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Sample records for bull spermatozoa express

  1. Protein expression pattern of PAWP in bull spermatozoa is associated with sperm quality and fertility following artificial insemination.

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    Kennedy, Chelsey E; Krieger, Kari Beth; Sutovsky, Miriam; Xu, Wei; Vargovič, Peter; Didion, Bradley A; Ellersieck, Mark R; Hennessy, Madison E; Verstegen, John; Oko, Richard; Sutovsky, Peter

    2014-05-01

    Post-acrosomal WW-domain binding protein (PAWP) is a signaling molecule located in the post-acrosomal sheath (PAS) of mammalian spermatozoa. We hypothesized that the proper integration of PAWP in the sperm PAS is reflective of bull-sperm quality and fertility. Cryopreserved semen samples from 298 sires of acceptable, but varied, fertility used in artificial insemination services were analyzed using immunofluorescence microscopy and flow cytometry for PAWP protein. In normal spermatozoa, PAWP fluorescence formed a regular band around the proximal PAS. Anomalies of PAWP labeling in defective spermatozoa were reflected in flow cytometry by varied intensities of PAWP-induced fluorescence. Distinct sperm phenotypes were also identified, including morphologically normal and some defective spermatozoa with moderate levels of PAWP; grossly defective spermatozoa with low/no PAWP; and defective spermatozoa with high PAWP. Analysis by ImageStream flow cytometry confirmed the prevalence of abnormal sperm phenotypes in the spermatozoa with abnormal PAWP content. Live/dead staining and video recording showed that some abnormal spermatozoa are viable and capable of progressive motility. Conventional flow-cytometric measurements of PAWP correlated significantly with semen quality and fertility parameters that reflect the sires' artificial insemination fertility, including secondary sperm morphology, conception rate, non-return rate, and residual value. A multiplex, flow-cytometric test detecting PAWP, aggresomes (ubiquitinated protein aggregates), and acrosomal integrity (peanut-agglutinin-lectin labeling) had a predictive value for conception rate, as demonstrated by step-wise regression analysis. We conclude that PAWP correlates with semen/fertility parameters used in the cattle artificial insemination industry, making PAWP a potential biomarker of bull fertility.

  2. Zinc and magnesium in bull and boar spermatozoa.

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    Arver, S; Eliasson, R

    1980-11-01

    Mean +/- s.e.m. concentrations (nmol/10(8) cells) of zinc and magnesium in bull spermatozoa were 30.6 +/- 6.6 and 119 +/- 28.8, respectively. Corresponding values for boar spermatozoa were 16.9 +/- 1.98 and 57.1 +/- 4.3. Bull spermatozoa washed twice in a standard buffered salt solution, pH 7.75, lost 72.6% of their zinc and 46.5% of their magnesium. Boar spermatozoa lost 40% of Zn and 18% of Mg, respectively. Addition of albumin (4% final concentration) to the washing solution did not increase the loss of ions from bull spermatozoa but increased the loss of zinc and magnesium from boar spermatozoa to 52% and 41%, respectively.

  3. 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

  4. Effect of some permeating cryoprotectants on CASA motility results in cryopreserved bull spermatozoa.

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    Awad, M M

    2011-02-01

    Computer-assisted sperm analyzers (CASA) have become the standard tool for evaluating sperm motility because they provide objective results for thousands of mammalian spermatozoa. Mammalian spermatozoa experience osmotic stress when the glycerol is added to the cells prior to freezing and removal from the cells after thawing. In order to minimize osmotic damage, cryoprotectants having lower molecular weights and greater membrane permeability than glycerol, were evaluated to determine their effectiveness for cryopreserving bull spermatozoa. The aim of this study was to compare the cryopreservation effects of low molecular weight cryoprotectants (ethylene glycol and methanol) to glycerol, on post-thaw CASA sperm parameters. Bull semen was diluted with tris-egg yolk extender containing 3% glycerol, 3, 2 and 1% ethylene glycol or 3, 2 and 1% methanol. Bull semen was frozen in 0.5 straws. Bull spermatozoa exhibited higher percentages (p<0.01) for total (Mot, 72.4%) and progressively (Prog, 29.5%) motilities when frozen in extender containing 3% glycerol compared to 3, 2 and 1% ethylene glycol or 3, 2 and 1% methanol. In conclusion, no advantages were found in using ethylene glycol or methanol to replace glycerol in bull semen freezing. Glycerol provided the best sperm characteristics for bull spermatozoa after freezing and thawing. The possibility of using ethylene glycol or methanol as permeating cryoprotectants for bull semen deserves further investigation, and these cryoprotectants should also be evaluated in extenders that contain disaccharides or cholesterol.

  5. The complete amino acid sequence of the basic nuclear protein of bull spermatozoa

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    Coelingh, J.P.; Monfoort, Cornelis H.; Rozijn, Thomas H.; Gevers Leuven, Jan A.; Schiphof, R.; Steyn-Parvé, Elizabeth P.; Braunitzer, Gerhard; Schrank, Barbara; Ruhfus, Annette

    1972-01-01

    The complete amino acid sequence of the basic nuclear protein of bull spermatozoa has been established. The sequence was partially deduced by characterization of peptides isolated from thermolysine and chymotryptic digests of the reduced and S-aminoethylated protein. The complete sequence of the fir

  6. Acrosome membrane integrity and cryocapacitation are related to cholesterol content of bull spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Srivastava N; Srivastava SK; Ghosh SK; Amit Kumar; Perumal P; Jerome A

    2013-01-01

    Objective: To evaluate the cryoinjury prediction of spermatozoa in relation to its cholesterol content at fresh and frozen-thaw stages. Methods: Ejaculates (n=12) were processed for cryopreservation, acrosome integrity (fluorescent and giemsa stains), cryoinjury (distribution of non capacitated, capacitated and acrosome reacted, pattern F, B and AR, respectively of Chlortetracycline, CTC assay), in vitro fertiltiy (IVF) and cholesterol content of spermatozoa at fresh, pre-freeze and frozen-thaw stages were evaluated. Values were fitted in prediction equation to predict acrosome integrity (AI) and cryoinjury. Results: Study indicated that cholesterol content of fresh spermatozoa can be used to predict cholesterol content of spermatozoa at pre-freeze and frozen-thaw stages of cryopreservation protocol with medium to high level of accuracy (P<0.05 and P<0.01, respectively). Cholesterol content of fresh spermatozoa can be used to predict AI, pattern B and AR and Penetration Index (PI) of IVF with medium level of accuracy (P<0.05) at frozen-thaw but not at pre-freeze stage. Similarly cholesterol content of frozen-thaw spermatozoa can be used to predict AI and pattern AR of frozen-thaw spermatozoa with medium level of accuracy (P<0.05). Conclusion: This study revealed strong evidence that cholesterol content of fresh as well as frozen-thaw bull spermatozoa can be a good predictor of level of cryoinjury following preservation at ultra low temperature.

  7. Effect of sex sorting on CTC staining, actin cytoskeleton and tyrosine phosphorylation in bull and boar spermatozoa.

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    Bucci, D; Galeati, G; Tamanini, C; Vallorani, C; Rodriguez-Gil, J E; Spinaci, M

    2012-04-01

    Sperm sorting is a useful technology that permits sex preselection. It presents some troubles because of low fertility after the process. The main aim of this work was to analyze the putative existence of capacitation-like changes in both boar and bull sperm subjected to sex sorting that could lead to a detriment of semen quality. The parameters used were CTC staining patterns, actin cytoskeleton organization and tyrosine phosphorylation patterns; the last two were determined by both western blotting and immunofluorescence. Sex sorted spermatozoa were compared with fresh, in vitro capacitated and in vitro acrosome reacted sperm. In both species, sex sorted sperm showed a CTC staining pattern similar to that observed after in vitro capacitation. The actin pattern distribution after sperm sorting also tended to be similar to that observed after in vitro capacitation, but this effect was more pronounced in bull than in boar spermatozoa. However, actin expression analysis through western blot did not show any change in either species. The tyrosine phosphorylation pattern in boar sperm was practically unaltered after the sex sorting process, but in bulls about 40% of spermatozoa had a staining pattern indicative of capacitation. Additionally, western blotting analysis evidenced some differences in the expression of protein tyrosine phosphorylation among fresh, capacitated, acrosome reacted and sex sorted sperm cells in both species. Our results indicate that not all the sex-sorted-related modifications of the studied parameters were similar to those occurring after "in vitro" capacitation, thus suggesting that sex sorting-induced alterations of sperm function and structure do not necessarily indicate the achievement of the capacitated status of sorted sperm.

  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

    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. Selected proteins of "prostasome-like particles" from epididymal cauda fluid are transferred to epididymal caput spermatozoa in bull.

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    Frenette, Gilles; Lessard, Carl; Sullivan, Robert

    2002-07-01

    During epididymal transit, spermatozoa acquire selected proteins secreted by epithelial cells. We recently showed that P25b, a protein with predictive properties for bull fertility, is transferred from prostasome-like particles present in the cauda epididymal fluid (PLPCd) to the sperm surface. To further characterize the interactions between PLPCd and epididymal spermatozoa, PLPCd were prepared by ultracentrifugation of bull epididymal fluid, then surface-exposed proteins were biotinylated and coincubated in different conditions with caput epididymal spermatozoa. Western blot analysis revealed that only selected proteins are transferred from PLPCd to spermatozoa. MALDI-TOF analysis revealed that these transferred proteins are closely related. The pattern of distribution of the PLPCd transferred varied from one sperm cell to the other, with a bias toward the acrosomal cap. This transfer appeared to be temperature sensitive, being more efficient at 32-37 degrees C than at 22 degrees C. Transfer of PLPCd proteins to spermatozoa was also pH dependant, the optimal pH for transfer being 6.0-6.5. The effect of divalent cations on PLPCd protein transfer to caput spermatozoa was investigated. Whereas Mg(2+) and Ca(2+) have no effect on the amount of proteins remaining associated with spermatozoa following coincubation, Zn(2+) had a beneficial effect. These results are discussed with regard to the function of PLPCd in epididymal sperm maturation.

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

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    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 soya-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 soya-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.

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

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

    2015-06-01

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

  14. Effect of different thawing procedures on the quality and fertility of the bull spermatozoa

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    Andrey Lyashenko

    2015-01-01

    Objective: To improve the indicators of motility, survival and fertilizing ability of spermatozoa by optimizing temperature factors and the duration of exposure at unfreezing straws. Methods:Straws by volume 0.25 mL were thawed at water bath temperatures at 65℃, 67℃ and 70℃ for 6-7 seconds and at 75℃for 4-6 seconds. Impact of exposure time and temperature thawing in the water bath on motility and survival of spermatozoa were studied. Results:Studies indicate that for the procedure of defrost water bath straws in seven seconds for temperature conditions of 65℃, 67℃ and 70℃, indicators of progressive motility and absolute survival rate were significantly higher than for the control group an average on 11.4 % (P<0.01). Optimum exposure time (6-7 seconds) and temperature range (65-70 ℃) defrosting semen doses were defined. Conclusions: Owing obtained the positive result, method of thawing was developed which increases the indicators of motility, survival and fertilizing capacity of bull semen.

  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. Concentration of copper, iron, zinc, cadmium, lead, and nickel in bull and ram semen and relation to the occurrence of pathological spermatozoa.

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    Massányi, P; Trandzik, J; Nad, P; Koreneková, B; Skalická, M; Toman, R; Lukac, N; Halo, M; Strapak, P

    2004-01-01

    In this study the concentration of copper, iron, zinc, cadmium, lead, and nickel in bull and ram semen and relation of these metals to spermatozoa morphology was investigated. Analysis by atomic absorption spectrophotometry showed that copper concentration was significantly higher (pzinc concentration was higher in bull semen in comparison with ram semen. The iron and cadmium concentrations in the semen were similar. Higher concentration of lead was found in ram semen. Higher levels of nickel were found in ram semen in comparison with bulls. In bull semen 11.79+/-4.88% of pathological spermatozoa was found. Higher occurrence of pathological spermatozoa was in ram semen (17.17+/-3.76) in comparison with the semen of bulls. Separated tail, tail torso, and knob twisted tail were the most frequent forms of pathological spermatozoa in both species. Correlation analysis in bulls showed high positive relation between iron and zinc (r = 0.72), nickel and separated tail (r = 0.76), separated tail and tail torso (r = 0.71), tail torso and total number of pathological spermatozoa (r=0.72), and between tail ball and total number of pathological spermatozoa (r = 0.78). In rams high positive correlation between cadmium and lead (r=0.98), nickel and separated tail (r=0.77), separated tail and total number of pathological spermatozoa (r=0.69), knob twisted tail and retention of cytoplasmic drop (r=0.78), and between knob twisted tail and other pathological spermatozoa (r = 0.71) was found. High negative correlation in ram semen was observed between copper and nickel (r=0.71), copper and separated tail (r=0.70), and between iron and tail torso (r=0.67). The results suggest that the studied metals have a direct effect on spermatozoa quality.

  17. Incorporation of selenium-75 into seminal plasma and spermatozoa of the bull.

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    Vanha-Perttula, T; Remes, E

    1990-01-01

    75Se was given intravenously into 5 bulls. Multiple blood and semen samples were taken and during slaughter 5, 10, 15, 20 and 80 days later samples of various reproductive and other organs were collected. After injection, 75Se in blood reached a peak at 6 h followed by a rapid decline. The label was mainly found in serum with very low levels in erythrocytes. Initially the serum 75Se was bound to a macromolecule with a mw of 80 kDa, but later a larger molecule (100 kDa) was observed. In semen 75Se was first mainly found in seminal plasma, where a plateau level was reached at 5 d followed by a gradual decline after 12 d. The total semen level, however, increased after 14 d and this increase was due to a rapid appearance of the label in spermatozoa. The sperm 75Se level reached a plateau at 20 d and remained high until 40 days, after which a gradual decline ensued. The seminal plasma 75Se eluted in gel filtration coincident with glutathione peroxidase. The highest levels of 75Se were found in the kidney followed by seminal vesicles and testicles. The seminal vesicle secretion was particularly rich in 75Se and its fractionation resembled that of the seminal plasma. 75Se appeared in the epididymal caput within 5 days and passed through the epididymis in 20 days. It is concluded that 75Se is actively incorporated in the bull seminal vesicles into GSH-Px, while in the testis it is incorporated into a structural sperm protein during spermatogenesis.

  18. Motility analysis of circularly swimming bull spermatozoa by quasi-elastic light scattering and cinematography.

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    Craig, T; Hallett, F R; Nickel, B

    1982-04-01

    The Rayleigh-Gans-Debye approximation is used to predict the electric field autocorrelation functions of light scattered from circularly swimming bull spermatozoa. Using parameters determined from cinematography and modeling the cells as coated ellipsoids of semiaxes a = 0.5 micrometers, b = 2.3 micrometers, and c = 9.0 micrometers, we were able to obtain model spectra that mimic the data exactly. A coat is found to be a necessary attribute of the particle. It is also clear that these model functions at 15 degrees may be represented by the relatively simple function used before by Hallett et al. (1978) to fit data from circularly swimming cells, thus giving some physical meaning to these functional shapes. Because of this agreement the half-widths of experimental functions can now be interpreted in terms of an oscillatory frequency for the movement of the circularly swimming cell. The cinematographic results show a trend to chaotic behavior as the temperature of the sample is increased, with concomitant decrease in overall efficiency. This is manifested by a decrease in oscillatory frequency and translational speed.

  19. Adverse effect of cake collapse on the functional integrity of freeze-dried bull spermatozoa.

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    Hara, Hiromasa; Tagiri, Miho; Hwang, In-Sul; Takahashi, Masato; Hirabayashi, Masumi; Hochi, Shinichi

    2014-06-01

    Under optimal freeze-drying conditions, solutions exhibit a cake-like porous structure. However, if the solution temperature is higher than the glass transition temperature of the maximally freeze-concentrated phase (Tg') during drying phase, the glassy matrix undergoes viscous flow, resulting in cake collapse. The purpose of the present study was to investigate the effect of cake collapse on the integrity of freeze-dried bull spermatozoa. In a preliminary experiment, factors affecting the Tg' of conventional EGTA buffer (consisting of Tris-HCl, EGTA and NaCl) were investigated in order to establish the main experimental protocol because EGTA buffer Tg' was too low (-45.0°C) to suppress collapse. Modification of the EGTA buffer composition by complete removal of NaCl and addition of trehalose (mEGTA buffer) resulted in an increase of Tg' up to -27.7°C. In the main experiment, blastocyst yields after ooplasmic injection of freeze-dried sperm preserved in collapsed cakes (drying temperature: 0 or -15°C) were significantly lower than those of sperm preserved in non-collapsed cake (drying temperature: -30°C). In conclusion, freeze-dried cake collapse may be undesirable for maintaining sperm functions to support embryonic development, and can be inhibited by controlling both Tg' of freeze-drying buffer and temperature during the drying phase.

  20. Phospholipid hydroperoxide glutathione peroxidase in bull spermatozoa provides a unique marker in the quest for semen quality analysis.

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    Stradaioli, G; Sylla, L; Monaci, M; Maiorino, M

    2009-07-01

    Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a selenoperoxidase accounting for most of the selenium content in mammalian testis, which has been found to be linked to fertility in humans. In this study, we addressed the issue whether PHGPx content in spermatozoa could be a predictive index of fertilization capacity for sire selection in bulls. Measurement of PHGPx in spermatozoa of 92 yearling bulls of three different Italian breeds (Chianina, Romagnola, and Marchigiana) revealed the presence of two quite well separated populations. A PHGPx activity of 130 mU/mg separated the high-PHGPx group (H-PHGPx, n=73) from the low-PHGPx group (L-PHGPx, n=19). Forward motility was markedly higher in the H-PHGPx group, which also contained a lower percentage of detached heads, abnormal midpiece, and proximal droplets. On the other hand, differently from the human studies, no correlation was observed between PHGPx activity and number of spermatozoa in the ejaculate. Apart from sperm count, which typically differed among breeds, and number of detached heads in the L-PHGPx group, which correlated with higher sperm count, no other significant difference in seminal parameters among breeds was apparent. The assay for sperm PHGPx activity therefore emerges as a unique tool to evaluate semen quality for sire selection.

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

    Directory of Open Access Journals (Sweden)

    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.

  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. Effect of equilibration times, freezing, and thawing rates on post-thaw quality of buffalo (Bubalus bubalis) bull spermatozoa.

    Science.gov (United States)

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

    2016-09-01

    The effects of equilibration times (E1, 2 h; E2, 4 h; E3, 6 h), freezing rates (FR1, manual, 5 cm above liquid nitrogen (LN2 ) for 10 min, plunging in LN2 ; FR2, programmable ultra-fast, holding at +4 °C for 2 min, from 4 to -10 °C at -10 °C/min, from -10 to -20 °C at -15 °C/min, from -20 to -120 °C at -60 °C/min, holding at -120 °C for 30 sec, plunging in LN2 ), and thawing rates (T1, 37 °C for 30 sec; T2, 50 °C for 15 sec; T3, 70 °C for 7 sec) were evaluated on quality of buffalo bull spermatozoa. Progressive motility (%), rapid velocity (%), average path velocity (VAP, μm/s), straight line velocity (VSL, μm/s), and mitochondrial transmembrane potential (%) were higher (p ultra-fast freezing rate, and rapid thawing at 70 °C for 7 sec in cryopreservation protocol improves the post-thaw quality of buffalo bull spermatozoa.

  4. 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.

  5. 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.

  6. Studies on effect of different seasons on expression of HSP70 and HSP90 gene in sperm of Tharparkar bull semen

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    J.S. Rajoriya

    2014-09-01

    Conclusion: It was concluded from the present study that there was no significant difference in the mRNA expression of HSP 70 and HSP 90 between the winter and summer season, presence of similar type of stress resistant spermatozoa in Tharparkar bull semen and the semen can be cryopreserved throughout the year in this prestigious Indian breed.

  7. Bovine Circadian Locomotor Output Cycles Kaput (CLOCK) and Clusterin (CLU) mRNA Quantitation in Ejaculated Crossbred Bull Spermatozoa.

    Science.gov (United States)

    Kumar, S; Deb, R; Singh, U; Ganguly, I; Mandal, D K; Tyagi, S; Kumar, M; Sengar, G; Sharma, S; Singh, R; Singh, R

    2015-06-01

    Mammalian circadian locomotor output cycles kaput (CLOCK) gene encodes a transcription factor that affects both the persistence and the period of circadian rhythms. Earlier reports suggested that CLOCK gene might be associated with male infertility in human. Present investigation, for the first time, reports that CLOCK gene expresses differentially between good and poor quality crossbred bull semen. The relative expression of CLOCK was significantly (p < 0.05) higher among good quality bull semen than motility-impaired ones. Clusterins (CLU) are series of genes associated with a variety of physiological activities including spermatogenesis, apoptosis and degenerative disease conditions. In the present context, we also investigated that the expression of CLU gene was significantly (p < 0.05) higher among motility-impaired crossbred bull semen compared to the good quality one.

  8. Alternative splicing, promoter methylation, and functional SNPs of sperm flagella 2 gene in testis and mature spermatozoa of Holstein bulls.

    Science.gov (United States)

    Guo, F; Yang, B; Ju, Z H; Wang, X G; Qi, C; Zhang, Y; Wang, C F; Liu, H D; Feng, M Y; Chen, Y; Xu, Y X; Zhong, J F; Huang, J M

    2014-02-01

    The sperm flagella 2 (SPEF2) gene is essential for development of normal sperm tail and male fertility. In this study, we characterized first the splice variants, promoter and its methylation, and functional single-nucleotide polymorphisms (SNPs) of the SPEF2 gene in newborn and adult Holstein bulls. Four splice variants were identified in the testes, epididymis, sperm, heart, spleen, lungs, kidneys, and liver tissues through RT-PCR, clone sequencing, and western blot analysis. Immunohistochemistry revealed that the SPEF2 was specifically expressed in the primary spermatocytes, elongated spermatids, and round spermatids in the testes and epididymis. SPEF2-SV1 was differentially expressed in the sperms of high-performance and low-performance adult bulls; SPEF2-SV2 presents the highest expression in testis and epididymis; SPEF2-SV3 was only detected in testis and epididymis. An SNP (c.2851G>T) in exon 20 of SPEF2, located within a putative exonic splice enhancer, potentially produced SPEF2-SV3 and was involved in semen deformity rate and post-thaw cryopreserved sperm motility. The luciferase reporter and bisulfite sequencing analysis suggested that the methylation pattern of the core promoter did not significantly differ between the full-sib bulls that presented hypomethylation in the ejaculated semen and testis. This finding indicates that sperm quality is unrelated to SPEF2 methylation pattern. Our data suggest that alternative splicing, rather than methylation, is involved in the regulation of SPEF2 expression in the testes and sperm and is one of the determinants of sperm motility during bull spermatogenesis. The exonic SNP (c.2851G>T) produces aberrant splice variants, which can be used as a candidate marker for semen traits selection breeding of Holstein bulls.

  9. l-Cysteine improves antioxidant enzyme activity, post-thaw quality and fertility of Nili-Ravi buffalo (Bubalus bubalis) bull spermatozoa.

    Science.gov (United States)

    Iqbal, S; Riaz, A; Andrabi, S M H; Shahzad, Q; Durrani, A Z; Ahmad, N

    2016-11-01

    The effects of l-cysteine in extender on antioxidant enzymes profile during cryopreservation, post-thaw quality parameters and in vivo fertility of Nili-Ravi buffalo bull spermatozoa were studied. Semen samples from 4 buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of l-cysteine (0.0, 0.5, 1.0, 2.0 and 3.0 mm) and frozen in 0.5-ml French straws. The antioxidative enzymes [catalase, super oxide dismutase and total glutathione (peroxidase and reductase)] were significantly higher (P cysteine as compared to other groups. Post-thaw total motility (%), progressive motility (%), rapid velocity (%), average path velocity (μm s(-1) ), straight line velocity (μm s(-1) ), curvilinear velocity (μm s(-1) ), beat cross frequency (Hz), viable spermatozoa with intact plasmalemma (%), acrosome and DNA integrity (%) were higher with the addition of 2.0 mm l-cysteine as compared to other groups (P cysteine than in the control. In conclusion, the addition of 2.0 mm l-cysteine in extender improved the antioxidant enzymes profile, post-thaw quality and in vivo fertility of Nili-Ravi buffalo bull spermatozoa.

  10. Comparative studies of elemental composition on ejaculated fowl, bull, rat, dog and boar spermatozoa by electron probe X-ray microanalysis.

    Science.gov (United States)

    Ashizawa, K; Ozawa, Y; Okauchi, K

    1987-01-01

    1. Comparative analyses of the concentrations of bulk and trace elements on the head, midpiece and tail regions of the ejaculated fowl, bull, rat, dog and boar spermatozoa were performed using X-ray microanalysis with scanning electron microscopy. 2. Although the pattern of distribution of elements on the surface of the three different subcellular regions was, in general, similar among all the species, there were substantial shifts in absolute concentrations. 3. Concentration of magnesium on the dog spermatozoa was significantly higher (about 2 times) than those of the other species. 4. Zinc, copper, iron and manganese concentrations were higher on fowl spermatozoa compared with those of the other species studied. 5. The Na-to-K ratios on the midpiece ranged from 1.46 (rat) to 2.26 (dog).

  11. Phospholipid hydroperoxide glutathione peroxidase: expression pattern during testicular development in mouse and evolutionary conservation in spermatozoa.

    Science.gov (United States)

    Nayernia, Karim; Diaconu, Mihaela; Aumüller, Gerhard; Wennemuth, Gunther; Schwandt, Iris; Kleene, Kenneth; Kuehn, Hartmut; Engel, Wolfgang

    2004-04-01

    Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a selenoprotein belonging to the family of glutathione peroxidases and has been implicated in antioxidative defense and spermatogenesis. PHGPx accounts for almost the entire selenium content of mammalian testis. In an attempt to verify the expression pattern of PHGPx, testes of mouse mutants with arrest at different stages of germ cell development and testes of mice at different ages were subjected to immunostaining with a monoclonal anti-PHGPx antibody. PHGPx was detected in Leydig cells of testes in all developmental stages. In the seminiferous tubuli, the PHGPx staining was first observed in testes of 21-day-old mice which is correlated with the appearance of the first spermatids. This result was confirmed when the testes of mutant mice with defined arrest of germ cell development were used. An immunostaining was observed in the seminiferous tubuli of olt/olt and qk/qk mice which show an arrest at spermatid differentiation. In Western blot analysis of proteins extracted from testes of mutant mice and from developing testes, two signals at 19- and 22-kDa were observed which confirm the existence of two PHGPx forms in testicular cells. In mouse spermatozoa, a subcellular localization of PHGPx and sperm mitochondria-associated cysteine-rich protein (SMCP) was demonstrated, indicating the localization of PHGPx in mitochondria of spermatozoa midpiece. For verifying the midpiece localization of PHGPx in other species, spermatozoa of Drosophila melanogaster, frog, fish, cock, mouse, rat, pig, bull, and human were used in immunostaining using anti-PHGPx antibody. A localization of PHGPx was found in the midpiece of spermatozoa in all species examined. In electronmicroscopical analysis, PHGPx signals were found in the mitochondria of midpiece. These results indicate a conserved crucial role of PHGPx during sperm function and male fertility.

  12. 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.

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

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    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

  14. 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

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    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

  15. Sex-sorting of spermatozoa affects developmental competence of in vitro fertilized oocytes in a bull-dependent manner

    OpenAIRE

    INABA, Yasushi; Abe, Reika; GESHI, Masaya; MATOBA, Satoko; Nagai, Takashi; Somfai, Tamás

    2016-01-01

    The aim of the present study was to clarify if flow-cytometric sex-sorting of bovine sperm affected in vitro blastocyst production in different bulls, either in terms of its ability to fertilize the oocyte or by interfering with post-fertilization embryo development. We performed in vitro fertilization (IVF) using both commercially available frozen-thawed X-sorted and non-sorted sperm of 4 Holstein bulls at 3 concentrations (1 × 106, 2 × 106, and 5 × 106 sperm/ml). When fertilization rates we...

  16. The effect of fenceline bull exposure on expression of oestrus in dairy cows

    NARCIS (Netherlands)

    Roelofs, J.B.; Soede, N.M.; Voskamp-Harkema, W.; Kemp, B.

    2008-01-01

    The objective of this study was to investigate whether dairy cows visit and interact with a fenceline-housed bull more during oestrus than outside oestrus and whether fenceline bull contact affects expression of oestrus. At one end of a free stall a fence with vertical open bars was placed behind wh

  17. Effect of low-density lipoproteins,spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4 ℃%低密度脂蛋白、精子浓度以及甘油对4℃下存储的公牛精子功能和活力的影响

    Institute of Scientific and Technical Information of China (English)

    Oscar Vera-Munoz; Lamia Amirat-Briand; Djemil Bencharif; Marc Anton; Serge Desherces; Eric Shmitt; Chantal Thorin; Daniel Tainturier

    2011-01-01

    An extender has been developed with low-density lipoproteins(LDLs)that eliminates the microbial risks associated with the use of whole egg yolk.The objective of this study was to assess the effects of substituting egg yolk with LDLs for use as an extender in sperm preservation at 4 ℃,as well as on spermatozoa motility,plasma membrane and acrosome integrity,at two different concentrations(80×106 and 240×106 sperm per ml)for 8 days and to evaluate glycerol toxicity in both extenders.A total of 12 ejaculates were collected from three bulls.Spermatozoa motility was examined using computer-assisted semen analysis.Plasma membrane integrity was determined using the hypo-osmotic swelling test and acrosome integrity with the fluorescein isothiocyanate-Pisum sativum agglutinin test.The semen was subsequently divided into four aliquots and diluted with Tris-egg yolk-glycerol(TEG),Tris-egg yolk without glycerol(TE),LDL with glycerol(LDL+)and LDL without glycerol(LDL-),at 80×106 and 240×106 sperm per ml.This study showed that the LDL+ and LDL- extenders were more effective at preserving spermatozoa motility,plasma membrane integrity and acrosome integrity than TEG and TE(P<0.05)during 8 days of incubation.After 3 days of incubation,a toxicity of glycerol was observed in TEG,whereas no significant difference was observed between LDL+ and LDL-.We can therefore conclude that the LDL extender can be used to refrigerate semen at 4 ℃ instead of TEG and TE at 80× 106 and 240×106 sperm per ml for elite bulls.This finding can be used to define a policy for the storage of high-quality bull semen.

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

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    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.

  19. Differential Genes Expression between Fertile and Infertile Spermatozoa Revealed by Transcriptome Analysis.

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    Sandeep Kumar Bansal

    Full Text Available It was believed earlier that spermatozoa have no traces of RNA because of loss of most of the cytoplasm. Recent studies have revealed the presence of about 3000 different kinds of mRNAs in ejaculated spermatozoa. However, the correlation of transcriptome profile with infertility remains obscure.Total RNA from sperm (after exclusion of somatic cells of 60 men consisting of individuals with known fertility (n=20, idiopathic infertility (normozoospermic patients, n=20, and asthenozoospermia (n=20 was isolated. After RNA quality check on Bioanalyzer, AffymetrixGeneChip Human Gene 1.0 ST Array was used for expression profiling, which consisted of >30,000 coding transcripts and >11,000 long intergenic non-coding transcripts.Comparison between all three groups revealed that two thousand and eighty one transcripts were differentially expressed. Analysis of these transcripts showed that some transcripts [ribosomal proteins (RPS25, RPS11, RPS13, RPL30, RPL34, RPL27, RPS5, HINT1, HSP90AB1, SRSF9, EIF4G2, ILF2] were up-regulated in the normozoospermic group, but down-regulated in the asthenozoospermic group in comparison to the control group. Some transcripts were specific to the normozoospermic group (up-regulated: CAPNS1, FAM153C, ARF1, CFL1, RPL19, USP22; down-regulated: ZNF90, SMNDC1, c14orf126, HNRNPK, while some were specific to the asthenozoospermic group (up-regulated: RPL24, HNRNPM, RPL4, PRPF8, HTN3, RPL11, RPL28, RPS16, SLC25A3, C2orf24, RHOA, GDI2, NONO, PARK7; down-regulated: HNRNPC, SMARCAD1, RPS24, RPS24, RPS27A, KIFAP3. A number of differentially expressed transcripts in spermatozoa were related to reproduction (n = 58 and development (n= 210. Some of these transcripts were related to heat shock proteins (DNAJB4, DNAJB14, testis specific genes (TCP11, TESK1, TSPYL1, ADAD1, and Y-chromosome genes (DAZ1, TSPYL1.A complex RNA population in spermatozoa consisted of coding and non-coding RNAs. A number of transcripts that participate in a host of

  20. Mice expressing aberrant sperm-specific protein PMIS2 produce normal-looking but fertilization-incompetent spermatozoa.

    Science.gov (United States)

    Yamaguchi, Ryo; Fujihara, Yoshitaka; Ikawa, Masahito; Okabe, Masaru

    2012-07-01

    Eight kinds of gene-disrupted mice (Clgn, Calr3, Pdilt, Tpst2, Ace, Adam1a, Adam2, and Adam3) show impaired sperm transition into the oviducts and defective sperm binding to the zona pellucida. All of these knockout strains are reported to lack or show aberrant expression of a disintegrin and metallopeptidase domain 3 (ADAM3) on the sperm membrane. We performed proteomic analyses of the proteins of these infertile spermatozoa to clarify whether the abnormal function is caused exclusively by a deficiency in ADAM3 expression. Two proteins, named PMIS1 and PMIS2, were missing in spermatozoa from Clgn-disrupted mice. To study their roles, we generated two gene-disrupted mouse lines. Pmis1-knockout mice were fertile, but Pmis2-knockout males were sterile because of a failure of sperm transport into the oviducts. Pmis2-deficient spermatozoa also failed to bind to the zona pellucida. However, they showed normal fertilizing ability when eggs surrounded with cumulus cells were used for in vitro fertilization. Further analysis revealed that these spermatozoa lacked the ADAM3 protein, but the amount of PMIS2 was also severely reduced in Adam3-deficient spermatozoa. These results suggest that PMIS2 might function both as the ultimate factor regulating sperm transport into the oviducts and in modulating sperm-zona binding.

  1. Penambahan Osteopontin dalam Pengencer Semen Beku Sapi Perah Friesian Holstein Meningkatkan Ekspresi B-Cell Cll/Lymphoma-2 Spermatozoa Postthawing (ADDITIONAL OSTEOPONTIN INTO FROZEN FRIESIAN-HOLSTEIN SEMEN DILUTER INCREASES THE EXPRESSION OF B-CELL CLL/L

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    Abdul Samik

    2015-05-01

    Full Text Available Post thawed frozen semen viability is one of the most important keys in artificial inseminationdepended on two major cell death mechanism, apoptosis and necrosis. It has been known that good fertilitydiary bull’s seminal plasma contains high concentration of osteopontin (OPN. Osteopontin also known ascell survival protein via inhibition to apoptotic cell death, and B-cell CLL/Lymphoma-2 (Bcl-2expressionmostly related to the ability of cell survival. The aim of this study was to investigate the influence ofadditional OPN into frozen semen diluter on post thawed sperm Bcl-2 expression. Fresh semen collectedfrom ±4 year Friesian Holstein bull. Treatment group divided into four groups i.e.: control group (P0:without OPN supplementation, (P1: fresh semen with OPN supplementation 5 ?g/5.107 spermatozoa, (P2:fresh semen with OPN supplementation 10 ?g/5.107 spermatozoa, (P3: fresh semen with OPNsupplementation 20 ?g/5.107spermatozoa. Bcl-2 sperm expression was determined usingimunocytochemistry. The result of this study showed that there was no significant difference betweengroup P0 and P1 (p>0,05, but both group P2 and group P3 showed a significant difference with P0 (p<0,05.Nevertheless, there was no significant difference between group P2to group P3 on post thawed FriesianHolstein sperm Bcl-2 expression (p>0,05. The conclusion of this study is osteopontin supplement in frozensemen diluter is capable to increase post thawed Friesian Holstein sperm Bcl-2 expression.

  2. Sexual polymorphisms of vomeronasal 1 receptor family gene expression in bulls, steers, and estrous and early luteal-phase heifers.

    Science.gov (United States)

    Kubo, Haruna; Otsuka, Midori; Kadokawa, Hiroya

    2016-02-01

    Vomeronasal 1 receptors (V1R) are a family of receptors for intraspecies chemosignals, including pheromones, and are expressed in the olfactory epithelium (OE) and vomeronasal organ (VO). Even in the well-studied rodents, it is unclear which members of the V1R family cause sexual polymorphisms, as there are numerous genes and it is difficult to quantify their expressions individually. Bovine species carry only 34 V1R homologs, and the OE and VOs are large enough to sample. Here, V1R expression was quantified in the OE and VOs of individual bovines. Based on the 34 gene sequences, we obtained a molecular dendrogram consisting of four clusters and six independent branches. Semi-quantitative RT-PCR was used to obtain gene expression profiles in the VOs and OE of 5 Japanese Black bulls, 5 steers, 7 estrous heifers and 6 early luteal-phase heifers. Ten genes showed significant between-group differences, and 22 showed high expression in VOs than in OE. The bulls showed higher expression of one gene more in OE and another in VOs (both Pexpressed more abundantly in steers than in bulls. The estrous heifers showed higher expression of a gene of the second cluster in OE, and a gene of the third cluster in VOs (both Pexpression exhibits sexual polymorphisms in cattle.

  3. Effects of long term storage of semen in liquid nitrogen on the viability, motility and abnormality of frozen thawed Frisian Holstein bull spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Abdul MALIK; Muhammad LAILY; Muhammad Irwan ZAKIR

    2015-01-01

    Objective: To evaluate effects of long term storage of semen in liquid nitrogen on the motility, concentration, viability, and abnormality of frozen-thawed. Methods: A total of four Friesian Holstein bulls were used for this study. One hundred forty semen straws with produced during period from 2008 to 2013 and stored in the liquid nitrogen at the AI center were used in the research. The sample straw was divided into six groups;each group consist 20 semen straws. For group one all straw semen was produced on the 2013 with storage in liquid nitrogen as long as one year, the group 2, 3, 4, 5, and 6 were produced on the 2012, 2011, 2010, 2009 and 2008 with storage in liquid nitrogen as long as 2, 3, 4, 5, and 6 year, respectively. Results:The viability of thawed sperm was not significantly different decreased (P>0.05) between storage on the 1 year and storage on the 2 years. Whereas, the viability was significantly different (P0.05) on the storage 1, 2 and 3 years. Whereas, the motility was significantly different (P0.05) on the storage 1, 2 and 3 years. Whereas, the abnormality was significantly increased (P0.05) during storage in liquid nitrogen as long as six years. Conclusions:Based on the results in these experiments, it may be concluded that concentration sperm during one year storage in liquid nitrogen resulted in similar concentration storage as long as six years. However, the viability and motility sperm thawed storage in liquid nitrogen during six years was lower than storage on the 1 and 2 years.

