or testosterone therapies, which are sometimes used in an attempt to improve male characteristics and bull libido, are not recommended for dromedary bulls in breeding work. Such steroids result in a decrease in testicular size and weight with fewer sperm per gram of testicular tissue being found and the sperm produced also have lower motility rates. Pain associated with the act of mating a she-camel, due to injuries or inflammation in the scrotum, testes, prepuce and sheath, can cause a permanent reduction in bull libido. Camel bulls achieving pregnancy rates more than 60% have had consistently higher spermatozoal concentrations and kinematic variables derived by the computerized cell motion analyzer (CMA) system. As far as physical capabilities are concerned, 3-year-old dromedary bulls, which have reached puberty, have been shown to be perfectly capable of fertilizing a female, but they have a limited sperm production to perform consistently

  12. Effect of a ceiling fan ventilation system on finishing young bulls' health, behaviour and growth performance.

    Science.gov (United States)

    Magrin, L; Brscic, M; Lora, I; Rumor, C; Tondello, L; Cozzi, G; Gottardo, F

    2017-06-01

    This research aimed at assessing the effects of a ceiling fan ventilation system on health, feeding, social behaviour and growth response of finishing young bulls fattened indoors during a mild summer season. A total of 69 Charolais young bulls were housed in six pens without any mechanical ventilation system (Control) and in six pens equipped with ceiling fans. The experimental period lasted 98 days from June until mid-September 2014. Four experimental days were considered in order to assess the effect of the ventilation system under two different microclimatic conditions: 2 alert days at monthly interval with temperature humidity index (THI) between 75 and 78, and 2 normal days with THI⩽74. Health and behaviour of the bulls were evaluated through 8-h observation sessions starting after morning feed delivery. The study was carried out during a rather cool summer with a climate average THI of 68.9 and 4 days with average THI>75. Despite these mild climate conditions, ceiling fans lowered litter moisture and acted as a preventive measure for bulls' dirtiness (odd ratio=47.9; 95% CI 19.6 to 117.4). The risk of abnormal breathing was increased for Control bulls (odd ratio=40.7; 95% CI 5.4 to 304.2). When exposed to alert THI conditions, respiration rate and panting scores increased and rumination duration dropped in Control bulls compared with bulls provided with a ceiling fan. During observations under alert THI, bulls spent less time eating, more time being inactive and consumed more water compared with normal THI conditions. Bulls' daily dry matter intake measured during the observation sessions decreased on alert compared with normal THI days (PCeiling fan treatment had no effect on bulls' growth performance or water consumption but these results most likely depended on the mild climate conditions. Ceiling fans proved to mitigate some of the negative effects of heat stress on bulls' behaviour (rumination, lying down and drinking water) and respiration rate

  13. Radiation damage to bull sperm motility. III. Further x-ray studies

    Energy Technology Data Exchange (ETDEWEB)

    Rikmenspoel, R.

    1975-08-01

    The results of previous radiation experiments, which indicated that the centriole serves as a control center for bull sperm motility, appear to be in conflict with experiments showing that the bull sperm flagellum is an autonomous oscillator. To resolve this conflict experiments were conducted to calibrate absolutely the dose-response curves for the radiation damage, and to measure the force production and the mechanochemical energy conversion after irradiation in bull sperm. The results indicate that the centriole acts as a mechanical anchor for the contractile fibers. (auth)

  14. Near-infrared reflectance bull's eye maculopathy as an early indication of hydroxychloroquine toxicity.

    Science.gov (United States)

    Wong, Keye L; Pautler, Scott E; Browning, David J

    2015-01-01

    In some patients, hydroxychloroquine ocular toxicity may progress even following cessation of therapy. Any leverage the clinician may use to allow earlier detection may avert significant vision loss. We report three cases suggesting that bull's eye maculopathy seen on near-infrared reflectance with a confocal scanning laser ophthalmoscope could be an early, objective manifestation of hydroxychloroquine ocular toxicity, and with progression of the disease this near-infrared "bull's eye" change may disappear. Alerting clinicians to this observation may allow a larger case series to corroborate the hypothesis that bull's eye maculopathy detected by near-infrared reflectance may represent an early sign of hydroxychloroquine toxicity.

  15. Duration Dependence in Stock Prices: An Analysis of Bull and Bear Markets

    DEFF Research Database (Denmark)

    Lunde, Asger; Timmermann, Allan

    2004-01-01

    This article studies time series dependence in the direction of stock prices by modeling the (instantaneous) probability that a bull or bear market terminates as a function of its age and a set of underlying state variables, such as interest rates. A random walk model is rejected both for bull...... and bear markets. Although it . ts the data better, a generalized autoregressive conditional heteroscedasticity model is also found to be inconsistent with the very long bull markets observed in the data. The strongest effect of increasing interest rates is found to be a lower bear market hazard rate...

  16. Content of heavy metals in the semen of bulls from various environments

    Energy Technology Data Exchange (ETDEWEB)

    Monkiewicz, J.; Jaczewski, S.; Dynarowicz, I.

    1975-01-01

    The content of metal ions (Cu, Pb, Zn) was examined in the semen of bulls kept under different environments. It was found that the content of Cu and Pb ions in the semen of bulls living near the copperworks and near to a highway was higher than that in the semen of bulls kept under normal conditions. The differences appeared to be highly significant statistically (P < 0.001). No significant statistical differences were found in the content of Zn. In addition, it was stated that the higher content of Pb influenced the survival of spermatozoons at highers temperature (46.5/sup 0/C). Further investigations will be performed under model conditions.

  17. Fertility Defects in Mice Expressing the L68Q Variant of Human Cystatin C

    Science.gov (United States)

    Whelly, Sandra; Serobian, Gaiane; Borchardt, Clinton; Powell, Jonathan; Johnson, Seethal; Hakansson, Katarina; Lindstrom, Veronica; Abrahamson, Magnus; Grubb, Anders; Cornwall, Gail A.

    2014-01-01

    Hereditary cystatin C amyloid angiopathy is an autosomal dominant disorder in which a variant form of cystatin C (L68Q) readily forms amyloid deposits in cerebral arteries in affected individuals resulting in early death. L68Q protein deposits in human cystatin C amyloid angiopathy patients have also been found in tissues outside of the brain including the testis, suggesting possible effects on fertility. Heterozygous transgenic mice (L68Q) that express the human L68Q variant of cystatin C under the control of the mouse cystatin C promoter were unable to generate offspring, suggesting the presence of L68Q cystatin C amyloid affected sperm function. In vitro studies showed that epididymal spermatozoa from L68Q mice were unable to fertilize oocytes and exhibited poor sperm motility. Furthermore, spermatozoa from L68Q mice exhibited reduced cell viability compared with wild type (WT) spermatozoa and often were detected in large agglutinated clumps. Examination of the epididymal fluid and spermatozoa from L68Q mice showed increased levels and distinct forms of cystatin C amyloid that were not present in WT mice. The addition of epididymal fluid from L68Q mice to WT spermatozoa resulted in a recapitulation of the L68Q phenotype in that WT spermatozoa showed reduced cell viability and motility compared with WT spermatozoa incubated in epididymal fluid from WT mice. L68Q epididymal fluid that was depleted of cystatin C amyloids, however, did not impair the motility of WT spermatozoa. Taken together these studies suggest that amyloids in the epididymal fluid can be cytotoxic to the maturing spermatozoa resulting in male infertility. PMID:24500719

  18. Re-evaluating the Contribution and Legacy of Hedley Bull

    Directory of Open Access Journals (Sweden)

    Emerson Maione Souza

    2008-06-01

    Full Text Available The article aims, in the first instance, to make a detailed analysis of the work of Hedley Bull, approaching the main themes and concepts developed by him. Secondly, it aims to re-evaluate the potential of the author’s contribution, given the new conditions of the post-Cold War period. With this in mind, the article critically analyses the most recent interpretations of this work, which seek to highlight its critical and normative potential, as well as to dissociate it from the realist tradition in international relations. These two facts differentiate the new commentators from older ones and reaffirm the continuing relevance of Hedley Bull’s work, the latter being the article’s chief conclusion.

  19. Effects of exposure of epididymal boar spermatozoa to seminal plasma on the binding of zona pellucida proteins during in vitro capacitation

    NARCIS (Netherlands)

    Harkema, W.; Colenbrander, B.; Engel, B.; Woelders, H.