  4. Differential methylation status of IGF2-H19 locus does not affect the fertility of crossbred bulls but some of the CTCF binding sites could be potentially important.

    Science.gov (United States)

    Jena, Subas C; Kumar, Sandeep; Rajput, Sandeep; Roy, Bhaskar; Verma, Arpana; Kumaresan, Arumugam; Mohanty, Tushar K; De, Sachinandan; Kumar, Rakesh; Datta, Tirtha K

    2014-04-01

    Associations between abnormal methylation of spermatozoan DNA with male infertility have been sought in recent years to identify a molecular explanation of differential spermatozoan function. The present work was undertaken to investigate the methylation profile of differentially methylated regions (DMRs) in the IGF2-H19 locus of Bos taurus X Bos indicus crossbred bull spermatozoa. Bulls having more than at least 100 insemination records over a period of 12 years were classified into two groups of five bulls each belonging to low- and high-fertility groups. The IGF2 and H19 DMR sequences in B. indicus cattle were observed to be in absolute homology with B. taurus cattle. The DNA of crossbred bull spermatozoa was isolated, bisulfite treated, and amplified for specific DMR regions using methylation-change-specific primers. The overall degree of methylation at IGF2-H19 DMRs was not found to be significantly different among two groups of bulls. The sixth CTCF binding site (CCCTC) identified in H19 DMR, however, had a significant methylation difference between the high- and low-fertility bulls. It was concluded that alteration of the methylation levels at IGF2-H19 DMRs might not be responsible for the fertility difference of crossbred bulls, although the role played by the specific CTCF binding sites at this locus, which could influence IGF2 expression during spermatogenesis and early embryonic development, deserves further attention.

  5. Identification and expression of GABAc receptor in rat testis and spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Shifeng Li; Yunbin Zhang; Haixiong Liu; Yuanchang Yan; Yiping Li

    2008-01-01

    Our previous studies showed that γ-aminobutyric acid (GABA)A and GABAB receptors are involved in rat sperm acrosome reaction induced by progesterone or GABA. Here,we report the presence of GABAc receptor in rat testis and spermatozoa.Full-length complementary DNA encoding the ρ1,ρ2 and ρ3 subunits of GABAc receptor were cloned from rat testis;their sequences are identical to those of rat GABAc receptor in retina.Reverse transcription-polymerase chain reaction analysis showed that during the development of rat testis,the transcript levels of the ρ1 and ρ2 subunits showed little change,while the expression of ρ3 was gradually up-regulated.Immunofluorescence analysis using an anti-ρ1 antibody revealed that GABAc receptor exists on the elongated spermatid and sperm.Using a chlortetracycline assay,we found that N(4)-chloroacetylcytosine arabinoside, a GABAc receptor agonist,triggered rat sperm acrosome reaction;whereas(1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid,a GABAc receptor antagonist, inhibited the ability of N(4)-chloroacetylcytosine arabinoside to induce acrosome reaction.These results suggested that GABAc receptors are also involved in rat sperm acrosome reaction.

  6. 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.

  7. Differential expression of mRNA aromatase in ejaculated spermatozoa from infertile men in relation to either asthenozoospermia or teratozoospermia.

    Science.gov (United States)

    Said, L; Saad, A; Carreau, S

    2014-03-01

    Oestrogen biosynthesis in ejaculated spermatozoa is an autonomous process, which may influence sperm functions. The purpose of this study was to evaluate the relationship between the expression of aromatase, sperm quality and seminal neutral α-glucosidase marker in semen of Tunisian infertile men: asthenozoospermia (A; n = 16), teratozoospermia (T; n = 12) and asthenoteratozoospermia (AT; n = 11) in comparison with 18 normozoospermic ones. Aromatase mRNA levels estimated by real-time PCR were reduced in groups T (52%) and AT (67%) compared to controls and inversely correlated with the percentage of normal forms. A higher coefficient of correlation was noted in presence of microcephaly or acrosome malformations (r = -0.64). The asthenozoospermic group was divided into two subgroups according to the relative amount of aromatase. The subgroup (A2) with higher aromatase transcript level was associated with an increased seminal pH, a decreased sperm viability, low sperm percentage motility and low neutral α-glucosidase semen levels. Our data highlight the involvement of aromatase in motility and morphology of spermatozoa. Thus, this enzyme could bring new insights about quality and fertilising capacity of human spermatozoa.

  8. Abnormal expression of centrosome protein (centrin) in spermatozoa of male human infertility

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    To study the relations between male infertility and centrosome protein (centrin) and the functions of centrin in spermatogenesis, the matured spermatozoa of 10 normal male people and 18 male infertility patients were stained by immunofluorescence labeling antibody against centrin. The results showed that two fluorescence signal dots appeared in the normal male spermatozoa and were located at the base of flagellum. They are proximal centriole and distal centriole. However, in some spermatozoa of the male infertility, centrin protein was located abnormally at the base of flagellum and its staining signals were spread, the normal proximal and distal centrioles were confused and could not be recognized separately. These results suggest that abnormality of centrosome protein may be related to male infertility. This discovery may be used as a marker of abnormal sperm and male infertility.

  9. 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...

  10. Bull Fertility and Its Relation with Density Gradient Selected Sperm

    Directory of Open Access Journals (Sweden)

    Allouche Lynda ,

    2017-01-01

    Full Text Available Background Sperm selection method is usually used to collect these cells for in vitro-assisted reproduction. Few studies reported the relationship of in vivo fertility and semen parameters after sperm selection; hence, the present study attempted to assess different semen parameters after post-thaw or sperm selection, using density gradient separation BoviPure®, to predict in vivo fertility. Materials and Methods In this experimental study, frozen semen quality of four Montbeliarde bulls were assessed after post-thaw (PT or after sperm selection (SSp, using density gradient separation BoviPure®, to predict the fertility rate in vivo. In addition to PT or SSp, semen was examined for concentration, motility, morphology abnormalities, viability, acrosome and plasma membrane integrities. Fertility was measured as non-return rates within 56 days after the first insemination (NRR or as corrected NRR, expressed as CNRR, to the factors influencing fertility using linear mixed model. Non-parametric Kruskal-Wallis test was performed to compare semen parameter variables. Fertility rates were compared using Chi-square test. Pearson correlation analysis was used to test the relationship between CNRR and semen parameters. Data was analysed using SPSS package program, version 21.0. Results Most of the examined bulls exhibited a high fertility rate (3/4 bulls, 62.1- 81.8% for NRR or 67.2-98.5% for CNRR. Fertility rate, expressed as CNRR, was significantly related to semen parameters after SSp, but not after PT. Thus, CNRR was increased with decrease of total motility, progressive spermatozoa and abaxial implantation frequencies after SSp (r=-0.999, P=0.001; r=-0.990, P=0.010; r=-0.988, P= 0.012, respectively; while, CNRR was decreased with decrease of SSp immotile spermatozoa (r=+0.995, P=0.005, underlying that maximal limit of determined immotile spermatozoa is 47%. Conclusion High frequencies of total and progressive motility spermatozoa, and abaxial

  11. Identification of differentially expressed proteins in fresh and frozen-thawed boar spermatozoa by iTRAQ-coupled 2D LC-MS/MS.

    Science.gov (United States)

    Chen, Xiaoli; Zhu, Huabin; Hu, Chuanhuo; Hao, Haisheng; Zhang, Junfang; Li, Kunpeng; Zhao, Xueming; Qin, Tong; Zhao, Kan; Zhu, Huishan; Wang, Dong

    2014-03-01

    Cryodamage is a major problem in semen cryopreservation, causing changes in the levels of proteins that influence the function and motility of spermatozoa. In this study, protein samples prepared from fresh and frozen-thawed boar spermatozoa were compared using the isobaric tags for relative and absolute quantification (iTRAQ) labeling technique coupled to 2D LC-MS/MS analysis. A total of 41 differentially expressed proteins were identified and quantified, including 35 proteins that were present at higher levels and six proteins that were present at lower levels in frozen-thawed spermatozoa by at least a mean of 1.79-fold (Psperm premature capacitation, adhesions, energy supply, and sperm-oocyte binding and fusion. The expression of four of these proteins, SOD1, TPI1, ODF2, and AKAP3, was verified by western blot analysis. We propose that alterations in these identified proteins affect the quality of cryopreserved semen and ultimately lower its fertilizing capacity. This is the first study to compare protein levels in fresh and frozen-thawed spermatozoa using the iTRAQ technology. Our preliminary results provide an overview of the molecular mechanisms of cryodamage in frozen-thawed spermatozoa and theoretical guidance to improve the cryopreservation of boar semen.

  12. Low long non-coding RNA HOTAIR expression is associated with down-regulation of Nrf2 in the spermatozoa of patients with asthenozoospermia or oligoasthenozoospermia.

    Science.gov (United States)

    Zhang, Lixin; Liu, Zhineng; Li, Xiaokang; Zhang, Ping; Wang, Jia; Zhu, Dandan; Chen, Xinping; Ye, Lihua

    2015-01-01

    HOTAIR, a long noncoding RNA, regulates development and progression of tumor cells and function of normal stem cells. However, the role and the molecular mechanism of HOTAIR in the spermatozoa of patients with asthenozoospermia and oligoasthenozoospermia are still unclear. Herein, 45 healthy control, 45 asthenozoospermic patients and 45 oligoasthenozoospermic patients were enrolled. Initially, through analyzing HOTAIR expression, we observed a decreased level of HOTAIR expression in patients. Subsequently, we found that there was a positive correlation between HOTAIR expression and Nrf2 expression in patients. The low expression of HOTAIR was also observed to be associated with specific sperm function parameters, including motility and vitality. In the ejaculated spermatozoa from patients, low level of histone H4 acetylation of the Nrf2 gene promoter was observed. Finally, we found that downregulation of HOTAIR expression reduced histone H4 acetylation in Nrf2 promoter and Nrf2 expression. Therefore, this study demonstrated that HOTAIR expression was low in the spermatozoa of patients with asthenozoospermia and oligoasthenozoospermia, which resulted in down-regulation of Nrf2 expression. Our data suggested the decrease of HOTAIR expression led to ROS related defects in sperm function.

  13. Viable and morphologically normal boar spermatozoa alter the expression of heat-shock protein genes in oviductal epithelial cells during co-culture in vitro.

    Science.gov (United States)

    Yeste, Marc; Holt, William V; Bonet, Sergi; Rodríguez-Gil, Joan E; Lloyd, Rhiannon E

    2014-09-01

    The principal aim of this study was to determine if boar spermatozoa influence the expression of four selected chaperone and heat-shock protein (HSP) genes-namely clusterin (CLU), HSP90AA1, HSPA5, and HSPA8-in oviductal epithelial cells (OECs) during in vitro co-culture. All corresponding proteins of these genes were previously identified in a sperm-interacting, 70-kDa soluble fraction derived from apical plasma membranes of OECs. The present study also sought to determine whether or not: (i) spermatozoa must directly bind to OEC for an effect on gene expression to be elicited and (ii) reproductive and nonreproductive epithelial cell types (LLC-PK1, pig kidney) respond equivalently, in terms of alterations in chaperone and HSP gene expression, during co-culture with sperm. Spermatozoa induced a significant upregulation (P sperm-binding index and on the viability and morphological quality of the bound sperm population. In conclusion, the upregulation of HSP genes caused by direct contact between spermatozoa and OECs, rather than nonreproductive epithelial cells, suggests HSPs could play an integral role in the modulation of sperm function in the oviductal reservoir.

  14. The expression of cysteine-rich secretory protein 2 (CRISP2) and its specific regulator miR-27b in the spermatozoa of patients with asthenozoospermia.

    Science.gov (United States)

    Zhou, Jun-Hao; Zhou, Qi-Zhao; Lyu, Xiao-Ming; Zhu, Ting; Chen, Zi-Jian; Chen, Ming-Kun; Xia, Hui; Wang, Chun-Yan; Qi, Tao; Li, Xin; Liu, Cun-Dong

    2015-01-01

    Cysteine-rich secretory protein 2 (CRISP2) is an important sperm protein and plays roles in spermatogenesis, modulation of flagellar motility, acrosome reaction, and gamete fusion. Clinical evidence shows a reduced CRISP2 expression in spermatozoa from asthenozoospermic patients, but the molecular mechanism underlying its reduction remains unknown. Herein, we carried out a study focusing on the CRISP2 reduction and its roles in asthenozoospermia. Initially, through analyzing CRISP2 expression and methylation on CRISP2 promoter activity in sperm, we observed a decreased expression of CRISP2 protein rather than its mRNA in the ejaculated spermatozoa from asthenozoospermic patients and no methylation in the CRISP2 promoter, suggesting CRISP2 expression may be regulated in the sperm at the posttranscriptional level. Subsequently, we found that microRNA 27b (miR-27b), predicted as a candidate regulator of CRISP2 using bioinformatics, was highly expressed in the ejaculated spermatozoa from asthenozoospermic patients. Luciferase reporter assay and transfection experiments disclosed that this microRNA could target CRISP2 by specifically binding its 3' untranslated region, suppressing CRISP2 expression. Extended clinical observation further confirmed a highly expressed miR-27b and its obviously negative correlation with CRISP2 protein expression in ejaculated spermatozoa samples from asthenozoospermic patients. Finally, we conducted a retrospective follow-up study to support that either high miR-27b expression or low CRISP2 protein expression was significantly associated with low sperm progressive motility, abnormal morphology, and infertility. Thus, this study provides the first preliminary insight into the mechanism leading to the reduced CRISP2 expression in asthenozoospermia, offering a potential therapeutic target for treating male infertility or for male contraception.

  15. 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.

  16. Microphotokinesigraphic analysis of buffalo spermatozoa II. Photokinesigraphic characteristics of buffalo vibrating spermatozoa

    Directory of Open Access Journals (Sweden)

    M. Boichev

    2010-02-01

    Full Text Available The study concerns images of vibrating spermatozoa in the ejaculation of buffalo bulls from Bulgarian murrah breed through the method of Tchakarov and Natchev [1962], obtained at 5 sec exposures, optimal for this particular species. The negative registration material has been developed with high-equalising developer by Windisch, which reduces the glow due to overexposure around highaktinic parts of spermatozoa. On the basis of the positive graphic material – kinesigrams, we have established and analysed the main movement patterns of vibrating spermatozoa: with vibrating tail; with vibrating head; with a vibrating tail and head; with whole bilateral vibration; vibrating in circle; chaotically vibrating. We have described the differences in photokinesigraphic images of spermatozoa within given subclass.

  17. AB035. The expression of cysteine-rich secretory protein 2 (CRISP2) and its specific regulator mir-27b in the spermatozoa of patients with asthenozoospermia

    Science.gov (United States)

    Zhou, Jun-Hao; Zhou, Qi-Zhao; Lyu, Xiao-Ming; Zhu, Ting; Chen, Zi-Jian; Chen, Ming-Kun; Xia, Hui; Wang, Chun-Yan; Qi, Tao; Li, Xin; Liu, Cun-Dong

    2016-01-01

    Background Cysteine-rich secretory protein 2 (CRISP2) is an important sperm protein and plays roles in spermatogenesis, modulation of flagellar motility, acrosome reaction, and gamete fusion. Clinical evidence shows a reduced CRISP2 expression in spermatozoa from asthenozoospermic patients, but the molecular mechanism underlying its reduction remains unknown. Herein, we carried out a study focusing on the CRISP2 reduction and its roles in asthenozoospermia Methods Spermatozoa were isolated from 90 study subjects’ ejaculated semen samples. DNA methylation was evaluated using bisulfite-sequencing PCR and methylation-specific PCR. The CRISP2 mRNA and protein expression levels were examined in the ejaculated spermatozoa by qRT-PCR and Western blot respectively. miRNA expression was detected by qRT-PCR. The direct regulatory effect of miR-27b on CRISP2 was predicted computationally and validated via luciferase reporter assay and in vitro experiments in which miR-27b mimic or inhibitor was transfected into 293T cells. Respective correlations of miR-27b and CRISP2 protein expression with clinical features were analyzed using Spearman’s correlation coefficient. Results Initially, low expression of CRISP2 protein rather than its mRNA was observed in the ejaculated spermatozoa from asthenozoospermic patients relative to normozoospermic males. Meanwhile, methylation was not found in CRISP2 promoter. These data suggest a possible post-transcriptional regulation of CRISP2 in asthenozoospermia. Subsequently, bioinformatics prediction, luciferase reporter assay and miR-27b transfection experiments revealed that miR-27b could specifically target CRISP2 by binding its 3’-UTR, suppressing CRISP2 expression post-transcriptionally. Further evidence was provided by the clinical observation of a high miR-27b expression in the ejaculated spermatozoa from asthenozoospermic patients and a negative correlation between miR-27b and CRISP2 protein expression. Finally, a retrospective

  18. The bull sperm microRNAome and the effect of fescue toxicosis on sperm microRNA expression.

    Science.gov (United States)

    Stowe, Heather M; Calcatera, Samantha M; Dimmick, Marcy A; Andrae, John G; Duckett, Susan K; Pratt, Scott L

    2014-01-01

    Tall fescue [Schedonorus phoenix (Scop.) Holub] accounts for nearly 16 million hectares of pasture in the Southeastern and Mid-Atlantic U.S. due to its heat, drought, and pest resistance, conferred to the plant by its symbiotic relationship with the endophyte Neotyphodium coenophialum. The endophyte produces ergot alkaloids that have negative effects on the growth and reproduction of animals, resulting in the syndrome known as fescue toxicosis. The objectives of our study were to identify microRNA (miRNA) present in bovine sperm and to evaluate the effects of fescue toxicosis on sperm miRNA expression. Angus bulls were assigned to treatments of either toxic or non-toxic fescue seed diets. Semen was collected and subjected to RNA isolation. Three samples from each treatment group were chosen and pooled for deep sequencing. To compare miRNA expression between treatment groups, a microarray was designed and conducted. For each of the top ten expressed miRNA, target prediction analysis was conducted using TargetScan. Gene ontology enrichment was assessed using the Database for Annotation, Visualization and Integrated Discovery. Sequencing results elucidated the presence of 1,582 unique small RNA present in sperm. Of those sequences, 382 were known Bos taurus miRNA, 22 were known but novel to Bos taurus, and 816 were predicted candidate miRNA that did not map to any currently reported miRNA. Of the sequences chosen for microarray, twenty-two showed significant differential expression between treatment groups. Gene pathways of interest included: regulation of transcription, embryonic development (including blastocyst formation), Wnt and Hedgehog signaling, oocyte meiosis, and kinase and phosphatase activity. MicroRNA present in mature sperm appears to not only be left over from spermatogenic processes, but may actually serve important regulatory roles in fertilization and early developmental processes. Further, our results indicate the possibility that environmental

  19. The bull sperm microRNAome and the effect of fescue toxicosis on sperm microRNA expression.

    Directory of Open Access Journals (Sweden)

    Heather M Stowe

    Full Text Available Tall fescue [Schedonorus phoenix (Scop. Holub] accounts for nearly 16 million hectares of pasture in the Southeastern and Mid-Atlantic U.S. due to its heat, drought, and pest resistance, conferred to the plant by its symbiotic relationship with the endophyte Neotyphodium coenophialum. The endophyte produces ergot alkaloids that have negative effects on the growth and reproduction of animals, resulting in the syndrome known as fescue toxicosis. The objectives of our study were to identify microRNA (miRNA present in bovine sperm and to evaluate the effects of fescue toxicosis on sperm miRNA expression. Angus bulls were assigned to treatments of either toxic or non-toxic fescue seed diets. Semen was collected and subjected to RNA isolation. Three samples from each treatment group were chosen and pooled for deep sequencing. To compare miRNA expression between treatment groups, a microarray was designed and conducted. For each of the top ten expressed miRNA, target prediction analysis was conducted using TargetScan. Gene ontology enrichment was assessed using the Database for Annotation, Visualization and Integrated Discovery. Sequencing results elucidated the presence of 1,582 unique small RNA present in sperm. Of those sequences, 382 were known Bos taurus miRNA, 22 were known but novel to Bos taurus, and 816 were predicted candidate miRNA that did not map to any currently reported miRNA. Of the sequences chosen for microarray, twenty-two showed significant differential expression between treatment groups. Gene pathways of interest included: regulation of transcription, embryonic development (including blastocyst formation, Wnt and Hedgehog signaling, oocyte meiosis, and kinase and phosphatase activity. MicroRNA present in mature sperm appears to not only be left over from spermatogenic processes, but may actually serve important regulatory roles in fertilization and early developmental processes. Further, our results indicate the possibility that

  20. Semen and reproductive profiles of genetically identical cloned bulls.

    Science.gov (United States)

    Tecirlioglu, R Tayfur; Cooney, Melissa A; Korfiatis, Natasha A; Hodgson, Renee; Williamson, Mark; Downie, Shara; Galloway, David B; French, Andrew J

    2006-06-01

    In this comparative study, reproductive parameters and semen characteristics of cloned bulls (n = 3) derived from somatic cell nuclear transfer (SCNT) were compared to their original cell donor Holstein-Friesian (n = 2) bulls from the same enterprise to assess the differences in reproductive potential between a donor bull and its clones. The parameters evaluated included motility of fresh, frozen-thawed and Percoll-treated frozen-thawed spermatozoa, as well as in vitro fertilization (IVF) ability, embryo quality, birth and survival of calves following IVF and embryo transfer with frozen-thawed semen. With fresh semen, spermatozoa from one cloned bull had lower motility than its donor. Cloned bulls had higher velocity parameters in fresh semen, but those effects were not obvious in frozen-thawed or frozen-thawed semen selected with a Percoll gradient. Semen collected from cloned bulls had significantly higher IVF rates compared to donors; however, embryo development per cleaved embryo or quality of blastocysts did not differ between donors and cloned bulls. Pregnancy and live offspring rates from one donor and its cloned bull did not differ between fresh (40%, 16/40 versus 46%, 17/37) and vitrified/thawed (13%, 2/16 versus 25%, 4/16) embryo transfer following IVF. A total of 26 calves were obtained from genotypically identical donor and cloned bulls with no signs of phenotypical abnormalities. These preliminary results suggested that the physiology of surviving postpubertal cloned bulls and quality of collected semen had equivalent reproductive potential to their original cell donor, with no evidence of any deleterious effects in their progeny.

  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. Identification and evaluation of reference genes for accurate gene expression normalization of fresh and frozen-thawed spermatozoa of water buffalo (Bubalus bubalis).

    Science.gov (United States)

    Ashish, Shende; Bhure, S K; Harikrishna, Pillai; Ramteke, S S; Muhammed Kutty, V H; Shruthi, N; Ravi Kumar, G V P P S; Manish, Mahawar; Ghosh, S K; Mihir, Sarkar

    2017-04-01

    The quantitative real time PCR (qRT-PCR) has become an important tool for gene-expression analysis for a selected number of genes in life science. Although large dynamic range, sensitivity and reproducibility of qRT-PCR is good, the reliability majorly depend on the selection of proper reference genes (RGs) employed for normalization. Although, RGs expression has been reported to vary considerably within same cell type with different experimental treatments. No systematic study has been conducted to identify and evaluate the appropriate RGs in spermatozoa of domestic animals. Therefore, this study was conducted to analyze suitable stable RGs in fresh and frozen-thawed spermatozoa. We have assessed 13 candidate RGs (BACT, RPS18s, RPS15A, ATP5F1, HMBS, ATP2B4, RPL13, EEF2, TBP, EIF2B2, MDH1, B2M and GLUT5) of different functions and pathways using five algorithms. Regardless of the approach, the ranking of the most and the least candidate RGs remained almost same. The comprehensive ranking by RefFinder showed GLUT5, ATP2B4 and B2M, MDH1 as the top two stable and least stable RGs, respectively. The expression levels of four heat shock proteins (HSP) were employed as a target gene to evaluate RGs efficiency for normalization. The results demonstrated an exponential difference in expression levels of the four HSP genes upon normalization of the data with the most stable and the least stable RGs. Our study, provides a convenient RGs for normalization of gene-expression of key metabolic pathways effected during freezing and thawing of spermatozoa of buffalo and other closely related bovines.

  3. 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

  4. Red Bull Crisis

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    Additives make Red Bull the first company to test China’s new Food Safety Law "Drink Red Bull when you feel sleepy or tired" is a famous advertising slogan well-known to Chinese. But a recent German

  5. 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

  6. Increased conception rates in beef cattle inseminated with nanopurified bull semen.

    Science.gov (United States)

    Odhiambo, John F; DeJarnette, J M; Geary, Thomas W; Kennedy, Chelsey E; Suarez, Susan S; Sutovsky, Miriam; Sutovsky, Peter

    2014-10-01

    Aberrant sperm phenotypes coincide with the expression of unique sperm surface determinants that can be probed by objective, biomarker-based semen analysis and targeted as ligands for semen purification. This study evaluated a nanoparticle-based magnetic purification method that removes defective spermatozoa (∼30% of sample) from bull semen and improves sperm sample viability and fertilizing ability in vitro and in vivo. Two types of nanoparticles were developed: a particle coated with antibody against ubiquitin, which is present on the surface of defective spermatozoa, and a particle coated with the lectin peanut agglutinin, which binds to glycans exposed by acrosomal damage. In a 2 yr artificial insemination field trial with 798 cows, a conception rate of 64.5% ± 3.7% was achieved with a 10 × 10(6) sperm dose of peanut agglutinin-nanopurified spermatozoa, comparable to a control nonpurified full dose of 20 × 10(6) spermatozoa per dose (63.3% ± 3.2%) and significantly higher than a 10 × 10(6) sperm dose of nonpurified control semen (53.7% ± 3.2%; P < 0.05). A total of 466 healthy calves were delivered, and no negative side effects were observed in the inseminated animals or offspring. Because the method is inexpensive and can be fully integrated in current protocols for semen cryopreservation, it is feasible for use in the artificial insemination industry to improve fertility with reduced sperm dosage inseminations. Spermatology will benefit from nanopurification methodology by gaining new tools for the identification of candidate biomarkers of sperm quality such as binder of sperm protein 5 (BSP5), described in the present study.

  7. Seminal Plasma Characteristics and Expression of ATP-binding Cassette Transporter A1 (ABCA1) in Canine Spermatozoa from Ejaculates with Good and Bad Freezability.

    Science.gov (United States)

    Schäfer-Somi, S; Palme, N

    2016-04-01

    The composition of seminal plasma and the localization of the ATP-binding cassette transporter A1 (ABCA1) in spermatozoa from good and bad freezers were compared to frozen-thawed spermatozoa from the same dog. Ejaculates were obtained from 31 stud dogs, and the sperm-rich fraction (SRF) was kept for analysis. One aliquot was used for the analysis of concentration, progressive motility (P; CASA), viability (V; CASA) and leucocyte count, and the analysis was performed by flow cytometry (FITC-PNA/PI), SCSA and HOST. In seminal plasma, concentration of albumin, cholesterol, calcium, inorganic phosphate, sodium, potassium, zinc and copper was measured. Semen smears were prepared and evaluated for the expression of ABCA1. The remainder of each ejaculate was frozen. After thawing, the quality assessment was repeated and further smears were prepared. According to post-thaw semen quality, dogs were assigned to good freezers (n = 20) or bad freezers (n = 11), the latter were defined as 40% morphologically abnormal sperm and/or Bad freezers were older than good freezers (5.3 vs 3.4 years, p bad freezers, the percentage of sperm with ABCA1 signal in the acrosome was lower (26.3% vs 35.7%, p good freezers (p bad freezer sperm, an increase in acrosome damages coincided with an increased loss of cholesterol transporters and cell death, and a lower cholesterol concentration in seminal plasma. Follow-up studies revealed whether a relation exists between these findings.

  8. Dense spermatozoa in stallion ejaculates contain lower concentrations of mRNAs encoding the sperm specific calcium channel 1, ornithine decarboxylase antizyme 3, aromatase, and estrogen receptor alpha than less dense spermatozoa.

    Science.gov (United States)

    Ing, N H; Forrest, D W; Love, C C; Varner, D D

    2014-07-15

    Stallions are unique among livestock in that, like men, they commonly receive medical treatment for subfertility. In both species, about 15% of individuals have normal semen parameters but are subfertile, indicating a need for novel analyses of spermatozoa function. One procedure for improving fertilizing capability of stallions and men is isolation of dense spermatozoa from an ejaculate for use in artificial insemination. In the current study, dense and less dense spermatozoa were purified by density gradient centrifugation from individual ejaculates from seven reproductively normal adult stallions. The RNA isolated from the spermatozoa seemed to be naturally fragmented to an average length of 250 bases, consistent with reports of spermatozoa RNA from other species. The DNAse treatment of RNA prepared from spermatozoa removed any genomic DNA contamination, as assessed by PCR with intron spanning primers for the protamine 1 (PRM1) gene. Concentrations of seven mRNAs in spermatozoa, correlated with the fertility of men and bulls, were quantified by reverse transcription polymerase chain reaction in dense and less dense spermatozoa. Concentrations of four mRNAs were two- to four-fold lower in dense spermatozoa compared with less dense spermatozoa: Encoding the spermatozoa-specific calcium channel (P 0.1). These results identify new differences in mRNA concentrations in populations of spermatozoa with dissimilar densities.

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

    OpenAIRE

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

    2015-01-01

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

  10. Microphotokinesigraphic analysis of buffalo spermatozoa I. Characteristic features and significance of the immobile buffalo spermatozoa

    Directory of Open Access Journals (Sweden)

    M. Boichev

    2010-02-01

    Full Text Available The article discusses the results of a study of immobile spermatozoa of buffalo bulls from Bulgarian murrah breed, based on photo images registered in various displays in conditions of dark background photokinesis. The analysis showed the way developer affects the brightness of kinesigrams. The study is based on samples of semen of six buffalos analysed by means of the microphotokinetic method developed by Tchakarov and Natchev. Brightness parameters of the cells and their photokinetic images have been established photometrically; their metric parameters have been defined by means of an eyepiece and object micrometer. It has been found out that the optimal exposure time for a photokinetic analysis of buffalo spermatozoa was(is 5 sec, and the recommended developer for treating of negative registration material is the pyrocatechin developer by Windisch.

  11. A g.-1256 A>C in the promoter region of CAPN1 is associated with semen quality traits in Chinese Holstein bulls.

    Science.gov (United States)

    Cui, Xiaohui; Sun, Yan; Wang, Xiuge; Yang, Chunhong; Ju, Zhihua; Jiang, Qiang; Zhang, Yan; Huang, Jinming; Zhong, Jifeng; Yin, Miao; Wang, Changfa

    2016-07-01

    The micromolar calcium-activated neutral protease gene (CAPN1) is a physiological candidate gene for sperm motility. However, the molecular mechanisms involved in regulating the expression of the CAPN1 gene in bulls remain unknown. In this study, we investigated the expression pattern of CAPN1 in testis, epididymis, and sperm at the RNA and protein levels by qRT-PCR, western blot, immunohistochemistry, and immunofluorescence assay. Results revealed that the expression of CAPN1 levels was higher in the sperm head compared with that in other tissues. Moreover, we identified a novel single-nucleotide polymorphism (g.-1256 A>C, ss 1917715340) in the noncanonical core promoter of the CAPN1 gene between base g.-1306 and g.-1012. Additionally, we observed greater sperm motility in bulls with the genotype CC than in those with the genotype AA (PC was revealed by transient transfection in MLTC-1 cells and luciferase report assay. Finally, CAPN1 was highly expressed in the spermatozoa with the CC genotype compared with that with the AA genotype by qRT-PCR. This study is the first report on genetic variant g.-1256 A>C in the promoter region of CAPN1 gene association with the semen quality of Chinese Holstein bulls by influencing its expression. g.-1256 A>C can be a functional molecular marker in cattle breeding.

  12. Sperm morphometry: a tool for detecting biophysical changes associated with viability in cryopreserved bovine spermatozoa.

    Science.gov (United States)

    García-Herreros, M; Leal, C L V

    2014-09-01

    The aim of this study was to determine whether computerised sperm head morphometric analysis can be used as a diagnostic tool for detecting biophysical changes associated with sperm viability in frozen-thawed bovine spermatozoa. Ejaculates from five bulls (4 ejaculates/bull) were pooled and processed for computerised morphometric analysis, and SYBR-14 green/ethidium homodimer-1 fluorescence-based live/dead viability assay was used simultaneously to confirm the viability index of frozen-thawed spermatozoa. Sperm samples were assigned to three experimental groups. The first group was enriched in live spermatozoa (after a double Percoll selection), the second group was enriched in dead spermatozoa (after a refreeze-thaw procedure), and the last group was a 50 : 50 pool of live/dead spermatozoa (from first and second group samples). There were significant differences (P sperm morphometric dimensional parameters among the three groups analysed, being the lowest overall sperm head dimension found in the second (dead spermatozoa) group. In conclusion, sperm head morphometry can be used as a potential diagnostic tool for detecting biophysical changes associated with sperm viability in frozen-thawed bovine spermatozoa.

  13. Seasonal and Ageing-Depending Changes of Aquaporins 1 and 9 Expression in the Genital Tract of Buffalo Bulls (Bubalus bubalis).

    Science.gov (United States)

    Arrighi, S; Bosi, G; Accogli, G; Desantis, S

    2016-08-01

    The presence of Aquaporins 1 (AQP1) and 9 (AQP9), integral membrane water channels that facilitate rapid passive movement of water and solutes, was immunohistochemically detected in the excurrent ducts collected from sexually mature buffalo bulls of proven fertility during the mating (late autumn-winter) and non-mating (late spring to the beginning of autumn) seasons. Furthermore, the research was performed also on the epididymal cauda of a senile buffalo bull with inactive testis. Aquaporins 1 and 9 were immunolocalized at distinct levels. In the efferent ducts, AQP1 immunoreactivity was strongly evidenced at the apical surface of the non-ciliated cells and weakly along the basal membrane of the epithelial cells. The latter reactivity disappeared during the non-mating season. No AQP1 immunoreactivity was detected in the epithelium of epididymis and vas deferens, whereas AQP1 was expressed in the smooth muscle layer of the vas deferens. Aquaporin 1 was present in the blood vessels and in small nerve bundles all along the genital tract. The supranuclear zone of the epididymal principal cells was AQP9 immunoreactive, limited to the corpus and cauda regions, and vas deferens. The samples collected in the two reproductive seasons showed a weaker AQP9 immunoreactivity during the non-mating season. A typical AQP9 immunoreactivity was noticed in the old buffalo examined. The tested AQP molecules showed a different expression pattern in comparison with laboratory mammals, primates, equine, dog and cat. In addition, seasonal differences were noticed which are possibly useful in regard to the comprehension of the morphophysiology of reproduction in the bubaline species, which are still a matter of debate.

  14. Adapting Bulls to Florida

    Science.gov (United States)

    The adaptation of bulls used for natural breeding purposes to the Gulf Coast region of the United States including all of Florida is an important topic. Nearly 40% of the U.S. cow/calf population resides in the Gulf Coast and Southeast. Thus, as A.I. is relatively rare, the number of bulls used for ...

  15. BullsEye

    DEFF Research Database (Denmark)

    Klokmose, Clemens Nylandsted; Kristensen, Janus Bager; Bagge, Rolf

    2014-01-01

    implemented primarily in shaders on the GPU. The techniques are realized in the BullsEye computer vision software. We demonstrate experimentally that BullsEye provides sub-pixel accuracy down to a tenth of a pixel, which is a significant improvement compared to the commonly used reacTIVision software....

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

    NARCIS (Netherlands)

    Duijn, C. van

    1962-01-01

    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 of

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

    Directory of Open Access Journals (Sweden)

    Bonet Sergi

    2011-06-01

    Full Text Available Abstract 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

  18. 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...

  19. 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...

  20. AB087. The expression of cysteine-rich secretory protein 2 (CRISP2) and its specific regulator miR-27b in the spermatozoa of patients with asthenozoospermia

    Science.gov (United States)

    Zhou, Junhao; Zhou, Qizhao; Lyu, Xiaoming; Zhu, Ting; Chen, Zijian; Chen, Mingkun; Xia, Hui; Wang, Chunyan; Qi, Tao; Li, Xin; Liu, Cundong

    2016-01-01

    Objective Cysteine-rich secretory protein 2 (CRISP2) is an important sperm protein and plays roles in spermatogenesis, modulation of flagellar motility, acrosome reaction, and gamete fusion. Clinical evidence shows a reduced CRISP2 expression in spermatozoa from asthenozoospermic patients, but the molecular mechanism underlying its reduction remains unknown. Herein, we carried out a study focusing on the CRISP2 reduction and its roles in asthenozoospermia. Methods Spermatozoa were isolated from 90 study subjects’ ejaculated semen samples. DNA methylation was evaluated using bisulfite-sequencing PCR and methylation-specific PCR. The CRISP2 mRNA and protein expression levels were examined in the ejaculated spermatozoa by qRT-PCR and Western blot respectively. MiRNA expression was detected by qRT-PCR. The direct regulatory effect of miR-27b on CRISP2 was predicted computationally and validated via luciferase reporter assay and in vitro experiments in which miR-27b mimic or inhibitor was transfected into 293T cells. Respective correlations of miR-27b and CRISP2 protein expression with clinical features were analyzed using Spearman’s correlation coefficient. Results Initially, low expression of CRISP2 protein rather than its mRNA was observed in the ejaculated spermatozoa from asthenozoospermic patients relative to normozoospermic males. Meanwhile, methylation was not found in CRISP2 promoter. These data suggest a possible post-transcriptional regulation of CRISP2 in asthenozoospermia. Subsequently, bioinformatics prediction, luciferase reporter assay and miR-27b transfection experiments revealed that miR-27b could specifically target CRISP2 by binding its 3’-UTR, suppressing CRISP2 expression post-transcriptionally. Further evidence was provided by the clinical observation of a high miR-27b expression in the ejaculated spermatozoa from asthenozoospermic patients and a negative correlation between miR-27b and CRISP2 protein expression. Finally, a retrospective

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

    Science.gov (United States)

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

    2016-01-01

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

  2. Selenium in bovine spermatozoa.

    Science.gov (United States)

    Niemi, S M; Kuzan, F B; Senger, P L

    1981-05-01

    This study investigated the association of selenium with ejaculated bovine spermatozoa. Over 75% of the radioactive spermatozoa. Over 75% of the radioactive selenium-75 was released after 30 min of incubation in 2 X 10(-3) dithiothreitol. Of the selenium-75 released by dithiothreitol, 85% was associated with spermatozoal protein. Protein containing selenium-75 was found predominantly in a single band after polyacrylamide gel electrophoresis. Molecular weight was approximately 21,500 daltons.

  3. Identification of MicroRNAs in Zebrafish Spermatozoa.

    Science.gov (United States)

    Jia, Kun-Tong; Zhang, Jing; Jia, Peng; Zeng, Lin; Jin, Yilin; Yuan, Yongming; Chen, Jieying; Hong, Yunhan; Yi, Meisheng

    2015-12-01

    MicroRNAs (miRNAs) participate in almost all biological processes. Plenty of evidences show that some testis- or spermatozoa-specific miRNAs play crucial roles in the process of gonad and germ cell development. In this study, the spermatozoa miRNA profiles were investigated through a combination of illumina deep sequencing and bioinformatics analysis in zebrafish. Deep sequencing of small RNAs yielded 11,820,680 clean reads. By mapping to the zebrafish genome, we identified 400 novel and 204 known miRNAs that could be grouped into 104 families. Furthermore, we selected the six highest expressions of known miRNAs to detect their expression patterns in different tissues by stem-loop quantitative real-time polymerase chain reaction. We found that among the six miRNAs, dre-miR-202-5p displayed specific and high expression in zebrafish spermatozoa and testis. Fluorescence in situ hybridization analysis indicated that dre-miR-202-5p was predominantly expressed in all kind of germ cells at different spermatogenetic stages, including spermatogonia and spermatozoa, but barely expressed in the germ cells in the ovary. This sex-biased expression pattern suggests that dre-miR-202-5p might be related to spermatogenesis and the functioning of spermatozoa. The identification of miRNAs in zebrafish spermatozoa and germ cells offers new insights into the spermatogenesis and spermatozoa in the teleost and other vertebrates.

  4. 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.

  5. Identification of bovine prolactin in seminal fluid, and expression and localization of the prolactin receptor and prolactin-inducible protein in the testis and epididymis of bulls exposed to ergot alkaloids.

    Science.gov (United States)

    Pratt, S L; Calcatera, S M; Stowe, H M; Dimmick, M A; Schrick, F N; Duckett, S K; Andrae, J G

    2015-03-01

    The objectives of this study were to determine (1) the presence and expression levels of bovine prolactin receptor (PRLR) and prolactin-inducible protein (PIP) in bovine testis and epididymis, and (2) the presence and concentrations of prolactin (PRL) present in seminiferous fluid in bulls consuming diets with (E+) or without (E-) ergot alkaloids. Bulls (n = 8) were sacrificed after 126 days (group A) of E+ or E- treatment or 60 days after all bulls (n = 6) were switched to the E- ration (group B). End point and real-time quantitative reverse transcription-polymerase chain reaction and immunohistochemistry were conducted on testis and epididymis samples to establish the presence and relative expression of PRLR and PIP. Seminal fluid samples obtained from bulls consuming E- and E+ diets were subjected to RIA for PRL. Both PIP and PRLR were present in testis and epididymis as determined by reverse transcription-polymerase chain reaction and immunohistochemistry. Prolactin-inducible protein mRNA abundance was affected by time of slaughter in testis and epididymis head, respectively (P Prolactin receptor mRNA expression was affected by time of slaughter in the epididymis (P < 0.05) and differed in testis samples because of treatment (P < 0.05). Radioimmunoassay establishes the presence of PRL in seminal fluid; however, differences in the concentration of PRL over two separate studies were inconsistent, possibly because of differences in diet. The presence and localization of the PRLR are consistent with expression data reported for other species, and the presence of PIP and PRL in seminal fluid is consistent with data generated in humans.

  6. 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].

  7. SPERM MORPHOLOGICAL ASSESSMENTS OF FRIESIAN HOLSTEIN BULL SEMEN COLLECTED FROM THREE ARTIFICIAL INSEMINATION CENTERS IN INDONESIA

    Directory of Open Access Journals (Sweden)

    B. Purwantara

    2014-10-01

    Full Text Available Morphologically abnormal sperm in semen has been associated with the sub fertility and sterilityfor many years. This study assessed the sperm morphology of Friesian Holstein bull semen which wascollected from three Artificial Insemination centers in Indonesia. Total of 22 bulls were used in thisstudy; an ejaculate from each bull was examined. Three to four glass slides were prepared for each bullsample; a drop of semen was placed on each glass slide, smeared, and air-dried. The smeared sampleswere stained with carbolfuchsin-eosin (Williams stain. Morphological abnormality types were recordedfrom total of 500 spermatozoa. Results demonstrated that all 22 bulls had low sperm abnormality(<10%. Pearshaped was the most frequently type of sperm abnormality found in the samples(0.81±0.93%; while detached head was the lowest (0.01±0.04%.

  8. The Effect of Sperm Morphology and Sire Fertility on Calving Rate of Finnish Ayrshire AI Bulls.

    Science.gov (United States)

    Attia, S; Katila, T; Andersson, M

    2016-02-01

    Good-quality semen is a prerequisite for successful and profitable artificial insemination (AI) of modern dairy cattle. Fertility of the bulls is evaluated with andrological examinations and semen analyses, such as morphology. However, little attention has been paid to the inheritance of bull fertility. In this study, we correlated sperm morphology, birth year and station of 695 AI bulls with calving rate (CR). Sperm morphology was clearly associated with CR underlining the usefulness of morphological examination in the assessment of fertility. The correlation between the proportion of normal spermatozoa and CR was significant (p bulls with the CR of their 27 sires to study the inheritance of fertility. Sire's CR did not correlate with the CR of the sons (p = 0.218). This result indicates that at least when sires of acceptable CR are used to produce sons for use in AI the inheritance of CR is not significantly correlated.

  9. Candidate gene markers for sperm quality and fertility in bulls

    Directory of Open Access Journals (Sweden)

    Chinmoy Mishra

    2013-10-01

    Full Text Available Fertility is one of the primary traits of reproduction in bulls. Decrease in fertility is a multifactorial condition and is verydifficult to diagnose. Among various causes genetic abnormality holds a major share. By identifying various genes that haveeffects on fertility the genetic cause behind subferility can be explored and also other non genetic factors can be identified.Advancement of molecular genetic tools now easily enables us to explore individual genes in animals. Identification of thesegenes will eventually lead to genome assembly and development of novel tools for analysing complex genetic traits. Thispaper gives a brief idea about the candidate genes for bull fertility, including genes encoding hormones and their receptors,proteins of the seminal plasma, proteins involved in spermatozoa-ovum binding and genes influencing sexual development.The chromosomal location and gene structure are described, based on the bovine genome assembly.

  10. Occurrence and functional significance of the transcriptome in bovine (Bos taurus) spermatozoa

    Science.gov (United States)

    Selvaraju, Sellappan; Parthipan, Sivashanmugam; Somashekar, Lakshminarayana; Kolte, Atul P; Krishnan Binsila, B.; Arangasamy, Arunachalam; Ravindra, Janivara Parameshwaraiah

    2017-01-01

    Mammalian spermatozoa deliver various classes of RNAs to the oocyte during fertilization, and many of them may regulate fertility. The objective of the present study was to determine the composition and abundance of spermatozoal transcripts in fresh bull semen. The entire transcriptome of the spermatozoa from bulls (n = 3) was sequenced using two different platforms (Ion Proton and Illumina) to identify the maximum number of genes present in the spermatozoa. The bovine spermatozoa contained transcripts for 13,833 genes (transcripts per million, TPM > 10). Both intact and fragmented transcripts were found. These spermatozoal transcripts were associated with various stages of spermatogenesis, spermatozoal function, fertilization, and embryo development. The presence of intact transcripts of pregnancy-associated glycoproteins (PAGs) in the spermatozoa suggest a possible influence of sperm transcripts beyond early embryonic development. The specific regions (exon, intron, and exon-intron) of the particular spermatozoal transcripts might help regulate fertilization. This study demonstrates that the use of two different RNA-seq platforms provides a comprehensive profile of bovine spermatozoal RNA. Spermatozoal RNA profiling may be useful as a non-invasive method to delineate possible causes of male infertility and to predict fertility in a manner that is more effective than the conventional methods. PMID:28276431

  11. Effects of spermatozoa-oviductal cell coincubation time and oviductal cell age on spermatozoa-oviduct interactions.

    Science.gov (United States)

    Aldarmahi, Ahmed; Elliott, Sarah; Russell, Jean; Fazeli, Alireza

    2014-01-01

    The oviduct plays a crucial role in sperm storage, maintenance of sperm viability and sperm transport to the site of fertilisation. The aim of the present study was to investigate the effects of oviductal cell culture passage number, oviductal cell age and spermatozoa-oviduct coincubation times on gene expression in oviductal cells. Immortalised oviductal epithelial cells (OPEC) obtained from two different cell passages (36 and 57) were subcultured three times with and without spermatozoa for 24 h (control group). In a second study, OPEC were cocultured with spermatozoa for different time intervals (0, 4, 12 and 24 h). Expression of adrenomedullin (ADM), heat shock 70 kDa protein 8 (HSPA8) and prostaglandin E synthase (PGES) in OPEC was measured by quantitative polymerase chain reaction. The expression of ADM and HSPA8 was decreased significantly in OPEC cells from Passage 57, particularly in the later subculture group. These effects on HSPA8, but not ADM, expression in OPEC were further altered after coculture with spermatozoa for 24 h. We also demonstrated that spermatozoa-oviduct coculture for 12 and 24 h resulted in significantly higher expression of ADM, HSPA8 and PGES in OPEC. Overall, the data suggest that the OPEC lose some of their properties as a result of oviductal cell aging and that there are spermatozoa-oviduct interactions leading to increased oviductal cell gene expression.

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

    Science.gov (United States)

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

    2015-01-01

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

  13. 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.

  14. 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 carcass traits; B x A bulls had the heaviest (P carcass weight, greatest (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.

  15. Effects of acrosomal conditions of frozen-thawed spermatozoa on the results of artificial insemination in Japanese Black cattle.

    Science.gov (United States)

    Kishida, Kazumi; Sakase, Mitsuhiro; Minami, Kenta; Arai, Miyuki M; Syoji, Reiko; Kohama, Namiko; Akiyama, Takayuki; Oka, Akio; Harayama, Hiroshi; Fukushima, Moriyuki

    2015-01-01

    The purposes of this study were to examine the relationship between male artificial insemination (AI) fertility and sperm acrosomal conditions assessed by new and conventional staining techniques and to identify possible reproductive dysfunctions causing low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions in Japanese Black bulls. We investigated individual differences among bulls in the results concerning (1) acrosomal conditions of frozen-thawed spermatozoa as assessed by not merely peanut agglutinin-lectin staining (a conventional staining technique) but also immunostaining of acrosomal tyrosine-phosphorylated proteins (a new staining technique), (2) routine AI using frozen-thawed spermatozoa as assessed by pregnancy diagnosis, (3) in vivo fertilization of frozen-thawed spermatozoa and early development of fertilized eggs as assessed by superovulation/AI-embryo collection tests and (4) in vitro fertilization of frozen-thawed spermatozoa with oocytes. The percentages of frozen-thawed spermatozoa with normal acrosomal conditions assessed by the abovementioned staining techniques were significantly correlated with the conception rates of routine AI, rates of transferable embryos in superovulation/AI-embryo collection tests and in vitro fertilization rates. These results are consistent with new suggestions that the distribution of acrosomal tyrosine-phosphorylated proteins as well as the acrosomal morphology of frozen-thawed spermatozoa are AI fertility-associated markers that are valid for the prediction of AI results and that low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions result from reproductive dysfunctions in the processes between sperm insemination into females and early embryo development, probably failed fertilization of frozen-thawed spermatozoa with oocytes.

  16. 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...

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

    Science.gov (United States)

    Westring, Christian G; Wiuf, Morten; Nielsen, S Jock; Fogleman, James C; Old, Jennifer B; Lenz, Camilla; Reich, Karl A; Morling, Niels

    2014-09-01

    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 samples, spermatozoa were identified in 45.7% with SPERM HY-LITER™ in Copenhagen, in 54.3% in the laboratory of the manufacturer of SPERM HY-LITER™, and 40.0% of the samples with Baecchi's staining method. When differences occurred between the two methods, it was significantly more often that SPERM HY-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 methods for the two years (27.9% and 32.1% respectively). Analysis of acid phosphatase (ACP) activity for the positive results obtained for these two years does not support the use of a negative ACP result as a prescreen for microscopic analysis for spermatozoa.

  18. 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.

  19. Acute BRSV infection in young AI bulls: effect on sperm quality.

    Science.gov (United States)

    Alm, K; Koskinen, E; Vahtiala, S; Andersson, M

    2009-06-01

    Bovine respiratory syncytial virus (BRSV) infection is an important part of the calf pneumonia complex, occasionally affecting even adult cattle. However, the pathogenicity of BRSV in animals older than 6 months is often neglected. Finland is free of many contagious diseases in farm animals, and this gives a good opportunity to study the effects of specific pathogens on bovine reproduction. This report describes the deteriorating effects of BRSV epizootics on sperm morphology and fertility of young dairy bulls (n = 79) at a bull station. More than half of the young bulls had a clinical respiratory disease caused by BRSV during their quarantine when they were 6 months old. Four of seven subsequent quarantine groups were affected. Six months later, when these seropositive bulls (n = 54) came into semen production, they had poorer sperm morphology, and the proportion of normal spermatozoa was 74.1% in BRSV-seropositive animals compared with 81.2% in seronegative bulls (n = 25) (p = 0.035). Field fertility was also slightly affected, the 60-day non-return rates were 75.2% and 76.8% for BRSV seropositive and seronegative bulls respectively (p = 0.014). Potential reasons for lowered sperm quality are discussed here.

  20. Aneuploidy in human spermatozoa.

    Science.gov (United States)

    Templado, C; Vidal, F; Estop, A

    2011-01-01

    We reviewed the frequency and distribution of disomy in spermatozoa obtained by multicolor-FISH analysis on decondensed sperm nuclei in (a) healthy men, (b) fathers of aneuploid offspring of paternal origin and (c) individuals with Klinefelter syndrome and XYY males. In series of healthy men, disomy per autosome is approximately 0.1% but may range from 0.03 (chromosome 8) to 0.47 (chromosome 22). The great majority of authors find that chromosome 21 (0.18%) and the sex chromosomes (0.27%) have significantly elevated frequencies of disomy although these findings are not universal. The total disomy in FISH studies is 2.26% and the estimated aneuploidy (2× disomy) is 4.5%, more than double that seen in sperm karyotypes (1.8%). Increased disomy levels of low orders of magnitude have been reported in spermatozoa of some normal men (stable variants) and in men who have fathered children with Down, Turner and Klinefelter syndromes. These findings suggest that men with a moderately elevated aneuploidy rate may be at a higher risk of fathering paternally derived aneuploid pregnancies. Among lifestyle factors, smoking, alcohol and caffeine have been studied extensively but the compounding effects of the 3 are difficult to separate because they are common lifestyle behaviors. Increases in sex chromosome abnormalities, some autosomal disomies, and in the number of diploid spermatozoa are general features in 47,XXY and 47,XYY males. Aneuploidy of the sex chromosomes is more frequent than aneuploidy of any of the autosomes not only in normal control individuals, but also in patients with sex chromosome abnormalities and fathers of paternally derived Klinefelter, Turner and Down syndromes.

  1. Effects of in vitro selenium addition to the semen extender on the spermatozoa characteristics before and after freezing in water buffaloes (Bubalus bubalis)

    OpenAIRE

    Kamran Dorostkar; Sayed Mortaza Alavi-Shoushtari; Aram Mokarizadeh

    2012-01-01

    The aim of the present study was to investigate the effect of in vitro supplementation of selenium on fresh and frozen spermatozoa quality of buffalo (Bubalus bubalis) bulls. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 0.5, 1, 2, 4 and 8 μg mL-1 sodium selenite and the sperm motility and viability were evaluated at 0 (T0) (immediately after dilution), 60 (T1) and 120 (T2) min after dil...

  2. 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.

  3. Effects of bovine spermatozoa preparation on embryonic development in vitro

    Directory of Open Access Journals (Sweden)

    Cergolj Marijan

    2006-11-01

    Full Text Available Abstract The aim of our research was to examine the ability of density gradient preparation BoviPure® and swim up method on bull sperm separation and in vitro embryo production (IVP systems. Frozen/thawed semen from six Simmental bulls was pooled and treated using both methods. The sperm motility, concentration, membrane activity, membrane integrity and acrosomal status were evaluated and compared before and after sperm processing using BoviPure® and swim up methods. We also evaluated and compared cleavage rates, embryo yield and quality between the methods. There were significant differences (P ®, but not after swim up method. However, there were significant differences for sperm results among those two mentioned methods. A total of 641 oocytes were matured and fertilized in vitro and cultured in SOFaaBSA. The percentage of cleavage (Day 2 and the percentage of hatched embryos (Day 9 were similar for both methods. However, embryo production rate (Day 7 was significantly higher using BoviPure® method (P ® treated sperm displayed higher quality embryos compared to swim up method (P ® method has an enhanced capacity in sperm selection for in vitro embryo production when compared with swim up method. So, we concluded that BoviPure® could be considered as a better alternative to swim up method for separating bull spermatozoa from frozen/thawed semen for IVP of bovine embryos.

  4. 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.

  5. 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 D (1,25(OH)2D3) in spermatozoa from normal and infertile...

  6. Vitamin D is positively associated with sperm motility and increases intracellular calcium in human spermatozoa

    DEFF Research Database (Denmark)

    Blomberg Jensen, Martin; Bjerrum, Poul J; Jessen, Torben E;

    2011-01-01

    BACKGROUND The vitamin D receptor (VDR) is expressed in human spermatozoa, and VDR-knockout mice and vitamin D (VD) deficiency in rodents results in impaired fertility, low sperm counts and a low number of motile spermatozoa. We investigated the role of activated VD (1,25(OH)(2)D(3)) in human...

  7. Expression of clusterin in spermatozoa of idiopathic asthenozoospermic men%聚集素在特发性弱精子症患者精子中的表达

    Institute of Scientific and Technical Information of China (English)

    唐敏; 刘边疆; 王尚乾; 李永飞; 李鹏超; 张炜

    2013-01-01

    Objective To investigate the role of clusterin in the pathogenesis of idiopathic asthenozoospermia.Methods We separated and purified the ejaculated spermatozoa from 22 idiopathic asthenozoospermia patients and 24 normozoospermic men by Percoll discontinuous density gradients,and detected the distribution and the expressions of clusterin mRNA and protein by using real-time fluorescent quantitatire polymerase chain reaction (FQ-PCR) and Western blotting separately.Results FQ-PCR revealed that the expression of clusterin mRNA was significantly lower in the spermatozoa of patients with idiopathic asthenozoospermia than in those of the normozoospermic men (0.11 ± 0.07 vs 0.98 ± 0.26,t =9.15,P <0.001),and Western blotting confirmed the results of FQ-PCR (0.66 ± 0.21 vs 1.44 ± 0.38,t =6.29,P <0.05).Conclusion The expression of clusterin is significantly decreased in the ejaculated spermatozoa of patients with idiopathic asthenozoospermia,which might be one of the causes of idiopathic asthenozoospermia.%目的 观察聚集素(Clusterin)在特发性弱精子症发生机制中的作用.方法 采用非连续密度梯度离心法分离纯化22例特发性弱精子症和24例正常男性精子,通过实时荧光定量聚合酶链反应(FQ-PCR)和Western blot方法,从基因和蛋白水平分别检测Clnsterin的表达差异.结果 FQ-PCR结果表明,Clusterin mRNA在特发性弱精子症患者精子中的表达较正常男性显著降低(0.11 ±0.07比0.98±0.26,t=9.15,P <0.001);Western blot结果示,Clusterin蛋白在特发性弱精子症患者精子中的表达较正常男性亦显著降低(0.66±0.21比1.44 ±0.38,t =6.29,P<0.05).结论 特发性弱精子症患者精子中Clusterin表达量显著降低,可能是导致弱精子症发生的重要环节之一.

  8. EFFECT OF TWO DIFFERENT ANTIBIOTIC COMBINATIONS ON FERTILITY OF FROZEN BUFFALO AND SAHIW AL BULL SEMEN

    Directory of Open Access Journals (Sweden)

    S.M.H. Andrabi, N. Ahmad, A. Abbas and M. Anzar

    2001-09-01

    Full Text Available This study was carried out to identify the suitable antibiotic combinations in semen extender for improvement in fertility of frozen semen of buffalo and cow (Sahiwal bulls to obtain better pregnancy rate through artificial insemination (AI. For this study eight first ejaculates, four each from a buffalo and a cow (Sahiwal bull were used. The ejaculates were split-sampled and diluted with Tris-citric acid extender (at 37°C; 50x 106 spermatozoa/mI, containing either SP (streptomycin 1000 ~g/ml and penicillin 1000 IU/ml or GTLS (gentamycin 500 µg/ml, Tylosin 100 µg/ml and linco-spectin 300/600 µg/ml. There was no difference in post-thaw motility for these samples. Fertility test based on 75-days first service pregnancy rate was determined under field conditions. A total of 400 inseminations were recorded, 200 for each buffalo and cow (Sahiwal with J 00 of each antibiotic combination, respectively. Fertility rates for SP-based frozen semen of buffalo bull were 41.66% and were 55.2% for GTLS-containing frozen semen, respectively. The results for GTLS were higher (P<0.0001 than SP. Similarly, fertility rates were higher (P<0.0001 for GTLS-based frozen semen of Sahiwal bull (78.78% than SP-containing frozen semen (69.6% of the same specie. Fertility rates also differed due to species of donor bulls. They were better (P<0.0001 for the frozen Sahiwal bull semen than that of the buffalo bull in both SP and GTLS- based frozen semen samples, respectively. In conclusion. seminal quality measured by field fertility trial indicated GTLS combination of antibiotics added to the semen extender was better for improvement in the fertility of frozen buffalo and Sahiwal bull semen, by yielding better pregnancy rates through AI.

  9. Assessment of semen quality in pure and crossbred Jersey bulls

    Directory of Open Access Journals (Sweden)

    Umesh Kumar

    2015-10-01

    Full Text Available Aim: To compare the seminal attributes of neat, pre-freeze (at equilibration, and post-freeze (24 h after freezing semen in pure and crossbred Jersey bulls. Materials and Methods: Total 36 ejaculates (3 ejaculates from each bull were collected from 6 pure Jersey and 6 crossbred Jersey bulls and evaluated for various seminal attributes during neat, pre-freeze, and post-freeze semen. Results: The mean (±standard error [SE] values of neat semen characteristics in pure and crossbred Jersey bulls were recorded such as volume (ml, color, consistency, mass activity (scale: 0-5, and sperm concentration (millions/ml. The extended semen was further investigated at pre-freeze and post-freeze stages and the mean (±SE values recorded at neat, pre-freeze, and post-freeze semen were compared between pure and crossbred Jersey bulls; sperm motility (80.55±1.70%, 62.77±1.35%, 46.11±1.43% vs. 80.00±1.80%, 65.00±1.66%, 47.22±1.08%, live sperm count (83.63±1.08%, 71.72±1.09%, 58.67±1.02% vs. 80.00±1.08%, 67.91±1.20%, 51.63±0.97%, total abnormal sperm count (8.38±0.32%, 12.30±0.39%, 16.75±0.42% vs. 9.00±0.45%, 12.19±0.48%, 18.11±0.64%, hypo-osmotic swelling (HOS reacted spermatozoa (71.88±0.77%, 62.05±0.80%, 47.27±1.05% vs. 72.77±1.02%, 62.11±0.89%, 45.94±1.33%, acrosome integrity (89.05±0.83%, 81.33±0.71%, 71.94±0.86% vs. 86.55±0.57%, 78.66±0.42%, 69.38±0.53%, and DNA integrity (99.88±0.07%, 100, 99.66±0.11% vs. 99.94±0.05%, 100, 99.44±0.18%,. The volume, color, consistency, sperm concentration, and initial motility in pure and crossbred Jersey bulls did not differ significantly (p>0.05. The mass activity was significantly (p0.05 was observed in abnormal sperm; HOS reacted spermatozoa and DNA integrity percentage among breeds. Conclusion: It may be concluded that the quality of pure Jersey bull semen was comparatively better than the crossbred Jersey bulls.

  10. 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...

  11. SEMEN CHARACTERISTICS OF CROSSBRED (FRIESIAN X SAHIW AL AND SAHIWAL YOUNG BULLS MAINT AINED UNDER SUB-TROPICAL CONDITIONS OF PUNJAB

    Directory of Open Access Journals (Sweden)

    Ahsan-ul-Haq, R.H. Mirza and I.A. Zahid

    2003-04-01

    Full Text Available Comparative studies of semen characteristics were carried out in 50% crossbred (Friesian X Sahiwal and purebred Sahiwal young bulls maintained under sub-tropical conditions at Research Institute of Physiology of Animal Reproduction, Bhunikey and Semen Production Unit. Qadirabad, respectively. Semen was collected on weekly intervals for a period of one year. The semen characteristics (mean+SE, i.e., volume per ejaculate (ml, mass motility (0-5 score, individual motility of spermatozoa (%, dead and abnormal spermatozoa (% were 5.62+0.14, 1.25+0.04, 50.50+0.93, 25.78+0.91 and 27.15+0.97, respectively in crossbred bulls. The corresponding values for these semen characteristics in Sahiwal bulls were 3.64+0.09, 1.36+0.04, 60.55+ 0.33, 27.73+0.87 and 15.41+0.86, respectively. These semen parameters differed significantly (P<0.05 between crossbred and Sahiwal bulls. It may be concluded from this study that the semen characteristics in crossbred (Friesian x Sahiwal bulls ~'ere poorer than purebred Sahiwal bulls.

  12. Adriamycin induces H2AX phosphorylation in human spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Zhong-Xiang Li; Ting-Ting Wang; Yan-Ting Wu; Chen-Ming Xu; Min-Yue Dong; Jian-Zhong Sheng; He-Feng Huang

    2008-01-01

    Aim: To investigate whether adriamycin induces DNA damage and the formation of γH2AX (the phosphorylated form of histone H2AX) foci in mature spermatozoa. Methods: Human spermatozoa were treated with adriamycin at different concentrations. γH2AX was analyzed by immunofluorescent staining and flow cytometry and double- strand breaks (DSB) were detected by the comet assay. Results: The neutral comet assay revealed that the treatment with adriamycin at 2 μg/mL for different times (0.5, 2, 8 and 24 h), or for 8 h at different concentrations (0.4, 2 and 10 μg/mL), induced significant DSB in spermatozoa. Immunofluorent staining and flow cytometry showed that the expression of γH2AX was increased in a dose-dependent and time-dependant manner after the treatment of adriamycin. Adriamycin also induced the concurrent appearance of DNA maintenance/repair proteins RAD50 and 53BP1 with γH2AX in spermatozoa. Wortmannin, an inhibitor of the phosphatidylinositol 3-kinase (PI3K) family, abolished the co-appearance of these two proteins with γH2AX. Conclusion: Human mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/repair proteins as somatic cells.

  13. 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

  14. EFFECT OF BULL AND SPERM PREPARATION METHOD ON IN VITRO FERTILIZATION OF BUFFALO OOCYTES

    Directory of Open Access Journals (Sweden)

    H. JAMIL, H. A. SAMAD, Z. I. QURESHI, N. REHMAN AND L. A. LODHI

    2007-01-01

    Full Text Available The present study was designed to compare fertilization rates following oocyte exposure to spermatozoa from different buffalo bulls, using three sperm preparation methods i.e. percoll density gradient, swim-up (modified Ca2 free Tyrode’s medium and TALP medium and sodium citrate washing prior to co-incubation with oocytes. Buffalo oocytes were aspirated from ovarian follicles within 1.5 to 2 hours after slaughter. They were matured in TCM-199 supplemented with 20% oestrus buffalo serum drops under mineral oil in CO2 incubator at 39C for 24 hours. Matured oocytes were transferred to the fertilization droplets and inseminated with 1x106 capacitated sperms prepared by different experimental methods. Data collected on recovered sperm motility immediately after treatment and 24 hours after incubation (at 37C and cleavage rate of co-incubated oocytes were subjected to statistical analysis. The percentage of motile spermatozoa was significantly higher (P<0.05 in semen samples prepared by swim-up method in Ca2 free Tyrode’s medium compared to other experimental techniques. Bull wise comparison showed significantly higher (P<0.05 motility in bull B1 (50.50 ± 5.92%, followed by bull B2 (46.46 ± 5.99% and B3 (45.96 ± 5.79%. Fertilization rate was also significantly (P<0.05 higher for spermatozoa prepared by Swim-up method in Ca2 free Tyrode’s medium (63.75 ± 2.81%, followed by sodium citrate (26.70 ± 5.08%, swim-up TALP (29.14 ± 3.74% and Percoll gradient density (23.89 ± 3.88%. Fertilization rate was significantly higher (P<0.05 in oocytes inseminated with semen from bull B1 (43.43 ± 8.59%, followed by B2 (33.38 ± 9.95% and B3 (30.80 ± 9.56%. The results of present study indicate that bulls and sperm preparation methods differ in their contribution to in vitro fertilization rate. Further studies are suggested to ascertain the factors responsible for such specific effects.

  15. Expression of TEKT3 in spermatozoa of idiopathic asthenozoospermia pa-tients%TEKT3在特发性弱精子症患者精子中的表达

    Institute of Scientific and Technical Information of China (English)

    李玉山; 陈河涛; 杨夕阳; 王全先; 刘军杰; 王琳凯; 苏彦华; 孙琳

    2015-01-01

    Aim:To investigate the function of TEKT 3 protein in the pathogenesis of patients with idiopathic astheno-zoospermia .Methods:Semen samples were collected from 35 normal controls and 35 idiopathic asthenozoospermia patients and purified using Percoll discontinuous density gradients , then the expression of TEKT 3 mRNA and TEKT3 protein were detected by RT-PCR and Western blot .Results: Compared with normal controls , the expressions of TEKT3 mRNA and protein in spermatozoa of idiopathic asthenozoospermia group were obviously decreased (t=9.610,10.090,P<0.001). Correlation analysis revealed that the expressions of TEKT 3 mRNA and protein were positively correlated with spermatozoa progressive motility(r=0.684,P=0.001;r=0.483,P=0.013).Conclusion: The reduced expression of TEKT3 may play an important role in the pathogenesis of idiopathic asthenozoospermia .%目的:探讨特发性弱精子症患者精子中TEKT3表达的特点。方法:采用非连续密度梯度离心法分离、纯化正常对照组和弱精子症组精子标本各35份,分别采用RT-PCR和Western blot检测精子中TEKT3 mRNA和蛋白的表达。结果:与正常对照组比较,弱精子症组TEKT3 mRNA 和蛋白表达均显著降低(t =9.610、10.090,P<0.001),相关性分析显示弱精子症组精子TEKT3 mRNA和蛋白表达水平与前向运动精子率呈正相关(r=0.684, P=0.001;r=0.483,P=0.013)。结论:TEKT3表达降低可能在特发性弱精子症发病中具有重要作用。

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

    Science.gov (United States)

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

    2017-02-07

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

  17. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  19. 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.

  20. PARK7 protein translocating into spermatozoa mitochondria in Chinese asthenozoospermia.

    Science.gov (United States)

    Sun, Yi; Zhang, Wen-Jia; Zhao, Xin; Yuan, Ren-Pei; Jiang, Hui; Pu, Xiao-Ping

    2014-09-01

    PARK7 (DJ1) is a multifunctional oxidative stress response protein that protects cells against reactive oxygen species (ROS) and mitochondrial damage. PARK7 defects are known to cause various physiological dysfunctions, including infertility. Asthenozoospermia (AS), i.e. low-motile spermatozoa in the ejaculate, is a common cause of human male infertility. In this study, we found that downregulation of PARK7 resulted in increased levels of lipid peroxide and ROS, decreased mitochondrial membrane potential, and reduced mitochondrial complex I enzyme activity in the spermatozoa from AS patients. Furthermore, it was observed that PARK7 was translocated into the mitochondria of damaged spermatozoa in AS. Finally, we examined the oxidative state of PARK7 and the results demonstrated the enhancement of oxidation, expressed by increased sulfonic acid residues, the highest form of oxidation, as the sperm motility decreased. Taken together, these results revealed that PARK7 deficiency may increase the oxidative stress damage to spermatozoa. Our present findings open new avenues of therapeutic intervention targeting PARK7 for the treatment of AS.

  1. Team swimming in ant spermatozoa.

    Science.gov (United States)

    Pearcy, Morgan; Delescaille, Noémie; Lybaert, Pascale; Aron, Serge

    2014-06-01

    In species where females mate promiscuously, competition between ejaculates from different males to fertilize the ova is an important selective force shaping many aspects of male reproductive traits, such as sperm number, sperm length and sperm-sperm interactions. In eusocial Hymenoptera (bees, wasps and ants), males die shortly after mating and their reproductive success is ultimately limited by the amount of sperm stored in the queen's spermatheca. Multiple mating by queens is expected to impose intense selective pressure on males to optimize the transfer of sperm to the storage organ. Here, we report a remarkable case of cooperation between spermatozoa in the desert ant Cataglyphis savignyi. Males ejaculate bundles of 50-100 spermatozoa. Sperm bundles swim on average 51% faster than solitary sperm cells. Team swimming is expected to increase the amount of sperm stored in the queen spermatheca and, ultimately, enhance male posthumous fitness.

  2. VIABILITAS SPERMATOZOA PETAURUS BREVICEPS PAPUANUS T.

    OpenAIRE

    NI MADE RAI SUARNI; I GUSTI AYU MANIK ERMAYANTI

    2014-01-01

    Experiment to determine the spermatozoa viability of Petaurus berviceps papuanus T. has been performed. Observation was done under microscope and trypan blue was used as stain. Viable spermatozoa were counted from a total of 100 spermatozoa. It was found that 4,75 % of the spermazoa could be maintained for 7.5 hours in the DMEM (Dulbecco's modified Eagle's medium) at 35oC.

  3. VIABILITAS SPERMATOZOA PETAURUS BREVICEPS PAPUANUS T.

    Directory of Open Access Journals (Sweden)

    NI MADE RAI SUARNI

    2014-08-01

    Full Text Available Experiment to determine the spermatozoa viability of Petaurus berviceps papuanus T. has been performed. Observation was done under microscope and trypan blue was used as stain. Viable spermatozoa were counted from a total of 100 spermatozoa. It was found that 4,75 % of the spermazoa could be maintained for 7.5 hours in the DMEM (Dulbecco's modified Eagle's medium at 35oC.

  4. SEMEN CHARACTERISTICS OF CROSSBRED (FRIESIAN X SAHIW AL) AND SAHIWAL YOUNG BULLS MAINT AINED UNDER SUB-TROPICAL CONDITIONS OF PUNJAB

    OpenAIRE

    Ahsan-ul-Haq, R.H. Mirza and I.A. Zahid

    2003-01-01

    Comparative studies of semen characteristics were carried out in 50% crossbred (Friesian X Sahiwal) and purebred Sahiwal young bulls maintained under sub-tropical conditions at Research Institute of Physiology of Animal Reproduction, Bhunikey and Semen Production Unit. Qadirabad, respectively. Semen was collected on weekly intervals for a period of one year. The semen characteristics (mean+SE), i.e., volume per ejaculate (ml), mass motility (0-5 score), individual motility of spermatozoa (%),...

  5. 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

  6. Season-induced variation in lipid composition is associated with semen quality in Holstein bulls.

    Science.gov (United States)

    Argov-Argaman, N; Mahgrefthe, K; Zeron, Y; Roth, Z

    2013-05-01

    Season-induced variation in fatty acid and cholesterol composition in bovine semen has been associated with semen quality. Given the specific roles of the various semen compartments (seminal fluids, sperm head, and sperm tail) in fertilization, we hypothesized that environmental-stress-induced alterations in the lipid composition of a specific compartment might impair semen quality and sperm function. Semen samples were collected from five mature Holstein-Friesian bulls during the summer (August to September) and winter (December to January). Semen was evaluated by computerized sperm-quality analyzer, calibrated for bulls' semen, and centrifuged to separate the spermatozoa from the seminal fluids. The spermatozoal fraction was sonicated to separate the sperm head and tail compartments. Cold lipid extraction was performed with chloroform:methanol (2:1, vol/vol). Lipids were identified and quantified by gas chromatography. Seasonal variation was found in both physiological and structural parameters. The proportion of spermatozoa defined as morphologically normal was higher in the winter, with higher motility, progressive motility, and velocity relative to summer samples. Lipid composition within fractions varied between seasons with prominent impairment in the tail compartment, characterized by high saturated fatty acid, low polyunsaturated fatty acid, and low cholesterol concentrations during the summer. Given the association between alterations in lipid composition and reduced sperm motility and velocity during the summer, it is suggested that lipid composition might serve to predict sperm quality.

  7. 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.

  8. Aquaporins 7 and 11 in boar spermatozoa: detection, localisation and relationship with sperm quality.

    Science.gov (United States)

    Prieto-Martínez, Noelia; Vilagran, Ingrid; Morató, Roser; Rodríguez-Gil, Joan E; Yeste, Marc; Bonet, Sergi

    2016-04-01

    Aquaporins (AQPs) are integral membrane water channels that allow transport of water and small solutes across cell membranes. Although water permeability is known to play a critical role in mammalian cells, including spermatozoa, little is known about their localisation in boar spermatozoa. Two aquaporins, AQP7 and AQP11, in boar spermatozoa were identified by western blotting and localised through immunocytochemistry analyses. Western blot results showed that boar spermatozoa expressed AQP7 (25kDa) and AQP11 (50kDa). Immunocytochemistry analyses demonstrated that AQP7 was localised in the connecting piece of boar spermatozoa, while AQP11 was found in the head and mid-piece and diffuse labelling was also seen along the tail. Despite differences in AQP7 and AQP11 content between boar ejaculates, these differences were not found to be correlated with sperm quality in the case of AQP7. Conversely, AQP11 content showed a significant correlation (Psperm membrane integrity and fluidity and sperm motility. In conclusion, boar spermatozoa express AQP7 and AQP11, and the amounts of AQP11 but not those of AQP7 are correlated with sperm motility and membrane integrity.

  9. Expression and Subcellular Localization of Retinoic Acid Receptor-α (RARα) in Healthy and Varicocele Human Spermatozoa: Its Possible Regulatory Role in Capacitation and Survival.

    Science.gov (United States)

    Perrotta, Ida; Perri, Mariarita; Santoro, Marta; Panza, Salvatore; Caroleo, Maria C; Guido, Carmela; Mete, Annamaria; Cione, Erika; Aquila, Saveria

    2015-01-01

    Varicocele, an abnormal tortuosity and dilation of veins of the pampiniform plexus, is the most common identifiable and correctable cause of male infertility. It is now becoming apparent that signaling through vitamin A metabolites, such as all-trans retinoic acid (ATRA), is indispensable for spermatogenesis and disruption of retinoic acid receptor-α (RARα) function may result in male sterility and aberrant spermatogenesis. Herein, we investigated by Western blot and immunogold electron microscopy the expression profiles and subcellular localization of RARα in healthy and varicocele human sperm; in addition, we analyzed the effects of ATRA on cholesterol efflux and sperm survival utilizing enzymatic colorimetric CHOD-PAP method and Eosin Y technique, respectively. In varicocele samples, a strong reduction of RARα expression was observed. Immunogold labeling evidenced cellular location of RARα also confirming its reduced expression in "varicocele" samples. Sperm responsiveness to ATRA treatment was reduced in varicocele sperm. Our study showed that RARα is expressed in human sperm probably with a dual role in promoting both cholesterol efflux and survival. RARα might be involved in the pathogenesis of varicocele as its expression is reduced in pathologic samples. Thus, ATRA administration in procedures for artificial insemination or dietary vitamin A supplementation might represent a promising therapeutic approach for the management of male infertility.