    2004-01-01

    The purpose of the investigation was to determine whether seminal plasma plays a role in the increase during in vitro capacitation of the number of boar spermatozoa with enhanced binding of zona pellucida proteins. Ejaculated spermatozoa and spermatozoa collected from the caudae epididymides of

  20. ASSESSMENT OF CHAROLAIS BULLS POPULATION STRUCTURE BASED ON SNPs ANALYSES

    Directory of Open Access Journals (Sweden)

    Nina Moravčíková

    2014-02-01

    Full Text Available The aim of this study was identification of SNPs in leptin (LEP, leptin receptor (LEPR, growth hormone (GH and specific pituitary transcription factor (Pit-1 genes in order to analyze genetic structure of Charolais bulls’ population. The total numbers of genomic DNA samples were taken from 52 breeding bulls and analyzed by PCR-RFLP method. After digestion with restriction enzymes were detected in bulls’ population alleles with frequency: LEP/Sau3AI A 0.83 and B 0.17 (±0.037; LEPR/BseGI C 0.95 and T 0.05 (±0.021, GH/AluI L 0.62 and V 0.38 (±0.048 and Pit1/HinfI A 0.40 and B 0.60 (±0.048. Based on the observed vs. expected genotypes frequencies population across loci were in Hardy-Weinberg equilibrium (P>0.05, only in case of Pit-1 locus was detected disequilibrium. Predominant were in analyzed breeding bulls LEP/Sau3AIAA (0.69, LEPR/T945MCC (0.90, GH/AluILL (0.43 and Pit-1/HinfIAB (0.65 genotypes. The observed heterozygosity of SNPs was also transferred to the low (LEP/Sau3AI/0.248 and LEPR/T945M/0.088 or median polymorphic information content (GH/AluI/0.366 and Pit-1/HinfI/0.370. Within genetic variability estimating negative (LEPR/T945M and Pit-1/HinfI and positive values (LEP/Sau3AI and GH/AluI of fixation indexes FIS indicating slight heterozygote excess or deficiency based on analyzed genetic marker were observed.

  1. Performance of continuous biodigestors supplied by young bull waste

    Energy Technology Data Exchange (ETDEWEB)

    Costa, Monica Sarolli S. de M. [Universidade Estadual do Oeste do Parana (UNIOESTE), Cascavel, PR (Brazil)], E-mail: monicas@unioeste.br; Lucas Junior, Jorge de [Universidade Estadual Paulista (FCA/UNESP), Jaboticabal, SP (Brazil). Fac. de Ciencias Agrarias e Veterinarias; Pivetta, Laercio Augusto [Universidade Estadual do Oeste do Parana (UNIOESTE), Marechal C. Rondon, PR (Brazil); Costa, Luiz A. de Mendonca

    2008-07-01

    The various systems of livestock farming in the industrial model promote physical and chemical changes on waste. In the fattening of cattle in the model of confinement for young bulls has been a reversal in the proportion between roughage and concentrate, or the animals receive a higher amount of protein compared to the traditional system of confinement. This change of the waste characteristics involves modification in the system of treatment used. In this work, it was aimed to evaluate the performance of batch biodigestors operated in continuous system, supplied by young bulls waste which received two differentiated diets by the proportion between roughage and concentrate, whether or not containing inoculum in the substrate composition and subjected to three levels of temperature (25, 35 and 40 deg C). The parameters evaluated were: reduction of total solids (TS) and volatile (VS), and the potential for biogas production. The results showed a reduction of TS higher in the treatment which was not used inoculum for diet 1 and 2 with the exception of the treatment which the substrate was referred to temperature of 40 deg C on diet 2. For the reduction of VS there was no interference from the use of inoculum on diet 1. On diet 2, the largest reductions were observed without the use of inoculum, with the exception of the 40 deg C temperature. For the potential for biogas production the treatment where they used waste derived from animals fed with diet 2, with the use of inoculum, in the temperature of 40 deg C showed the greatest value, or 0.53 m{sup 3} of biogas per kg TS added. (author)

  2. Development and evaluation of a bioenergetics model for bull trout

    Science.gov (United States)

    Mesa, Matthew G.; Welland, Lisa K.; Christiansen, Helena E.; Sauter, Sally T.; Beauchamp, David A.

    2013-01-01

    We conducted laboratory experiments to parameterize a bioenergetics model for wild Bull Trout Salvelinus confluentus, estimating the effects of body mass (12–1,117 g) and temperature (3–20°C) on maximum consumption (C max) and standard metabolic rates. The temperature associated with the highest C max was 16°C, and C max showed the characteristic dome-shaped temperature-dependent response. Mass-dependent values of C max (N = 28) at 16°C ranged from 0.03 to 0.13 g·g−1·d−1. The standard metabolic rates of fish (N = 110) ranged from 0.0005 to 0.003 g·O2·g−1·d−1 and increased with increasing temperature but declined with increasing body mass. In two separate evaluation experiments, which were conducted at only one ration level (40% of estimated C max), the model predicted final weights that were, on average, within 1.2 ± 2.5% (mean ± SD) of observed values for fish ranging from 119 to 573 g and within 3.5 ± 4.9% of values for 31–65 g fish. Model-predicted consumption was within 5.5 ± 10.9% of observed values for larger fish and within 12.4 ± 16.0% for smaller fish. Our model should be useful to those dealing with issues currently faced by Bull Trout, such as climate change or alterations in prey availability.

  3. A two-tailed Comet assay for assessing DNA damage in spermatozoa.

    Science.gov (United States)

    Enciso, Maria; Sarasa, Jonas; Agarwal, Ashok; Fernández, Jose Luis; Gosálvez, Jaime

    2009-05-01

    DNA fragmentation is considered an important parameter of semen quality, and of significant value as a predictor of male fertility. Poor quality chromatin is closely associated with, and highly indicative of, some fertility problems. Many methodologies to assess DNA fragmentation in spermatozoa are available, but they are all unable to differentiate between single-stranded DNA breaks (SSB) and double-stranded DNA breaks (DSB) in the same sperm cell. The two-tailed Comet assay (2T-Comet) protocol overcomes this limitation. A modification of the original Comet assay was developed for the simultaneous evaluation of DNA SSB and DSB in human spermatozoa. The 2T-Comet assay is a fast, sensitive, and reliable procedure for the quantification and characterization of DNA damage in spermatozoa. It is an innovative method for assessing sperm DNA integrity, which has important implications for human fertility and andrological pathology.

  4. In vitro effects of radiofrequency electromagnetic waves on bovine spermatozoa motility.

    Science.gov (United States)

    Lukac, Norbert; Massanyi, Peter; Roychoudhury, Shubhadeep; Capcarova, Marcela; Tvrda, Eva; Knazicka, Zuzana; Kolesarova, Anna; Danko, Jan

    2011-01-01

    In this study the effects of 1800 MHz GSM-like radiofrequency electromagnetic waves (RF-EMW) exposure on bovine semen was monitored. The experimental samples were analyzed in vitro in four time periods (0, 30, 120 and 420 min) and compared with unexposed samples (control). Spermatozoa motility was determined by computer assisted semen analyzer (CASA). Evaluation of the percentage of motile spermatozoa showed significant (P radiation suggested that the parameters of path and velocity at the beginning of the culture significantly increase, but after longer culture (420 minutes) a significant decrease occur in the experimental group as compared to control. In general, results of this experiment indicate a negative time-dependent effect of 1800 MHz RF-EMW radiation on bovine spermatozoa motility.

  5. Molecular changes and signaling events occurring in spermatozoa during epididymal maturation.

    Science.gov (United States)

    Gervasi, M G; Visconti, P E

    2017-03-01

    After leaving the testis, spermatozoa have not yet acquired the ability to move progressively and are unable to fertilize oocytes. To become fertilization competent, they must go through an epididymal maturation process in the male, and capacitation in the female tract. Epididymal maturation can be defined as those changes occurring to spermatozoa in the epididymis that render the spermatozoa the ability to capacitate in the female tract. As part of this process, sperm cells undergo a series of biochemical and physiological changes that require incorporation of new molecules derived from the epididymal epithelium, as well as post-translational modifications of endogenous proteins synthesized during spermiogenesis in the testis. This review will focus on epididymal maturation events, with emphasis in recent advances in the understanding of the molecular basis of this process. © 2017 American Society of Andrology and European Academy of Andrology.