  10. Selection of physiological spermatozoa during intracytoplasmic sperm injection.

    Science.gov (United States)

    Torki-Boldaji, B; Tavalaee, M; Bahadorani, M; Nasr-Esfahani, M H

    2017-02-01

    Sperm genomic integrity has a significant effect on intracytoplasmic sperm injection (ICSI) outcomes, especially post-implantation. Spermatozoa selected based on motility and morphology do not guarantee the genomic integrity of spermatozoa. Nearly fifty percentage of spermatozoa in infertile men with normal morphology present different degrees of DNA fragmentation. However, capacitated or hyperactivated spermatozoa show lower degrees of DNA fragmentation. Therefore, selection of hyperactivated spermatozoa may improve ICSI outcome. Routinely, for ICSI, fast-moving spermatozoa with A or B motility pattern are mainly selected for injection. The result of this study shows that in processed semen samples, hyperactivated spermatozoa are mainly observed in B motility pattern while, in viscous medium like polyvinylpyrrolidone (PVP), hyperactivated spermatozoa are mainly present in spermatozoa with C pattern of motility (nonprogressive). Therefore, we propose spermatozoa with C motility pattern which contains the main population of physiological or hyperactivated spermatozoa should be selected for ICSI.

  11. The development of beef breeding bulls.

    Science.gov (United States)

    Engelken, T J

    2008-08-01

    Management of the bull battery will have a dramatic impact on profitability of the cow/calf enterprise. It is critical that young bulls be selected and developed to maximize longevity and productivity for the eventual buyer. Bulls must be structurally sound, healthy, and have adequate libido in order to service the required number of females. Once bulls complete their first breeding season, special care must be taken in order to ensure that they recover and regain needed body condition and pass a bull breeding soundness examination (BBSE). Mature bulls that have reached their genetic potential for growth require less intensive management, but the health program and annual BBSE cannot be overlooked. Mature bulls are also more likely to carry venereal disease and should be screened according to local disease incidence and state regulations. All bulls, regardless of age, should be observed early during the breeding season to ensure that they are physically capable of mounting and servicing females. The establishment of a complete management program, especially for young bulls, is essential to ensure that ranch resources are used efficiently, including maintenance of a high level of reproductive performance of the cow herd.

  12. On Chip Spermatozoa and Leucocytes Counter

    NARCIS (Netherlands)

    Segerink, Loes; Sprenkels, Ad; Vermes, István; Berg, van den Albert; Kim, Tae Song; Lee, Yoon-Sik; Chung, Taek-Dong; Jeon, Noo Li; Lee, Sang-Hoon; Suh, Kahp-Yang; Choo, Jaebum; Kim, Yong-Kweon

    2009-01-01

    In this project we aim to develop a semen quality test system based on a lab-on-a-chip. Since an important quality parameter of semen is the concentration of spermatozoa, we focus on counting the spermatozoa by using impedance measurements in a microchannel. However, semen contains also an interferi

  13. Effects of MboII and BspMI polymorphisms in the gonadotropin releasing hormone receptor (GnRHR) gene on sperm quality in Holstein bulls.

    Science.gov (United States)

    Yang, Wu-Cai; Tang, Ke-Qiong; Yu, Jun-Na; Zhang, Chun-Yan; Zhang, Xiao-Xia; Yang, Li-Guo

    2011-06-01

    The hypothalamic gonadotropin-releasing hormone receptor (GnRHR) plays an essential physiological role in reproductive function, which triggers the synthesis and release of luteinizing hormone and follicle stimulating hormone in the pituitary. The objective of this study was to investigate the effects of polymorphisms of GnRHR gene on the quality of fresh and frozen semen in Holstein bulls. The PCR-RFLP method was applied to detect G286A and T340C transitions determining MboII and BspMI polymorphisms, respectively, in the exon I of bovine GnRHR gene and evaluated its associations with sperm quality traits in 131 Holstein bulls. In polymorphic locus 286, bulls with the GA genotype had significantly higher sperm motility in frozen semen (FMOT) than GG genotype (P bulls with heterozygote CT genotype had significantly higher sperm motility (MOT), semen volume per ejaculate (VOL), and lower abnormal spermatozoa rate (ASR) than homozygote TT genotype (P Bulls contained one A allele or C allele had a favorable, positive effect on sperm quality traits. These results indicate that GnRHR gene can be a potential marker for improving sperm quality traits, and imply that bulls with GA or CT genotype should be selected in breeding program.

  14. Proteomic changes in mammalian spermatozoa during epididymal maturation

    Institute of Scientific and Technical Information of China (English)

    R. John Aitken; Brett Nixon; Minjie Lin; Adam J. Koppers; Yun H. Lee; Mark A. Baker

    2007-01-01

    Epididymal maturation is associated with the activation of a cAMP-induced tyrosine phosphorylation cascade,which is ultimately associated with the expression of capacitation-dependent sperm functions, such as hyperactivated movement and acrosomal exocytosis. As spermatozoa progress through the epididymis they first acquire the capacity to phosphorylate tyrosine on targets on the principal piece, followed by the midpiece. By the time these cells have reached the cauda epididymidis they can phosphorylate the entire tail from neck to endpiece. This particular pattern of phosphorylation is associated with the ontogeny of fully functional spermatozoa that are capable of fertilizing the oocyte. Proteomic analyses indicate that this change is associated with the phosphorylation of several mitochondrial proteins, creation of a mitochondrial membrane potential and activation of mitochondrial free radical generation. At least in rodent species, activation of sperm mitochondria appears to be a particularly important part of epididymal maturation.

  15. Structure of chromatin in spermatozoa.

    Science.gov (United States)

    Björndahl, Lars; Kvist, Ulrik

    2014-01-01

    The specialized structure of the sperm chromatin has a dual function - first to protect the DNA from damage during storage and transport to the oocyte, and then to enable a rapid and complete unpacking of the undamaged paternal genome in the ooplasm. It is evident that zinc has a pivotal role in maintaining the structural stability and in enabling a rapid decondensation at the appropriate time. It is important for the sperm chromatin structure that the spermatozoa are ejaculated together with the zinc-rich prostatic secretion. Early exposure to zinc-binding seminal vesicular fluid can deplete the sperm chromatin of zinc and most likely induce surplus formation of disulfide bridges, likely to cause incomplete and delayed decondensation of the sperm chromatin in the oocyte. A premature decrease in sperm chromatin structure stability is likely to increase the risk for damage to the DNA due to increased access to the genome for DNA damaging compounds. The status of the sperm chromatin structure can vary in vitro depending on the exposure to zinc-depleting conditions when spermatozoa are stored in semen after ejaculation. When sperm DNA damage tests are evaluated and validated, it is therefore essential to also take into account the dynamics of zinc-dependent and zinc-independent sperm chromatin stability.

  16. Chloride channels are involved in sperm motility and are downregulated in spermatozoa from patients with asthenozoospermia.

    Science.gov (United States)

    Liu, Shan-Wen; Li, Yuan; Zou, Li-Li; Guan, Yu-Tao; Peng, Shuang; Zheng, Li-Xin; Deng, Shun-Mei; Zhu, Lin-Yan; Wang, Li-Wei; Chen, Li-Xin

    2016-06-03

    Human spermatozoa encounter an osmotic decrease from 330 to 290 mOsm l-1 when passing through the female reproductive tract. We aimed to evaluate the role of chloride channels in volume regulation and sperm motility from patients with asthenozoospermia. Spermatozoa were purified using Percoll density gradients. Sperm volume was measured as the forward scatter signal using flow cytometry. Sperm motility was analyzed using computer-aided sperm analysis (CASA). When transferred from an isotonic solution (330 mOsm l-1 ) to a hypotonic solution (290 mOsm l-1 ), cell volume was not changed in spermatozoa from normozoospermic men; but increased in those from asthenozoospermic samples. The addition of the chloride channel blockers, 4,4'-diisothiocyanatostilbene-2,2'- isulfonic acid (DIDS) or 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) to the hypotonic solution caused the normal spermatozoa to swell but did not increase the volume of those from the asthenozoospermic semen. DIDS and NPPB decreased sperm motility in both sets of semen samples. The inhibitory effect of NPPB on normal sperm motility was much stronger than on spermatozoa from the asthenozoospermic samples. Both sperm types expressed ClC-3 chloride channels, but the expression levels in the asthenozoospermic samples were much lower, especially in the neck and mid-piece areas. Spermatozoa from men with asthenozoospermia demonstrated lower volume regulating capacity, mobility, and ClC-3 expression levels (especially in the neck) than did normal spermatozoa. Thus, chloride channels play important roles in the regulation of sperm volume and motility and are downregulated in cases of asthenozoospermia.

  17. Sperm macromolecules associated with bull fertility.

    Science.gov (United States)

    Kaya, Abdullah; Memili, Erdoğan

    2016-06-01

    Bull fertility, ability of the sperm to fertilize and activate the egg that sustain embryo development, is vitally important for effective and efficient production of cattle. Fertility is a complex trait with low heritability. Despite recent advances in genomic selection and possibility of enormous paternal benefits to profitable cattle production, there exist no reliable tests for evaluating semen quality and predicting bull fertility. This review focuses on sperm macromolecules such as transcripts, proteins and the epigenome, i.e., the functional genome that are associated with bull fertility. Generating new information in these systems is important beyond agriculture because such progress advances the fundamental science of the mammalian male gamete while at the same time introduces biotechnology into livestock production. Sperm macromolecules and epigenome markers associated with bull fertility can be used alone or in combination with the current SNP microarrays to determine sperm quality and to indicate bull fertility.

  18. Apoptosis and DNA damage in human spermatozoa

    Institute of Scientific and Technical Information of China (English)

    R John Aitken; Adam J Koppers

    2011-01-01

    DNA damage is frequently encountered in spermatozoa of subfertile males and is correlated with a range of adverse clinical outcomes including impaired fertilization, disrupted preimplantation embryonic development, increased rates of miscarriage and an enhanced risk of disease in the progeny. The etiology of DNA fragmentation in human spermatozoa is closely correlated with the appearance of oxidative base adducts and evidence of impaired spermiogenesis. We hypothesize that oxidative stress impedes spermiogenesis,resulting in the generation of spermatozoa with poorly remodelled chromatin. These defective cells have a tendency to default to an apoptotic pathway associated with motility loss, caspase activation, phosphatidylserine exteriorization and the activation of free radical generation by the mitochondria. The latter induces lipid peroxidation and oxidative DNA damage, which then leads to DNA fragmentation and cell death. The physical architecture of spermatozoa prevents any nucleases activated as a result of this apoptotic process from gaining access to the nuclear DNA and inducing its fragmentation. It is for this reason that a majority of the DNA damage encountered in human spermatozoa seems to be oxidative. Given the important role that oxidative stress seems to have in the etiology of DNA damage, there should be an important role for antioxidants in the treatment of this condition. If oxidative DNA damage in spermatozoa is providing a sensitive readout of systemic oxidative stress, the implications of these findings could stretch beyond our immediate goal of trying to minimize DNA damage in spermatozoa as a prelude to assisted conception therapy.

  19. 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.

  20. The influence of gamete co-incubation length on the in vitro fertility and sex ratio of bovine bulls with different penetration speed.

    Science.gov (United States)

    Sattar, A; Rubessa, M; Di Francesco, S; Longobardi, V; Di Palo, R; Zicarelli, L; Campanile, G; Gasparrini, B

    2011-12-01

    The objectives of this work were to evaluate whether the sperm penetration speed is correlated to the in vitro fertility and whether adapting the gamete co-incubation length to the kinetics of the bull improves in vitro fertility and affects the sex ratio. In vitro matured oocytes were co-incubated with spermatozoa from four different bulls (A-D). At various post-insemination (p.i.) times (4, 8, 12, 16 and 20 h), samples of oocytes were fixed and stained with DAPI for nuclei examination, while the remaining ones were transferred into culture to evaluate embryo development. The blastocysts produced were sexed by PCR. Two bulls (A and B) had faster kinetics than the others (C and D), as shown by the higher penetration rates recorded at 4 h p.i. (43%, 30%, 11% and 6%, respectively for bulls A, B, C and D; pbulls did not reflect their in vitro fertility. The incidence of polyspermy was higher for faster penetrating bulls (36%, 24%, 16% and 4%, respectively for bulls A, B, C and D; pbulls may be improved by adapting the co-incubation length to their penetration speed. A sperm-oocyte co-incubation length of 8 h ensured the greatest blastocyst yields for the two faster penetrating bulls. On the contrary, 16 h co-incubation was required to increase (pbulls. Bulls with a faster kinetics did not alter the embryo sex ratio towards males. The female/male (F/M) ratios recorded were 2.1, 1.4, 1.2, 1.3 and 1.6, respectively at 4, 8, 12, 16 and 20 h p.i.

  1. 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.

  2. Differential expression of VASA gene in ejaculated spermatozoa from normozoospermic men and patients with oligozoospermia%生殖细胞特异基因VASA在正常和少精子症患者精子中的表达研究

    Institute of Scientific and Technical Information of China (English)

    Xin Guo; Yao-Ting Gui; Ai-Fa Tang; Li-Hua Lu; Xin Gao; Zhi-Ming Cai

    2007-01-01

    Aim: To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods: Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction (RT-PCR), quantitative RT-PCR (QRT-PCR), immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels. Results: VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR. Conclusion: The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.%目的:探讨生殖细胞高度特异性基因VASA在正常和少精子症患者精子中的表达,明确该基因在精子发生中的作用和相关机制.方法:正常和少精子症患者精液通过Percoll梯度分离获得精子.采用RT-PCR、realtime PCR、Western blot和免疫荧光方法分析该基因转录产物和蛋白表达.结果:VASA mRNA在射出精子中表达,正常精子VASA mRNA水平是少精子症精子的5倍;免疫荧光显示VASA蛋白主要定位在精子头部质膜和尾部,其表达强度正常组明显高于病变组,Western blot结果与免疫荧光结果相一致.结论:少

  3. 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....... METHODS: 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. RESULTS: 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...

  4. Management and Breeding Soundness of Mature Bulls.

    Science.gov (United States)

    Palmer, Colin W

    2016-07-01

    Mature bulls must be fed a balanced ration, vaccinated appropriately, and undergo a breeding soundness evaluation to ensure they meet what is required of a short, but intense breeding season. To be classified as a satisfactory potential breeder, minimum standards for physical soundness, scrotal circumference, sperm motility, and sperm morphology must be achieved using an accepted bull-breeding soundness evaluation format. Sperm production requires approximately 70 days. Heat and stress are the most common insults to spermatogenesis, causing an increase in morphologic abnormalities with obesity-associated scrotal fat accumulation being the most frequent cause of elevated testicular temperature in mature bulls.

  5. Detrimental effects of non-functional spermatozoa on the freezability of functional spermatozoa from boar ejaculate.

    Directory of Open Access Journals (Sweden)

    Maria J Martinez-Alborcia

    Full Text Available 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 (p<0.01 as the proportion of functional spermatozoa in the semen samples before freezing decreased, irrespective of the semen donor. However, the magnitude of the effect differed (p<0.01 among boars. Moreover, semen samples with the largest non-functional sperm subpopulation before freezing showed the highest (p<0.01 levels of MDA after thawing. The thawed viable spermatozoa of semen samples with a high proportion of non-functional spermatozoa before freezing were also functionally different from those of samples with a low proportion of non-functional spermatozoa. These differences consisted of higher (p<0.01 levels of intracellular ROS generation (assessed with 5-(and-6 chloromethyl-20,70-dichlorodihydrofluorescein diacetate acetyl ester; CM-H(2DCFDA and increased (p<0.01 membrane 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.

  6. Computing mating bull fertility from DHI nonreturn data.

    Science.gov (United States)

    Clay, J S; McDaniel, B T

    2001-05-01

    Animal model methodology was used to compute yearly measures of relative fertility of Holstein AI mating bulls based upon 70-d nonreturn of first breedings as reported to U.S. DHIA from 1988 through 1997. Estimated Relative Conception Rates (ERCR) were computed for bulls with a minimum of 50 first breedings in a single year using variance ratios 45.5 for mating bull, 45.5 for animal genetic effects, and 31 for permanent environment. The model assumed repeatability across lactations of 0.05 and included fixed effects of herd-year-month bred and classes of parity, early lactation energy-corrected milk and days open when bred. Estimates of fertility were greater for breedings to cows that were young, had low early lactation production, and were in late stages of lactation. ERCR were expressed as difference in nonreturn from the average AI mating bull of herdmates. Values ranged from -18 to +13. For ERCR computed from a minimum of 1000 breedings, 90% were within four units of zero. Early ERCR computed from a few breedings in a single year were tested for ability to predict later ERCR computed from a minimum of 1000 different breedings. Early ERCR computed from 300 or more matings accurately predicted later independent ERCR. For yearly estimates each based upon a minimum of 1000 breedings, 8% changed more than three units, and 4% declined more than three units. Correlations between ERCR and predicted transmitting abilities protein and type production index were significant but accounted for little variance. Correlations between ERCR and other traits were not significant.

  7. An assessment of dairy herd bulls in southern Australia: 1. Management practices and bull breeding soundness evaluations.

    Science.gov (United States)

    Hancock, A S; Younis, P J; Beggs, D S; Mansell, P D; Stevenson, M A; Pyman, M F

    2016-12-01

    In the pasture-based, seasonally calving dairy herds of southern Australia, the mating period usually consists of an initial artificial insemination period followed by a period of natural service using herd bulls. Bull breeding soundness evaluations (BBSE) were performed on 256 bulls from 32 dairy herds in southwest Victoria, using guidelines produced by the Australian Cattle Veterinarians, before and immediately after a single natural mating period. At the same time, herd managers were questioned regarding the management of the bulls. The objectives of this study were to describe the management practices of dairy herd bulls; to describe the causes of increased risk of reduced fertility in dairy herd bulls, as measured by a standard BBSE; and to describe the reasons for bull removal by herd managers during mating. At the premating BBSE, 19.5% of bulls were classified as high risk of reduced fertility, mostly due to physical abnormalities and reduced semen quality. At the postmating BBSE, 36.5% of bulls were classified as high risk of reduced fertility, mostly due to physical abnormalities, primarily lameness. Of the bulls used, 15.9% were removed from normal mating use by the herd manager, predominantly due to lameness and injuries. A premating BBSE is recommended in dairy herd bulls to identify bulls at risk of reduced fertility. Lameness is the most common problem in dairy herd bulls during the natural mating period, and risk factors associated with lameness in these bulls should be identified to better manage herd bulls.

  8. Ion channels that control fertility in mammalian spermatozoa.

    Science.gov (United States)

    Navarro, Betsy; Kirichok, Yuriy; Chung, Jean-Ju; Clapham, David E

    2008-01-01

    Whole-cell voltage clamp of mammalian spermatozoa was first achieved in 2006. This technical advance, combined with genetic deletion strategies, makes unambiguous identification of sperm ion channel currents possible. This review summarizes the ion channel currents that have been directly measured in mammalian sperm, and their physiological roles in fertilization. The predominant currents are a Ca2+-selective current requiring expression of the 4 mCatSper genes, and a rectifying K+ current with properties most similar to mSlo3. Intracellular alkalinization activates both channels and induces hyperactivated motility.

  9. Analysis of reactive oxygen species generating systems in rat epididymal spermatozoa.

    Science.gov (United States)

    Vernet, P; Fulton, N; Wallace, C; Aitken, R J

    2001-10-01

    Epididymal sperm maturation culminates in the acquisition of functional competence by testicular spermatozoa. The expression of this functional state is dependent upon a redox-regulated, cAMP-mediated signal transduction cascade that controls the tyrosine phosphorylation status of the spermatozoa during capacitation. Analysis of superoxide anion (O2(-.)) generation by rat epididymal spermatozoa has revealed a two-component process involving electron leakage from the sperm mitochondria at complexes I and II and a plasma membrane NAD(P)H oxidoreductase. Following incubation in a glucose-, lactate-, and pyruvate-free medium (-GLP), O2(-.) generation was suppressed by 86% and 96% in caput and cauda spermatozoa, respectively. The addition of lactate, malate, or succinate to spermatozoa incubated in medium -GLP stimulated O2(-.) generation. This increase could be blocked by rotenone and oligomycin (R/O) in the presence of malate or lactate but not succinate. Stimulation with all three substrates, as well as spontaneous O2(-.) production in +GLP medium, was blocked by the flavoprotein inhibitor, diphenylene iodonium. Diphenylene iodonium, but not R/O, suppressed NAD(P)H-induced lucigenin-dependent chemiluminescence. This NAD(P)H-dependent enzyme resided in the sperm plasma membrane and its activity was regulated by zinc and uncharacterized cytosolic factors. Reverse transcription-polymerase chain reaction analysis indicated that the sperm NAD(P)H oxidoreductase complex is quite distinct from the equivalent leukocyte system.

  10. Acid extrusion from human spermatozoa is mediated by flagellar voltage-gated proton channel.

    Science.gov (United States)

    Lishko, Polina V; Botchkina, Inna L; Fedorenko, Andriy; Kirichok, Yuriy

    2010-02-05

    Human spermatozoa are quiescent in the male reproductive system and must undergo activation once introduced into the female reproductive tract. This process is known to require alkalinization of sperm cytoplasm, but the mechanism responsible for transmembrane proton extrusion has remained unknown because of the inability to measure membrane conductance in human sperm. Here, by successfully patch clamping human spermatozoa, we show that proton channel Hv1 is their dominant proton conductance. Hv1 is confined to the principal piece of the sperm flagellum, where it is expressed at unusually high density. Robust flagellar Hv1-dependent proton conductance is activated by membrane depolarization, an alkaline extracellular environment, endocannabinoid anandamide, and removal of extracellular zinc, a potent Hv1 blocker. Hv1 allows only outward transport of protons and is therefore dedicated to inducing intracellular alkalinization and activating spermatozoa. The importance of Hv1 for sperm activation makes it an attractive target for controlling male fertility.

  11. CORRELATION BETWEEN HYPO-OSMOTIC SWELLING TEST AND VARIOUS CONVENTIONAL SEMEN EVALUATION PARAMETERS IN FRESH NILI-RAVI BUFFALO AND SAHIWAL COW BULL SEMEN

    Directory of Open Access Journals (Sweden)

    L. A. LODHI, M. ZUBAIR, Z. I. QURESHI, I. AHMAD AND H. JAMIL

    2008-12-01

    Full Text Available The present study was conducted to determine the correlation of hypo-osmotic swelling test with conventional semen evaluation parameters of fresh semen collected from two Nili-Ravi buffalo and two Sahiwal cow bulls. A total of 10 pooled samples (each comprising two consecutive ejaculates from each bull were collected. Each semen sample was divided into two parts. One part was used for the evaluation of semen by conventional method, while the other part was subjected to hypo-osmotic swelling test by using 150 mOsm/L sodium citrate fructose solution. The mean sperm positive to HOS test was 85.25% both in Nili-Ravi buffalo and Sahiwal cow bull semen. Statistical analysis of the data revealed a significant (P<0.05 positive correlation between progressive motility, morphologically normal spermatozoa, sperm viability and percentage of HOS test positive spermatozoa for both species. It was inferred that HOS test could be a valuable method for routine evaluation of semen for artificial insemination.

  12. STUDY OF ESTROGEN BINDING SITE ON HUMAN EJACULATED SPERMATOZOA

    Institute of Scientific and Technical Information of China (English)

    CHUJin-Shong; WANGYi-Fei

    1989-01-01

    The specific estrogen binding site for 17β-estradiol has been investigated on human spermatozoa by electron microscopec autoradiography. The results show that the binding sites were distributed over the surface of human spermatozoa: acrosomal cap, equatorial

  13. Profiling of sperm gene transcripts in crossbred (Bos taurus x Bos indicus) bulls.

    Science.gov (United States)

    H M, Yathish; Kumar, Subodh; Dubey, Prem P; Modi, Rajendra P; Chaudhary, Rajni; A, Siva Kumar; Ghosh, Subrata K; Sarkar, Mihir; B, Sivamani

    2017-02-01

    Crossbred cattle in some sectors of the world have a significant role in enhancing milk production thereby enhancing the per capita milk availability as a human food source. However, there are certain constraints associated with crossbred animals, such as disease susceptibility, increased reproductive problems, repeat breeding and poor seminal quality. The semen of crossbred bulls has a poor freezing capacity, increased cryo-damage, poor mass cell motility, greater percentages of dead/abnormal sperm and poor initial and post-freeze cell motility. The rejection rate of crossbred bulls for cryostorage of semen has been reported to be as great as 50% as a result of unacceptable semen quality. The identification of superior bulls using molecular technologies is needed which necessitates identification of the genes having a role in sperm function. The present study was, therefore, conducted to gain information on identification and expression of genes having a role in sperm motility in crossbred bulls. The gene transcripts in bulls with sperm of superior and inferior quality were profiled in Vrindavani crossbred cattle by microarray analyses and the results were verified by real time-quantitative PCR. Microarray analyses revealed 19,454 genes which were differentially expressed. At a two-fold cut off, 305 genes were differentially (Pbulls with superior motility.

  14. Incorporation of nanoparticles within mammalian spermatozoa using in vitro capacitation

    Science.gov (United States)

    There is still much unknown about the journey of spermatozoa within the female genital tract. Recent studies have investigated mammalian spermatozoa labeling with fluorescent quantum dot nanoparticles (QD) for non-invasive imaging. Furthermore, the incorporation of these QD within the spermatozoa ma...

  15. Influence of chemical elements on mammalian spermatozoa.

    Science.gov (United States)

    Marzec-Wróblewska, U; Kamiński, P; Lakota, P

    2012-01-01

    Exposure to heavy metals is the most important risk factor in the assessment of spermatogenesis. About 30-40 % cases of infertility are caused by the male factor, and most of them are due to the small quantity of spermatozoa or to inferior spermatozoa quality. The negative impact on sperm motility, morphology and concentration of such chemical elements as Al, Cr, Cd, Pb or Fe was observed, while positive influence was noticed for Zn, Mg, and Ca. The influence of Mn, Cu, Ni or Se on spermatozoa is ambiguous. Chemical elements known as necessary for capacitation and acrosome reaction are Zn, Mg and Ca, while Cd and Pb disturb initiation and progress of the acrosome reaction. The positive effect of chemical elements Al, Cd, Cr, Cu, Ni, Pb, Se, and Zn, lies in their protection against oxidative stress. On the other hand, Al, Cu and Ni induce structural changes in the testes and epididymis or influence interactions with other chemical elements.

  16. The nitric oxide synthase of mouse spermatozoa.

    Science.gov (United States)

    Herrero, M B; Goin, J C; Boquet, M; Canteros, M G; Franchi, A M; Perez Martinez, S; Polak, J M; Viggiano, J M; Gimeno, M A

    1997-07-01

    Nitric oxide synthase (NOS) was evidenced in mature mouse spermatozoa by means of biochemical techniques and Western blot. During 120 min of incubation, 10(7) spermatozoa synthesized 7 +/- 2 pmol of L-[14C]citrulline. Besides, L-citrulline formation depended on the incubation time and on the concentration of L-arginine present in the incubation medium. Different concentrations of N(G)-nitro-L-arginine methyl ester (L-NAME) but not aminoguanidine, inhibited L-[14C]citrulline formation. Western-blot analysis of solubilized sperm proteins revealed a unique band of M(r)=140 kDa with the neural, endothelial and inducible NOS antisera tested. These results provide evidence that mature mouse sperm contains a NOS isoform and that spermatozoa have the potential ability to synthesize NO, suggesting a role for endogenous NO on mammalian sperm function.

  17. RNASET2 in human spermatozoa and seminal plasma: a novel relevant indicator for asthenozoospermia.

    Science.gov (United States)

    Liu, Y; Chen, G; Lu, L; Sun, H; Guo, Q; Xue, K; Fan, Y; Ding, Z

    2013-01-01

    Adequate sperm motility is requisite for human fertilization, whereas the underlying causes or mechanisms of impaired sperm motility, asthenozoospermia, still remain largely unknown. RNASET2 (Ribonuclease T2) may be one of the effectors modulating human sperm motility. We determined if there is a correlation between RNASET2 expression levels in human semen from asthenozoospermia and fertile individuals. Thus, RNASET2 expression levels in spermatozoa and seminal plasma of healthy and asthenozoospermia individuals were evaluated using Western blot, laser scanning confocal microscope analysis, ELISA and flow cytometry. The results revealed that RNASET2 expression was identified in both human spermatozoa and seminal plasma. In spermatozoa from fertile individuals, it was localized to the acrosome, neck and the middle piece of tail regions. However, in spermatozoa from asthenozoospermia individuals (n = 67), RNASET2 staining was especially more frequent and evident in the neck and middle piece than that in fertile individuals (n = 59, p asthenozoospermia than in fertile individuals (p asthenozoospermia individuals may contribute to sperm motility impairment.

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

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    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.

  19. Effect of incubation on freezability of cholesterol-loaded cyclodextrin treated buffalo (Bubalus bubalis spermatozoa

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    S. A. Lone

    2016-02-01

    Full Text Available Aim: The aim of this study was to investigate the effect of incubation on freezability of cholesterol loaded cyclodextrin (CLC treated buffalo spermatozoa. Materials and Methods: Semen samples with mass motility of 3+ and greater, collected from Murrah buffalo bulls were utilized. Immediately after collection, four equal groups of semen sample were made. Group I was kept as control and diluted with Tris upto concentration of 60×106 sperm/ml, where as Groups II, III, and IV were treated with CLC at 3 mg/120× 106 spermatozoa, incubated at 37°C for action of CLC for 10, 15 and 20 min, respectively, and diluted with tris upto concentration of 60×106 sperm/ml. All groups were subjected to equilibration and freezing. The evaluation of semen samples from all groups was carried out at fresh, pre-freeze and post-thaw stage for progressive motility, viability and hypo-osmotic swelling response (HOS response. Results: At the pre-freeze stage, significantly (p<0.05 higher percentage of progressive motility and viability was observed in treatment groups as compared to control with no significant difference among treatment groups. HOS response was significantly (p<0.05 higher in treatment groups as compared to control at pre-freeze stage. At post-thaw stage, significantly (p<0.05 higher percentage of progressive motility, viability and HOS response was recorded in Group II as compared to control and other treatment groups (III and IV. Group II retained significant post-thaw motility and viability at various post-thaw incubation periods. Conclusion: Incubation period of 10 min for CLC treated buffalo spermatozoa yielded significantly higher results in terms of freezability as compared to incubation for 15 and 20 min.

  20. The effect of oviductal fluid on protein tyrosine phosphorylation in cryopreserved boar spermatozoa differs with the freezing method.

    Science.gov (United States)

    Kumaresan, A; Johannisson, A; Saravia, F; Bergqvist, A S

    2012-02-01

    Sperm capacitation takes place in the oviduct and protein tyrosine phosphorylation of sperm proteins is a crucial step in capacitation and acquisition of fertilizing potential. Cryopreserved spermatozoa show altered expression of protein tyrosine phosphorylation in the oviduct. The present study compared two freezing methods (conventional-conventional freezing (CF) and simplified-simplified freezing (SF) methods) for their effect on the ability of boar spermatozoa to undergo protein tyrosine phosphorylation in response to oviductal fluid (ODF). Cryopreserved boar-spermatozoa were incubated with pre- and post-ovulatory ODF for 6 h at 38 °C under 5% CO(2). Aliquots of sperm samples were taken at hourly intervals and analyzed for kinematics and protein tyrosine phosphorylation. Global protein tyrosine phosphorylation in spermatozoa was measured using flow cytometry and different patterns of phosphorylation were assessed using confocal microscopy. Immediately after thawing, no significant difference was observed in post-thaw sperm motility, velocity and global tyrosine phosphorylation between the two methods of freezing although the freezing method significantly (P sperm phosphorylation increased in response to both preovulatory (EODF) and postovulatory oviductal fluid. However, if the SF method was used, a significant increase in these patterns was noticed only in the EODF treated group. The present study demonstrates that preovulatory isthmic ODF induce tyrosine phosphorylation in a higher proportion of boar spermatozoa compared to the post-ovulatory fluid and that the method of freezing significantly influences the response of post-thaw spermatozoa to porcine ODF.

  1. Inactivation of AMPKα1 induces asthenozoospermia and alters spermatozoa morphology.

    Science.gov (United States)

    Tartarin, Pauline; Guibert, Edith; Touré, Aminata; Ouiste, Claire; Leclerc, Jocelyne; Sanz, Nieves; Brière, Sylvain; Dacheux, Jean-Louis; Delaleu, Bernadette; McNeilly, Judith R; McNeilly, Alan S; Brillard, Jean-Pierre; Dupont, Joëlle; Foretz, Marc; Viollet, Benoit; Froment, Pascal

    2012-07-01

    AMP-activated protein kinase (AMPK), a key regulator of cellular energy homeostasis, is present in metabolic tissues (muscle and liver) and has been identified as a modulator of the female reproductive functions. However, its function in the testis has not yet been clearly defined. We have investigated the potential role of AMPK in male reproduction by using transgenic mice lacking the activity of AMPK catalytic subunit α1 gene [α1AMPK knockout (KO)]. In the testis, the α1AMPK subunit is expressed in germ cells and also in somatic cells (Sertoli and Leydig cells). α1AMPK KO male mice show a decrease in fertility, despite no clear alteration in the testis morphology or sperm production. However, in α1AMPK(-/-) mice, we demonstrate that spermatozoa have structural abnormalities and are less motile than in control mice. These spermatozoa alterations are associated with a 50% decrease in mitochondrial activity, a 60% decrease in basal oxygen consumption, and morphological defects. The α1AMPK KO male mice had high androgen levels associated with a 5- and 3-fold increase in intratesticular cholesterol and testosterone concentrations, respectively. High concentrations of proteins involved in steroid production (3β-hydroxysteroid dehydrogenase, cytochrome steroid 17 alpha-hydroxylase/17,20 lysate, and steroidogenic acute regulatory protein) were also detected in α1AMPK(-/-) testes. In the pituitary, the LH and FSH concentrations tended to be lower in α1AMPK(-/-) male mice, probably due to the negative feedback of the high testosterone levels. These results suggest that total α1AMPK deficiency in male mice affects androgen production and quality of spermatozoa, leading to a decrease in fertility.

  2. 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...

  3. 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.

  4. 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 spermatozo

  5. Seasonal variations of cauda epididymal spermatozoa of bucks

    Directory of Open Access Journals (Sweden)

    Dilip Kumar Swain

    2016-09-01

    Full Text Available Objective: The study was conducted to evaluate the influence of season on cauda epididymal spermatozoa isolated from bucks. Materials and methods: Testes of 30 mature bucks were collected from local slaughter house, and were processed for the retrieval of cauda epididymal spermatozoa for evaluation. Testes were collected in three seasons (winter, summer and rainy, and each season was having 10 pairs of testicles. Recovered spermatozoa from the cauda epididymis were processed immediately for evaluation of semen attributes (Spermatozoa motility, viability, plasma membrane integrity, acrosomal status and DNA integrity. Results: Physiological effect of season was observed on progressive motility, percent of live spermatozoa, spermatozoal membrane integrity (HOST, acrosomal integrity, capacitation status and DNA integrity. Progressive motility, percent live spermatozoa, HOST positive spermatozoa, were found significantly (P<0.05 high in summer season, whereas, significantly (P<0.05 lower comet positive spermatozoa were found in summer season as compared to rainy and winter. Compromised acrosomal status was seen in winter and rainy seasons as compared to summer. Conclusion: Compromised acrosome along with plasma membrane and higher percentage of spermatozoa with damaged DNA in cauda spermatozoa were observed during winter and rainy seasons as compared to summer season. Summer season was found to be the most suitable season for collection of cauda epididymal spermatozoa and can effectively be used for assisted reproduction with further investigations of associated mechanisms. [J Adv Vet Anim Res 2016; 3(3.000: 263-267

  6. Increased frequency of aneuploidy in long-lived spermatozoa.

    Science.gov (United States)

    You, Young-Ah; Park, Yoo-Jin; Kwon, Woo-Sung; Yoon, Sung-Jae; Ryu, Buom-Yong; Kim, Young Ju; Pang, Myung-Geol

    2014-01-01

    Aneuploidy commonly causes spontaneous abortions, stillbirths, and aneuploid births in humans. Notably, the majority of sex chromosome aneuploidies in live births have a paternal origin. An increased frequency of aneuploidy is also associated with male infertility. However, the dynamics and behavior of aneuploid spermatozoa during fertilization in humans have not been studied in detail. Therefore, we compared the frequency of aneuploidy and euploidy in live spermatozoa from normozoospermic men over a 3-day period. To assess the dynamics and behavior of aneuploid spermatozoa, we simultaneously evaluated sperm viability using the hypo-osmotic swelling test and sperm aneuploidy using fluorescence in situ hybridization. Whereas the frequency of viable euploid spermatozoa significantly decreased over 3 days, the frequency of viable spermatozoa with aneuploidy interestingly showed a time-dependent increase. In addition, spermatozoa with abnormal sex chromosomes survived longer. To compared with spermatozoa with other swelling patterns, those with tail-tip swelling patterns had a lower frequency of aneuploidy at all time points. This study revealed the novel finding that the frequency of aneuploid spermatozoa with fertilization capability significantly increased compared to that of euploid spermatozoa over 3 days, suggesting that aneuploid spermatozoa can survive longer than euploid spermatozoa and have a greater chance of fertilizing oocytes.

  7. Increased frequency of aneuploidy in long-lived spermatozoa.

    Directory of Open Access Journals (Sweden)

    Young-Ah You

    Full Text Available Aneuploidy commonly causes spontaneous abortions, stillbirths, and aneuploid births in humans. Notably, the majority of sex chromosome aneuploidies in live births have a paternal origin. An increased frequency of aneuploidy is also associated with male infertility. However, the dynamics and behavior of aneuploid spermatozoa during fertilization in humans have not been studied in detail. Therefore, we compared the frequency of aneuploidy and euploidy in live spermatozoa from normozoospermic men over a 3-day period. To assess the dynamics and behavior of aneuploid spermatozoa, we simultaneously evaluated sperm viability using the hypo-osmotic swelling test and sperm aneuploidy using fluorescence in situ hybridization. Whereas the frequency of viable euploid spermatozoa significantly decreased over 3 days, the frequency of viable spermatozoa with aneuploidy interestingly showed a time-dependent increase. In addition, spermatozoa with abnormal sex chromosomes survived longer. To compared with spermatozoa with other swelling patterns, those with tail-tip swelling patterns had a lower frequency of aneuploidy at all time points. This study revealed the novel finding that the frequency of aneuploid spermatozoa with fertilization capability significantly increased compared to that of euploid spermatozoa over 3 days, suggesting that aneuploid spermatozoa can survive longer than euploid spermatozoa and have a greater chance of fertilizing oocytes.

  8. PERUBAHAN VIABILITAS DAN STRUKTUR SUBSELULER SPERMATOZOA DOMBA SETELAH PENGERINGBEKUAN

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    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.

  9. Initial analysis of sperm DNA methylome in Holstein bulls

    Science.gov (United States)

    Aberrant DNA methylation patterns have been associated with abnormal semen parameters, idiopathic male infertility and early embryonic loss in mammals. Using Holstein bulls with high (Bull1) or low (Bull2) fertility rates, we created two representative sperm DNA methylomes at a single-base resolutio...

  10. Activity of antioxidative enzymes in fresh and frozen thawed buffalo (Bubalus bubalis) spermatozoa in relation to lipid peroxidation and semen quality

    Institute of Scientific and Technical Information of China (English)

    G Kadirvel; SatishKumar; S K Ghosh; P Perumal

    2014-01-01

    Objective:To investigate the activity of antioxidative enzymes in fresh and frozen thawed spermatozoa in relation to lipid peroxidation and semen quality in buffalo(Bubalus bubalis) bulls.Methods:Forty two semen ejaculates from seven buffalo bulls were collected by artificial vagina method and were used for the study.Sperm motility, livability, plasma membrane and acrosomal integrity, buffalo cervical mucous penetration test were assessed in fresh and frozen thawed semen.Intracellular antioxidative enzymatic activity such as super oxide dismutase (SOD), catalase(CAT), glutathione peroxidase(GSHPx) and reduced glutathione(GSH), reactive oxygen species(ROS) and lipid peroxidation(LPO) were estimated in fresh and frozen thawed semen.Results:A significant(P<0.01) reduction in activity of antioxidative enzymes(SOD by 47.7%,GSHPx by62.7% andGSH by58.6%) in frozen thawed spermatozoa as compared to fresh spermatozoa was found.Although the catalase activity was varied from0 to3.8IU/109sperm in fresh semen, but after freezing and thawing this activity was not detectable.These enzyme activities had a strong positive association with sperm motility, membrane integrity and distance traveled by vanguard spermatozoa in buffalo cervical mucus and negative correlation withLPO andROS. However, no significant correlation with acrosomal integrity was found.Conclusion:It was concluded that loss of activity of intracellular antioxidative enzymes was evident after freezing and thawing and there was a strong association between the antioxidative enzyme activities,ROS, lipid peroxidation and sperm function in buffalo semen.

  11. Changes in the expression of transient receptor potential canonical type 1 channel mRNA in asthenospermia ejaculated spermatozoa and its significance%弱精子症患者精子中TRPC1基因表达的变化及意义

    Institute of Scientific and Technical Information of China (English)

    李世林; 王怀鹏; 蒲小勇

    2012-01-01

    Objective To investigate the relationship between the expression of transient receptor potential canonical type 1 channel (TRPC1) mRNA in asthenospermia ejaculated spermatozoa and asthenospermia. Methods Semen samples were obtained from the healthy volunteers and patients with asthenospermia. Ejaculated spermatozoa were separated by discontinuous Percoll Gradient centrifugation. The movement parameters of spermatozoa were measured by computer-aided sperm analysis (CASA). Expressions of TRPClmRNA in the sperm of healthy men and asthenospermia patients were detected using the reverse transcription polymerase chain reaction (RT-PCR) and realtime quantitative RT-PCR. Results The relative expression of TRPC1 mRNA in the sperm of healthy men and asthenospermia patients were (0.91±0.15) and (0.32±0.08) respectively, which showed a significant difference (P<0.01). The movement parameters of spermatozoa in asthenospermia decreased obviously compared with that of the healthy group. The Pearson correlation analysis showed the expression of TRPC1 mRNA in spermatozoa was strongly and positively correlated with the linear velocity(r=0.895, PO.01), whipping frequency (r=0.742, P<0.01), forwardness (r=0.782, P<0.01), linearity (r=0.721, P<0.05), and the amplitude of the side pendulum of spermatozoa(r=0.809, P<0.01), whereas it was weakly and positively relative to the curve velocity(r=0.254, P<0.05) and average path velocity (r=0.364, P<0.05). Conclusion The abnormal expression of TRPC1 mRNA might be one of the causes of asthenospermia. It has provided us with a new approach for asthenospermia study.%目的 探讨弱精子症与瞬时感受器电位C1型(transient receptor potential canonical type 1,TRPC1)基因表达的关系.方法 收集40例A、B级精子活力低于20%的弱精子症患者(实验组)和40例精子活力正常健康成年男性(对照组)的精液,采用Percoll非连续梯度离心法分离精液精子,计算机辅助精液分析(CASA

  12. 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.

  13. 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.

  14. The expression variations of IL-1 beta and TNF-alpha mRNA in human spermatozoa%细胞因子IL-1β和TNF-α mRNA在人精子表达的变化及意义

    Institute of Scientific and Technical Information of China (English)

    白文俊; 朱积川; 邓庆平; 闫征; 何培英

    2001-01-01

    目的 了解IL-1β和TNF-α在人精子表达的变化及意义。方法 应用逆转录聚合酶链式反应(RT-PCR)检测13例正常生育者、30例不育者及30例慢性前列腺炎患者精子IL-1β和TNF-αmRNA表达。结果 不育者精子中IL-1β和TNF-αmRNA表达均显著低于正常生育者(P=0.018和P=0.046)。慢性前列腺炎患者精子中TNF-αmRNA的表达显著低于正常生育者(P=0.018),而IL-1βmRNA表达与正常生育者相比差别无显著性意义(P=0.42)。IL-1β和TNF-αmRNA在不育者与慢性前列腺炎患者精子中的表达差别无显著性意义(P=0.49和P=0.62)。结论 某些因素可能干扰不育症患者精子IL-1β和TNF-α的合成,导致精液质量下降及不育。生殖道炎症也可能影响精子TNF-α表达,其机制和意义有待进一步研究证实。%Objective To investigate the expression variations of IL-1 betaand TNF-alpha in human spermatozoa.Methods IL-1β and TNF-α mRNA expressions were studied by means of reverse transcription polymerase reaction (RT-PCR)in 13 fertile men,in 30 infertiles and in 30 chronic prostatitis patients.Results The expressions of IL-1β and TNF-α mRNA in infertile men were significantly lower than those in fertile men(P=0.018 and P=0.050),and the expressions TNF-α mRNA in chronic prostatitis patients were significantly lower than those in fertile men(P=0.018 ),but the expressions of IL-1β in these patients were not significantly lower than in fertile men(P=0.420). The expression differences of IL-1β and TNF-α mRNA between the infertile men and chronic prostatitis patients were not significantly(P=0.488 and P=0.616).Conclusions Some unknown factors might impair the production of IL-1β and TNF-α in infertile spermatozoa, and that may be responsible for the poor quality of semen and the infertile status in these patients. Furthermore, urogenital inflammation could also disturb the expression of TNF-α in human spermatozoa.

  15. Genotype-independent transmission of transgenic fluorophore protein by boar spermatozoa.

    Directory of Open Access Journals (Sweden)

    Wiebke Garrels

    Full Text Available Recently, we generated transposon-transgenic boars (Sus scrofa, which carry three monomeric copies of a fluorophore marker gene. Amazingly, a ubiquitous fluorophore expression in somatic, as well as in germ cells was found. Here, we characterized the prominent fluorophore load in mature spermatozoa of these animals. Sperm samples were analyzed for general fertility parameters, sorted according to X and Y chromosome-bearing sperm fractions, assessed for potential detrimental effects of the reporter, and used for inseminations into estrous sows. Independent of their genotype, all spermatozoa were uniformly fluorescent with a subcellular compartmentalization of the fluorophore protein in postacrosomal sheath, mid piece and tail. Transmission of the fluorophore protein to fertilized oocytes was shown by confocal microscopic analysis of zygotes. The monomeric copies of the transgene segregated during meiosis, rendering a certain fraction of the spermatozoa non-transgenic (about 10% based on analysis of 74 F1 offspring. The genotype-independent transmission of the fluorophore protein by spermatozoa to oocytes represents a non-genetic contribution to the mammalian embryo.

  16. PIXE analysis of human spermatozoa isolated from seminal plasma

    Science.gov (United States)

    Maeda, K.; Sasa, Y.; Kusuyama, H.; Yoshida, K.; Uda, M.

    1990-04-01

    PIXE has been applied to the multielemental and microanalysis of human spermatozoa. This is the first attempt to determine the chemical compositions of the motile spermatozoa free from contaminations of seminal plasma without loss of component elements during washing. The spermatozoa were isolated from semen by letting them swim into a kind of physiological saline, Tyrode's solution. Relative concentrations of P, K, Ca, Ti, Fe, Zn and Br in motile spermatozoa were determined by the use of the chlorine K X-ray peak intensity for evaluating the amount of Tyrode's solution contained in the sample targets. The concentrations of calcium and iron in spermatozoa were considerably higher than in seminal plasma. The concentrations of P, K, Zn and Br in spermatozoa were not so different from those in seminal plasma.

  17. Non-motile tetraploid spermatozoa of Misgurnus loach hybrids

    OpenAIRE

    Zhao, Yan; Fujimoto, Takafumi; Psenicka, Martin; Saito, Taiju; Arai, Katsutoshi

    2016-01-01

    We have compared various properties of spermatozoa from the wild diploid male pond loach Misgurnus anguillicaudatus to those from the interspecific male hybrid of the cross between a female M. anguillicaudatus and a male mud loach M. mizolepis. Our results show that spermatozoa from this interspecific hybrid had poor motility, low viability, abnormal morphology, a larger volume of mitochondrial mass per cell and higher ATP content of spermatozoa with tetraploid DNA content, and they were pres...

  18. Cryopreservation of amphioxus ( Branchiostoma belcheri) spermatozoa

    Institute of Scientific and Technical Information of China (English)

    XU Yuyan; ZHANG Shicui; VERAPONG Vuthiphandchai; SUBUNTITH Nimrat

    2009-01-01

    Amphioxus, Branchiostoma belcheri, a closest relative of vertebrates, is at a high risk of extinction due to a combination of low effective population size, altered native habitats and environmental pollution, yet little is known about cryopreservation of its gametes. This study deals with the cryopreservation of amphioxus semen. The main findings are that (1) the extender of Yao et al. is the best one among the four extenders examined; (2) the appropriate ratio of semen to extender of Yao et al. plus cryoprotectant is from 1:5 to 1:7; (3) dimethyl sulfoxide (DMSO) and methanol are the better cryoprotectants than glycerol, with DMSO giving the best results; (4) the eggs fertilized with post-thaw spermatozoa are capable of developing to at least hatching stage, and the highest hatching rate is (12.4±3.0)%. This is the first report on freezing and thawing of amphioxus spermatozoa,providing a simple and practical protocol for cryopreservation of amphioxus spermatozoa and laying a foundation for safeguarding this endangered species.

  19. The Enzymatic Antioxidant System of Human Spermatozoa

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    Cristian O’Flaherty

    2014-01-01

    Full Text Available The ejaculated spermatozoon, as an aerobic cell, must fight against toxic levels of reactive oxygen species (ROS generated by its own metabolism but also by other sources such as abnormal spermatozoa, chemicals and toxicants, or the presence of leukocytes in semen. Mammalian spermatozoa are extremely sensitive to oxidative stress, a condition occurring when there is a net increase in ROS levels within the cell. Opportunely, this specialized cell has a battery of antioxidant enzymes (superoxide dismutase, peroxiredoxins, thioredoxins, thioredoxins reductases, and glutathione s-transferases working in concert to assure normal sperm function. Any impairment of the antioxidant enzymatic activities will promote severe oxidative damage which is observed as plasma membrane lipid peroxidation, oxidation of structural proteins and enzymes, and oxidation of DNA bases that lead to abnormal sperm function. Altogether, these damages occurring in spermatozoa are associated with male infertility. The present review contains a description of the enzymatic antioxidant system of the human spermatozoon and a reevaluation of the role of its different components and highlights the necessity of sufficient supply of reducing agents (NADPH and reduced glutathione to guarantee normal sperm function.

  20. Hypo-osmotic test in cat spermatozoa.

    Science.gov (United States)

    Comercio, E A; Monachesi, N E; Loza, M E; Gambarotta, M; Wanke, M M

    2013-10-01

    The hypo-osmotic (HOS) test has been used in other species as an indicator of the fertilising capacity of spermatozoa. The aims of this study were to assess the response of domestic cat spermatozoa to the hypo-osmotic test, to determine the type of solution, concentration and time of incubation needed to obtain a maximum percentage of swelling, to correlate the selected combination with the percentages of progressive motility and to evaluate whether dilution of the ejaculate alters the results. Incubation for 30 and 45 min in solutions of fructose and of citrate of 50 and 100 mOsmol kg⁻¹ was evaluated. The highest percentage of swelling was obtained using the 50 mOsmol kg⁻¹ solution, and no significant differences were observed between the times of exposure to the solutions. A positive correlation was observed between the percentage of individual progressive motility and the percentage of sperm swelling in a 50 mOsmol kg⁻¹ fructose solution, with no significant differences being observed between raw and diluted semen samples. The results of this study suggest that the HOS test could be useful for evaluating membrane function in domestic cat spermatozoa, both in raw semen and in samples diluted in the EZ Mixin® commercial extender, and thus could be incorporated into routine semen evaluation protocols.

  1. Zinc content in epididymal spermatozoa of metoclopramide-treated rats.

    Science.gov (United States)

    Kaminska, B; Rozewicka, L; Dominiak, B; Mielnicka, M; Mikulska, D

    1987-01-01

    Zinc content was determined separately in spermatozoa taken from epididymal caput and cauda in rats. It was revealed that spermatozoa transported from the epididymal caput to the cauda reduce about 54% of zinc. This reduction is significantly inhibited in spermatozoa of rats receiving metoclopramide. That is also accompanied by a fall of testosterone level in blood serum and of delta 5, 3 beta-HSD activity in Leydig cells. It was found out that the reduction of zinc in spermatozoa at the time of their passage through the epididymis is the process that depends on androgens.

  2. Simple vitrification for small numbers of human spermatozoa.

    Science.gov (United States)

    Endo, Yuji; Fujii, Yoshitaka; Shintani, Kasumi; Seo, Momoyo; Motoyama, Hiroaki; Funahashi, Hiroaki

    2012-03-01

    Conventional freezing procedures and containers are not appropriate for spermatozoa from the testis because of their low number and poor in-situ motility, and various types of container have been utilized to freeze small numbers of spermatozoa. This study tried to develop a vitrification method for small numbers of spermatozoa using the Cell Sleeper, which is a closed type of cell-cryopreservation container. The container with spermatozoa were cooled in liquid nitrogen vapour and then stored in a cryotank. Sperm motility parameters improved significantly (P < 0.05) by vitrification in oil-free droplets rather than in droplets covered with oil. After vitrification of five spermatozoa per container, all spermatozoa were recovered and the viable sperm rate was significantly higher when spermatozoa were vitrified in a 3.5-ll droplet rather than in 0.5 ll (72.0% versus 38.0%; P < 0.01). Recovery, motility and viability rates of vitrified–warmed spermatozoa were similar between the Cell Sleeper and the CryoTop groups. In conclusion, the Cell Sleeper is a highly effective tool for the cryopreservation of small numbers of spermatozoa and limited cells can be vitrified quickly and simply without significant loss.

  3. State of Sexual Maturity of Nelore Bulls

    OpenAIRE

    Camilo José Ramírez López; Clara Cecilia Rugeles Pinto; Faider Alberto Castaño Villadiego; Víctor Enrique Gómez León; Tamires Miranda Neto; José Domingo Guimarães

    2015-01-01

    This research aimed to determine the state of sexual maturity of young Nelore bulls and its relation to scrotal circumference and seminal characteristics. 1985 animals (aged between 19 and 23 months), fed with tropical pastures (Brachiaria brizantha, Brachiaria decumbes, and Panicum maximum), were evaluated through andrological examination. Physical characteristics of the ejaculate, sperm morphology and scrotal circumference (SC) were examined. After the andrological examination, animals were...

  4. State of Sexual Maturity of Nelore Bulls

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    Camilo José Ramírez López

    2015-12-01

    Full Text Available This research aimed to determine the state of sexual maturity of young Nelore bulls and its relation to scrotal circumference and seminal characteristics. 1985 animals (aged between 19 and 23 months, fed with tropical pastures (Brachiaria brizantha, Brachiaria decumbes, and Panicum maximum, were evaluated through andrological examination. Physical characteristics of the ejaculate, sperm morphology and scrotal circumference (SC were examined. After the andrological examination, animals were classified as suitable for reproduction (andrological class 1; suitable for reproduction in natural mating system (class 2; temporarily unsuitable for reproduction (class 3; and discarded (class 5. To compare the measurements found in different andrological classes, two tests were used: the Tukey test with 5% probability of error, and simple Pearson correlations to verify the relationships between the studied characteristics. 84.5% of the bulls were found to be sexually mature. 39.75% of the animals suitable for reproduction presented a SC greater than 34 cm, and only 0.71% of the study population showed a scrotal circumference less than 28 cm. Favorable high correlations between SC and the physical characteristics of semen were evidenced. The article concludes that scrotal circumference is an excellent characteristics for evaluating and selecting young Nelore bulls.

  5. Triangle zone beer and bull May 11, 1994

    Energy Technology Data Exchange (ETDEWEB)

    Newson, A. C.; Kubli, T. E.; Dechesne, R. G.; MacKay, P. E.; Reid, J. P.; Varsek, J. L.

    1996-06-01

    A review and a summing up of the papers contributed to this special issue of the Bulletin of Canadian Petroleum Geology on triangle zones was prepared in an informal `beer and bull` session by an ad hoc panel of contributors. There was no consensus as to the precise meaning, or the accuracy of the term `triangle zone`, although there was some agreement that if one wants to define structures with a roughly triangular shape, based on seismic expression alone, the term `wedge` might be more appropriate, since the focus of triangle zone development is tectonic activity creating a wedge that delaminates crust as it propagates towards the foreland. It was generally agreed (for now at least) that triangle zones are just one type of thrust front structure when viewed in the larger context of mountain belts around the world. In the classical sense, triangle zones appear to be restricted to the Alberta and Montana segment of the Cordilleran thrust belt. 1 ref.

  6. Beneficial Effects of Nitric Oxide Induced Mild Oxidative Stress on Post-Thawed Bull Semen Quality

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    Mohsen Sharafi

    2015-07-01

    Full Text Available Background: Cryopreservation of semen requires optimized conditions to minimize the harmful effects of various stresses. The main approach for protection of sperm against stress is based on the use of antioxidants and cryoprotectants, which are described as defensive methods. Recently, the application of controlled mild stressors has been described for activation of a temporary response in oocyte, embryo and somatic cells. In this study a sub-lethal oxidative stress induced by precise concentrations of nitric oxide (NO has been evaluated for sperm during cryopreservation. Materials and Methods: In this experimental study, we used different concentrations of NO [0 μM (NO-0, 0.01 μM (NO-0.01, 0.1 μM (NO-0.1, 1 μM (NO-1, 10 μM (NO-10 and 100 μM (NO-100] during cryopreservation of bull semen. Their effects on post-thawed sperm quality that included motility and velocity parameters, plasma membrane functionality, acrosome integrity, apoptosis status, mitochondrial activity and lipid peroxidation after freezing-thawing were investigated. Results: Exposure of sperm before freezing to NO-1 significantly increased total motility (88.4 ± 2.8%, progressive motility (50.4 ± 3.2% and average path velocity (VAP, 53.8 ± 3.1 μm/s compared to other extenders. In addition, NO-1 significantly increased plasma membrane functionality (89.3 ± 2.9% compared to NO-0 (75.3 ± 2.9%, NO-0.01 (78.3 ± 2.9%, NO-0.1 (76.4 ± 2.9%, NO-10 (64 ± 2.9% and NO-100 (42 ± 2.9%. Sperm exposed to NO-1 produced the highest percentage of viable (85.6 ± 2.3% and the lowest percentage of apoptotic (10.8 ± 2.4% spermatozoa compared to the other extenders. Also, NO-100 resulted in a higher percentage of dead spermatozoa (27.1 ± 2.7% compared to the other extenders. In terms of mitochondrial activity, there was no significant difference among NO-0 (53.4 ± 3.2, NO-0.01 (52.1 ± 3.2, NO-0.1 (50.8 ± 3.2 and NO-1 (53.1 ± 3.2. For acrosome integrity, no significant different

  7. Effect of zeranol on sexual development of crossbred bulls.

    Science.gov (United States)

    Godfrey, R W; Randel, R D; Rouquette, F M

    1989-07-01

    Three groups of 1/2 Simmental X 1/4 Brahman X 1/4 Hereford bull calves were used during two different years to study effects of zeranol on sexual development. At 154 d of age, half the calves were implanted with 36 mg zeranol and half, not implanted, served as controls. Implanted calves were reimplanted at 90-d intervals throughout the trial (9 mo) each year. Trial 1 was conducted with 24 calves and Trial 2 was conducted the following year with 10 bulls. Twenty-four days after weaning (200 d of age) and at 28-d intervals thereafter, bulls in drylot in Trial 1 were weighted, scrotal circumference (SC) was measured and an ejaculate of semen was collected by electroejaculation to determine puberty. At these times, bulls were given 200 micrograms of GnRH i.m. and blood was collected at 0, 1, 2, 3, 4 and 5 h after GnRH. Serum concentrations of LH and testosterone (TEST) were determined. At slaughter, testis weight, length and circumference and pubertal status were recorded. Bulls implanted with zeranol had smaller SC than control bulls during the entire 9-mo period (P less than .0001). More control bulls reached puberty than did implanted bulls (82.4 vs 23.5%, respectively; P less than .001). Control bulls had larger testis measurements at slaughter (P less than .0001). Implants did not alter total weight gain or ADG (P greater than .10).(ABSTRACT TRUNCATED AT 250 WORDS)

  8. Evaluation of Spermatological Parameters of Frozen Bull Semen with Conventional and CASA (Computer-Aided Sperm Analysis Method

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    Necmettin TEKİN

    2016-01-01

    Full Text Available The aim of this study was to compare the assessments made by the CASA system and subjective method (by using phase-contrast microscope for spermatological examinations of imported and locally produced semen. Frozen semen (imported and local production belonging to 20 different bulls was examined by phase contrast light microscope method (conventional method and method supported by computer (CASA method for evaluating the principle semen characteristics. For imported semen, considering the average values of samples examined by the two methods, significant differences (P<0.05 were observed in the values of motility and concentration as well as the rates of abnormal and dead spermatozoa. For domestic semen, significant differences (P<0.05 were observed for the concentration of samples by the conventional evaluations and for the assessment of motility and concentration by the CASA method. It was observed that there were significantly (P<0.05 higher data only for the concentration assessed by both methods, while no such differences between the values of motility as well as the rates of abnormal and dead spermatozoa were found when considering the general average rates. By using the two methods, findings from the examination of pre-determined parameters were compared and their reliabilities were displayed herein

  9. Effect of cysteine and glutamine added to extender on post-thaw sperm functional parameters of buffalo bull.

    Science.gov (United States)

    Topraggaleh, T R; Shahverdi, A; Rastegarnia, A; Ebrahimi, B; Shafiepour, V; Sharbatoghli, M; Esmaeili, V; Janzamin, E

    2014-09-01

    Amino acids seem to be crucial components for semen freezing extender due to antioxidant properties. Therefore, this study aimed to assess motility parameters, membrane integrity, intracellular reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and DNA damage to detect the optimum concentrations of cysteine and glutamine for buffalo semen cryopreservation. Twenty ejaculates of four buffalo bulls were diluted in tris-egg yolk extender and divided into seven equal groups consisting of cysteine (5, 7.5 and 10 mmol), glutamine (10, 15 and 20 mmol) and no additive. Supplementation of 5 and 7.5 mmol cysteine and 15 mmol glutamine in cryopreservation extender significantly increased post-thaw motility and plasma membrane integrity of spermatozoa with significant reduction in intracellular ROS when compared with control groups (P Cysteine at 7.5 mmol concentration elevated progressive motility and MMP, compared with control (P cysteine and 15 mmol glutamine in semen cryopreservation extender has more potential to decrease intracellular ROS, and subsequently elevate motility and membrane integrity of buffalo frozen-thawed spermatozoa.

  10. Treatment with zinc, d-aspartate, and coenzyme Q10 protects bull sperm against damage and improves their ability to support embryo development.

    Science.gov (United States)

    Gualtieri, R; Barbato, V; Fiorentino, I; Braun, S; Rizos, D; Longobardi, S; Talevi, R

    2014-09-01

    Reactive oxygen species (ROS) are physiologically generated during mitochondrial respiration and are involved in several signaling mechanisms. However, under pathological conditions, the concentration of ROS may exceed the antioxidant scavenging systems and subsequently lead to cell damage. High ROS levels have been proven to be detrimental to spermatozoa and furthermore compromise sperm function through lipid peroxidation, protein damage, and DNA strand breakage. Although the oral administration of antioxidants has been demonstrated to improve the semen quality in subfertile men, it is still a matter of debate if it can positively influence fertilization outcome and embryo developmental competence. Studies carried out in suitable animal models could resolve these fundamental questions. Hence, the main aims of the present study were to evaluate: (1) the effects of zinc, d-aspartate, and coenzyme Q10, included in the dietary supplement Genadis (Merck Serono), on bull sperm motility and DNA fragmentation; and (2) whether treated spermatozoa have a superior competence in fertilization and in supporting the development of healthy embryos. Our data indicate that this treatment prevents the loss of sperm motility and the rise in sperm DNA fragmentation over time. Moreover, blastocyst rate was found to be significantly higher in oocytes fertilized by treated spermatozoa, and these blastocysts harbored a significantly lower percentage of apoptotic cells.

  11. Morphological characteristics of spermatozoa before and after renal transplantation

    Institute of Scientific and Technical Information of China (English)

    Long-Gen Xu; Shi-Fang Shi; Xiao-Ping Qi; Xiao-Feng Huang; Hui-Ming Xu; Qi-Zhe Song; Xing-Hong Wang; Zong-Fu Shao; Jun-Rong Zhang

    2005-01-01

    Aim: To investigate the changes of the spermatozoa ultrastructures before and after renal transplantation in uremic patients. Methods: The sperm of five uremic patients before and after transplantation and four healthy volunteers were collected and examined by scanning electron microscopy. Results: Abnormal spermatozoa were found in patients pre-transplantation; abnormalities included deletion of the acrosome, absence of the postacrosomal and postnuclear ring, dumbbell-like changes of the head, tail curling, and absence of the mitochondrial sheath in the midsegment. After renal transplantation, most of the spermatozoa became normal. Conclusion: There are many abnormalities with regard to the appearance and structure of the head, acrosome, mitochondria and tail of the spermatozoa in uremic patients. The majority of the spermatozoa returned to normal after renal transplantation, but a few still presented some abnormalities possibly relating to the administration of immunosuppressants.

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

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    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.

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

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    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.

  14. Growth and muscle development characteristics of purebred Angus and Brahman bulls.

    Science.gov (United States)

    Solomon, M B; West, R L; Hentges, J F

    1986-01-01

    Thirty-eight purebred bulls (10 to 17 mo of age) were used to determine the effects of breed (Angus or Brahman) and slaughter weight (60, 80, 90 or 100% of the average mature dam's weight for the respective breed) on growth and muscle development characteristics. Angus bulls grazed summer forage after weaning whereas Brahman bulls were fed to simulate gains achieved on forage by Angus. Bulls were then placed in a confinement feedlot for finishing to their appropriate slaughter weight (293, 369, 411, and 469 kg for Angus and 307, 427, 464 and 520 kg for Brahman). No major differences due to breed were found for predicted carcass composition. The LD muscle from Brahman bulls contained more total DNA (2.27 v. 1.19 g), more total protein (768.22 v. 593.59 g) and generally less total lipid (70.56 v. 101.26 g) when expressed on a total muscle (wet tissue) basis. The percentages and areas for all three muscle fiber types were not affected by breed. As carcass weight increased, muscle weights, total protein, lipid, protein:DNA and muscle fiber size for the three fiber types increased. Total DNA content increased only up to the 90% weight group and then leveled off. The percentage of alpha R fibers decreased while the percentage of alpha W fibers increased with increasing carcass weight. These data suggest that slaughtering animals based on a percentage of their dam's mature weight seems to be a practical method for making comparisons of animals on an equivalent compositional basis. Moreover, it appears that histochemical and biochemical evaluations of skeletal muscle can successfully identify what point in the growth cycle an animal is in.

  15. Effects of a GnRH administration on testosterone profile, libido and semen parameters of dromedary camel bulls.

    Science.gov (United States)

    Monaco, Davide; Fatnassi, Meriem; Padalino, Barbara; Aubé, Lydiane; Khorchani, Touhami; Hammadi, Mohamed; Lacalandra, Giovanni Michele

    2015-10-01

    GnRH treatment has been suggested to increase testosterone levels temporarily and to stimulate libido in stallions, but its use has not fully ascertained in dromedary camels. The aim of this work was to study the effects of administering 100 μg of GnRH on testosterone profile, libido and semen parameters in dromedary camels. The same bulls were used as self-controls and experimental group. Blood samples were collected every 20 min (T0-T12) for 4h, and semen collections were performed over a 2-hour period after T12. GnRH was administered immediately after T0. In GnRH-treated bulls, testosterone levels showed an upward trend, peaking after 140 min, and then slowly decreasing. GnRH administration also led to a decrease in mating time and an increase in spermatozoa concentration. Overall, it seems that administration of 100 μg GnRH might increase testosterone levels temporarily and enhance camel reproduction performance.

  16. A new case of reciprocal translocation in a young bull: rcp(11;21)(q28;q12).

    Science.gov (United States)

    Molteni, L; Perucatti, A; Iannuzzi, A; Di Meo, G P; De Lorenzi, L; De Giovanni, A; Incarnato, D; Succi, G; Cribiu, E; Eggen, A; Iannuzzi, L

    2007-01-01

    Routine cytogenetic investigations of the Chianina cattle (BTA) breed revealed the presence of longer and smaller chromosomes than the largest (BTA1) and smallest (BTA29) chromosomes in the cells of a young, normal-looking bull used for reproduction. Application of both RBA-banding and Ag-NOR techniques, as well as the use of the FISH technique and specific molecular markers of both BTA11 (IL1B, ASS and LGB) and BTA21 (SERPINA and D21S45) established that these two abnormal chromosomes were the product of a reciprocal translocation between BTA11 and BTA21. Both der(11) and der(21) were C-band positive and the chromosome regions affected were rcp(11;21)(q28;q12). The young bull had a normal body conformation, including external genitalia, normal levels of testosterone (as in the control) and non-detectable levels of both 17 beta-estradiol and progesterone (as in the control). The animal never showed libido in the presence of both males and females in oestrus. After slaughter at 18 months, histological evaluation revealed normal organized testes, seminiferous tubules and epididymis but with poor proliferative germ cells consisting mainly of spermatogonia, middle pachytene spermatocytes and early spermatids with late spermatids and spermatozoa being very rare.

  17. Protocol for determining bull trout presence

    Science.gov (United States)

    Peterson, James; Dunham, Jason B.; Howell, Philip; Thurow, Russell; Bonar, Scott

    2002-01-01

    The Western Division of the American Fisheries Society was requested to develop protocols for determining presence/absence and potential habitat suitability for bull trout. The general approach adopted is similar to the process for the marbled murrelet, whereby interim guidelines are initially used, and the protocols are subsequently refined as data are collected. Current data were considered inadequate to precisely identify suitable habitat but could be useful in stratifying sampling units for presence/absence surveys. The presence/absence protocol builds on previous approaches (Hillman and Platts 1993; Bonar et al. 1997), except it uses the variation in observed bull trout densities instead of a minimum threshold density and adjusts for measured differences in sampling efficiency due to gear types and habitat characteristics. The protocol consists of: 1. recommended sample sizes with 80% and 95% detection probabilities for juvenile and resident adult bull trout for day and night snorkeling and electrofishing adjusted for varying habitat characteristics for 50m and 100m sampling units, 2. sampling design considerations, including possible habitat characteristics for stratification, 3. habitat variables to be measured in the sampling units, and 3. guidelines for training sampling crews. Criteria for habitat strata consist of coarse, watershed-scale characteristics (e.g., mean annual air temperature) and fine-scale, reach and habitat-specific features (e.g., water temperature, channel width). The protocols will be revised in the future using data from ongoing presence/absence surveys, additional research on sampling efficiencies, and development of models of habitat/species occurrence.

  18. 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.

  19. Localization of porcine seminal plasma (PSP) proteins in the boar reproductive tract and spermatozoa.

    Science.gov (United States)

    Manásková, P; Jonáková, V

    2008-06-01

    Spermadhesins are proteins containing a characteristic CUB domain, originally isolated from seminal plasma and ejaculated spermatozoa in domestic animals. Boar spermadhesins are multifunctional proteins exhibiting ligand-binding abilities with various endogenous ligands present in the male and female reproductive tracts and may play a role in the reproduction process. Porcine spermadhesins (AQN, AWN, PSP protein families) are secreted mainly by the seminal vesicles, but their mRNAs have been found also in the cauda epididymis and prostate. Unlike AQN and AWN spermadhesins, localization of porcine seminal plasma (PSP) proteins in the boar reproductive tract has not been completely resolved. This work has focused on PSP protein expression and localization in the boar reproductive organs and on spermatozoa. Using specific rabbit polyclonal antibodies (anti-PSP I and anti-PSP II), PSP I and PSP II proteins were immunodetected in tissue extracts and in secretory tissues of cauda epididymis, prostate, seminal vesicles and Cowper's glands on the blots and by an indirect immunofluorescence technique, respectively. Moreover, the ability of PSP proteins to bind to epididymal spermatozoa indicated their presence on cauda epididymal and ejaculated spermatozoa. Porcine seminal plasma proteins bind to the sperm surface at ejaculation and may modulate several aspects of sperm activity during reproduction. PSP proteins are produced not only by seminal vesicles and prostate, but also by epididymis. However, their prospective role in sperm epididymal maturation is not clear. Further characterization of seminal plasma protein forms expressed in the individual reproductive organs will help to understand their subsequent role in the reproduction process.

  20. Analysis of Beta-Lactoglobuline Gene (LGB Polymorphism in Different Breeds of Bulls by High Resolution Melting

    Directory of Open Access Journals (Sweden)

    Martina Miluchová

    2012-05-01

    Full Text Available The goal of the paper was to identify  - lactoglobulin gene polymorphism in bulls. The  - lactoglobulin (LGB is expressed in milk and is important in the evaluation of milk production potential and butterfat and protein content. LGB is localized on bovine chromosome 11. The AA genotype of LGB is associated with higher milk yield, the BB genotype with higher fat and casein content and is more desirable for cheese making. The material involved 46 bulls (Slovak spotted breed – 41 bulls, Pinzgau breed – 3 bulls and Holstein breed – 2 bulls. Bovine genomic DNA was isolated from sperm using commercial kit NucleoSpin Tissue and used in order to estimate LGB genotypes by means of PCR RFLP method and high resolution melting analysis (HRMA. In the population of Slovak spotted breed we detected all genotypes AA, AB and BB with frequency 0.3415, 0.4390 and 0.2195, subsequently. In Pinzgau breed was detected homozygote genotypes AA and BB with frequency 0.3333 and 0.6667. In Holstein breed was observed only heterozygote genotype AB with frequency 1.

  1. Cupuassu cake in feeding feedlot Nellore young bulls

    Directory of Open Access Journals (Sweden)

    Diego Azevedo Mota

    2014-10-01

    Full Text Available This study aimed to evaluate the intake, digestibility and feeding behavior of Nellore young bulls, fed with diets containing different levels of cupuassu cake. Five animals with an average body weight (BW of 186.8 ± 19.5 kg and approximately 13 months of age were used. The animals were divided into five treatments with different levels of cupuassu cake (CC: 0.0%; 5.0%; 10%; 15% and 20%, in substitution of corn and soybean meal and were subjected to the experiment 5 × 5 latin square, composed of five animals and five periods. Each of the five experimental periods lasted 15 days: 10 days for diet adaptation and five days for the assessment of nutrient digestibility. The feeding behavior (visual observation was measured at the end of each experimental period (15th day for 24 h. The dry matter (DMI, ether extract (EEI and crude protein intake (CPI, all expressed in kg/day and %BW, presented a quadratic effect (P<0.10 related to the inclusion of different levels of cupuassu cake, where it was observed that the major consumption of these fractions occurred in the treatment with 5% CC. For digestibility of dry matter (DMD; crude protein (DPB; ether extract (EED; neutral detergent fiber (NDFD; acid detergent fiber (ADFD; This study aimed to evaluate the intake, digestibility and feeding behavior of Nellore young bulls, fed with diets containing different levels of cupuassu cake. Five animals with an average body weight (BW of 186.8 ± 19.5 kg and approximately 13 months of age were used. The animals were divided into five treatments with different levels of cupuassu cake (CC: 0.0%; 5.0%; 10%; 15% and 20%, in substitution of corn and soybean meal and were subjected to the experiment 5 × 5 latin square, composed of five animals and five periods. Each of the five experimental periods lasted 15 days

  2. Sperm fucosyltransferase-5 mediates spermatozoa-oviductal epithelial cell interaction to protect human spermatozoa from oxidative damage.

    Science.gov (United States)

    Huang, Venus Wenxin; Lee, Cheuk-Lun; Lee, Yin-Lau; Lam, Kevin K W; Ko, Jennifer K Y; Yeung, William S B; Ho, Pak-Chung; Chiu, Philip C N

    2015-06-01

    Oxidative damage by reactive oxygen species (ROS) is a major cause of sperm dysfunction. Excessive ROS generation reduces fertilization and enhances DNA damage of spermatozoa. Interaction between spermatozoa and oviductal epithelial cells improves the fertilizing ability of and reduces chromatin damage in spermatozoa. Our previous data showed that oviductal epithelial cell membrane proteins interact with the human spermatozoa and protect them from ROS-induced reduction in sperm motility, membrane integrity and DNA integrity. Sperm fucosyltransferase-5 (sFUT5) is a membrane carbohydrate-binding protein on human spermatozoa. In this study, we demonstrate for the first time that sFUT5 is involved in human spermatozoa-oviduct interaction and the beneficial effects of such interaction on the fertilizing ability of human spermatozoa. Anti-sFUT5 antibody-treated spermatozoa had reduced binding to oviductal membrane proteins. It is consistent with the result that affinity-purified sFUT5 is bound to the epithelial lining of human oviduct and to the immortalized human oviductal epithelial cell line, OE-E6/E7. Pretreatment of spermatozoa with anti-sFUT5 antibody and oviductal membrane proteins with sFUT5 suppressed the protective action of oviductal membrane proteins against ROS/cryopreservation-induced oxidative damage in spermatozoa. Asialofetuin, a reported sFUT5 substrate, can partly mimic the protective effect of oviductal epithelial cell membrane proteins on sperm motility, membrane and DNA integrity. The results enhance our understanding on the protective mechanism of oviduct on sperm functions.