  6. Effect of Air Space in Storage Vials on Motility of Spermatozoa in Chilled Buck Semen

    Directory of Open Access Journals (Sweden)

    Magnus Paul K and Lali F Anand 1

    Full Text Available This study was conducted in order to find out the effect of air space on the top of glass vial in which semen is stored, on the motility of spermatozoa. 45 samples collected from two bucks over a span of 6 months were used for experiment. Goat milk extender was the diluent used. Two ml each of diluted semen after noting their initial motility was stored in 2 ml and 5 ml vials. Samples were stored at 5°C and motility of spermatozoa noted at 24 and 48 hours. Semen without air space was found to preserve the motility better than semen with air space on 24 and 48 hours of incubation. This could be better attributed to reactive oxygen species production by the spermatozoa, but further investigation is needed in this aspect to confirm it. [Veterinary World 2010; 3(9.000: 421-423

  7. Characterization of NAADP-mediated calcium signaling in human spermatozoa

    Energy Technology Data Exchange (ETDEWEB)

    Sánchez-Tusie, A.A. [Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos (Mexico); Vasudevan, S.R.; Churchill, G.C. [Department of Pharmacology, University of Oxford, Oxford OX1 3QT, England (United Kingdom); Nishigaki, T. [Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos (Mexico); Treviño, C.L., E-mail: ctrevino@ibt.unam.mx [Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos (Mexico)

    2014-01-10

    Highlights: •Human sperm cells synthesize NAADP. •NAADP-AM mediates [Ca{sup 2+}]{sub i} increases in human sperm in the absence of [Ca{sup 2+}]{sub o}. •Human sperm have two acidic compartments located in the head and midpiece. -- Abstract: Ca{sup 2+} signaling in spermatozoa plays a crucial role during processes such as capacitation and release of the acrosome, but the underlying molecular mechanisms still remain unclear. Nicotinic acid adenine dinucleotide phosphate (NAADP) is a potent Ca{sup 2+}-releasing second messenger in a variety of cellular processes. The presence of a NAADP synthesizing enzyme in sea urchin sperm has been previously reported, suggesting a possible role of NAADP in sperm Ca{sup 2+} signaling. In this work we used in vitro enzyme assays to show the presence of a novel NAADP synthesizing enzyme in human sperm, and to characterize its sensitivity to Ca{sup 2+} and pH. Ca{sup 2+} fluorescence imaging studies demonstrated that the permeable form of NAADP (NAADP-AM) induces intracellular [Ca{sup 2+}] increases in human sperm even in the absence of extracellular Ca{sup 2+}. Using LysoTracker®, a fluorescent probe that selectively accumulates in acidic compartments, we identified two such stores in human sperm cells. Their acidic nature was further confirmed by the reduction in staining intensity observed upon inhibition of the endo-lysosomal proton pump with Bafilomycin, or after lysosomal bursting with glycyl-L-phenylalanine-2-naphthylamide. The selective fluorescent NAADP analog, Ned-19, stained the same subcellular regions as LysoTracker®, suggesting that these stores are the targets of NAADP action.

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

    Directory of Open Access Journals (Sweden)

    José Luis Ros-Santaella

    Full Text Available 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; p<0.001. Our results clearly show that a higher volume of the sperm principal piece results in poor freezability, and highlights the key role of flagellum size in sperm cryopreservation success.

  9. [From spermatid to spermatozoa : what changes are necessary for development?].

    Science.gov (United States)

    Testart, J

    1996-01-01

    The male gamete modifications during spermiogenesis were summarized in view to answer the question : "Is it a risk for a baby conceived by fertilization with a spermatid to be different of a baby conceived with a spermatozoon?" Cytoplasmic changes in spermiogenesis mainly concern the Golgi apparatus (acrosome vesicle), centrosomal material (flagelium), mitochondria (periaxonemal ring) and ooplasm volume (drastic reduction). These changes are not necessary in case of microfertilization (ICSI) and their absence could be without consequences on post fertilization development. Nuclear changes affect chromatin with the substitution of histones by protamines to insure sperm ADN stability across male and female genital tracts. Spermatid gene transduction brings new proteins which are related to fertilization (acrosomal enzymes, flagellum proteins, protamines...) and seem no longer useful after gametes fusion. There is no report of genomic imprinting occurring during spermiogenesis in mammals. Moreover, the important sperm changes during epididymal maturation (formation of disulfure bonds, acquisition of motility and of molecules for oocyte recognition, methylation of certain genes...) were found not necessary for embryo development in case of ICSI with epididymal or testicular spermatozoa. Normal and fertile animals (mice and rabbits) were born from ICSI with spermatids originated from normal males; an animal model with sterile males delivering apparently normal spermatids is lacking. Although the risk for defectuous babies from fertilization with spermatids is highly subjective, caution is needed and requires team competence, complete informed consent by the patients and exhaustive analysis of the methods used and results obtained. Moreover the evident risk to "reproduce" the father infertility must be compared with the psychosocial injury induced by using the alternative option of donor insemination.

  10. Optimization of a protocol for cryopreservation of mouse spermatozoa using cryotubes.

    Science.gov (United States)

    Hasegawa, Ayumi; Yonezawa, Kazuya; Ohta, Akihiko; Mochida, Keiji; Ogura, Atsuo

    2012-01-01

    The rapid increase in the number of genetically modified mouse strains has produced a high demand for their frozen spermatozoa from laboratories and mouse banking facilities. Historically, plastic straws have been used preferentially as containers for frozen mammalian spermatozoa because spermatozoa frozen in plastic straws have a high survival rate after thawing. However, plastic straws are more fragile and are used less often than the cryotubes used for conventional cell freezing. In this study, we sought to develop a new protocol for sperm freezing using cryotubes as the container to increase the accessibility of mouse sperm cryopreservation. Epididymal spermatozoa were collected from mature ICR or C57BL/6J (B6) males and were suspended in 18% raffinose and 3% skim milk solution. We then optimized the following conditions using the sperm survival rate as an index: 1) distance of cryotubes from the surface of the liquid nitrogen at freezing, 2) volume of the sperm suspension in the cryotube and 3) temperature of warming sperm during thawing. The best result was obtained when cryotubes containing 10 µl of sperm suspension were immersed 1 cm below the surface of the liquid nitrogen and then thawed at 50 C. The fertilization rates using spermatozoa frozen and thawed using this method were 63.1% in ICR mice and 28.2% in B6 mice. The latter rate was increased to 62.3% by adding reduced glutathione to the fertilization medium. After embryo transfer, 68% and 62% of the fertilized oocytes developed into normal offspring in the ICR and B6 strains, respectively. These results show that cryotubes can be used for cryopreservation of mouse spermatozoa under optimized conditions. This protocol is easy and reproducible, and it may be used in laboratories that do not specialize in sperm cryopreservation.

  11. Identification of the inorganic pyrophosphate metabolizing, ATP substituting pathway in mammalian spermatozoa.

    Directory of Open Access Journals (Sweden)

    Young-Joo Yi

    Full Text Available Inorganic pyrophosphate (PPi is generated by ATP hydrolysis in the cells and also present in extracellular matrix, cartilage and bodily fluids. Fueling an alternative pathway for energy production in cells, PPi is hydrolyzed by inorganic pyrophosphatase (PPA1 in a highly exergonic reaction that can under certain conditions substitute for ATP-derived energy. Recombinant PPA1 is used for energy-regeneration in the cell-free systems used to study the zymology of ATP-dependent ubiquitin-proteasome system, including the role of sperm-borne proteasomes in mammalian fertilization. Inspired by an observation of reduced in vitro fertilization (IVF rates in the presence of external, recombinant PPA1, this study reveals, for the first time, the presence of PPi, PPA1 and PPi transporter, progressive ankylosis protein ANKH in mammalian spermatozoa. Addition of PPi during porcine IVF increased fertilization rates significantly and in a dose-dependent manner. Fluorometric assay detected high levels of PPi in porcine seminal plasma, oviductal fluid and spermatozoa. Immunofluorescence detected PPA1 in the postacrosomal sheath (PAS and connecting piece of boar spermatozoa; ANKH was present in the sperm head PAS and equatorial segment. Both ANKH and PPA1 were also detected in human and mouse spermatozoa, and in porcine spermatids. Higher proteasomal-proteolytic activity, indispensable for fertilization, was measured in spermatozoa preserved with PPi. The identification of an alternative, PPi dependent pathway for ATP production in spermatozoa elevates our understanding of sperm physiology and sets the stage for the improvement of semen extenders, storage media and IVF media for animal biotechnology and human assisted reproductive therapies.

  12. Acetyl-L-carnitine: An effective antioxidant against cryo-damage on human spermatozoa with asthenospermia.

    Science.gov (United States)

    Zou, Yu-Jie; Yang, Jing; Chang, Shuo; Xu, Wang-Ming; Yin, Tai-Lang; Long, Wen

    2017-12-01

    A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process. The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen (ROS) level after freezing-thawing process. The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia. The cryopreservation of human spermatozoa treated with acetyl-L-carnitine at different concentrations (group B: 2.5 mmol/L, group C: 7.5 mmol/L, group D: 15 mmol/L) was compared with control (group A: no acetyl-L-carnitine given). For the frozen-thawed spermatozoa, the viability, motility and DNA integrity were measured by comet assay, acrosome integrity by FITC-PNA staining and ROS level was determined in each group. The results showed that there were no significant differences in motility and viability between group A and group B, while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A. As compared with group A, the values for DNA integrity parameters including comet rate (CR), tail DNA percentage (TD), tail length (TL) and Oliver tail moment (OTM) were significantly reduced in group C and group D. Group C and group D also displayed a higher proportion of intact acrosome than group A. No significant difference in ROS level was found between group A and group B, while with the increase in acetyl-L-carnitine concentration, the ROS level in groups C and D was significantly reduced as compared with that in group A. In conclusion, acetyl-L-carnitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa.