  3. 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

  4. Short communication: Use of young bulls in the United States

    Science.gov (United States)

    The availability of genomic evaluations since 2008 has resulted in many changes to dairy cattle breeding programs. One such change has been the increased contribution of young bulls (0.8 to 3.9 yr old) to those programs. The increased use of young bulls was investigated using pedigree data and b...

  5. 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.

  6. 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.

  7. Influence of Macrophages on the Rooster Spermatozoa Quality.

    Science.gov (United States)

    Kuzelova, L; Vasicek, J; Chrenek, P

    2015-08-01

    The goal of this study was to evaluate the occurrence of macrophages in rooster semen and to investigate their impact on the spermatozoa quality. Ross 308 breeder males (n = 30) with no evidence of genital tract infections were used to determine the concentration of macrophages using fluorescently conjugated acetylated low-density lipoprotein (AcLDL). Subsequently, the roosters were divided into two groups on the basis of semen macrophage concentration, and semen quality was compared in two heterospermic samples. We applied computer-assisted semen analysis (CASA) system to determine motility parameters. Fluorescence microscopy and flow cytometry were used to evaluate occurrence of apoptotic and dead spermatozoa. Spermatozoa fertility potential was examined after intravaginal artificial insemination of hens. Eighteen roosters (control group) contained 0.2-3% of macrophages within spermatozoa population and ten roosters (macrophage group) had 10-15% of macrophages. Males from macrophage group had lower (p < 0.05) motility parameters (total and progressive movement, velocity curved line) and increased concentration of dead spermatozoa detected by flow cytometry and fluorescence microscopy (p < 0.001 and p ˂ 0.05, respectively). Differences (p < 0.05) between fluorescent microscopy and flow cytometry in results on spermatozoa apoptosis and viability were observed. No significant difference was found between groups in fertility of spermatozoa. In conclusion, the higher presence of macrophages in rooster semen may have a negative effect on some parameters of rooster spermatozoa evaluated in vitro. Furthermore, our study suggests that flow cytometry allows more precise examination of spermatozoa viability and apoptosis in a very short time compared with the fluorescent microscopy.

  8. Resorption of the element zinc from spermatozoa by the epididymal epithelium.

    Science.gov (United States)

    Henkel, R; Baldauf, C; Schill, W-B

    2003-04-01

    In this study, elimination of the element zinc from spermatozoa during epididymal maturation was investigated. Testes and epididymides from 40 bulls were collected; epididymal fluid was flushed, pooled, labelled with 0.5 MBq 65Zn2+ per sample and proteins were separated on a Sephacryl S-200 HR and zinc chelate column chromatography. To follow the resorption of zinc in the epididymal epithelial lining, an autometallographic technique (AMG) was performed in tissue from caput, corpus, cauda and vas deferens. The results showed a zinc-binding protein fraction with an apparent molecular weight of 150-160 kDa, which was enriched after chelate column chromatography. Specific labelling of 65Zn was about five times higher in the caput than in the cauda epididymidis. AMG revealed no detectable zinc in the caput, but a significant increase of zinc resorption from the corpus to the cauda and vas deferens. Controls showed that the detectable zinc was located within the principal cells. In conclusion, our study proves that zinc present in the sperm flagellum starts to be mobilized in the caput epididymidis and is resorbed by the epididymal epithelium as from the corpus. This zinc elimination is a mandatory step in sperm maturation to obtain motility.

  9. Probability to retrieve testicular spermatozoa in azoospermic patients

    Institute of Scientific and Technical Information of China (English)

    H.-J.Glander; L.-C.Horn; W.Dorschner; U.Paasch; J.Kratzsch

    2000-01-01

    Aim: The degree of probability to retrieve spermatozoa from testicular tissue for intracytoplasmic sperm injection into oocytes is of interest for counselling of infertility patients. We investigated the relation of sperm retrieval to clinical data and histological pattern in testicular biopsies from azoospermic patients. Methods: In 264 testicular biopsies from 142 azoospermic patients, the testicular tissue was shredded to separate the spermatozoa, histological semi - thin sections of which were then evaluated using Johnsen score. Results: The retrieval of spermatozoa correlated significantly ( P 18 U/L, testicular volume < 5 mL, mean Johnsen score<5, and maximum Johnsen score < 7.

  10. EFFECT OF SPERM CONCENTRATION ON EJACULATE FOR MORPHOMETRIC TRAITS OF SPERMATOZOAS OF THE PIETRAIN BREED BOARS

    Directory of Open Access Journals (Sweden)

    Dorota BANASZEWSKA

    2010-06-01

    Full Text Available An attempt to evaluate the effect of spermatozoa concentration in one ejaculate on their measurements, shape, frequency of occurrence of morphological abnormalities in spermatozoa and physical traits of boar ejaculates in Pietrain breed was made. It was concluded that there was a slight dependence between the content of spermatozoa in one ejaculate and morphometrical traits of spermatozoa. In semen with lower content of spermatozoa (I and II group, the spermatozoa had slightly longer heads (by about 0.18 μm than in semen with large spermatozoa concentration (III group. Spermatozoa in ejaculates with the lowest spermatozoa concentration were characterized by the longest flagellum and the largest total length. The total length of spermatozoa was decreasing in groups of larger concentration, which was caused by both lower length of heads and flagella. Some differences in spermatozoa shape in relation to their concentration in one ejaculate were found. Spermatozoa in ejaculates, which were classified into II group, seemed to have less prolate shape than spermatozoa in ejaculates of I and III groups. It was stated that the content of spermatozoa in one ejaculate affected the frequency of spermatozoa with morphological changes. Semen assigned to II group was distinguished by the best quality.

  11. POSSIBILITY OF COMPUTER EXPERIMENT IN STUDY OF ANIMAL SPERMATOZOA HETEROGENEITY L. V. Gorbunov, Y. M. Mazharova

    Directory of Open Access Journals (Sweden)

    L. V.

    2016-04-01

    Full Text Available A simulation model for evaluating the survival and fertilizing capacity of animal spermatozoa was developed, taking into account the initial condition of the sperm and the effectiveness of cryopreservation stages. The model is based on an analytical expression that reflects the main reasons for the survival of reproductive cells in onto-, techno- and phylogenesis. The decrease in spermatozoa resistance depends on a number of biological factors — the animal species, physiological conditions of sperm donor and recipient, the ejaculate quality, and technological factors — the effectiveness of the methods of cell cryopreservation and egg insemination. The discrepancy between the results of cell motility obtained by calculation and experimental methods amounted to less than 2% as a result of our own experiments and to less than 5% for the data taken from literature. A feature of the model is the complete independence of the effectiveness of studied techniques from the heterogeneity of animal sperm. The conducted computer experiment showed that the difference between the values of initial motility and fertilizing capacity of sperm varies from 50 to 100% depending on the difference of biological parameters, while the index of the effectiveness of selected technique creates an error of about 1%. Comparative analysis of alternative technologies of spermatozoa cryopreservation showed the maximum efficiency of the stages of cryoprotectant use, freeze mode, survival and fertilizing capacity of the object. The use of computer modeling allows to greatly reduce the spread in spermatozoa preservation values that were obtained in different experiments, and thus to reduce the time and costs it takes to obtain reliable results.

  12. Sequential development of flagellar defects in spermatids and epididymal spermatozoa of selenium-deficient rats.

    Science.gov (United States)

    Olson, Gary E; Winfrey, Virginia P; Hill, Kristina E; Burk, Raymond F

    2004-03-01

    In this study cauda epididymal spermatozoa of rats maintained on a selenium-deficient diet for 5 and 7 months exhibited an array of flagellar defects. Spermatids and spermatozoa were analyzed by light and electron microscopy to define the appearance of flagellar abnormalities during spermiogenesis and post-testicular sperm development. Late spermatids of selenium-deficient rats displayed normal structural organization of the flagellar plasma membrane, axoneme, outer dense fibers, fibrous sheath and annulus, but they exhibited a premature termination of the mitochondrial sheath. A comparison of late spermatids and caput epididymal spermatozoa revealed that a late step in flagellar differentiation was the structural remodeling of the annulus and its accompanying fusion with both the fibrous sheath and the mitochondrial sheath. In selenium-deficient animals, however, the annulus failed to fuse with the mitochondrial sheath, generating an apparent weak point in the flagellum. After epididymal passage, cauda epididymal spermatozoa of selenium-deficient animals also exhibited extensive flagellar disorganization resulting from the apparent sliding and extrusion of specific outer dense fiber-doublet microtubule complexes from the proximal and the distal ends of the mitochondrial sheath and the accompanying loss of the midpiece plasma membrane. Only fiber complex number 4 was extruded proximally, whereas fibers 4, 5, 6 and 7 were extruded from the mitochondrial sheath-deficient posterior midpiece. Axonemal fibers 8, 9, 1, 2 and 3 retained their normal geometric relationships. These data suggest that the known loss of male fertility in selenium deficiency results from the sequential development of sperm defects expressed during both spermiogenesis and maturation in the epididymis.

  13. Differentiation of murine male germ cells to spermatozoa in a soft agar culture system

    Institute of Scientific and Technical Information of China (English)

    Mahmoud Abu Elhija; Eitan Lunenfeld; Stefan Schlatt; Mahmoud Huleihel

    2012-01-01

    Establishment of an in vitro system that allows the development of testicular germ cells to sperm will be valuable for studies of spermatogenesis and future treatments for male infertility.In the present study,we developed in vitro culture conditions using three-dimensional agar culture system (SACS),which has the capacity to induce testicular germ cells to reach the final stages of spermatogenesis,including spermatozoa generation.Seminiferous tubules from testes of 7-day-old mice were enzymatically dissociated,and intratubular cells were cultured in the upper layer of the SACS in RPMI medium supplemented with fetal calf serum (FCS).The lower layer of the SACS contained only RPMI medium supplemented with FCS.Colonies in the upper layer were isolated after 14 and 28 days of culture and were classified according to their size.Immunofluorescence and real-time PCR were used to analyse specific markers expressed in undifferentiated and differentiated spermatogonia (Vasa,Dazl,OCT-4,C-Kit,GFR- α-1,CD9 and α-6-integrin),meiotic cells (LDH,Crem-1 and Boule) and post-meiotic cells (Protamine-1,Acrosin and SP-10).Our results reveal that it is possible to induce mouse testicular pre-meiotic germ cell expansion and induce their differentiation to spermatozoa in SACS.The spermatozoa showed normal morphology and contained acrosomes.Thus,our results demonstrate that SACS could be used as a novel in vitro system for the maturation of pre-meiotic mouse germ cells to post-meiotic stages and morphologically-normal spermatozoa.

  14. Localization of Ureaplasma Urealyticum on Human Spermatozoa

    Institute of Scientific and Technical Information of China (English)

    徐晨; 王一飞

    1994-01-01

    Semen specimens were collected from 20 normal fertile men and 20 unexplained infertile men with U reaplasma urealyticum (U.U.) in semen. Spermatozoa of both groups were examined by immunogold technique and immunofluorescence test. A number of gold particles of the U. U. adhered to the sperm surface of infertile men were observed. Strong specific fluorescence was noticed on the sperm surface of infertile men, mostly on the midpiece and/ or postacrosomal region. A significant increase of sperm morphological abnormality, especially the swollen midpiece, the coiled tail, and head-tail angalation sperms was observed in infertile group. The sperm motility in infertile group is dramatically lower than that infertile group, It is hypothesized that teratospermia, poor motility and interference with sperm-ovum interaction might be the possible mechanisms for male infertility caused by Ureaplasma urealyticum.

  15. Fertility management of bulls to improve beef cattle productivity.

    Science.gov (United States)

    Thundathil, Jacob C; Dance, Alysha L; Kastelic, John P

    2016-07-01

    Global demand for animal proteins is increasing, necessitating increased efficiency of global food production. Improving reproductive efficiency of beef cattle, especially bull fertility, is particularly critical, as one bull can breed thousands of females (by artificial insemination). Identifying the genetic basis of male reproductive traits that influence male and female fertility, and using this information for selection, would improve herd fertility. Early-life selection of elite bulls by genomic approaches and feeding them to optimize postpubertal reproductive potential are essential for maximizing profitability. Traditional bull breeding soundness evaluation, or systematic analysis of frozen semen, eliminates bulls or semen samples that are grossly abnormal. However, semen samples classified as satisfactory on the basis of traditional approaches differ in fertility. Advanced sperm function assays developed for assessing compensatory and noncompensatory (submicroscopic) sperm traits can predict such variations in bull fertility. New knowledge on epigenetic modulations of sperm DNA, messenger RNA, and proteins is fundamental to refine and expand sperm function assays. Sexed semen, plus advanced reproductive technologies (e.g., ovum pickup and in vitro production of embryos) can maximize the efficiency of beef cattle production. This review is focused on genetic considerations for bull selection, physiology of reproductive development, breeding soundness evaluation, recent advances in assessing frozen semen, and existing and emerging uses of sexed semen in beef cattle production.

  16. Reproductive behaviour in bulls raised under tropical and subtropical conditions.

    Science.gov (United States)

    Galina, C S; Horn, M M; Molina, R

    2007-06-01

    The present review describes the behavioral characteristics of bulls raised under tropical and subtropical conditions and emphasizes the difficulties associated with adequately monitoring their performance in the field to predict reproductive potential. Most of the information generated for improving our understanding of bull behavior under range conditions has been generated in Bos taurus bulls. The limited information available in Bos indicus indicates that males searching for cows in estrus display different sexual patterns when compared to B. taurus bulls and a poor selection of a sire utilized in range conditions can have an important impact in cattle production. Screening and selecting [cg1] bulls for desirable reproductive traits and high libido is known to improve the reproductive performance of the herd. The reproductive and genetic potential of a bull is influenced by factors such as management, age, nutrition and problems related to the female such as embryonic death and anestrus. However, behavioral characteristics of bulls when detecting and serving cows in estrus is poorly understood.

  17. Spermatozoa in the sperm-peak-fraction of the boar ejaculate show a lower flow of Ca(2+) under capacitation conditions post-thaw which might account for their higher membrane stability after cryopreservation.

    Science.gov (United States)

    Hossain, Md Sharoare; Johannisson, Anders; Siqueira, Amanda Pimenta; Wallgren, Margareta; Rodriguez-Martinez, Heriberto

    2011-10-01

    Boar spermatozoa collected in the ejaculate sperm peak-portion (P1, first 10 mL of the sperm-rich fraction, SRF), had shown a higher resilience to freezing and thawing compared to spermatozoa from the rest of the ejaculate (2nd portion of the SRF plus the post-sperm-rich fraction, PSRF), even when using a simplified freezing technique, as long as spermatozoa were incubated in their own seminal plasma (SP). This experiment studied the stability of P1- and SRF-P1 boar spermatozoa frozen in MiniFlatPacks (MFP), post-thaw, using flow cytometry. Since spermatozoa from either portion showed similar cryosurvival and low proportions of unstable membranes (<3%, annexin-V/propidium iodide staining), and only a tendency for SRF-P1 live spermatozoa to depict acrosome exocytosis (FITC-PNA/PI/H33342); they were explored for Ca(2+) contents using a Fluo-4 probe under in vitro capacitating conditions (mBO+ medium), as well they were tested for their ability to sustain a short Ca(2+)-ionophore (A23187) in vitro challenge. The proportions of live spermatozoa depicting high Ca(2+)-levels were initially <2% but increased over incubation time, particularly in SRF-P1(P<0.05), while proportions of live spermatozoa with low Ca(2+)-levels were basically constant over incubation time (~11-14%), for either portion. Incubation in capacitation medium did not modify the proportions of low-Ca(2+) but dramatically increased the proportions of high-Ca(2+) spermatozoa (P<0.001) already after 15 min exposure, highest for SRF-P1 spermatozoa. While the proportion of live spermatozoa with intact acrosome was significantly decreased among SRF-P1 (P<0.001), that of P1-spermatozoa remained unchanged, probably owing to the lowest relative content of cytosolic Ca(2+). The results suggest that spermatozoa in the P1-portion are more resilient to express acrosome exocytosis post-thaw compared to those bathing in the rest of the SRF-fraction when cryopreserved using a simplified technique, in MFPs.

  18. 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

  19. Evaluation of epididymal function through specific protein on spermatozoa.

    Science.gov (United States)

    Del Río, A G; De Sánchez, L Z; Sirena, A

    1984-01-01

    Investigations were focused on the characterization of specific epididymal proteins on the human spermatozoa as a representative parameter for epididymal function. An easy and attainable method, suitable for investigators and clinical use, is proposed in this article.

  20. 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 ...

  1. 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.

  2. Storage of sexed boar spermatozoa: Limits and perspectives.

    Science.gov (United States)

    Spinaci, M; Perteghella, S; Chlapanidas, T; Galeati, G; Vigo, D; Tamanini, C; Bucci, D

    2016-01-01

    Despite the great potential application of sex-sorted spermatozoa in swine, the technology is not practiced in the pig industry because of technical factors and species-specific issues. The susceptibility of boar spermatozoa to stresses induced by the sorting procedure, the relative slowness of the sex-sorting process together with the high sperm numbers required for routine artificial insemination in pig are some of the main factors limiting the commercial application of this technology in pigs. This review briefly describes the damage to spermatozoa during sex sorting, focusing on an additional limiting factor: increased susceptibility of sexed boar spermatozoa to injuries induced by liquid storage and cryopreservation that, in turn, impairs sperm quality leading to unsatisfactory results in vivo. Strategies to extend the lifespan of sex-sorted boar spermatozoa and to improve their fertilizing ability after liquid storage or cryopreservation need to be implemented before this technology can be used in pig farms. In this regard, encapsulation in barium alginate membranes could be a promising technique to optimize the in vivo use of sexed boar spermatozoa, by protecting, targeting, and controlling the release of sperm into the female genital tract.

  3. Expression of TCTE3 in spermatozoa of idiopathic asthenozoospermic pa-tients%特发性弱精子症患者精子中 TCTE3的表达

    Institute of Scientific and Technical Information of China (English)

    李玉山; 吉晓菲; 王全先; 杨险峰; 潘周辉; 胡耀龙

    2014-01-01

    目的:探讨T复合体相关睾丸表达3( TCTE3)与特发性弱精子症发生的关系。方法:采用RT-PCR和Western blot方法检测正常成年男性(正常对照)和特发性弱精子症患者各30例精子标本中TCTE3 mRNA和蛋白的表达。结果:TCTE3 mRNA和蛋白在特发性弱精子症组精子中的相对表达量均低于正常对照组( t=3.038和2.460,P=0.005和0.017)。结论:TCTE3表达的降低可能在弱精子症的发生中起着重要作用。%Aim:To explore the relationship of T-complex associated testis expressed 3(TCTE3) with idiopathic asthe-nozoospermia .Methods:The mRNA and protein expressions of TCTE 3 in 30 cases of normal adult male and 30 cases of idi-opathic asthenozoospermia semen samples were detected by using RT-PCR and Western blot .Results:The relative expres-sion levels of TCTE3 mRNA and protein in the idiopathic asthenozoospermia group were significantly lower than those in the normal control group(t=3.038 and 2.460,P=0.005 and 0.017).Conclusion: The reduced expression of TCTE3 may play an important role in the incidence of idiopathic asthenozoospermia .

  4. 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...

  5. Molecular, cellular, and physiological determinants of bull fertility

    OpenAIRE

    PARISI, Alexis Marie; THOMPSON, Shannon Kate; Kaya, Abdullah; MEMİLİ, Erdoğan

    2014-01-01

    Bull fertility, defined as the ability of the spermatozoon to fertilize and activate the ovum and then support embryonic and even fetal development, is a crucial factor influencing animal reproduction and production efficiency. Despite significant influences of fertility on herd genetic improvement and efficient cattle production, the mechanisms regulating this phenotype and reliable biomarkers associated with bull fertility are poorly defined. Specifically, there is a broad lack of knowledge...

  6. Yamaha BT1100 Bull Dog 斗牛

    Institute of Scientific and Technical Information of China (English)

    李鹏飞; 卓鹏(摄影)

    2010-01-01

    提到雅马哈的重型街车大家首先想到的是XJR1300、FZ1000等,今天我们要说的是被大家所忽略的BT1100 Bull Dog,Bull Dong翻译成中文大概意思是斗牛犬,所以俗称斗牛犬。

  7. 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...

  8. Caracterización funcional y localización del receptor GM-CSF en espermatozoides bovinos Functional characterization and localization of GM-CSF receptor in bovine spermatozoa

    Directory of Open Access Journals (Sweden)

    L. T. VILANOVA

    2003-12-01

    Full Text Available El Factor Estimulador de Colonias de Macrófagos y Granulocitos (GM-CSF es una citoquina pleiotrópica que tiene como principal función regular la proliferación y diferenciación celular de los precursores de células mieloides, así como también estimular el funcionamiento de granulocitos mononucleares maduros y fagocitos. Su receptor es una glicoproteína compuesta por dos subunidades, a y ß, que se expresan en células mieloides precursoras y maduras, así como también en otras células no hematopoyéticas. Nosotros hemos demostrado recientemente que los espermatozoides bovinos expresan receptores de GM-CSF funcionales que señalizan un aumento del transporte de glucosa y vitamina C. En este estudio se determinó la presencia de este receptor en espermatozoides epididimarios y eyaculados, localizándose la subunidad a en la región acrosómica y en la cola de los espermatozoides, y la subunidad ß en la cola espermática. Mediante análisis computarizado del movimiento espermático se encontró que el GM-CSF aumenta el patrón de movimiento espermático en la mayoría de las variables seminales estudiadas en espermatozoides capacitados en presencia de fructosa. Estos hallazgos sugieren que GM-CSF es una molécula clave para el mejor entendimiento de la fisiología espermáticaThe granulocyte-macrophage colony stimulating factor (GM-CSF is a pleiotropic cytokine with the main function of regulating the proliferation and differentiation of myeloid precursor cells as well as to stimulate the functioning of mature mononuclear granulocytes and phagocytes. Its receptor is a glycoprotein formed by two subunits, a and ß, and it is expressed in precursor and mature myeloid cells, as well as in some nonhematopoietic cells. We have recently demonstrated that bull spermatozoa express functional GM-CSF receptors that signal an increased glucose and vitamin C uptake. The presence of GM-CSF receptor in epididymal and ejaculated spermatozoa was

  9. 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...... 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...

  10. Stallion spermatozoa: putative target of estrogens; presence of the estrogen receptors ESR1, ESR2 and identification of the estrogen-membrane receptor GPER.

    Science.gov (United States)

    Arkoun, Brahim; Gautier, Camille; Delalande, Christelle; Barrier-Battut, Isabelle; Guénon, Isabelle; Goux, Didier; Bouraïma-Lelong, Hélène

    2014-05-01

    Among mammals, the stallion produces the largest amount of testicular estrogens. These steroid hormones are produced mainly by Leydig and Sertoli cells in the testis and also in the epididymis. Their role in horse testicular physiology and their ability to act on spermatozoa are still unknown. In order to determine if spermatozoa are targets for estrogens, the presence of estrogen receptors in mature ejaculated spermatozoa has been investigated. The presence of a single isoform of ESR1 (66kDa) and ESR2 (61kDa) was found by Western-blot analysis in samples from seven stallions. Confocal analysis mainly showed a flagellar localization for both receptors. Immuno-TEM experiments revealed that they are mostly located near the membranes, which are classically associated with rapid, non-genomic, effects. Moreover, we evidenced the expression of the seven transmembrane estradiol binding receptor GPER in colt testis. The protein was also localized at the connecting piece in mature spermatozoa. In conclusion, our results suggest that horse spermatozoa are a target for estrogens, which could act on several receptors either during the epididymal transit and/or in the female genital tract.

  11. Expression of TEKT4 protein decreases in the ejaculated spermatozoa of idiopathic asthenozoospermic men%TEKT4蛋白在特发性弱精子症患者精子中的表达

    Institute of Scientific and Technical Information of China (English)

    武文斌; 李玉山; 吉晓菲; 王全先; 高学敏; 杨险峰; 潘周辉; 冯晓霞

    2012-01-01

    目的:探讨TEKT4蛋白在特发性弱精子症发生机制中的作用. 方法:采用非连续密度梯度离心分离和纯化特发性弱精子症患者和正常男性精子,采用RT-PCR和Western印迹方法,从mRNA和蛋白水平检测TEKT4的表达及其差异. 结果:RT-PCR结果表明,TEKT4 mRNA在特发性弱精子症患者精子中的表达水平与正常男性相比显著降低(0.59±0.13 vs 0.75±0.15,t=4.325,P<0.05);Western印迹结果与RT-PCR结果一致,TEKT4蛋白在特发性弱精子症患者精于中的表达水平与正常男性相比亦显著降低(0.48±0.14 vs 0.69±0.13,t=5.939,P<0.05). 结论:特发性弱精子症患者精子中TEKT4表达水平显著降低,可能是导致弱精子症发生的重要环节之一.%Objective: To investigate the role of the TEKT4 protein in the pathogenesis of idiopathic asthenozoospermia. Methods; We separated and purified the ejaculated sperm from idiopathic asthenozoospermia patients and normozoospermic men by Percoll discontinuous density gradients, and detected the distribution and the expressions of TEKT4 mRNA and TEKT4 protein by RT-PCR and Western blot. Results-. RT-PCR revealed that the expression of TEKT4 mRNA was significantly lower in the sperm of the idiopathic asthenozoospermia patients than in those of the normozoospermic men (0.59 ±0.13 isO. 75 ±0.15, t=4.325, P<0.05), and Western blot confirmed the results of RT-PCR (0.48 ±0.14 iv 0.69 ±0.13, (=5.939, P<0.05). Conclusion: The expression of TEKT4 is significantly decreased in the ejaculated sperm of idiopathic asthenozoospermia patients, which might be one of the causes of idiopathic asthenozoospermia.

  12. Cryopreservation of Spermatozoa in Veterinary Medicine

    Directory of Open Access Journals (Sweden)

    Pesch S

    2007-01-01

    Full Text Available Semen cryopreservation and artificial insemination (AI are the most important biotechnological techniques presently applied in animal breeding. Its large-scale introduction in cattle breeding some 60 years ago aimed at the prevention of genital infections transmittable via natural mating as well as breeding progress and success by rapid spreading of valuable genes. Relating to non-food (pet animals, the avoidance of travel and quarantine restrictions and the conservation of genetic resources are additional advantages. A strict legal framework guarantees sire identity and health innocuousness. Similarly strict guidelines regulate the quality of fresh semen, its processing for cryopreservation and its quality after freezing and thawing. Successful application of AI, particularly when using frozen and thawed semen, requires a proven breeding soundness of both, the semen donor and the semen recipient. Satisfactory results matching those of natural mating can then be obtained. It can be expected that the combined use of AI and sexed spermatozoa in distinct breeding programs will further boost breeding progress.

  13. Immunolocalisation of ghrelin and obestatin in human testis, seminal vesicles, prostate and spermatozoa.

    Science.gov (United States)

    Moretti, E; Vindigni, C; Tripodi, S A; Mazzi, L; Nuti, R; Figura, N; Collodel, G

    2014-01-01

    The role of ghrelin and obestatin in male reproduction has not completely been clarified. We explored ghrelin and obestatin localisation in the male reproductive system. Polyclonal antibodies anti-ghrelin and anti-obestatin were used to detect the expression of these hormones in human testis, prostate and seminal vesicles by immunocytochemistry, while in ejaculated and swim up selected spermatozoa by immunofluorescence. Sertoli cells were positive for both peptides and Leydig cells for ghrelin; germ cells were negative for both hormones. Mild signals for ghrelin and obestatin were observed in rete testis; efferent ductules were the most immune reactive region for both peptides. Epididymis was moderately positive for ghrelin; vas deferens and seminal vesicles showed intense obestatin and moderate ghrelin labelling; prostate tissue expressed obestatin alone. Ejaculated and selected spermatozoa were positive for both peptides in different head and tail regions. This study confirms ghrelin localisation in Leydig and Sertoli cells; the finding that ghrelin is expressed in rete testis, epididymis, vas deferens and seminal vesicles is novel, as well as the localisation of obestatin in almost all tracts of the male reproductive system. This research could offer insights for stimulating other studies, particularly on the role of obestatin in sperm physiology, which is still obscure.

  14. Immunomagnetic removal of cryo-damaged human spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Uwe Paasch; Sonja Grunewald; Katja Wuendrich; Torsten Jope; Hans-Jurgen Glander

    2005-01-01

    Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMP, △ψm) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the efficiency of immunomagnetic cell separation (MACS) of these spermatozoa via annexin Ⅴ-binding. Methods: The mTMP and aCP in fresh and cryopreserved spermatozoa were detected by fluorescence microscopy and by Western blots. The sperm suspensions were divided into two sperm fractions (with intact and deteriorated membranes) by magnetic cell separation (MiniMACS, Miltenyi Biotec, Bergisch Gladbach, Germany) in dependence on their binding to superparamagnetic annexin Ⅴ-microbeads (AN-MB). Results: The cryopreservation decreased the portion of spermatozoa with intact mTMP from 80.1% ± 7.2 % to 53.5 % ± 13.1% and increased the spermatozoa with activated pancaspases (aCP) from 21.8 % ± 2.6 % to 47.7 % ± 5.8 % (n = 10; mean ± SEM; P < 0.01). The activation of caspases 1, 3, 8, and 9 in the cryopreserved spermatozoa was confirmed by Western blots (n = 22). MACS reduced significantly the percentage of cryopreserved spermatozoa with dissipated mTMP to 8.1 ± 3.9 (P < 0.01) and also those with aCP to 9.3%±2.2%. Western blot analyses confirmed the increase of the activated caspase3, 9, and 8 in the AN-MB-positive fraction (P<0.05) compared with the AN-MB-negative fraction. The MACS separation effect was confirmed by anti-annexin Ⅴ-antibodies. There was no significant influence of the separation column and the magnetic field on the sperm functions. Conclusion: The cryopreservation impaired the mTMP and enhanced the activation status of caspases in human spermatozoa. The immunomagnetic sperm separation via binding of AN-MB could deplete low quality spermatozoa from cryopreserved semen samples.

  15. 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

  16. DNA fragmentation of spermatozoa and assisted reproduction technology.

    Science.gov (United States)

    Henkel, Ralf; Kierspel, Eva; Hajimohammad, Marjam; Stalf, Thomas; Hoogendijk, Christiaan; Mehnert, Claas; Menkveld, Roelof; Schill, Wolf-Bernhard; Kruger, Thinus F

    2003-01-01

    Despite the ever-increasing knowledge of the fertilization process, there is still a need for better understanding of the causes of sperm DNA fragmentation and its impact on fertilization and pregnancy. For this reason, human sperm DNA fragmentation was investigated by means of the terminal deoxynucleotidyl transferase-mediated dUDP nick-end labelling (TUNEL) assay and the production of reactive oxygen species (ROS) in the ejaculate and in the spermatozoa themselves. These data were correlated with fertilization and pregnancy data from IVF and intracytoplasmic sperm injection (ICSI) patients. Sperm DNA fragmentation did not correlate with fertilization rate, but there was a significantly reduced pregnancy rate in IVF patients inseminated with TUNEL-positive spermatozoa. ICSI patients exhibited the same tendency. This implies that spermatozoa with damaged DNA are able to fertilize an oocyte, but at the time the paternal genome is switched on, further development stops. The determination of ROS in the ejaculate and the percentage of ROS-producing spermatozoa revealed markedly stronger correlations between sperm functions (i.e. motility) and the percentage of ROS-producing spermatozoa. The influence of seminal leukocytes, known to produce large amounts of oxidants, on sperm DNA fragmentation should not be neglected.

  17. Effect of mitochondrial calcium uniporter blocking on human spermatozoa.

    Science.gov (United States)

    Bravo, A; Treulen, F; Uribe, P; Boguen, R; Felmer, R; Villegas, J V

    2015-08-01

    Calcium (Ca(2+) ) regulates a number of essential processes in spermatozoa. Ca(2+) is taken up by mitochondria via the mitochondrial calcium uniporter (mCU). Oxygen-bridged dinuclear ruthenium amine complex (Ru360) has been used to study mCU because it is a potent and specific inhibitor of this channel. In bovine spermatozoa, it has been demonstrated that mitochondrial calcium uptake inhibition adversely affects the capacitation process. It has been demonstrated in human spermatozoa that mCU blocking, through Ru360, prevents apoptosis; however, the contribution of the mCU to normal human sperm function has not been studied. Therefore, the aim of this study was to evaluate the effect of mCU blocking on human sperm function. Spermatozoa obtained from apparently healthy donors were incubated with 5 and 10 μm Ru360 for 4 h at 37 °C. Viability was assessed using propidium iodide staining; motility was determined by computer-aided sperm analysis, adenosine triphosphate (ATP) levels using a luminescence-based method, mitochondrial membrane potential (ΔΨm) using JC-1 staining and reactive oxygen species (ROS) production using dihydroethidium dye. Our results show that mCU blocking significantly reduced total sperm motility and ATP levels without affecting sperm viability, ΔΨm and ROS production. In conclusion, mCU contributes to the maintenance of sperm motility and ATP levels in human spermatozoa.

  18. Nuclear status of immature and mature stallion spermatozoa.

    Science.gov (United States)

    Dias, G M; Retamal, C A; Tobella, L; Arnholdt, A C V; López, M L

    2006-07-15

    'The highly packed chromatin of mature spermatozoa results from replacement of somatic-like histones by highly basic arginine- and cysteine-rich protamines during spermatogenesis, with additional conformational changes in chromatin structure during epididymal transit. The objective of the present study was to compare the nuclear characteristics of immature and mature epididymal stallion spermatozoa, using a variety of experimental approaches. Resistance to in vitro decondensation of chromatin, following exposure to SDS-DTT and alkaline thioglycolate, increased significantly in mature spermatozoa. Evaluation of the thiol-disulfide status (monobromobimane labeling) demonstrated that immature cells obtained from ductulli efferentes contained mostly thiol groups, whereas these groups were oxidized in mature cells collected from the cauda epididymidis. Based on atomic absorption spectrophotometry, maturation of stallion spermatozoa was accompanied by a 60% reduction in the Zn(2+) content of sperm cells, concomitant with increased concentrations of this ion in epididymal fluid. Furthermore, the degree of disulfide bonding was inversely correlated with susceptibility of chromatin to acid denaturation (SCSA). Collectively, these data were consistent with the hypothesis that maturation of stallion spermatozoa involves oxidation of sulphydryl groups to form intra- and intermolecular disulfide links between adjacent protamines, with loss of zinc as an integral feature. These changes endow mechanical and chemical resistance to the nucleus, ensuring efficient transmission of the paternal genome at fertilization.

  19. 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.

  20. Proteome analysis of round-headed and normal spermatozoa by 2-D fluorescence difference gel electrophoresis and mass spectrometry%以荧光差异凝胶双向电泳和质谱技术对圆头精子和正常精子的蛋白质组学分析

    Institute of Scientific and Technical Information of China (English)

    Ting-Ting Liao; Zhen Xiang; Wen-Bing Zhu; Li-Qing Fan

    2009-01-01

    Globozoospermia is a severe form of teratozoospermia characterized by round-headed spermatozoa with an absent acrosome, an aberrant nuclear membrane and midpiece defects. Globozoospermia is diagnosed by the presence of 100% round-headed spermatozoa on semen analysis, and patients with this condition are absolutely infertile. The objective of this study was to investigate the differences in protein expression between human round-headed and normal spermatozoa. Two-dimensional (2-D) fluorescence difference gel electrophoresis (DIGE) coupled with mass spectrometry (MS) was used in this study. Over 61 protein spots were analysed in each paired normal/round-headed comparison, using DIGE technology along with an internal standard. In total, 35 protein spots identified by tandem mass spectrometry (MS/MS) exhibited significant changes (paired t-test, P < 0.05) in the expression level between normal and round-headed spermatozoa. A total of nine proteins were found to be upregulated and 26 proteins were found to be downregulated in round-headed spermatozoa compared with normal spermatozoa. The differentially expressed proteins that we identified may have important roles in a variety of cellular processes and structures, including spermatogenesis, cell skeleton, metabolism and spermatozoa motility.

  1. Autophagy-related proteins are functionally active in human spermatozoa and may be involved in the regulation of cell survival and motility

    Science.gov (United States)

    Aparicio, I. M.; Espino, J.; Bejarano, I.; Gallardo-Soler, A.; Campo, M. L.; Salido, G. M.; Pariente, J. A.; Peña, F. J.; Tapia, J. A.

    2016-01-01

    Macroautophagy (hereafter autophagy) is an evolutionarily highly conserved cellular process that participates in the maintenance of intracellular homeostasis through the degradation of most long-lived proteins and entire organelles. Autophagy participates in some reproductive events; however, there are not reports regarding the role of autophagy in the regulation of sperm physiology. Hence, the aim of this study was to investigate whether autophagy-related proteins are present and functionally active in human spermatozoa. Proteins related to autophagy/mitophagy process (LC3, Atg5, Atg16, Beclin 1, p62, m-TOR, AMPKα 1/2, and PINK1) were present in human spermatozoa. LC3 colocalized with p62 in the middle piece of the spermatozoa. Autophagy activation induced a significant increase in motility and a decrease in PINK1, TOM20 expression and caspase 3/7 activation. In contrast, autophagy inhibition resulted in decreased motility, viability, ATP and intracellular calcium concentration whereas PINK1, TOM20 expression, AMPK phosphorylation and caspase 3/7 activation were significantly increased. In conclusion our results show that autophagy related proteins and upstream regulators are present and functional in human spermatozoa. Modification of mitochondrial proteins expression after autophagy activation/inhibition may be indicating that a specialized form of autophagy named mitophagy may be regulating sperm function such as motility and viability and may be cooperating with apoptosis. PMID:27633131

  2. 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.

  3. 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.

  4. Zinc content of maturing spermatozoa in oestrogen treated rats.

    Science.gov (United States)

    Srivastava, A; Chowdhury, A R; Setty, B S

    1983-02-01

    Zinc content of spermatozoa collected from the caput and cauda portions of the rat epididymis was determined by atomic absorption spectroscopy. The results showed about 60% reduction in the spermatozoal zinc content by the time they reach the cauda epididymis. This reduction was inhibited in rats receiving micro dose oestrogen which induced 'functional' sterility. It appears that the fall in zinc content of spermatozoa during their transport in the epididymis is related to sperm maturation and that oestrogen treatment interferes with this reduction in sperm zinc content.

  5. Toxicity and uptake of heavy metals by human spermatozoa

    Energy Technology Data Exchange (ETDEWEB)

    Battersby, S.; Chandler, J.A.; Morton, M.S.

    1982-02-01

    The effects of metallic copper on human spermatozoa are described. Incubation with the metal caused a fall on the percentage of motile sperm, which was directly related to the surface area of copper employed and to the copper content of whole semen. These changes were accompanied by a decrease in semen zinc levels and an uptake of copper by individual sperm cells, as determined by X-ray microanalysis. Low concentrations of ionic copper caused a less marked fall in sperm motility, although the metal was generally more toxic than zinc or cadmium ions. All three metals were accumulated by spermatozoa on incubation, with cadmium causing a decrease in sperm nuclear zinc concentrations.

  6. PREDICTION OF GROSS FEED EFFICIENCY IN ITALIAN HOLSTEIN FRIESIAN BULLS

    Directory of Open Access Journals (Sweden)

    Raffaella Finocchiaro

    2015-09-01

    Full Text Available The aim of this study was to predict gross feed efficiency of Italian Holstein Friesian bulls selected for production, functional and type traits. A total of 12,238 bulls, from the April 2015 genetic evaluation, were used. Predicted daily gross feed efficiency (pFE was obtained as ratio between milk yield (MY and predicted dry matter intake (pDMI. Phenotypic trend for MY, predicted body weight (pBW and pFE were calculated by the bull birth year. The results suggest that pFE can be successfully selected to increase profitability of dairy cattle using the current milk recording system. Direct measurements on DMI should be considered to confirm results of pFE obtained in the present study.

  7. Uroperitoneum attributable to ruptured urachus in a yearling bull.

    Science.gov (United States)

    Baxter, G M; Zamos, D T; Mueller, P O

    1992-02-15

    Ruptured urachus was found to result in uroperitoneum in a yearling Beefmaster bull. The uroperitoneum was initially believed to be attributable to ruptured bladder secondary to urolithiasis; however, catheter decompression of the bladder through an ischiatic urethrotomy did not resolve the uroperitoneum. The persistent urachus was diagnosed and removed through caudal right flank laparotomy with the bull standing. The urachus was attached to the umbilicus, communicated with the lumen of the bladder, and had a mucosal lining. Ruptured urachus is an unusual cause of uroperitoneum, but can cause clinical signs identical to those of ruptured bladder. Persistent urachus is a congenital abnormality in many species, but may be hereditary in Beefmaster cattle. In addition, the bull in this report developed hyperkalemia, which is considered an unusual finding in cattle with uroperitoneum.

  8. Effects of in vitro selenium addition to the semen extender on the spermatozoa characteristics before and after freezing in water buffaloes (Bubalus bubalis

    Directory of Open Access Journals (Sweden)

    Kamran Dorostkar

    2012-12-01

    Full Text Available The aim of the present study was to investigate the effect of in vitro supplementation of selenium on fresh and frozen spermatozoa quality of buffalo (Bubalus bubalis bulls. Five healthy buffalo bulls (5 ejaculates from each bull were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control, 0.5, 1, 2, 4 and 8 μg mL-1 sodium selenite and the sperm motility and viability were evaluated at 0 (T0 (immediately after dilution, 60 (T1 and 120 (T2 min after diluting semen. In the second step, semen samples were diluted with tris-egg yolk-glycerol extender containing the same amounts of sodium selenite, cooled to 4 ˚C, equilibrated and semen parameters (motility, viability, membrane integrity and DNA damage were estimated. Then, the semen was packed in 0.5 mL French straws and frozen in liquid nitrogen. Later, the semen was thawed and analyzed for the same parameters, as well as total antioxidant capacity. Results showed that addition of 1 and 2 μgmL-1 selenium to the semen extender significantly increased the sperm motility of fresh and equilibrated semen compared to the control without affecting other parameters. However, in frozen-thawed semen, extenders containing 1 and 2 μg mL-1 selenium significantly improved sperm motility, viability, membrane integrity and semen total antioxidant capacity and also resulted in lower DNA damaged sperms. In this study selenium supplementation of semen extender of 4 and 8 μg mL-1 had deleterious effects on sperm parameters as early as the samples were prepared for freezing.

  9. Effects of in vitro selenium addition to the semen extender on the spermatozoa characteristics before and after freezing in water buffaloes (Bubalus bubalis).

    Science.gov (United States)

    Dorostkar, Kamran; Alavi-Shoushtari, Sayed Mortaza; Mokarizadeh, Aram

    2012-01-01

    The aim of the present study was to investigate the effect of in vitro supplementation of selenium on fresh and frozen spermatozoa quality of buffalo (Bubalus bubalis) bulls. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 0.5, 1, 2, 4 and 8 µg mL(-1) sodium selenite and the sperm motility and viability were evaluated at 0 (T0) (immediately after dilution), 60 (T1) and 120 (T2) min after diluting semen. In the second step, semen samples were diluted with tris-egg yolk-glycerol extender containing the same amounts of sodium selenite, cooled to 4 ˚C, equilibrated and semen parameters (motility, viability, membrane integrity and DNA damage) were estimated. Then, the semen was packed in 0.5 mL French straws and frozen in liquid nitrogen. Later, the semen was thawed and analyzed for the same parameters, as well as total antioxidant capacity. Results showed that addition of 1 and 2 µgmL(-1) selenium to the semen extender significantly increased the sperm motility of fresh and equilibrated semen compared to the control without affecting other parameters. However, in frozen-thawed semen, extenders containing 1 and 2 µg mL(-1) selenium significantly improved sperm motility, viability, membrane integrity and semen total antioxidant capacity and also resulted in lower DNA damaged sperms. In this study selenium supplementation of semen extender of 4 and 8 µg mL(-1) had deleterious effects on sperm parameters as early as the samples were prepared for freezing.

  10. Effects of roads on bull trout (Salvelinus confluentus), a federally threatened species

    OpenAIRE

    Teachout, Emily; Quan, Jennifer

    2001-01-01

    The bull trout (Salvelinus confluentus) is federally listed as threatened under the Endangered Species Act. Bull trout are apex predators requiring a large prey base and a large home range, and are known to move throughout and between basins in search of prey. However, bull trout are dependent upon very cold, clean waters for spawning (below 9 degrees Celsius) and are typically characterized as spawning in the upper-most reaches of watersheds. Bull trout have four life history forms: resident...

  11. Sources of spermatozoa loss during collection and artificial insemination of horses.

    Science.gov (United States)

    Côté, Michelle A; Blum, Katherine M; Burd, Matthew A

    2011-07-01

    During artificial insemination of horses, it is important to accurately estimate the number of spermatozoa in each insemination dose. However, little research exists regarding sources of spermatozoa loss during collection and artificial insemination. Therefore, spermatozoal losses were quantified in the dismount loss (187.6×10(6)±62.5×10(6)spermatozoa), gel fraction (179.8×10(6)±61.7×10(6)spermatozoa), and the collection receptacle (136.1×10(6)±26.9×10(6)spermatozoa). Spermatozoal losses were examined in the centrifuge tube (25.8×10(6)±2.1×10(6)spermatozoa), AI pipette during the air removal (90.9×10(6)±8.5×10(6)spermatozoa), and spermatozoa remaining in the AI pipette after insemination (342.9×10(6)±21.4×10(6)spermatozoa). The average cumulative loss was 14.2±2.9% of the total spermatozoa ejaculated with approximately half of the loss due to the process of semen collection and half due to the process of artificial insemination. Spermatozoa retained in the AI pipette, after insemination with extended semen, represented the greatest source of loss.

  12. Pneumomediastinum and subcutaneous emphysema in a rabid bull

    Directory of Open Access Journals (Sweden)

    Clairton Marcolongo-Pereira

    2015-01-01

    Full Text Available This report describes pneumomediastinum and subcutaneous emphysema in a bull with paralytic rabies. The bull displayed severe swelling in the head, neck, chest and forelimbs extending to the ventrum, including the prepuce, 7 days after the development of neurological signs. At necropsy, extensive subcutaneous emphysema was observed in the neck region, abdominal wall and proximal forelimbs. The lungs failed to collapse and had severe emphysema with bullae. Rabies was suspected due to an ongoing outbreak at the farm and was confirmed by histologic lesions and immunohistochemistry. Similar to humans, pneumomediastinum and subcutaneous emphysema should be considered as an unusual complication of rabies in cattle.

  13. 75 FR 2269 - Endangered and Threatened Wildlife and Plants; Revised Designation of Critical Habitat for Bull...

    Science.gov (United States)

    2010-01-14

    ... Breeding, Reproduction, or Rearing (or Development) of Offspring Bull trout have more specific habitat... Designation of Critical Habitat for Bull Trout in the Coterminous United States; Proposed Rule #0;#0;Federal... Designation of Critical Habitat for Bull Trout in the Coterminous United States AGENCY: Fish and...

  14. 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 gener

  15. 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...

  16. Resazurin reduction and other tests of semen quality and fertility of bulls

    Institute of Scientific and Technical Information of China (English)

    RobertH.Foote

    1999-01-01

    Aim: This study was undertaken to compare the reduction in color of two dyes methylene blue (MBRT) anddye (RRT) with other tests of bull semen quality and to examine their relationship to fertility. Methods: One hundredsixty-four ejaculates from 59 bulls were examined, processed, fertility, and semen from these bulls averaged

  17. 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.

  18. Methods and Effects of Hongshan Cock Spermatozoa Cryopreservation

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The survival rates, acrosomic integrity rates of frozen semen from Hongshan cock and fertilities of the frozen-thawed spermatozoa in both pellets and straws were studied. The result shows that the average survival rate of spermatozoa in straws was 0.451±0.056, and 0.390±0.040 in pellets, the former was significantly higher than the later (n=20, p<0.01). The average acrosomic integrity rate of the sperms was 0.613±0.049 in straws, and 0.476±0.057 in pellets. The former was significantly higher than the later (n=20, p<0.05). The fertility rates (%) of sperms in pellets, straws and freshly collected semen at the third day after artificial insemination (AI) were 85.32±2.32, 87.73±1.00 and 90.77±1.68, respectively. The fertility rates of the three types of semen at the fourth and fifth days were lower than the third day's. Variance analysis shows that the fertilities of the spermatozoa in pellets and straws were significantly lower than freshly collected semen (n=3, p<0.01), and the fertilities of the spermatozoa in straws were significantly higher than those in pellets (n=3, p<0.01).

  19. Application of antioxidants and centrifugation for cryopreservation of boar spermatozoa.

    Science.gov (United States)

    Zhang, Wei; Yi, Kangle; Chen, Chao; Hou, Xiaofeng; Zhou, Xu

    2012-06-01

    Although cryopreserved boar semen has been available since 1975, a major breakthrough in commercial application has not yet occurred due to the high susceptibility of boar spermatozoa to damage during cryopreservation and the complicated process required for deep freezing. In recent years, the application of antioxidants during the cryopreservation of boar semen has been the subject of considerable research aimed at improving the quality of post-thaw semen. Centrifugation is necessary before using cryopreservation protocols for freezing boar spermatozoa. Studies of the effect of different centrifugation regimens on boar sperm recovery, yield and cryosurvival have made significant contributions. Therefore this review elucidates results of recent applications of various antioxidants and centrifugation regimens used in efforts to improve cryopreservation of boar spermatozoa. This review is intended to enhance understanding of the roles of these antioxidants and centrifugation regimens with respect to mechanisms that increase resistance to cryodamage of boar spermatozoa. In addition, the discussion addresses the need for developing an objective evaluation of effectiveness and estimating the prospect of application of new techniques for the cryopreservation of boar semen and its use in artificial insemination.

  20. Motility of liquid stored ram spermatozoa is altered by dilution rate independent of seminal plasma concentration.

    Science.gov (United States)

    Mata-Campuzano, M; Soleilhavoup, C; Tsikis, G; Martinez-Pastor, F; de Graaf, S P; Druart, X

    2015-11-01

    The fertility after use of liquid stored ram semen following cervical AI rapidly decreases if semen is stored beyond 12h. The dilution of seminal plasma is often cited as a key contributor to the diminished motility and fertility of ram spermatozoa subjected to liquid preservation. Two experiments were conducted to assess the effect of spermatozoa concentration (i.e. dilution rate) and percentage of seminal plasma on the motility and viability of liquid stored ram spermatozoa. In Experiment 1, semen was diluted to one of seven concentrations ranging from 0.2 to 1.4×10(9)spermatozoa/ml with milk and assessed for motility after 3 or 24h of storage at 15°C. In Experiment 2, semen was collected and washed to remove seminal plasma before re-dilution to 0.2-1.4×10(9)spermatozoa/ml with milk containing 0%, 20% or 40% (final v/v ratio) seminal plasma and assessed for viability and motility after 3 or 24h of storage at 15°C. Whereas motility was not affected by spermatozoa concentration after 3h of storage, the proportion of progressive spermatozoa decreased after 24h of storage when spermatozoa concentration was greater than 1.0×10(9)spermatozoa/ml. The duration of preservation and the spermatozoa concentration affected spermatozoa motility but had no impact on spermatozoa viability. This negative effect of greater spermatozoa concentrations on motility was independent of the presence and the concentration of seminal plasma. The seminal plasma at both concentrations (20% and 40%) had a protective effect on spermatozoa motility after 24h of storage. These findings have the potential to improve the efficiency of cervical AI with liquid stored ram semen.

  1. The Quality of Spermatozoa of Gembrong Goats during Cryopreservation Process

    Directory of Open Access Journals (Sweden)

    F. A. Pamungkas

    2014-08-01

    Full Text Available Gembrong goat is an Indonesia local goat having specific characteristic that is currently categorized as a breed that is at risk of extinction. In this context, the cryopreservation of gametes is important to support a genome resource bank for storage of gametes for an indefinite period of time. Evaluation of semen and spermatozoa quality was performed to determine the survival of spermatozoa and this information will be used as a reference in the cryopreservation of semen and spermatozoa. The aim of this experiment was to study the characteristics of Gembrong goat’s semen and spermatozoa during cryopreservation process. Once a week, semen from three Gembrong goats (ages about 2-3 years old was collected using artificial vagina and then frozen with TRIS extender. After freezing, the semen was thawed. Macro- and microscopic parameters of semen and spermatozoa were assessed in fresh and frozen-thawed semen. Results showed that in the fresh semen, the volume was 0.5 mL, sperm abnormalities was 5.74%, sperm concentration was 6731 x 106/mL, the sperm motility was 78.33%, live sperm was 83.17%, and sperm membrane integrity was 78.53%. After-thawing observation showed that sperm motility decreased to 49% (P<0.05 that was lower as compared to that in the fresh and post-equilibration semen. Similarly, the percentage of sperm viability and membrane integrity during cryopreservation showed a similar pattern with the sperm motility. In conclusion, the fresh semen of Gembrong goat had a good quality and met the requirement for further cryopreservation process. Similarly, the quality of frozen-thawed semen of Gembrong goat is eligible for artificial insemination (AI or in vitro embryo production.

  2. How to Identify and Forecast Bull and Bear Markets?

    NARCIS (Netherlands)

    H.J.W.G. Kole (Erik); D.J.C. van Dijk (Dick)

    2013-01-01

    textabstractThe state of the equity market, often referred to as a bull or a bear market, is of key importance for financial decisions and economic analyses. Its latent nature has led to several methods to identify past and current states of the market and forecast future states. These methods encom

  3. The White Bull effect: abusive coauthorship and publication parasitism.

    Science.gov (United States)

    Kwok, L S

    2005-09-01

    Junior researchers can be abused and bullied by unscrupulous senior collaborators. This article describes the profile of a type of serial abuser, the White Bull, who uses his academic seniority to distort authorship credit and who disguises his parasitism with carefully premeditated deception. Further research into the personality traits of such perpetrators is warranted.

  4. The effect of selected staining techniques on bull sperm morphometry.

    Science.gov (United States)

    Banaszewska, Dorota; Andraszek, Katarzyna; Czubaszek, Magdalena; Biesiada-Drzazga, Barbara

    2015-08-01

    Sperm morphometry has some value as an indicator of reproductive capacity in males. In laboratory practice a variety of slide-staining methods are used during morphological evaluation of semen to predict male fertility. The aim of this study was to determine the effect of staining of semen using four different techniques on the morphometry of the bull sperm cell. The material for the study consisted of semen collected from test bulls of the Black-and-White variety of Holstein-Friesians. The results obtained in the study indicate differences in the dimensions of bull sperm heads when different slide staining techniques were used. The most similar results for sperm head dimensions were obtained in the case of SpermBlue(®) and eosin+gentian violet complex, although statistically significant differences were found between all the staining techniques. Extreme values were noted for the other staining techniques - lowest for the Papanicolaou and highest for silver nitrate, which may indicate more interference in the cell by the reagents used in the staining process. However, silver nitrate staining was best at identifying the structures of the sperm cell. Hence it is difficult to determine which of the staining methods most faithfully reveals the dimensions and shape of the bull sperm.

  5. Consequences of hazardous dietary calcium deficiency for fattening bulls

    Directory of Open Access Journals (Sweden)

    Näkki Päivi

    2006-12-01

    Full Text Available Abstract Background Deficient mineral supplementation on a feedlot farm resulted in severe clinical manifestations in fattening bulls. Animals mistakenly received only 60–70% of the recommended calcium intake, while simultaneously receiving twice the amount of phosphorus recommended. Thus, the dietary Ca/P ratio was severely distorted. After approximately six months on such a diet, four fattening bulls were euthanized because of severe lameness and 15% of other animals on the farm were having clinical leg problems. Veterinary consultation revealed the mistake in mineral supplementation. Methods Fattening bulls were divided into three groups depending on the time of their arrival to the farm. This enabled the effect of mineral imbalance at different growth phases to be examined. After slaughtering, the bones of both front and hind limbs were macroscopically evaluated. Results Over 80% of the animals with a calcium-deficient diet had at least one severe osteoarthritic lesion. The economic impact of the calcium deficiency was statistically significant. Conclusion Calcium deficiency with distorted Ca/P ratio yielded a severe outbreak of osteoarthritis in fattening bulls. Calcium deficiency caused a more serious lesions in age group 5–12 months than age group 12–18 months. Besides causing obvious economic losses osteoarthritis is also a welfare issue for feedlot animals.

  6. Exploring Essential Conditions: A Commentary on Bull et al. (2008)

    Science.gov (United States)

    Borthwick, Arlene; Hansen, Randall; Gray, Lucy; Ziemann, Irina

    2008-01-01

    The editorial by Bull et al. (2008) on connections between informal and formal learning made explicit one element of solving what Koehler and Mishra (2008) termed a "wicked problem." This wicked (complex, ill-structured) problem involves working with teachers for effective integration of technology in support of student learning. The…

  7. 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.

  8. 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

  9. Effect of sperm concentration in an ejaculate on morphometric traits of spermatozoa in Duroc boars.

    Science.gov (United States)

    Kondracki, S; Wysokińska, A; Iwanina, M; Banaszewska, D; Sitarz, D

    2011-01-01

    The experimental material consisted of 75 ejaculates collected form 8 Duroc boars. The ejaculates were divided into three groups according to sperm concentration in an ejaculate. An ejaculate was obtained from each boar monthly and it was used to make microscopic preparations to examine spermatozoa morphology. In each preparation morphometric measurements were taken of fifteen randomly selected spermatozoa characterized by normal morphology. The following measurements of spermatozoa were taken: length and width of the spermatozoa head, head area, length of the flagellum, perimeter of the spermatozoon head and total spermatozoon length. The results were used to calculate indicators of spermatozoa morphology. Moreover, assessments were made of frequency of morphological defects to isolate spermatozoa with primary and secondary abnormalities following the Blom classification system. It was found that the concentration of spermatozoa in the ejaculate influenced the morphometric characteristics of spermatozoa. Ejaculates with low sperm concentrations are characterized by larger spermatozoa as compared to ejaculates with high sperm concentrations. However, sperm concentration in the ejaculate does not much influence the shape of spermatozoa.

  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. Low temperature storage of rhesus monkey spermatozoa and fertility evaluation by intracytoplasmic injection.

    Science.gov (United States)

    Yeoman, Richard R; Mitalipov, Shoukhrat; Gerami-Naini, Behzad; Nusser, Kevin D; Wolf, Don P

    2005-06-01

    The objective was to develop a sperm freezing procedure suitable for use in the propagation of valuable founder animals by assisted reproductive technologies. Here, we report a comparison of processing methods by measuring the motility of fresh and frozen-thawed rhesus monkey spermatozoa and fertility via intracytoplasmic spermatozoa injection (ICSI) of sibling oocytes. Washed spermatozoa were frozen in straws or in pellets using different cryoprotective media and processed post-thaw with or without a density gradient centrifugation step. Among the four study series, motility post-thaw was improved with density gradient centrifugation (17-24% versus 75%, P<0.01) achieving levels similar to fresh spermatozoa. Spermatozoa injected oocytes (total n=377) were co-cultured on BRL cells and observed for fertilization and development. With spermatozoa frozen in straws in liquid nitrogen vapors, the fertilization rate after ICSI was lower than with fresh spermatozoa (40-44% versus 77-86%, P<0.05), even with the Percoll-enriched fraction that exhibited robust motility. In contrast, somewhat slower freezing of spermatozoa in pellets on dry ice supported fertilization rates (73%) that were similar to the fresh counterpart. Developmental rates of fertilized eggs were similar in all experiments. A total of 106 embryo transfers has resulted in the first primate born after ICSI with F/T ejaculated spermatozoa plus 22 other infants to date. Additionally, a 3-4 h incubation after thawing improved the fertilization rate with spermatozoa from a male with poor post-thaw recovery of sperm motility. In conclusion, an acceptable fertilization rate after ICSI with motile, frozen-thawed primate spermatozoa was observed comparable to that obtained with fresh spermatozoa allowing small quantities of competent spermatozoa to be used with ICSI to facilitate propagation of desirable primate genotypes.

  12. 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.

  13. 泰国湿地水牛(Bubalus bubalis)精液质量的季节变化%Seasonal variation in semen quality of swamp buffalo bulls (Bubalus bubalis) in Thailand

    Institute of Scientific and Technical Information of China (English)

    S.Koonjaenak; V.Chanatinart; S.Aiumlamai; T.Pinyopumimintr; H.Rodriguez-Martinez

    2007-01-01

    Aim:To test the hypothesis that season affects the semen quality of swamp buffalo (Bubalus bubalis) bulls used for artificial insemination (AI) under tropical conditions in Thailand, as it does in Bos taurus and Bos indicus. Methods: Clinical and andrological examinations, and monitoring of semen production and quality were carried out on five mature, healthy swamp buffalo AI bulls in Thailand from July 2004 to the end of June 2005. Sperm output, motility, morphology and plasma membrane integrity (PMI) were compared between three seasons of the year (rainy, I.e. July-October; winter, I.e. November-February; and summer, I.e. March-June) with distinct ambient temperature and humidity. Results: All bulls were diagnosed as clinically healthy and with good libido throughout the study. Ejaculate volume, pH, sperm concentration, total sperm number and initial sperm motility did not differ between seasons, whereas PMI and the relative proportion of morphologically normal spermatozoa were highest in summer and lowest in winter (P < 0.05). Buffalo age, week of collection and season influenced sperm morphology (P < 0.05-0.001). Among morphological abnormalities,only proportions of tail defects were affected by season, being highest in the rainy season and lowest in summer (P<0.001). In conclusion, climatic changes did not seem to largely affect semen sperm output or viability. Although the proportions of PMI and tail abnormalities were affected by season, they were always below what is considered unacceptable for AI bull sires. Conclusion: Seasonal changes did not appear to cause deleterious changes in sperm quality in swamp buffalo AI-sires in tropical Thailand.

  14. 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

  15. Reproductive characteristics of grass-fed, luteinizing hormone-releasing hormone-immunocastrated Bos indicus bulls.

    Science.gov (United States)

    Hernandez, J A; Zanella, E L; Bogden, R; de Avila, D M; Gaskins, C T; Reeves, J J

    2005-12-01

    Two field trials were conducted in Brazil to evaluate LHRH immunocastration of Bos indicus bulls (d 0 = 2 yr of age). In Study I, 72 bulls were assigned randomly to one of three treatment groups: LHRH0-immunized, castrated, and intact. Immunized animals (n = 25) received a primary and two booster injections of ovalbumin-LHRH-7 and thioredoxin-LHRH-7 fusion proteins on d 0, 141, and 287. Twenty-three bulls were surgically castrated on d 141, and 24 served as intact controls. All animals were slaughtered on d 385, at approximately 3 yr of age. In Study II, 216 bulls were assigned randomly to the same three treatments as in Study I; however, because of a drought in the area, bulls were kept on pasture an additional year, and a fourth treatment was added, in which one-half the LHRH-immunized bulls received an additional booster on d 639 (fourth immunization). All animals in Study II were slaughtered on d 741 (4 yr of age). Luteinizing hormone-releasing hormone antibodies increased following each immunization for immunized bulls, but they were not detectable in castrate or intact animals in either study. Consequently, scrotal circumference was suppressed in immunized bulls compared with intact controls in both studies. By d 287, serum concentrations of testosterone in LHRH-immunized bulls were decreased compared with intact controls (P bulls (173 +/- 22 and 26 +/- 2 g, respectively) and fourth immunization bulls (78 +/- 23 and 20 +/- 2 g, respectively; Study II). At the end of each study, BW was greater (P bulls than for castrated and LHRH-immunized animals. In these two studies, the efficacy of the LHRH fusion proteins to induce an effect similar to that of surgical castration was considered 92 and 93%, respectively. These data support the concept that immunocastration of bulls at 2 yr of age was successful and that it has practical application as a tool for producing grass-fattened bulls in Brazil.

  16. Understanding the mechanisms of ATPase beta family genes for cellular thermotolerance in crossbred bulls

    Science.gov (United States)

    Deb, Rajib; Sajjanar, Basavaraj; Singh, Umesh; Alex, Rani; Raja, T. V.; Alyethodi, Rafeeque R.; Kumar, Sushil; Sengar, Gyanendra; Sharma, Sheetal; Singh, Rani; Prakash, B.

    2015-12-01

    Na+/K+-ATPase is an integral membrane protein composed of a large catalytic subunit (alpha), a smaller glycoprotein subunit (beta), and gamma subunit. The beta subunit is essential for ion recognition as well as maintenance of the membrane integrity. Present study was aimed to analyze the expression pattern of ATPase beta subunit genes (ATPase B1, ATPase B2, and ATPase B3) among the crossbred bulls under different ambient temperatures (20-44 °C). The present study was also aimed to look into the relationship of HSP70 with the ATPase beta family genes. Our results demonstrated that among beta family genes, transcript abundance of ATPase B1 and ATPase B2 is significantly ( P P < 0.01) with HSP70, representing that the change in the expression pattern of these genes is positive and synergistic. These may provide a foundation for understanding the mechanisms of ATPase beta family genes for cellular thermotolerance in cattle.

  17. External morphology of spermatozoa and spermatozeugmata of the freshwater mussel Truncilla truncata (Mollusca: Bivalvia: Unionidae)

    Science.gov (United States)

    Waller, D.L.; Lasee, B.A.

    1997-01-01

    Truncilla truncata males release spherical aggregates of spermatozoa, called spermatozeugmata, at spawning. Sperm aggregates from other bivalve species have been described, but few detailed studies exist of the morphology of unionid spermatozeugmata and spermatozoa. We provide the first description of the external morphology of spermatozeugmata and spermatozoa of T. Truncata. The spermatozeugmata had an inside diameter of 76 mu m and contained 8000-9000 spermatozoa. Heads of spermatozoa were directed toward the center of the sphere into a translucent shell; tails were arranged radially and caused the spermatozeugmata to rotate. Spermatozoa of T. Truncata measured 3.3 mu m in length (excluding tail) and each had a head, a midpiece and a flagellum. We also documented the release of spermatozeugmata in two additional unionid species, Lampsilis cardium and Amblema plicata plicata.

  18. Diet-induced obesity alters kinematics of rat spermatozoa

    Institute of Scientific and Technical Information of China (English)

    IP Oyeyipo; PJ Maartens; SS duPlessis

    2015-01-01

    Objective:To investigate the effect of DIO on the kinematics and viability of spermatozoa in an albino rat model.Methods:Sperm suspensions from normal (Control) and diet-induced obese (DIO) Wistar rats were collected and incubated for various times (30, 60, 120 or 180 min at 37℃). Motility parameters were analyzed with computer-aided sperm analysis (CASA), while viability was assessed by means of a dye exclusion staining technique (eosin/nigrosin).Results: Results reveal that there was a significant time dependent decrease (P<0.05) in progressive motility, curvilinear velocity and beat cross frequency after 60 min, while amplitude of lateral head displacement and sperm viability was significantly reduced (P<0.05) after 120 min in the DIO group compared to control spermatozoa.Conclusions: These results provided evidence that obesity is detrimental to sperm parameter in rats possibly through increased testicular temperature as a result of a rise in fat deposition.

  19. Search for a small egg by spermatozoa in restricted geometries.

    Science.gov (United States)

    Yang, J; Kupka, I; Schuss, Z; Holcman, D

    2016-08-01

    The search by swimmers for a small target in a bounded domain is ubiquitous in cellular biology, where a prominent case is that of the search by spermatozoa for an egg in the uterus. This is one of the severest selection processes in animal reproduction. We present here a mathematical model of the search, its analysis, and numerical simulations. In the proposed model the swimmers' trajectories are rectilinear and the speed is constant. When a trajectory hits an obstacle or the boundary, it is reflected at a random angle and continues the search with the same speed. Because hitting a small target by a trajectory is a rare event, asymptotic approximations and stochastic simulations are needed to estimate the mean search time in various geometries. We consider searches in a disk, in convex planar domains, and in domains with cusps. The exploration of the parameter space for spermatozoa motion in different uterus geometries leads to scaling laws for the search process.

  20. Inositol tri-phosphate inhuman and ascidian spermatozoa.

    Science.gov (United States)

    Tosti, E; Palumbo, A; Dale, B

    1993-05-01

    Using a specific protein binding assay we have shown that a spermatozoon of the ascidian Ciona intestinalis contains 1.58 +/- 0.74 x 10(-19) moles of inositol 1,4,5-tri-phosphate (InsP3), while a human spermatozoon contains 6.4 +/- 0.14 x 10(-19) moles. Induction of the acrosome reaction (AR) in both species, by exposure to the calcium ionophore A23187, does not significantly alter levels of InsP3, suggesting that phosphatidylinositol (PI) turnover is not necessary for the calcium ionophore induced AR. Furthermore, PI turnover in ascidian spermatozoa appears to be insensitive to lithium and phorbol ester. The high intracellular concentration of InsP3 in spermatozoa, corresponding to 50-200 microM, suggests it may play a role in egg activation.

  1. 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

  2. Behavior and inflammation of the rumen and cecum in Holstein bulls fed high-concentrate diets with different concentrate presentation forms with or without straw supplementation.

    Science.gov (United States)

    Devant, M; Penner, G B; Marti, S; Quintana, B; Fábregas, F; Bach, A; Arís, A

    2016-09-01

    Twenty-four individually housed Holstein bulls (395 ± 7.3 kg BW and 252 ± 3.1 d age) were exposed to a 2 × 2 factorial design (meal vs. pellets; with vs. without straw) to evaluate the effect of concentrate form and provision of straw in finishing diets on behavior and expression of rumen and cecum epithelium genes related to inflammation and behavior. Concentrate and straw consumption were recorded monthly and behavior (self-grooming, social, oral nonnutritive, tongue rolling, eating, drinking, ruminating, and lying) was recorded every two weeks. Bulls were slaughtered after 64 d of exposure to treatments, lesions on the rumen and liver were assessed, and samples of the rumen and cecum were collected. Straw supplementation tended ( = 0.08) to increase concentrate intake (8.0 vs. 7.4 ± 0.26 kg/d), increased ( straw. Provision of straw increased ruminal pH, but the magnitude of the change was greater when the concentrate was provided as meal compared with pellets (interaction, straw was not supplemented, all rumen samples had papillae fusion, whereas only 16.7% of bulls fed pellets and straw had papillae fusions (interaction, straw compared with bulls without straw. For the ruminal epithelium, straw provision tended to increase the relative expression ratio of (which stimulates peptide YY, PYY, and serotonin secretion; = 0.06) and α (which modulates immune reactions and behavior; = 0.09) and increased and (tight junction proteins; straw had increased ( straw supplementation in bulls fed high-concentrate diets modifies behavior and affects rumen macroscopic morphology and expression of epithelial genes that could be related to behavior and inflammation.

  3. 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

  4. Hydroxyflutamide alters the characteristics of live boar spermatozoa.

    Science.gov (United States)

    Zarzycka, Marta; Kotwicka, Malgorzata; Jendraszak, Magdalena; Skibinska, Izabela; Kotula-Balak, Malgorzata; Bilinska, Barbara

    2014-10-15

    Our previous study revealed that in vitro incubation of boar ejaculates with hydroxyflutamide (OH-Flu) causes changes in sperm plasma membrane integrity and its stability and sperm mitochondrial oxidative capability. To broaden the knowledge of cellular physiology of spermatozoa, we investigated direct effects of OH-Flu administered for 2 and 24 hours at concentrations of 5, 50, and 100 μg/mL, on sperm mitochondrial membrane potential and mitochondrial superoxide anion production using JC-1 dye and MitoSOX Red fluorescent probe, respectively. We further measured phosphatidylserine membrane translocation (PST) from the inner to the outer layer of the sperm plasma membrane using an annexin-V binding assay. To provide new information of direct effects of OH-Flu on cell signaling pathway, we measured sperm intracellular calcium ion dynamics using Fluo-3. Finally, we assessed sperm motility using a computer-assisted spermatozoa analysis system. Motile sperm were highlighted using the "C-Ruch" computer program for detailed analysis of the straight line velocity distribution. For each functional test, boar spermatozoa were examined and analyzed by flow cytometry and/or confocal microscopy. The results revealed a significant decrease (Psperm mitochondrial membrane potential and a concomitant increase (Psperm motility. Hydroxyflutamide significantly decreased (Psperm subpopulation percentage after 15 minutes and reduced the straight line velocity distribution (Psperm intracellular calcium ion concentration. Altogether, the altered in vitro characteristics of live boar spermatozoa provide new insight into direct effects of OH-Flu on sperm mitochondrial membrane potential, superoxide anion production, translocation of membrane phosphatidylserine, free calcium ion dynamics, and sperm motility.

  5. A redox-regulated tyrosine phosphorylation cascade in rat spermatozoa.

    Science.gov (United States)

    Lewis, B; Aitken, R J

    2001-01-01

    Rat spermatozoa from both the caput and cauda epididymidis were shown to generate superoxide anion (O2-.) both spontaneously and following stimulation with NAD(P)H. Caput spermatozoa gave a significantly greater O2- response to NADPH stimulation than caudal cells, whereas in both cell types the responses to exogenous NADPH and NADH were approximately equivalent. Analysis of H2O2 production revealed that this oxidant was generated only by caudal epididymal cells and only in these cells did the stimulation of reactive oxygen species (ROS) production with NADPH lead to an increase in tyrosine phosphorylation. Stimulation of ROS production with NADPH increased intracellular cyclic adenosine monophosphate (cAMP) levels in both caput and caudal epididymal cells, but only in caudal cells did cAMP stimulate tyrosine phosphorylation, in keeping with the NADPH results. On the basis of these findings we propose that tyrosine phosphorylation in rat spermatozoa is driven by ROS acting via 2 different but complementary mechanisms; O2-. stimulates tyrosine kinase activity indirectly through the elevation of intracellular cAMP while H2O2 acts directly on the kinase/phosphatase system, stimulating the former and inhibiting the latter. Zinc was examined as a potential regulator of this signal transduction cascade and was shown to suppress tyrosine phosphorylation in caput cells but to promote this activity in caudal spermatozoa, possibly through an inhibitory effect on tyrosine phosphatase activity. These results reveal the maturation of a redox-regulated, cAMP-mediated, signal transduction cascade during epididymal transit in the rat that is sensitive to zinc and plays a key role in the control of tyrosine phosphorylation events associated with capacitation.

  6. Effect of zinc oxide nanoparticles on viability of human spermatozoa

    OpenAIRE

    Abolfazl Barkhordari; Seyedhossein Hekmatimoghaddam; Ali Jebali; Mohammad Ali Khalili; Alireza Talebi; Marzieh Noorani

    2013-01-01

    Background: The extensive use of different nanoparticles has raised great concerns about their occupational and biological safety. Objective: The aim of this study was to evaluate the cytotoxic effect of zinc oxide nanoparticles (ZnO NPs) on viability of spermatozoa. Materials and Methods: Semen samples were obtained from 15 healthy persons, and were analyzed using WHO guidelines. Each semen sample was separately incubated with different concentrations of ZnO NPs (10, 100, 500, and 1000 µg/mL...

  7. In vitro Study of the Spermatozoa Motility in the Lizard Eutropis carinata

    Directory of Open Access Journals (Sweden)

    Divya Dosemane

    2015-01-01

    Full Text Available Reptilian epididymis is considered as an important excurrent duct system required for the sperm maturation. Reptilian epididymis synthesizes and secretes proteins which vary in different regions of the epididymis. Hence, to investigate the effect of the secretions of different regions of the epididymis on spermatozoa motility, an in vitro study was undertaken to observe the changes in the patterns of motility of the testicular spermatozoa incubated with luminal contents of different regions of the epididymis in the lizard Eutropis carinata for the first time. The non motile testicular spermatozoa from the testis exhibited different patterns of motility, when incubated with the luminal contents of different regions of the epididymis. The spermatozoa from the testis, different regions of the epididymis exhibited 8 different patterns of motility (a-h. Testicular spermatozoa incubated with the anterior and middle epididymal luminal contents showed the motility patterns almost similar to that of the spermatozoa of the anterior and middle epididymis respectively. In contrast to the spermatozoa of the posterior epididymis, none of the testicular spermatozoa showed any movement when incubated with the posterior epididymal luminal contents. This study throws light on the importance of each region of epididymis in the physiological maturation of spermatozoa.

  8. Hydrogen Sulfide and/or Ammonia Reduces Spermatozoa Motility through AMPK/AKT Related Pathways

    Science.gov (United States)

    Zhao, Yong; Zhang, Wei-Dong; Liu, Xin-Qi; Zhang, Peng-Fei; Hao, Ya-Nan; Li, Lan; Chen, Liang; Shen, Wei; Tang, Xiang-Fang; Min, Ling-Jiang; Meng, Qing-Shi; Wang, Shu-Kun; Yi, Bao; Zhang, Hong-Fu

    2016-11-01

    A number of emerging studies suggest that air pollutants such as hydrogen sulfide (H2S) and ammonia (NH3) may cause a decline in spermatozoa motility. The impact and underlying mechanisms are currently unknown. Boar spermatozoa (in vitro) and peripubertal male mice (in vivo) were exposed to H2S and/or NH3 to evaluate the impact on spermatozoa motility. Na2S and/or NH4Cl reduced the motility of boar spermatozoa in vitro. Na2S and/or NH4Cl disrupted multiple signaling pathways including decreasing Na+/K+ ATPase activity and protein kinase B (AKT) levels, activating Adenosine 5‧-monophosphate (AMP)-activated protein kinase (AMPK) and phosphatase and tensin homolog deleted on chromosome ten (PTEN), and increasing reactive oxygen species (ROS) to diminish boar spermatozoa motility. The increase in ROS might have activated PTEN, which in turn diminished AKT activation. The ATP deficiency (indicated by reduction in Na+/K+ ATPase activity), transforming growth factor (TGFβ) activated kinase-1 (TAK1) activation, and AKT deactivation stimulated AMPK, which caused a decline in boar spermatozoa motility. Simultaneously, the deactivation of AKT might play some role in the reduction of boar spermatozoa motility. Furthermore, Na2S and/or NH4Cl declined the motility of mouse spermatozoa without affecting mouse body weight gain in vivo. Findings of the present study suggest that H2S and/or NH3 are adversely associated with spermatozoa motility.