  13. Identification of the Inorganic Pyrophosphate Metabolizing, ATP Substituting Pathway in Mammalian Spermatozoa

    Science.gov (United States)

    Yi, Young-Joo; Sutovsky, Miriam; Kennedy, Chelsey; Sutovsky, Peter

    2012-01-01

    Inorganic pyrophosphate (PPi) is generated by ATP hydrolysis in the cells and also present in extracellular matrix, cartilage and bodily fluids. Fueling an alternative pathway for energy production in cells, PPi is hydrolyzed by inorganic pyrophosphatase (PPA1) in a highly exergonic reaction that can under certain conditions substitute for ATP-derived energy. Recombinant PPA1 is used for energy-regeneration in the cell-free systems used to study the zymology of ATP-dependent ubiquitin-proteasome system, including the role of sperm-borne proteasomes in mammalian fertilization. Inspired by an observation of reduced in vitro fertilization (IVF) rates in the presence of external, recombinant PPA1, this study reveals, for the first time, the presence of PPi, PPA1 and PPi transporter, progressive ankylosis protein ANKH in mammalian spermatozoa. Addition of PPi during porcine IVF increased fertilization rates significantly and in a dose-dependent manner. Fluorometric assay detected high levels of PPi in porcine seminal plasma, oviductal fluid and spermatozoa. Immunofluorescence detected PPA1 in the postacrosomal sheath (PAS) and connecting piece of boar spermatozoa; ANKH was present in the sperm head PAS and equatorial segment. Both ANKH and PPA1 were also detected in human and mouse spermatozoa, and in porcine spermatids. Higher proteasomal-proteolytic activity, indispensable for fertilization, was measured in spermatozoa preserved with PPi. The identification of an alternative, PPi dependent pathway for ATP production in spermatozoa elevates our understanding of sperm physiology and sets the stage for the improvement of semen extenders, storage media and IVF media for animal biotechnology and human assisted reproductive therapies. PMID:22485177

  14. The Testicular and Epididymal Expression Profile of PLCζ in Mouse and Human Does Not Support Its Role as a Sperm-Borne Oocyte Activating Factor

    Science.gov (United States)

    Aarabi, Mahmoud; Yu, Yang; Xu, Wei; Tse, Man Y.; Pang, Stephen C.; Yi, Young-Joo; Sutovsky, Peter; Oko, Richard

    2012-01-01

    Phospholipase C zeta (PLCζ) is a candidate sperm-borne oocyte activating factor (SOAF) which has recently received attention as a potential biomarker of human male infertility. However, important SOAF attributes of PLCζ, including its developmental expression in mammalian spermiogenesis, its compartmentalization in sperm head perinuclear theca (PT) and its release into the ooplasm during fertilization have not been established and are addressed in this investigation. Different detergent extractions of sperm and head/tail fractions were compared for the presence of PLCζ by immunoblotting. In both human and mouse, the active isoform of PLCζ was detected in sperm fractions other than PT, where SOAF is expected to reside. Developmentally, PLCζ was incorporated as part of the acrosome during the Golgi phase of human and mouse spermiogenesis while diminishing gradually in the acrosome of elongated spermatids. Immunofluorescence localized PLCζ over the surface of the postacrosomal region of mouse and bull and head region of human spermatozoa leading us to examine its secretion in the epididymis. While previously thought to have strictly a testicular expression, PLCζ was found to be expressed and secreted by the epididymal epithelial cells explaining its presence on the sperm head surface. In vitro fertilization (IVF) revealed that PLCζ is no longer detectable after the acrosome reaction occurs on the surface of the zona pellucida and thus is not incorporated into the oocyte cytoplasm for activation. In summary, we show for the first time that PLCζ is compartmentalized as part of the acrosome early in human and mouse spermiogenesis and is secreted during sperm maturation in the epididymis. Most importantly, no evidence was found that PLCζ is incorporated into the detergent-resistant perinuclear theca fraction where SOAF resides. PMID:22428063

  15. The testicular and epididymal expression profile of PLCζ in mouse and human does not support its role as a sperm-borne oocyte activating factor.

    Directory of Open Access Journals (Sweden)

    Mahmoud Aarabi

    Full Text Available Phospholipase C zeta (PLCζ is a candidate sperm-borne oocyte activating factor (SOAF which has recently received attention as a potential biomarker of human male infertility. However, important SOAF attributes of PLCζ, including its developmental expression in mammalian spermiogenesis, its compartmentalization in sperm head perinuclear theca (PT and its release into the ooplasm during fertilization have not been established and are addressed in this investigation. Different detergent extractions of sperm and head/tail fractions were compared for the presence of PLCζ by immunoblotting. In both human and mouse, the active isoform of PLCζ was detected in sperm fractions other than PT, where SOAF is expected to reside. Developmentally, PLCζ was incorporated as part of the acrosome during the Golgi phase of human and mouse spermiogenesis while diminishing gradually in the acrosome of elongated spermatids. Immunofluorescence localized PLCζ over the surface of the postacrosomal region of mouse and bull and head region of human spermatozoa leading us to examine its secretion in the epididymis. While previously thought to have strictly a testicular expression, PLCζ was found to be expressed and secreted by the epididymal epithelial cells explaining its presence on the sperm head surface. In vitro fertilization (IVF revealed that PLCζ is no longer detectable after the acrosome reaction occurs on the surface of the zona pellucida and thus is not incorporated into the oocyte cytoplasm for activation. In summary, we show for the first time that PLCζ is compartmentalized as part of the acrosome early in human and mouse spermiogenesis and is secreted during sperm maturation in the epididymis. Most importantly, no evidence was found that PLCζ is incorporated into the detergent-resistant perinuclear theca fraction where SOAF resides.

  16. Analysis of the effects of polyphenols on human spermatozoa reveals unexpected impacts on mitochondrial membrane potential, oxidative stress and DNA integrity; implications for assisted reproductive technology.

    Science.gov (United States)

    Aitken, R J; Muscio, L; Whiting, S; Connaughton, H S; Fraser, B A; Nixon, B; Smith, N D; De Iuliis, G N

    2016-12-01

    The need to protect human spermatozoa from oxidative stress during assisted reproductive technology, has prompted a detailed analysis of the impacts of phenolic compounds on the functional integrity of these cells. Investigation of 16 individual compounds revealed a surprising variety of negative effects including: (i) a loss of mitochondrial membrane potential (Δψm) via mechanisms that were not related to opening of the permeability transition pore but associated with a reduction in thiol expression, (ii) a decline in intracellular reduced glutathione, (iii) the stimulation of pro-oxidant activity including the induction of ROS generation from mitochondrial and non-mitochondrial sources, (iv) stimulation of lipid peroxidation, (v) the generation of oxidative DNA damage, and (vi) impaired sperm motility. For most of the polyphenolic compounds examined, the loss of motility was gradual and highly correlated with the induction of lipid peroxidation (r=0.889). The exception was gossypol, which induced a rapid loss of motility due to its inherent alkylating activity; one consequence of which was a marked reduction in carboxymethyl lysine expression on the sperm tail; a post-translational modification that is known to play a key role in the regulation of sperm movement. The only polyphenols that did not appear to have adverse effects on spermatozoa were resveratrol, genistein and THP at doses below 100μM. These compounds could, therefore, have some therapeutic potential in a clinical setting. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Effects of cadmium salt on phosphomonoesterases activity and fertilizing ability of fowl spermatozoa.

    Science.gov (United States)

    Mohan, J; Moudgal, R P; Panda, J N

    1992-03-01

    Daily administration of cadmium salt for 25 days (2.5 mg per Kg body weight) in the male domestic fowl caused the end of treatment period. An increased incidences of concentration. Fertility dropped to zero at the end of the treatment period. Activity of acid and alkaline phosphomonoesterases were also drastically reduced by the end of treatment period. An increased incidences of morphological abnormalities of spermatozoa were noticed in the treated birds. After 46 days cessation of the treatment, full recovery of the above measures was found. These alterations suggest the reversible type of effect of cadmium chloride on the spermatozoa of male domestic fowl.

  18. Major protein alterations in spermatozoa from infertile men with unilateral varicocele.

    Science.gov (United States)

    Agarwal, Ashok; Sharma, Rakesh; Durairajanayagam, Damayanthi; Ayaz, Ahmet; Cui, Zhihong; Willard, Belinda; Gopalan, Banu; Sabanegh, Edmund

    2015-02-22

    The etiology of varicocele, a common cause of male factor infertility, remains unclear. Proteomic changes responsible for the underlying pathology of unilateral varicocele have not been evaluated. The objective of this prospective study was to employ proteomic techniques and bioinformatic tools to identify and analyze proteins of interest in infertile men with unilateral varicocele. Spermatozoa from infertile men with unilateral varicocele (n=5) and from fertile men (control; n=5) were pooled in two groups respectively. Proteins were extracted and separated by 1-D SDS-PAGE. Bands were digested and identified on a LTQ-Orbitrap Elite hybrid mass spectrometer system. Bioinformatic analysis identified the pathways and functions of the differentially expressed proteins (DEP). Sperm concentration, motility and morphology were lower, and reactive oxygen species levels were higher in unilateral varicocele patients compared to healthy controls. The total number of proteins identified were 1055, 1010 and 1042 in the fertile group, and 795, 713 and 763 proteins in the unilateral varicocele group. Of the 369 DEP between both groups, 120 proteins were unique to the fertile group and 38 proteins were unique to the unilateral varicocele group. Compared to the control group, 114 proteins were overexpressed while 97 proteins were underexpressed in the unilateral varicocele group. We have identified 29 proteins of interest that are involved in spermatogenesis and other fundamental reproductive events such as sperm maturation, acquisition of sperm motility, hyperactivation, capacitation, acrosome reaction and fertilization. The major functional pathways of the 359 DEP related to the unilateral varicocele group involve metabolism, disease, immune system, gene expression, signal transduction and apoptosis. Functional annotations showed that unilateral varicocele mostly affected small molecule biochemistry and post-translational modification proteins. Proteins expressed uniquely in the

  19. The microRNA signature of mouse spermatozoa is substantially modified during epididymal maturation.

    Science.gov (United States)

    Nixon, Brett; Stanger, Simone J; Mihalas, Bettina P; Reilly, Jackson N; Anderson, Amanda L; Tyagi, Sonika; Holt, Janet E; McLaughlin, Eileen A

    2015-10-01

    In recent years considerable effort has been devoted to understanding the epigenetic control of sperm development, leading to an increased appreciation of the importance of RNA interference pathways, and in particular miRNAs, as key regulators of spermatogenesis and epididymal maturation. It has also been shown that sperm are endowed with an impressive array of miRNA that have been implicated in various aspects of fertilization and embryo development. However, to date there have been no reports on whether the sperm miRNA signature is static or whether it is influenced by their prolonged maturation within the male reproductive tract. To investigate this phenomenon, we employed next-generation sequencing to systematically profile the miRNA signature of maturing mouse spermatozoa. In so doing we have provided the first evidence for the posttesticular modification of the sperm miRNA profile under normal physiological conditions. Such modifications include the apparent loss and acquisition of an impressive cohort of some 113 and 115 miRNAs, respectively, between the proximal and distal epididymal segments. Interestingly, the majority of these changes occur late in maturation and include the uptake of novel miRNA species in addition to a significant increase in many miRNAs natively expressed in immature sperm. Because sperm are not capable of de novo transcription, these findings identify the epididymis as an important site in establishing the sperm epigenome with the potential to influence the peri-conceptual environment of the female reproductive tract, contribute to the inheritance of acquired characteristics, and/or alter the developmental trajectory of the resulting offspring. © 2015 by the Society for the Study of Reproduction, Inc.

  20. Comparison on accuracy of different nonlinear models in predicting growth of Podolica bulls

    National Research Council Canada - National Science Library

    Selvaggi, Maria; Laudadio, Vito; D'Alessandro, Angela Gabriella; Dario, Cataldo; Tufarelli, Vincenzo

    2017-01-01

    .... This study was carried out to estimate the parameters of logistic, Gompertz, Richards and von Bertalanffy growth curve models in a sample of Podolica young bulls to determine the goodness of fit...

  1. Bulls grazing Kentucky 31 tall fescue exhibit impaired growth, semen quality, and decreased semen freezing potential

    Science.gov (United States)

    Serum prolactin (PRL) and testosterone concentrations, body weight, body composition, semen quality, and semen freezing potential for bulls grazing the toxic tall fescue (Lolium arundinaceum [Schreb.] Darbysh. ¼ Schedonorous arundinaceum [Schreb.] Dumort.) cultivar Kentucky 31 (E+) compared with a n...

  2. BULL'S EYE MACULOPATHY WITH MUTATIONS IN RDS/PRPH2 AND ROM-1.

    Science.gov (United States)

    Essilfie, Juliet O; Sanfilippo, Christian J; Sarraf, David

    2017-11-16

    To report a case of bull's eye maculopathy associated with mutations in RDS/PRPH2 and ROM-1 genes. We present a case report of a patient with a characteristic maculopathy and describe the multimodal retinal imaging findings including spectral domain optical coherence tomography and fundus autofluorescence and full-field electrophysiology. The results of genetic testing are also reported. A 60-year-old woman presented with decreased vision and a remarkable bull's eye maculopathy with retinal examination. Fundus autofluorescence illustrated a striking pattern of speckled hyperautofluorescence and hypoautofluorescence that highlighted the bull's eye maculopathy in each eye and guided genetic testing, which confirmed a mutation of the RDS/PRPH2 gene and a novel mutation of the ROM-1 gene. Multimodal imaging including fundus autofluorescence may guide genetic testing in patients with a characteristic maculopathy. RDS/PRPH2 genetic mutation can be associated with a bull's eye maculopathy with a signature fundus autofluorescence presentation.

  3. Quality Traits of Meat from Young Limousin, Charolais and Hereford Bulls

    Directory of Open Access Journals (Sweden)

    Janina Pogorzelska*, Jan Miciński, Halina Ostoja1, Ireneusz M. Kowalski2, Józef Szarek3 and Emilia Strzyżewska3

    2013-01-01

    Full Text Available The objective of this study was to determine the effects of beef cattle breed and muscle type on the proximate chemical composition and quality traits of meat, including processing suitability. The experimental materials comprised samples of musculus longissimus dorsi (LD muscle and musculus semitendinosus (ST muscle collected from the carcasses of young Limousin, Charolais and Hereford bulls. The chemical composition, texture, hydration and color parameters of LD and ST muscles were determined. Meat from Limousin and Charolais bulls, characterized by higher body mass at slaughter contained more protein than meat from Hereford bulls. Meat from Hereford bulls had a higher fat content, compared with the other two breeds. Texture parameters, including hardness, gumminess and chewiness, varied depending on muscle type and cattle breed. An analysis of the maximum shear force values showed that the mechanical properties of beef also varied depending on cattle breed and muscle type.

  4. Bull Trout Life History, Genetics, Habitat Needs, and Limiting Fact in Central and Northeast Oregon. Annual Report 1999.

    Energy Technology Data Exchange (ETDEWEB)

    Hemmingsen, Alan R.; Gunckel, Stephanie L.; Howell, Philip J.

    2001-08-01

    This section describes work accomplished in 1999 that continued to address two objectives of this project. These objectives are (1) determine the distribution of juvenile and adult bull trout Salvelinus confluentus and habitats associated with that distribution, and (2) determine fluvial and resident bull trout life history patterns. Completion of these objectives is intended through studies of bull trout in the Grande Ronde, Walla Walla, and John Day basins. These basins were selected because they provide a variety of habitats, from relatively degraded to pristine, and bull trout populations were thought to vary from relatively depressed to robust. In all three basins we used radio telemetry to determine the seasonal movements of bull trout. In the John Day and Walla Walla basins we also used traps to capture migrant bull trout. With these traps, we intended to determine the timing of bull trout movements both upstream and downstream, determine the relative abundance, size and age of migrant fish, and capture bull trout to be implanted with radio transmitters. In the John Day basin, we captured adult and juvenile bull trout from the upper John Day River and its tributaries, Call Creek, Reynolds Creek, and Roberts Creek. In the Walla Walla basin, we captured adult and juvenile bull trout from Mill Creek.

  5. Detection of Paratuberculosis in Breeding Bulls at Pakistani Semen Production Units: A Continuous Source of Threat

    OpenAIRE

    Muhammad ABBAS; Munir, Muhammad; Khaliq, Syed Abdul; Haq, Muhammad Ikram Ul; Tanveer Khan, Muhammad; Qureshi, Zafar Ul Ahsan

    2011-01-01

    Paratuberculosis is a chronic bowel disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Its secretion through semen highlights the importance of paratuberculosis-free breeding bulls. The breeding and teaser bulls at three semen production units (SPUs) located in Punjab, Pakistan, were screened for the presence of antibodies against MAP. A total of 253 samples were collected from SPUs and a commercially available indirect screen ELISA (Is-ELISA) was applied. Is-EL...

  6. Gross testicular abnormalities in indigenous breeds of bulls in Eastern Ethiopia

    OpenAIRE

    Amare Eshetu Gemeda

    2017-01-01

    Objective: This study was conducted to assess the prevalence of different types of gross testicular disorders in bulls, and to evaluate the associations with sampling year, age, and body condition. Materials and methods: In this study, a total of 398 apparently healthy bulls were randomly selected that were brought from different parts of eastern Ethiopia to the Haramaya University abattoir for slaughtering during the period from June 2014 to September 2016. Ante- and post-mortem examinat...

  7. Relationships among temperament, behavior, and growth during performance testing of bulls.

    Science.gov (United States)

    Lockwood, S A; Kattesh, H G; Krawczel, P D; Kirkpatrick, F D; Saxton, A M; Rhinehart, J D; Wilkerson, J B

    2015-12-01

    Excitable cattle are dangerous to personnel and have reduced individual performance. The aim of this study was to 1) identify objective criteria for evaluating bull temperament and 2) examine relationships among temperament, behavior, and performance of bulls during an 84-d performance test. Angus bulls ( = 60) were reared in 6 pens based on BW and age. Pen scores (PS; 1 = docile and 5 = very aggressive) were assigned on d -1, 27, 55, and 83. Exit velocity (EV), BW, time to exit the chute, and order through the chute were recorded on d 0, 28, 56, and 84. The ADG was calculated for the 84-d test period, and ultrasound data and frame score calculations were recorded on d 84. Dataloggers measured steps taken, lying time, number of lying bouts, and lying bout duration of bulls ( = 27; 3 pens) from d 3 to 28 and d 59 to 84. Bulls with a d -1 PS of 1 or 2 were categorized as calm (PScalm; = 40), whereas bulls with a PS of 3 or 4 were categorized as excitable (PSexcitable; = 20). Bulls were separated into 2 groups based on the bottom 20 EV (EVcalm) and top 20 EV (EVexcitable) on d 0. Mixed model ANOVA (SAS 9.3) was used to compare groups for the two temperament assessment methods, behavior, and growth performance. Mean EV decreased ( testing period. Additionally, the potential lack of innate temperament variation may have attributed to the little difference seen among the behavioral and performance data. Therefore, temperament should be reassessed within a novel environment with new handlers to differentiate between the bull's true temperament and its ability to habituate.

  8. Cardiovascular and cerebrovascular effects in response to red bull consumption combined with mental stress.

    Science.gov (United States)

    Grasser, Erik Konrad; Dulloo, Abdul G; Montani, Jean-Pierre

    2015-01-15

    The sale of energy drinks is often accompanied by a comprehensive and intense marketing with claims of benefits during periods of mental stress. As it has been shown that Red Bull negatively impacts human hemodynamics at rest, we investigated the cardiovascular and cerebrovascular consequences when Red Bull is combined with mental stress. In a randomized cross-over study, 20 young healthy humans ingested either 355 ml of a can Red Bull or water and underwent 80 minutes after the respective drink a mental arithmetic test for 5 minutes. Continuous cardiovascular and cerebrovascular recordings were performed for 20 minutes before and up to 90 minutes after drink ingestion. Measurements included beat-to-beat blood pressure (BP), heart rate, stroke volume, and cerebral blood flow velocity. Red Bull increased systolic BP (+7 mm Hg), diastolic BP (+4 mm Hg), and heart rate (+7 beats/min), whereas water drinking had no significant effects. Cerebral blood flow velocity decreased more in response to Red Bull than to water (-9 vs -3 cm/s, p Red Bull; similar increases were also observed after water ingestion. In combination, Red Bull and mental stress increased systolic BP by about 10 mm Hg, diastolic BP by 7 mm Hg, and heart rate by 20 beats/min and decreased cerebral blood flow velocity by -7 cm/s. In conclusion, the combination of Red Bull and mental stress impose a cumulative cardiovascular load and reduces cerebral blood flow even under a mental challenge. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Analysis of organisation sport event of event Red Bull Crashed Ice 2009

    OpenAIRE

    Rak, Zdeněk

    2009-01-01

    Title: Analysis of organisation sport event of event Red Bull Crashed Ice 2009 Work goal: Analysis of organisation of event... Methods: Descriptive analysis, SWOT analysis, Interview with experts. Annotation: Organisation of Red Bull Crashed Ice includes possible areas of improvement with concrete suggestions to activities of organisational process of this sport event. Introduced proposals result from results of SWOT analysis and interviews with professionals. Results: Conclusion and advices ...

  10. Performance and methane emissions of grazing Nellore bulls supplemented with crude glycerin.

    Science.gov (United States)

    San Vito, E; Lage, J F; Messana, J D; Dallantonia, E E; Frighetto, R T S; Reis, R A; Neto, A J; Berchielli, T T

    2016-11-01

    Supplementation of grass-fed cattle with low-cost feeding alternatives may be an attractive way to improve efficiency of cattle production. We hypothesized that inclusion of crude glycerin (CG) in supplements provided to grass-fed cattle could improve feed conversion without negative effects on growth performance while reducing methane emissions. Our hypothesis was tested using Nellore bulls grazing tropical pasture ( = 50; initial BW of 427 ± 19.41 kg; age of 17 ± 2 mo) supplemented with increasing concentrations (0, 70, 140, 210, and 280 g/kg DM basis of supplement) of CG and corn gluten replacing corn grain. A second experiment was conducted using 10 ruminally cannulated Nellore steers (490.1 ± 47.8 kg BW; age of 25 mo) to assess the impact of different concentrations of glycerin in the supplement on ruminal VFA concentration. Inclusion of CG did not affect total DMI ( = 0.53), DMI of forage ( = 0.41), supplement DMI ( = 0.47), organic matter intake ( = 0.50), crude protein intake ( = 0.24), NDF intake ( = 0.49), GE intake ( = 0.50), NDF digestibility ( = 0.17), final BW ( = 0.17), LM area ( = 0.50), rib fat thickness ( = 0.87), or carcass gain ( = 0.13). The inclusion of CG in the supplement linearly increased ( < 0.001) the molar proportion of propionate, butyrate, and valerate; linearly decreased acetate ( = 0.001); and did not affect the molar proportion of isovalerate ( = 0.31) and isobutyrate ( = 0.63), thereby reducing the acetate to propionate ratio ( < 0.001). The increase of CG supplementation of young bulls in pasture had a quadratic effect on BW gain ( = 0.002), with lower BW gain with 140 g/kg DM of CG in the supplement and tended ( = 0.06) to improve G:F. Inclusion of CG did not affect ruminal CH emission expressed in kilograms per year ( = 0.74), grams per kilogram of DMI ( = 0.69), and grams per kilogram of carcass gain ( = 0.48). Crude glycerin supplementation was not effective as a strategy to reduce CH emission in grass-fed cattle. However

  11. Oviduct binding ability of porcine spermatozoa develops in the epididymis and can be advanced by incubation with caudal fluid.

    Science.gov (United States)

    Peña, Santiago; Summers, Phillip; Gummow, Bruce; Paris, Damien B B P

    2015-06-01

    The sperm reservoir is formed when spermatozoa bind to the epithelium of the uterotubal junction and caudal isthmus of the oviduct. It is an important mechanism that helps synchronize the meeting of gametes by regulating untimely capacitation and polyspermic fertilization. This study investigated the influence of epididymal maturation and caudal fluid on the ability of spermatozoa to bind to oviduct epithelium using a model porcine oviduct explant assay. Spermatozoa from the rete testis, middle caput (E2-E3), middle corpus (E6), and cauda (E8) of Large White or Large White × Landrace boars aged 10 to 14 months were diluted in modified Androhep solution and incubated with porcine oviduct explants. Results reported in this study support our hypothesis that testicular spermatozoa need to pass through the regions of the epididymis to acquire the ability to bind to the oviduct. There was a sequential increase in the number of spermatozoa that bound to oviduct explants from the rete testis to caudal epididymis. Binding of caudal spermatozoa to isthmic explants was the highest (15.0 ± 1.2 spermatozoa per 1.25 mm(2), mean ± standard error of the mean; P ≤ 0.05) and lowest by spermatozoa from the rete testis (2.0 ± 0.3 per 1.25 mm(2)), and higher to isthmus from sows compared to gilts (35.8 ± 6.7 per 1.25 mm(2) vs. 14.8 ± 3.0 per 1.25 mm(2); P ≤ 0.05). Binding of ejaculated spermatozoa to porcine isthmus was higher than that for caudal spermatozoa (26.3 ± 1.4 per 1.25 mm(2) vs. 15.0 ± 0.8 per 1.25 mm(2); P ≤ 0.05) and higher to porcine than to bovine isthmus (26.3 ± 2.3 per 1.25 mm(2) vs. 18.8 ± 1.9 per 1.25 mm(2); P ≤ 0.05). Incubation of spermatozoa from the caput and corpus in caudal fluid increased the ability of spermatozoa to bind to the oviduct epithelium (P ≤ 0.05). In conclusion, the capacity of testicular spermatozoa to bind to the oviduct epithelium increases during their maturation in the epididymis and can be advanced by components of the

  12. Bioenergetic evaluation of diel vertical migration by bull trout (Salvelinus confluentus) in a thermally stratified reservoir

    Science.gov (United States)

    Eckmann, Madeleine; Dunham, Jason; Connor, Edward J.; Welch, Carmen A.

    2018-01-01

    Many species living in deeper lentic ecosystems exhibit daily movements that cycle through the water column, generally referred to as diel vertical migration (DVM). In this study, we applied bioenergetics modelling to evaluate growth as a hypothesis to explain DVM by bull trout (Salvelinus confluentus) in a thermally stratified reservoir (Ross Lake, WA, USA) during the peak of thermal stratification in July and August. Bioenergetics model parameters were derived from observed vertical distributions of temperature, prey and bull trout. Field sampling confirmed that bull trout prey almost exclusively on recently introduced redside shiner (Richardsonius balteatus). Model predictions revealed that deeper (>25 m) DVMs commonly exhibited by bull trout during peak thermal stratification cannot be explained by maximising growth. Survival, another common explanation for DVM, may have influenced bull trout depth use, but observations suggest there may be additional drivers of DVM. We propose these deeper summertime excursions may be partly explained by an alternative hypothesis: the importance of colder water for gametogenesis. In Ross Lake, reliance of bull trout on warm water prey (redside shiner) for consumption and growth poses a potential trade-off with the need for colder water for gametogenesis.

  13. Are the Scaling Properties of Bull and Bear Markets Identical? Evidence from Oil and Gold Markets

    Directory of Open Access Journals (Sweden)

    Samet Günay

    2014-10-01

    Full Text Available In this study, the scaling properties of the oil and gold return volatilities have been analyzed in the context of bull and bear periods. In the determination of bull and bear turning points, we used the Modified Bry-Boschan Quarterly (MBBQ algorithm. Results showed that the business cycle phase shapes of the bear periods in the oil market are almost linear, whereas the bull and bear periods of the gold and bull period of the oil market are convex. This means that there are sharper declines in the bear period of the oil market. Following the detection of bull and bear periods, scaling exponent H analysis was performed via the aggregated variance, Higuchi’s statistic, Peng’s statistic, rescaled range, boxed periodogram and wavelet fit models, which are from the time, frequency and wavelet domains. As there are conflicts about the credibility of these methods in the literature, we have used the shuffling procedure in order to determine the most robust methods. According to the results, bear periods have higher volatility persistency than bull periods.

  14. Growth performance, carcass characteristics and meat quality of finishing bulls fed crude glycerin-supplemented diets

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    Mônica Chaves Françozo

    2013-04-01

    Full Text Available This study was conducted to evaluate the performance, carcass characteristics and chemical composition of Longissimusmuscle (LM of the bulls. Twenty-four Nellore bulls were used in a complete randomised design. The bulls were randomly assigned to one of the three diets containing 0, 5 or 12% glycerin. Final BW and ADG were similar (P>0.05 between the bulls fed with 5 or 12% of glycerin but were higher (P0.05 by glycerin level. Hot carcass weight increased (P0.05 the conformation, colour, texture, marbling and pH. There was difference (P>0.05 for moisture, ashes and crude protein among glycerin levels. Bulls fed 12% glycerin present the highest (P<0.03 total lipids on LM. The percentage of saturated fatty acids (SFA, monounsaturated acids (MUFA, polyunsaturated acids (PUFA, n-6 and n-3 fatty acids and PUFA/SFA and n-6:n-3 ratios of the LM were similar among the diets. In conclusion, glycerin level did not affect the animal performance and carcass characteristics of Nellore bulls finished in feedlot.

  15. Are brown trout replacing or displacing bull trout populations in a changing climate?

    Science.gov (United States)

    Al-Chokhachy, Robert K.; Schmetterling, David A.; Clancy, Chris; Saffel, Pat; Kovach, Ryan; Nyce, Leslie; Liermann, Brad; Fredenberg, Wade A.; Pierce, Ron

    2016-01-01

    Understanding how climate change may facilitate species turnover is an important step in identifying potential conservation strategies. We used data from 33 sites in western Montana to quantify climate associations with native bull trout (Salvelinus confluentus) and non-native brown trout (Salmo trutta) abundance and population growth rates (λ). We estimated λ using exponential growth state space models and delineated study sites based on bull trout use for either Spawning and Rearing (SR) or Foraging, Migrating, and Overwintering (FMO) habitat. Bull trout abundance was negatively associated with mean August stream temperatures within SR habitat (r = -0.75). Brown trout abundance was generally highest at temperatures between 12 and 14°C. We found bull trout λ were generally stable at sites with mean August temperature below 10°C but significantly decreasing, rare, or extirpated at 58% of the sites with temperatures exceeding 10°C. Brown trout λ were highest in SR and sites with temperatures exceeding 12°C. Declining bull trout λs at sites where brown trout were absent suggests brown trout are likely replacing bull trout in a warming climate.

  16. Reaction norms in weights at 365 days old in nellore bulls in northern Brazil

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    Jorge Luís Ferreira

    2015-12-01

    Full Text Available Genotype by environment interaction (GxE studies are of particular interest in Brazil because of the regional diversity of environmental effects and the wide variety of management systems. The present study evaluates GxE effects on 365 d weight (365W of Nellore cattle raised on pasture in northern Brazil. The analysis utilized random regression techniques to model the reaction norm. Fixed effects consisted of sex, contemporary group, and the covariate of age of cow at calving. The environmental gradient, defined by the concatenation of a bull and the state in which the calf was born, was modeled by second order Legendre polynomials. Direct additive genetic and residual effects were fit as random. Results showed differences in the magnitude of expression of genotype in proportion to decreasing favorability of the environment. As the environment became more unfavorable, the correlation of breeding value to different environments decreased. The correlations between the intercept and the level slope for 365W feature were of moderate magnitude, predominantly indicating the reclassification of sires in different environments. Reaction standard model was coherent from a technical and biological view point and enabled the perception of GxE in the genetic evaluation of Nellore cattle in the states of Maranhão, Pará and Tocantins. 

  17. Monitor and Protect Wigwam River Bull Trout for Koocanusa Reservoir; Skookumchuck Creek Juvenile Bull Trout and Fish Habitat Monitoring Program, Annual Report 2002.

    Energy Technology Data Exchange (ETDEWEB)

    Cope, R.

    2003-06-01

    The Skookumchuck Creek juvenile bull trout (Salvelinus confluentus) and fish habitat-monitoring program is a co-operative initiative of the British Columbia Ministry of Water, Land, and Air Protection and Bonneville Power Administration. This project was commissioned in planning for fish habitat protection and forest development within the Skookumchuck Creek watershed and was intended to expand upon similar studies initiated within the Wigwam River from 2000 to 2002. The broad intent is to develop a better understanding of juvenile bull trout and Westslope cutthroat trout recruitment and the ongoing hydrologic and morphologic processes, especially as they relate to spawning and rearing habitat quality. The 2002 project year represents the first year of a long-term bull trout-monitoring program with current studies focused on collecting baseline information. This report provides a summary of results obtained to date. Bull trout represented 72.4% of the catch. Fry dominated the catch because site selection was biased towards electrofishing sample sites which favored high bull trout fry capture success. The mean density of all juvenile bull trout was estimated to be 6.6 fish/100m{sup 2}. This represents one-half the densities reported for the 2002 Wigwam River enumeration program, even though enumeration of bull trout redds was an order of magnitude higher for the Wigwam River. Typically, areas with combined fry and juvenile densities greater than 1.5 fish per 100 m{sup 2} are cited as critical rearing areas. Trends in abundance appeared to be related to proximity to spawning areas, bed material size, and water depth. Cover components utilized by juvenile and adult bull trout and cutthroat trout were interstices, boulder, depth, overhead vegetation and LWD. The range of morphological stream types encompass the stable and resilient spectrum (C3(1), C3 and B3c). The Skookumchuck can be generalized as a slightly entrenched, meandering, riffle-pool, cobble dominated

  18. Validation of the sperm quality analyzer and the hypo-osmotic swelling test for frozen-thawed ram and minke whale (Balaenoptera bonarensis) spermatozoa.

    Science.gov (United States)

    Fukui, Yutaka; Togawa, Morihiko; Abe, Norihito; Takano, Yuuki; Asada, Masatsugu; Okada, Aki; Iida, Kenji; Ishikawa, Hajime; Ohsumi, Seiji

    2004-02-01

    The object of the present study was to investigate the validation of the sperm quality analyzer (SQA) and the hypo-osmotic swelling (HOS) test with standard sperm analysis methods in frozen-thawed ram and minke whale spermatozoa. In rams, highly significant correlations were observed in the percentage of motile spermatozoa (Psperm concentration (Pwhale spermatozoa, the SVI (sperm viability index) values by the standard method were significantly (Psperm motility index (SMI) values by SQA. The percentage of motile spermatozoa was also significantly correlated (Pwhale spermatozoa, and that the SQA and HOS test values are significantly correlated in ram spermatozoa. However, sperm concentration and morphologically normal spermatozoa are not assessed accurately by SQA in minke whales.

  19. Use of Bayesian Inference to Correlate In Vitro Embryo Production and In Vivo Fertility in Zebu Bulls

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    Mateus José Sudano

    2011-01-01

    Full Text Available The objective of this experiment was to test in vitro embryo production (IVP as a tool to estimate fertility performance in zebu bulls using Bayesian inference statistics. Oocytes were matured and fertilized in vitro using sperm cells from three different Zebu bulls (V, T, and G. The three bulls presented similar results with regard to pronuclear formation and blastocyst formation rates. However, the cleavage rates were different between bulls. The estimated conception rates based on combined data of cleavage and blastocyst formation were very similar to the true conception rates observed for the same bulls after a fixed-time artificial insemination program. Moreover, even when we used cleavage rate data only or blastocyst formation data only, the estimated conception rates were still close to the true conception rates. We conclude that Bayesian inference is an effective statistical procedure to estimate in vivo bull fertility using data from IVP.

  20. Proteomic analysis of boar spermatozoa and quantity changes of superoxide dismutase 1, glutathione peroxidase, and peroxiredoxin 5 during epididymal maturation.

    Science.gov (United States)

    Park, Kyunghee; Jeon, Seunghye; Song, Yoon-Jae; Yi, Lee S H

    2012-11-01

    Mammalian spermatozoa and their various proteins undergo various modifications during maturation in the epididymis. To characterize proteins that change in quantity during this maturational process, boar spermatozoa were collected from various regions of the epididymis, and extracts were analyzed by two-dimensional gel electrophoresis (2-DE). A number of proteins were identified as changing in quantity, and MALDI-MS analysis revealed that superoxide dismutase 1 (SOD1) from the acrosomal proteins of spermatozoa, and glutathione peroxidase (GPX) and peroxiredoxin 5 from the membranous fraction increased during the epididymal transit of spermatozoa. These proteins are antioxidants that remove reactive oxygen species (ROS); they are presumed to protect spermatozoa during epididymal transit and storage. Western blot analysis of SOD1, GPX and peroxiredoxin 5 showed that these protein levels increased as the spermatozoa traveled from the caput to the cauda epididymis. Activity analysis showed that total SOD activity also increased. Therefore, we conclude that several antioxidant proteins increase during the transit of boar spermatozoa through the epididymis, ultimately contributing to the maturation and/or survival of sperm. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. IN VITRO STUDY OF THE EFFECT OF 17β-ESTRADIOL AND 4-NONYLPHENOL ON BOVINE SPERMATOZOA

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    Jana Lukáčová

    2012-10-01

    Full Text Available Nonylphenol (NP, an environmental endocrine disruptor, is a final metabolite of nonylphenol ethoxylate (NPE that is able to interfere with hormonal system of numerous organisms. Estrogens play a central role in female reproduction, but also affect the male reproductive system. In males, stimulate mammalian spermatozoa capacitation, acrosome reaction and fertilizing ability. The aim of this in vitro study was to determine the effect of 17β-estradiol and nonylphenol (NP on the spermatozoa motility. Specifically, we examined the dose- and time-dependent effect of nonylphenol (1, 10, 100 and 200 µg/mL with the addition 1 µg/mL of 17β-estradiol on the motility and progressive motility of bovine spermatozoa during two time periods (0 h and 6 h. The spermatozoa motility was determined by CASA (Computer Assisted Semen Analyzer system using the Sperm VisionTM program. The results showed decreased average values of motility in all experimental groups during 0 h of in vitro cultivation. The motility and the progressive motility of bovine spermatozoa increased in the experimental groups using concentrations 10, 100 and 200 µg/mL after 6 h of cultivation and significant differences (P<0.05 were detected between these groups and the control group. The results suggest that the addition of 17β-estradiol could positively affect spermatozoa motility during the short-term cultivation of spermatozoa with NP.

  2. SRC family kinases in hamster spermatozoa: evidence for the presence of LCK.

    Science.gov (United States)

    Singh, Durgesh Kumar; Deshmukh, Rohit Kumar; Narayanan, Praveen Kumar; Shivaji, Sisinthy; Siva, Archana Bharadwaj

    2017-05-01

    Sperm capacitation is a prerequisite for successful fertilization. Increase in tyrosine phosphorylation is considered the hallmark of capacitation and attempts to understand its regulation are ongoing. In this regard, we attempted to study the role of SRC family kinases (SFKs) in the hamster sperm functions. Interestingly, we found the presence of the lymphocyte-specific protein tyrosine kinase, LCK, in mammalian spermatozoa and further characterized it in terms of its localization and function. LCK was found in spermatozoa of several species, and its transcript was identified in the hamster testis. Autophosphorylation of LCK at the Y394 residue increased as capacitation progressed, indicating an upregulation of LCK activity during capacitation. Inhibition of LCK (and perhaps the other SFKs) with the use of a specific inhibitor showed a significant decrease in protein tyrosine phosphorylation of several proteins, implying LCK/SFKs as key tyrosine kinase(s) regulating tyrosine phosphorylation during hamster sperm capacitation. Dihydrolipoamide dehydrogenase was identified as a substrate for LCK/SFK. LCK/SFKs inhibition significantly reduced the percentage fertilization ( in vitro ) but had no effect on sperm motility, hyperactivation and acrosome reaction. In summary, this is the first report on the presence of LCK, an SFK of hematopoietic lineage in spermatozoa besides being the first study on the role of SFKs in the spermatozoa of Syrian hamsters. © 2017 Society for Reproduction and Fertility.

  3. Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

    NARCIS (Netherlands)

    Nguyen, Quynh Thu; Wallner, Ulrike; Schmicke, Marion; Waberski, Dagmar; Henning, Heiko

    2016-01-01

    Mammalian spermatozoa utilize ATP as the energy source for key functions on the route to fertilization. ATP and its precursor nucleotides ADP and AMP are determined in many sperm physiology studies, mostly by bioluminescence assays. Assay results vary widely, mainly due to different efficiency in

  4. Radiolabeling of mammalian spermatozoa and their use to monitor sperm transport in females. [Sheep

    Energy Technology Data Exchange (ETDEWEB)

    Lorton, S.P.; Gatley, S.J.; Lieberman, L.M.; First, N.L.

    1980-03-15

    Radiolabeling of mammalian spermatozoa with /sup 131/I, /sup 67/Ga, /sup 111/In, and /sup 99m/Tc was investigated. Spermatozoa were labeled with /sup 99m/Tc for in vivo studies because of a high labeling yield (70 to 90%) combined with the lack of impairment of sperm motility. Ovariectomized sheep were brought into estrus by sequential administration of progesterone and estradiol cyprionate. Sheep were necropsied up to 6 hours after insemination with /sup 99m/Tc-labeled ram sperm and their reproductive tracts were resected and examined with a rectilinear scanner. Radioactivity was clearly observed in the fallopian tubes, with larger amounts in the vaginas, cervices, and uteri. In contrast, when /sup 99m/Tc-spermatozoa were replaced with /sup 99m/TcO/sub 4//sup -/, much less radioactivity remained in the reproductive tract at resection and this was evenly distributed. Some label left the /sup 99m/Tc-spermatozoa in vivo, but radioactivity remained on the cells long enough to consider attempting to monitor sperm transport in vivo in a suitable species.

  5. Comparison of glycerol, lactamide, acetamide and dimethylsulfoxide as cryoprotectants of Japanese white rabbit spermatozoa.

    Science.gov (United States)

    Kashiwazaki, Naomi; Okuda, Yasushi; Seita, Yasunari; Hisamatsu, Shin; Sonoki, Shigenori; Shino, Masao; Masaoka, Toshio; Inomata, Tomo

    2006-08-01

    The rabbit is considered to be a valuable laboratory animal. We compared glycerol, lactamide, acetamide, and dimethylsulfoxide (DMSO) as cryoprotectants in egg-yolk diluent of ejaculated Japanese white rabbit spermatozoa for improvement of sperm cryopreservation methods. Rabbit semen was frozen with 1.0 M glycerol, lactamide, acetamide, or DMSO in plastic straws. Forward progressive motility and plasma membrane integrity of the post-thaw spermatozoa were examined. The rate of forward progressive motile spermatozoa in lactamide (37.8 +/- 3.0%) was significantly (P<0.05) higher than in glycerol (17.0 +/- 3.3%). In addition, the rates of sperm plasma membrane integrity in lactamide and acetamide (35.9 +/- 3.3% and 30.2 +/- 3.0%, respectively) were significantly (P<0.05) higher than in glycerol (17.0 +/- 2.6%). The results indicate that 1.0 M lactamide and acetamide have higher cryoprotective effects than 1.0 M glycerol for cryopreservation of Japanese white rabbit spermatozoa.

  6. A carbohydrate-antioxidant hybrid polymer reduces oxidative damage in spermatozoa and enhances fertility.

    Science.gov (United States)

    Fleming, Craig; Maldjian, André; Da Costa, Daniel; Rullay, Attvinder K; Haddleton, David M; St John, Justin; Penny, Paul; Noble, Raymond C; Cameron, Neil R; Davis, Benjamin G

    2005-10-01

    Gamete-gamete interactions are critically modulated by carbohydrate-protein interactions that rely on the carbohydrate-selective recognition of polyvalent carbohydrate structures. A galactose-binding protein has been identified in mammalian spermatozoa that has similarity to the well-characterized hepatic asialoglycoprotein receptor. With the aim of exploiting the ability of this class of proteins to bind and internalize macromolecules displaying galactose, we designed hybrid carbohydrate-antioxidant polymers to deliver antioxidant vitamin E (alpha-tocopherol) to porcine spermatozoa. Treatment of sperm cells with one hybrid polymer in particular produced large increases in intracellular sperm levels of alpha-tocopherol and greatly reduced endogenous fatty acid degradation under oxidative stress. The polymer-treated spermatozoa had enhanced physiological properties and longer half-lives, which resulted in enhanced fertilization rates. Our results indicate that hybrid polymer delivery systems can prolong the functional viability of mammalian spermatozoa and improve fertility rates, and that our functionally guided optimization strategy can be applied to the discovery of active glycoconjugate ligands.

  7. 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. © 2015 Blackwell Verlag GmbH.