  9. Cauda Epididymis Spermatozoa: Cryopreservation and Utilization for Artificial Insemination and In Vitro Fertilization

    Directory of Open Access Journals (Sweden)

    Fitra Aji Pamungkas

    2012-12-01

    Full Text Available Genetic material either from animals of economical interest or from wildlife conservation can be lost anytime by unexpected death of the animal, low libido, or disorder at reproduction. In this case, an effort can be made occur to avoid the total lost of that genetic material by using an epididymis spermatozoa. Cauda epididymis spermatozoa generally motile, mature and can be used to fertilize oocytes as well as ejaculated spermatozoa. Some research indicated that cryopreservation of cauda epididymis spermatozoa for the purpose of artificial insemination and in vitro fertilization showed the ability to fertilize oocytes and produce offspring.

  10. Differential analysis of two-dimensional gel electrophoresis profiles of spermatozoa protein in human normal motility sperm and idiopathic asthenospermia

    Institute of Scientific and Technical Information of China (English)

    SHEN Shu-lin; HE Da-lin; LUO Yong; NING Liang

    2006-01-01

    Objective: To evaluate the application of two-dimensional electrophoresis in the research of differentially expressed proteins in the human asthenospermia. Methods: Two-dimensional gel electrophoresis was performed on 4 normal sperm samples from healthy men and 4 sperm samples from 4 asthenospermia patients. After silver staining, the differential expression proteins were analyzed by PDQuest 2D analysis software. Results: Six differential protein spots were identified. Four spots showed increased expression in the control gels compared with the patient gels. Conclusion: The protein profiles of differential expression between the normal spermatozoa and idiopathic asthenospermia were established and some differential proteins were found. The data of this study would establish the better fundament for further isolation and identification of differentially expressed proteins in human asthenospermia sperm.

  11. 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

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

    Science.gov (United States)

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

    2012-01-01

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

  13. 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

  14. Bull breeding soundness, semen evaluation and cattle productivity.

    Science.gov (United States)

    Chenoweth, P J; McPherson, F J

    2016-06-01

    The bull breeding soundness evaluation (BBSE) has evolved as a cost-effective veterinary procedure which provides benefits such as risk-reduction and improvements in strategic bull usage, herd fertility and economics. Semen evaluation is an important component of the BBSE when performed appropriately; a consideration that is increasingly addressed by third party andrology laboratories. The combination of competent physical/reproductive exams (including scrotal circumference measurements) and semen evaluations can contribute greatly to the fertility and economics of individual herds as well as adding to understanding of those factors which affect cattle fertility. Despite such advantages, there remain challenges in achieving full acceptance of BBSEs, particularly by the dairy industry and in developing countries.

  15. Analysis of High Reproductivity on Wild Yak Bull

    Institute of Scientific and Technical Information of China (English)

    陆仲璘

    2005-01-01

    Sperm concentration of wild yak bull is 2. 13 billion per milliliter. Survival time at 0-4 ℃ is 57hrs. After thawing survival time at 37 ℃ is 12 hrs. Resistance coefficient is 144,000. Abnormal sperm rate and acrosomal integrated rate of post-thawing is 9.17% and 87.53% respectively. Moving viscosity is 1. 169 cp. Total nitrogen is 1 437.7 mg/100 ml. Head of sperm is significantly shorter and end piece is significantly longer than that of domestic yak and yellow cattle. Activity of hyaluronidase is highly stronger than the domestic yak,and the activity of LDH is higher than the domestic yak by 48 %. Hence,the fertility of the wild yak bull is powerful and the artificial inseminated rate with domestic yak and yellow cattle is 88.9% and 71.58% respectively.

  16. 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.

  17. In vitro fertility assessment of Kundhi buffalo bull semen

    Directory of Open Access Journals (Sweden)

    Hamzo Khan Kunbhar,

    2011-02-01

    Full Text Available The study was conducted on in-vitro fertility assessment of frozen thawed semen collected from Kundhi buffalo bull maintained at Department of Animal Reproduction, Faculty of Animal Husbandry and Veterinary Sciences, Sindh Agriculture University Tandojam. Before freezing of semen, each ejaculate was assessed for volume, sperm concentration, mass activity and moss motility percentage. Twenty semen samples having motility 60% or above were frozen for post-thaw assessment. Frozen thawed semen was incubated at 250C for 5 hours and examined for progressive linear motility and live dead sperm count. The mean volume, mass activity, moss motility percentage sperm concentrations and pH of the semen were found to be 2.79±0.217 ml, 2.85±0.111, 71.75±2.621, 11.35±1.255 millions/ml and 5.8185±0.092 respectively of fresh semen. No significant difference was found between the parameters except pH, which was significantly different between the bulls. The mean sperm motility percentage and live dead sperm count % of Kundhi buffalo bull semen was found to be 20.46±1.62 and 6.9± 0.2% for frozen semen. A significant (P< 0.05 difference was found between the bulls for post-thaw motility percentage. It was found that at 01 hour incubation, 43.25±2.95% of sperms were motile having 11.78±0.28 % dead sperm count. It was gradually decline from 0 to 5 hours incubation, After 5 hours, all sperms were found dead. It is concluded that sperms maintaining long term motility and having less live dead sperms count were considered suitable for artificial insemination.

  18. Selection of bull dams in population of Simmental cattle

    OpenAIRE

    Pantelić V.; Novaković Ž.; Ostojić-Andrić D.

    2009-01-01

    Increase of production and improvement of the quality of milk, as well as of the intensity of fertility, are main prerequisites of modern cattle production. For the purpose of production of domestic Simmental bulls it is necessary to select the best cows from main herd. These heads as a rule represent approx. 1% of best cows in the controlled population primary in regard to production of milk and milk fat, but also in regard to body development, udder development and fertility traits. Fertili...

  19. Interrelationships Between Apoptosis and Fertility in Bull Sperm

    OpenAIRE

    Dogan, Sule; MASON, Melissa C.; Govindaraju, Aruna; BELSER, Lauren; Kaya, Abdullah; Stokes, John; ROWE, Dennis; Memili, Erdogan

    2012-01-01

    Abstract Male fertility, the ability of sperm to fertilize and activate the egg and support early embryogenesis, is vital for mammalian reproduction. Despite producing adequate numbers of sperm with normal motility and morphology, some males suffer from low fertility whose molecular mechanisms are not known. The objective was to determine apoptosis in sperm from high and low fertility bulls and its relationship with male fertility. DNA damage, phosphatidylserine (PS) translocation, and expres...

  20. Gender Metaphor for Colonialism in John Bull's Other Island

    Institute of Scientific and Technical Information of China (English)

    曹笑丽

    2015-01-01

    John Bull's Other Island is the only play of George Bernard Shaw dealing with the Irish problem.He manages to define and ridicule colonialism in it and satires English colonialist’s intention and nature.This paper studies colonialism in this play through sexual metaphor,to decipher how it works simultaneously as an intimation of the British colonial attitudes and a satire of economicexploitation.

  1. Bull selection and use in northern Australia. 5. Social behaviour and management.

    Science.gov (United States)

    Fordyce, G; Fitzpatrick, L A; Cooper, N J; Doogan, V J; De Faveri, J; Holroyd, R G

    2002-05-15

    Calf output of bulls was derived using DNA typing for paternity following multiple-sire mating at two sites in northern Australia. At Swan's Lagoon Beef Cattle Research Station, 12, mixed-age, Brahman cross bulls were continuously mated with an average of 325 females in a 22km2 open-savannah paddock. Water was available in two troughs. Behaviour of the bulls and location of cows were monitored. At Kamilaroi Station, 2- to 2.5-year-old Brahman bulls were introduced to the study. Twenty-four bulls (HIGH%) were mated in an 84km2 paddock for 3.5 months to 411 heifers in 1995/1995 and for 4.5 months to 350 heifers and 320 first-lactation cows in 1995/1996. A second group of 10 bulls (LOW%) selected on reproductive soundness was mated concurrently in a neighbouring 60km2 paddock to 411 heifers in 1995/1995 and to 350 heifers and 298 first-lactation cows in 1995/1996. In each paddock in both years, 300-350 females were expected to cycle during mating. Both paddocks were flat and semi-forested and water was available only at troughs. At both sites, detailed physical and reproductive examinations of all bulls were conducted prior to and post-mating.Calf output of individual bulls was highly variable but repeatable (r=0.6-0.7) between years. Up to 90% of the 270-380 calves resulting from each mating were sired by between 6 and 8 bulls. Reducing from 3.7 to 2.8% bulls:females at Swan's Lagoon did not delay conceptions. At Kamilaroi, reproductively sound bulls achieved an estimated 5-6 conceptions per week over the peak mating period when sufficient cycling females were available. Differences in pregnancy rates between paddocks appeared due to differences in nutrition and it appeared that conceptions were not delayed with LOW% vs. HIGH% bulls. Variance between bulls in calf output was substantially lower when fewer bulls were used. Bull attrition occurred each year in the HIGH% paddock but not in the LOW% paddock. Calf output was unrelated to body condition of bulls. Seven of

  2. Pregnancy rates in cattle with cryopreserved sexed spermatozoa: effects of laser intensity, staining conditions and catalase.

    Science.gov (United States)

    Schenk, J L; Seidel, G E

    2007-01-01

    The overall aim of this research was to improve fertility of cattle inseminated with sexed spermatozoa by improving sperm sorting procedures. Six field trials were conducted in which 4,264 heifers were inseminated into the uterine body with cryopreserved sexed or unsexed control spermatozoa. Pregnancy or calving rates with doses of 2 x 10(6) sexed spermatozoa ranged from 32 to 51%; these averaged 69% of the pregnancy rates with 20 x 10(6) unsexed, control spermatozoa (range 53 to 79% of controls). Fertility of sexed spermatozoa was especially low on farms where control fertility was low. Accuracy of sexing ranged from 86 to 91%. Laser power of 150 mW for interrogating spermatozoa did not result in lower pregnancy rates (43%) than when power was decreased as much as possible for a particular sorting batch (50 to 130 mW) to still achieve sexing accuracy (38% pregnant). Addition of catalase to fluids containing spermatozoa was beneficial when thawed spermatozoa were incubated in vitro for 2 h but had no effect on pregnancy rates. There also was no effect on pregnancy rates between two concentrations of Hoechst 33342 for staining spermatozoa. Freezing 2 x 10(6) sexed spermatozoa at 20 x 10(6)/ml resulted in a slightly higher rate of pregnancy (P < 0.05) than at 10 x 10(6)/ml. The information obtained in these trials, along with other improvements, notably lowering pressure in the sorting system from 50 to 40 psi, has been used to improve procedures for sexing spermatozoa commercially.

  3. Effects of different concentrations of sucrose or trehalose on the post-thawing quality of cattle bull semen

    Institute of Scientific and Technical Information of China (English)

    Reda I. El-Sheshtawy; Gamal A. Sisy; Walid S. El-Nattat

    2015-01-01

    Objective: To examine the effect of different concentrations of trehalose or sucrose (50 or 100 or 200 mM) on post-thawed quality of bull semen, cryo-preserved in Tris-citric acid-egg yolk-fructose (TCYF). Methods:Semen samples were diluted in TCYF extender, TCYF +trehalose (50, 100 and 150 mM/L) or TCYF+sucrose (50, 100 and 150 mM/L) to ensure 60 million motile spermatozoa mL-1, cooled slowly up to 5℃ and equilibrated for 4 h. Semen was packed into 0.25 mL polyvinyl French straws. The straws were placed horizontally on a rack and frozen in a vapor 4 cm above liquid nitrogen (LN2) for 10 minutes then dipped in liquid LN2. Frozen straws were thawed at 37℃ for 1 min. The parameters studied were sperm motility, sperm viability, sperm abnormality, sperm membrane integrity (HOST), percent of normal intact acrosome and DNA fragmentation. Results:The output data demonstrated that addition of 50–100 mM of trehalose or sucrose/L TCYF after chilling at 5℃ had significantly (P<0.0001) ameliorated motility, membrane integrity, viability, abnormal morphology, and acrosome integrity %compared to control diluted semen while 50 mM of trehalose/L, and 50-100 mM of sucrose/L to TCYF diluent had significantly (P<0.0001) improved after thawing motility (43.00,% 45.00%and 41.00%, respectively), membrane integrity (67.40%, 67.80%and 69.40%, respectively), life sperm % (70.20%, 69.40%and 71.40%respectively), and acrosome integrity percentages (56.40%, 58.80%and 55.80%respectively) compared to the control tris-base diluent, while diminishing the abnormal sperm morphology (6.20, 3.80 and 3.80 respectively) and DNA fragmentation (3.60%, 3.80%and 3.80%respectively). Besides, the addition of 100 mM of trehalose/L to tris-base diluent has also a promising effect when added to the tris-base diluent concerning the above parameters. Conclusion:It is finally concluded that the addition of 50- 100 mM trehalose or sucrose/L to TCYF have a beneficial effect in chilling diluted bull semen

  4. Changes in mean scrotal circumference in performance tested Swedish beef bulls over time

    OpenAIRE

    Eriksson Pernilla; Lundeheim Nils; Söderquist Lennart

    2012-01-01

    Abstract Background There is a growing interest in beef cattle breeding in Sweden. The majority of the females are bred naturally, which is why it is important to choose healthy fertile bulls to obtain good reproduction and profitability. The breeding soundness evaluation includes measurement of scrotal circumference (SC). Our aim was to analyze if the SC of performance tested beef bulls has changed over the years. In total, 1332 bulls (Angus, Charolais, Hereford and Simmental) from 13 batche...

  5. Joint disorder; a contributory cause to reproductive failure in beef bulls?

    Science.gov (United States)

    Persson, Ylva; Söderquist, Lennart; Ekman, Stina

    2007-11-05

    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.

  6. 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.

  7. Duration Dependence in Stock Prices: An Analysis of Bull and Bear Markets

    OpenAIRE

    Asger Lunde; Allan Timmermann

    2003-01-01

    This paper investigates the presence of bull and bear market states in stock price dynamics. A new definition of bull and bear market states based on sequences of stopping times tracing local peaks and troughs in stock prices is proposed. Duration dependence in stock prices is investigated through posterior mode estimates of the hazard function in bull and bear markets. We find that the longer a bull market has lasted, the lower is the probability that it will come to a termination. In contra...

  8. Effect of behaviour of Holstein Friesian and Simmental bulls on semen quality

    Directory of Open Access Journals (Sweden)

    Krzysztof Adamczyk

    2013-06-01

    Full Text Available The aim of the study was to determine the relationship between the behaviour of Holstein-Friesian and Simmental bulls and the quality of their semen. A total of 76 breeding bulls of the Holstein-Friesian and dual-purpose Simmental breeds were investigated. Analysis was made of the response of bulls to humans and other bulls, facial hair whorl position and length, scrotal circumference, and semen characteristics (mean ejaculate volume, mean sperm concentration, and sperm wave motion. The age and breed of the bulls had a statistically significant effect on semen quality, scrotal circumference and the animals response to an unfamiliar human (Plt,0.05, Plt,0.01. The coefficients of correlation between the bull s response to a handler and to other bulls averaged 0.73. In general, only weak correlations were found between behavioural traits of the bulls and quality of their semen. It is worth noting a good correlation (r=0.50; Plt;0.05 between hair whorl position and sperm concentration in Simmental bulls.

  9. Tarsal lameness of dairy bulls housed at two artificial insemination centers: 24 cases (1975-1987).

    Science.gov (United States)

    Bargai, U; Cohen, R

    1992-10-01

    Degenerative joint disease of the tarsi was diagnosed in 20 of 24 Holstein bulls with tarsal lameness at 2 artificial insemination centers from 1975 to 1987. Each of the 2 centers housed about 100 bulls/yr. Of the 24 bulls with tarsal lameness, 22 were from the artificial insemination center designated as A, and 2 were from the center designated as B. Examination of the housing and management procedures revealed that center A had concrete floors with cuboidal-shaped yards, whereas center B had deep sand flooring, with long, narrow yards. The only other difference between the 2 centers was that center A used 1- and 2-year-old bulls as teasers for older, heavier bulls to mount, whereas center B used bulls that were at least 6 years old to withstand the stress placed on their hind limbs by the weight of bulls undergoing semen collection. Radiographic lesions of tarsi of bulls from both centers ranged from distention of the tibiotarsal joint pouch to hypertrophic degenerative osteoarthritis of the distal, intertarsal, and tarsometatarsal joints. It was concluded that the concrete flooring and the semen collecting practices were responsible for the high prevalence of tarsal lameness and degenerative joint disease of the tarsi in bulls housed at center A.

  10. 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.

  11. 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.

  12. Application of quantum dot nanoparticles for potential non-invasive bio-imaging of mammalian spermatozoa

    Science.gov (United States)

    Various obstacles are encountered by mammalian spermatozoa during their journey through the female genital tract, and only few or none will reach the site of fertilization. Currently, there are limited technical approaches for non-invasive investigation of spermatozoa migration after insemination. A...

  13. 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.

  14. Effect of semen extender and density gradient centrifugation on the motility and fertility of turkey spermatozoa.

    Science.gov (United States)

    Morrell, J M; Persson, B; Tjellström, H; Laessker, A; Nilsson, H; Danilova, M; Holmes, P V

    2005-12-01

    In the absence of commercially viable methods for cryopreserving turkey spermatozoa, new processing methods are required to extend the functional life of stored turkey spermatozoa for artificial insemination. The present study evaluates the efficacy of a new extender (Turkey Semen Extend) and investigates the use of density gradient centrifugation in processing turkey spermatozoa for artificial insemination. The new extender is compared with two commercially available turkey semen extenders, Beltsville Poultry Semen Extender and Ovodyl. Turkey spermatozoa in Turkey Semen Extend were still motile 20 h after collection, representing a considerable improvement over the other semen extenders (40%, 0% and 8% for Turkey Semen Extend, Beltsville Poultry Semen Extender and Ovodyl, respectively). A field trial on a commercial turkey farm showed improved fertilization rates following insemination of turkey hens with semen extended in Turkey Semen Extend (89.7%) compared with Beltsville Poultry Semen Extender (86.9%). This difference is statistically significant (p < 0.05). Processing on a density gradient, optimized for turkey spermatozoa, also increased sperm survival (50% gradient-prepared spermatozoa still motile after 18 h compared with <10% non-processed spermatozoa). Preliminary studies indicate that gradient preparation of spermatozoa may aid survival during cryopreservation.

  15. 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 ...

  16. 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. Th

  17. Molecular determinants of nucleosome retention at CpG-rich sequences in mouse spermatozoa

    NARCIS (Netherlands)

    Erkek, S.; Hisano, M.; Liang, C.Y.; Gill, M.; Murr, R.; Dieker, J.W.C.; Schubeler, D.; Vlag, J. van der; Stadler, M.B.; Peters, A.H.

    2013-01-01

    In mammalian spermatozoa, most but not all of the genome is densely packaged by protamines. Here we reveal the molecular logic underlying the retention of nucleosomes in mouse spermatozoa, which contain only 1% residual histones. We observe high enrichment throughout the genome of nucleosomes at CpG

  18. Early apoptotic changes in human spermatozoa and their relationships with conventional semen parameters and sperm DNA fragmentation

    Institute of Scientific and Technical Information of China (English)

    Hao-Bo Zhang; Shao-Ming Lu; Chun-Yan Ma; Li Wang; Xiao Li; Zi-Jiang Chen

    2008-01-01

    Aim: To investigate whether early apoptotic changes in spermatozoa can be significant markers for sperm quality.Methods: Two early apoptotic changes in the semen of 56 men were assessed using Annexin V (AN)/propidium iodide (PI) staining for phosphatidylserine externalization and JC-1 staining for mitochondrial membrane potential (MMP). The results were compared with conventional semen parameters and DNA fragmentation identified using the TUNEL assay. Results: The different labeling patterns in the bivariate Annexin V/PI analysis identified four distinc-tive spermatozoa populations. The percentage of AN-/PI- spermatozoa positively correlated with conventional semen parameters and MMP, but negatively correlated with TUNEL (+) spermatozoa. As for the AN-/PI+ fraction, we found an opposite result in comparison to AN-/PI- spermatozoa. The level of early apoptotic AN+/PI- spermatozoa negatively correlated with MMP and sperm motility. The level of late apoptotic AN+/PI+ spermatozoa negatively correlated with conventional semen parameters and MMP, and positively correlated with TUNEL (+) spermatozoa. MMP positively correlated with conventional semen parameters, but negatively correlated with TUNEL (+) spermatozoa. Conclusion:Although early apoptotic AN+/PI- spermatozoa only negatively correlates with sperm motility, the differences in proportion of each subpopulation of spermatozoa (especially, the percentage of AN-/PI-spermatozoa), and decreased MMP might be significant markers for diagnosing male infertility. They possibly bring additional information to predict the outcome of in vitro fertilization.

  19. TINJAUAN PENAMBAHAN MINERAL ZN DALAM PAKAN TERHADAP KUALITAS SPERMATOZOA PADA SAPI FRISIAN HOLSTEIN JANTAN

    OpenAIRE

    Sus Derthi Widhyari; Anita Esfandiari; Agus Wijaya; Retno Wulansari; Setyo Widodo; Leni Maylina

    2015-01-01

    Bulls are expected to be able to produce a good quality and quantity of sperm. The objective of this experiment was to study the effect of Zn supplementation on the sperm quality, in Frisian holstein bulls. Ten bulls, 1618 months of age were used in this experiment. The experimental bulls were divided into two groups, i.e.,group without Zn supplementation (control) and group with 60 ppm of Zn supplementation. Zn supplementations were given everyday for a period of four months. Semen was coll...

  20. Evaluation of the damage in fish spermatozoa cryopreservation

    Institute of Scientific and Technical Information of China (English)

    LI Jun; LIU Qinghua; ZHANG Shicui

    2006-01-01

    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.

  1. [The application of eosin and propidium iodide in evaluation of vitality of human spermatozoa].

    Science.gov (United States)

    Ploskonos, М В

    2014-11-01

    The article analyzes comparative assessment of vitality of spermatozoa by condition of permeability of membranes for eosin and propidium iodide and comparison of results acquired using technique of light and fluorescent microscopy. The comparison of data of light microscopy with eosin staining with data of fluorescent microscopy with propidium iodide staining demonstrated that percentage of content of spermatozoa separated from ejaculates of 28 fertile males and stained with eosin was reliably higher (34.8 ± 3.2) than percentage of content of spermatozoa with stained with propidium iodide (2.1 ± 4.0). After incubation of spermatozoa under room temperature during 24 hours percentage of unviable cells with stained eosin also was higher than in case of propidium iodide staining correspondingly (44.5 ± 3.3% and 34.7 ± 3.6%). The analysis of vitality of spermatozoa under damaging effect of oxidative stress on cell membrane developed by 4 hours incubation with 200 mkM of hydrogen peroxide (H2O2) demonstrated that under staining of spermatozoa with propidium iodide significantly higher percentage of damaged cells is detected. In such cases, eosin staining is less suitable for detection of vitality of spermatozoa (73.6 ± 5.8% against 51.7 ± 6.4%). The carried out experiment demonstrates that in case of detected effects on spermatozoa (for example, effect of oxidative stress) the light microscopy insufficiently adequate reflects degree of damage of membranes of spermatozoa. The fluorescent microscopy detects a higher percentage of spermatozoa with damaged membrane.

  2. 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 di...... into mechanisms by which HFD transgenerationally reprograms the epigenome of sperm cells, thereby affecting metabolic tissues of offspring throughout two generations....... 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......1 male offspring showed common DNA methylation and small non-coding RNA expression signatures. Altered expression of sperm miRNA let-7c was passed down to metabolic tissues of the offspring, inducing a transcriptomic shift of the let-7c predicted targets. CONCLUSION: Our results provide insight...

  3. Distribution and Movement of Bull Trout in the Upper Jarbidge River Watershed, Nevada

    Science.gov (United States)

    Allen, M. Brady; Connolly, Patrick J.; Mesa, Matthew G.; Charrier, Jodi; Dixon, Chris

    2010-01-01

    In 2006 and 2007, we surveyed the occurrence of bull trout (Salvelinus confluentus), the relative distributions of bull trout and redband trout (Oncorhynchus mykiss), and stream habitat conditions in the East and West Forks of the Jarbidge River in northeastern Nevada and southern Idaho. We installed passive integrated transponder (PIT) tag interrogation systems at strategic locations within the watershed, and PIT-tagged bull trout were monitored to evaluate individual fish growth, movement, and the connectivity of bull trout between streams. Robust bull trout populations were found in the upper portions of the East Fork Jarbidge River, the West Fork Jarbidge River, and in the Pine, Jack, Dave, and Fall Creeks. Small numbers of bull trout also were found in Slide and Cougar Creeks. Bull trout were numerically dominant in the upper portions of the East Fork Jarbidge River, and in Fall, Dave, Jack, and Pine Creeks, whereas redband trout were numerically dominant throughout the rest of the watershed. The relative abundance of bull trout was notably higher at altitudes above 2,100 m. This study was successful in documenting bull trout population connectivity within the West Fork Jarbidge River, particularly between West Fork Jarbidge River and Pine Creek. Downstream movement of bull trout to the confluence of the East Fork and West Fork Jarbidge River both from Jack Creek (rkm 16.6) in the West Fork Jarbidge River and from Dave Creek (rkm 7.5) in the East Fork Jarbidge River was detected. Although bull trout exhibited some downstream movement during the spring and summer, much of their emigration occurred in the autumn, concurrent with decreasing water temperatures and slightly increasing flows. The bull trout that emigrated were mostly age-2 or older, but some age-1 fish also emigrated. Upstream movement by bull trout was detected less than downstream movement. The overall mean annual growth rate of bull trout in the East Fork and West Fork Jarbidge River was 36 mm

  4. SEMEN PRODUCTION AND PRODUCTIVE LIFE OF SAHIWAL BULLS: RELATIONSHIP WITH GENETIC WORTH

    Directory of Open Access Journals (Sweden)

    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.

  5. Selection of normal spermatozoa with a vacuole-free head (x6300) improves selection of spermatozoa with intact DNA in patients with high sperm DNA fragmentation rates.

    Science.gov (United States)

    Hammoud, I; Boitrelle, F; Ferfouri, F; Vialard, F; Bergere, M; Wainer, B; Bailly, M; Albert, M; Selva, J

    2013-06-01

    Intracytoplasmic morphologically selected sperm injection (IMSI, 6300× magnification with Nomarski contrast) of a normal spermatozoon with a vacuole-free head could improve the embryo's ability to grow to the blastocyst stage and then implant. However, the most relevant indications for IMSI remain to be determined. To evaluate the potential value of IMSI for patients with a high degree of sperm DNA fragmentation (n = 8), different types of spermatozoa were analysed in terms of DNA fragmentation. Motile normal spermatozoa with a vacuole-free head selected at 6300× magnification had a significantly lower mean DNA fragmentation rate (4.1 ± 1.1%, n = 191) than all other types of spermatozoa: non-selected spermatozoa (n = 8000; 26.1 ± 1.5% versus 4.1 ± 1.1%; P sperm DNA fragmentation rates, selection of normal spermatozoa with a vacuole-free head (6300×) yields the greatest likelihood of obtaining spermatozoa with non-fragmented DNA.

  6. 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.

  7. Identification and Localization of the Cyclic Nucleotide Phosphodiesterase 10A in Bovine Testis and Mature Spermatozoa

    Science.gov (United States)

    Goupil, Serge; Maréchal, Loïze; El Hajj, Hassan; Tremblay, Marie-Ève; Richard, François J.

    2016-01-01

    In mammals, adenosine 3’, 5’-cyclic monophosphate (cAMP) is known to play highly important roles in sperm motility and acrosomal exocytosis. It is known to act through protein phosphorylation via PRKA and through the activation of guanine nucleotide exchange factors like EPAC. Sperm intracellular cAMP levels depend on the activity of adenylyl cyclases, mostly SACY, though transmembrane-containing adenylyl cyclases are also present, and on the activity of cyclic nucleotide phosphodiesterases (PDE) whose role is to degrade cAMP into 5’-AMP. The PDE superfamily is subdivided into 11 families (PDE1 to 11), which act on either cAMP or cGMP, or on both cAMP and cGMP although with different enzymatic properties. PDE10, which is more effective on cAMP than cGMP, has been known for almost 15 years and is mostly studied in the brain where it is associated with neurological disorders. Although a high level of PDE10A gene expression is observed in the testis, information on the identity of the isoforms or on the cell type that express the PDE10 protein is lacking. The objective of this study was to identify the PDE10A isoforms expressed in the testis and germ cells, and to determine the presence and localization of PDE10A in mature spermatozoa. As a sub-objective, since PDE10A transcript variants were reported strictly through analyses of bovine genomic sequence, we also wanted to determine the nucleotide and amino acid sequences by experimental evidence. Using RT-PCR, 5’- and 3’-RACE approaches we clearly show that PDE10A transcript variants X3 and X5 are expressed in bovine testis as well as in primary spermatocytes and spermatids. We also reveal using a combination of immunological techniques and proteomics analytical tools that the PDE10A isoform X4 is present in the area of the developing acrosome of spermatids and of the acrosome of mature spermatozoa. PMID:27548062

  8. ASSESSMENT OF CHAROLAIS BULLS POPULATION STRUCTURE BASED ON SNPs ANALYSES

    OpenAIRE

    Nina Moravčíková; Tomáš Minarovič; Anna Trakovická

    2014-01-01

    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...

  9. Spermatozoa-like cell invaders (nuclear vlimata) in human neoplasia.

    Science.gov (United States)

    Logothetou-Rella, H

    1993-07-01

    Spermatozoa-like cells (nuclear vlimata) have been identified in malignant cell cultures and embryonic cells, also common in the cytology and histology of all types of human neoplasia even after chemotherapy. A new mechanism of invasion of malignant cells has been described, according to which neoplastic cells behave and function as parasites using host-cells to divide, survive and eventually produce nuclear vlimata (bullets). Nuclear vlimata are the end cell products of incomplete, unequal, assymetrical division of neoplastic cells. The nuclear vlima exhibits similar morphology to spermatozoa and virus (head with, or without, tail) and invades the cytoplasm and/or nucleus of surrounding host-cells by a similar mechanism to sperm-oocyte interaction (fertilization) or viral cell infection, in the events of nuclear vlima-->tumor-->nuclear vlima-->tumor. The nuclear vlima head contains and transfers DNA, and when incorporated into the host-nucleus is indistinguishable from nucleoli and when in the cytoplasm is similar to sperm pronucleus, observed after sperm penetration of the oocyte. Function of nuclear vlimata is directly dependent on the specific extracellular matrix produced by malignant cells, consisting of glycosaminoglycans-protease-membranes. This mechanism of invasion constitutes the link of all scientific information concerning human neoplasia.

  10. Zinc-protein from rat prostate fluid binds epididymal spermatozoa.

    Science.gov (United States)

    Sansone, G; Abrescia, P

    1991-09-01

    The detection and the isolation of a zinc-protein from the secretion of the rat dorsolateral prostate is described. The purification procedure, based on gel filtration and cationic exchange chromatography, allowed to separate a minor protein (Mr approximately 66,000) from free zinc ions and other secretory components. Two zinc ions were estimated to be associated with one molecule of isolated protein. The zinc-protein was labelled with 125I and then incubated at 37 degrees C with spermatozoa from rat epididymal cauda. Time-dependent in vitro binding of the radioactive protein to sperm cells was demonstrated. This binding was not affected by the presence of proteins from the seminal vesicle during the incubation, while it was blocked in the presence of an excess of unlabelled zinc-protein. After binding, the labelled spermatozoa were treated with a buffer containing 0.5% sodium deoxycholate and 40 mM EDTA; only very small amounts of label were removed from the cells, thus suggesting that the zinc-proteins were kept on the plasma membrane by interactions which do not involve merely hydrophobic bonds.

  11. Modification of spermatozoa quality in mature small ruminants.

    Science.gov (United States)

    Martin, G B; de St Jorre, T Jorre; Al Mohsen, F A; Malecki, I A

    2011-01-01

    This review is based largely, but not entirely, on the assumption that gamete quality is directly linked to sperm output and thus testicular mass, an approach made necessary by the absence of a large body of data on factors that affect gamete quality in ruminants. On the other hand, there is a change in the efficiency of sperm production per gram of testicular tissue when the testis is growing or shrinking, a clear indicator of changes in the rates of cell loss during the process of spermatogenesis, probably through apoptosis. We therefore postulate that the spermatozoa that do survive when the testis is shrinking are of a lower quality than those that are produced when the testis is growing and the rate of sperm survival is increasing. In adult small ruminants in particular, testicular mass and sperm production are highly labile and can be manipulated by management of photoperiod (melatonin), nutrition, genetics and behaviour ('mating pressure'). Importantly, these factors do not act independently of each other - rather, the outcomes in terms of sperm production are dictated by interactions. It therefore seems likely that spermatozoa quality will be affected by these same factors, but definitive answers await detailed studies.

  12. Effects of pedigree and exotic genetic inheritance on semen production traits of dairy bulls

    Institute of Scientific and Technical Information of China (English)

    VijethaBajjalliThippeswamy; SiddarthaShankar Layek; ArumugamKumaresan; TusharKumar Mohanty; Ashok Kumar Gupta; Atish Kumar Chakravarty; Ayyasamy Manimaran; Shiv Prasad

    2014-01-01

    Objective:To study the effects of different levels of exotic inheritance on ejaculate quality in bulls and its passage through different generations.Methods:Data on semen production traits and ejaculate quality were obtained for38 crossbred bulls and grandsire-sire-progeny relationship in relation to semen quality was studied.The bulls were classified into three groups based on the level of exotic inheritance viz.F1,50.0%-62.5% exotic germplasm and >75% exotic germplasm.Results:Results of the present study indicated that about40% of the ejaculates obtained from the crossbred bulls were rejected from further processing due to poor ejaculate quality.TheF1 bulls produced significantly higher proportions(57.00±10.00) of poor quality ejaculates compared to the interse mated bulls.The age at first semencollection in crossbred bulls ranged from567 to1010 days with an average of738.89±18.18 days while the mean age at first semen freezing was865.72±34.60 days.Conclusions:It may be inferred that the“acceptable quality semen producing ability” decreased from grandsire through sire to male progeny and among the increasing exotic genetic levels ofCB cattle,F1 bulls produced significantly higher“low grade ejaculates” that were unfit for cryopreservation.

  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. 7 CFR 59.102 - Mandatory daily reporting for cows and bulls.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Mandatory daily reporting for cows and bulls. 59.102... (CONTINUED) LIVESTOCK MANDATORY REPORTING Cattle Reporting § 59.102 Mandatory daily reporting for cows and bulls. (a) In General. The corporate officers or officially designated representatives of each cow...

  15. Evaluation of Semen Quality of Holstein Friesian and Jersey Bulls Maintained under Subtropical Environment

    Directory of Open Access Journals (Sweden)

    M. Fiaz*, R. H. Usmani1, M. Abdullah and T. Ahmad2

    2010-04-01

    Full Text Available Semen production data of Holstein-Friesian and Jersey bulls collected over a period of three years was analyzed to examine seasonal effects on quantity and quality of semen. Purebred breeding bulls of Holstein-Friesian and Jersey breeds (n = 18 for each breed, maintained under naturally ventilated open-sided sheds, were used for semen collection. Meteorological information on ambient temperature, relative humidity and rainfall was used to divide the calendar year into two stressful summer seasons viz; dry summer (April - June and wet summer (July–September and a stress free season (October - March. The number of ejaculates per bull recorded during three seasons were not significantly different (P>0.05 in both breeds. Friesian bulls produced lower ejaculatory volume during dry summer season, whereas Jersey bulls produced higher (P0.05 but in Jersey bulls it was lower during wet summer than other seasons. Wet summer resulted in reduced number of semen doses frozen per bull in both breeds. The results of this study indicated that wet summer season deteriorated semen quality in terms of mass motility, individual motility and number of doses in both breeds, except individual motility in semen of Friesian bulls.

  16. Associations between endotoxin-induced metabolic changes and temperament in Brahman bulls

    Science.gov (United States)

    The influence of temperament on the alteration of metabolic parameters in response to a lipopolysaccharide (LPS) challenge was investigated. Brahman bulls were selected for this study based on temperament score. Bulls were fitted with indwelling jugular catheters for serial sampling to evaluate peri...

  17. 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 corpus 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.

  18. 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.

  19. 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

  20. Species variation in osmotic, cryoprotectant, and cooling rate tolerance in poultry, eagle, and Peregrine Falcon spermatozoa

    Science.gov (United States)

    Blanco, J.M.; Gee, G.; Wildt, D.E.; Donoghue, A.M.

    2000-01-01

    Potential factors influencing spermatozoa survival to cryopreservation and thawing were analyzed across a range of the following avian species: domestic chicken (Gallus domesticus), domestic turkey (Meleagris gallopavo), golden eagle (Aquila chrysaetos), Bonelli's eagle (Hieraaetus fasciatus), imperial eagle (Aquila adalberti), and peregrine falcon (Falco peregrinus). Studies focused on spermatozoa tolerance to the following: 1) osmotic stress, 2) different extracellular concentrations of the cryoprotectant dimethylacetamide (DMA), 3) equilibration times of 1 versus 4 h, 4) equilibration temperature of 4 versus 21 degrees C, and 5) rapid versus slow cooling before cryopreservation and standard thawing. Sperm viability was assessed with the live/dead stain (SYBR14/ propidium iodine). Sperm viability at osmolalities >/=800 mOsm was higher (P: /=2.06 M), experienced decreased (P: < 0.05) spermatozoa survival in all species, except the golden eagle and peregrine falcon. Number of surviving spermatozoa diminished progressively with increasing DMA concentrations in all species. Increased equilibration temperature (from 4 to 21 degrees C) markedly reduced (P: < 0.05) spermatozoa survival in all species except the Bonelli's eagle and turkey. Rapid cooling was detrimental (P: < 0.05) to spermatozoa from all species except the imperial eagle and the chicken. These results demonstrate that avian spermatozoa differ remarkably in response to osmotic changes, DMA concentrations, equilibration time, temperature, and survival after fast or slow freezing. These differences emphasize the need for species-specific studies in the development and enhancement of assisted breeding for poultry and endangered species.

  1. Sperm Hy-Liter™: an effective tool for the detection of spermatozoa in sexual assault exhibits.

    Science.gov (United States)

    De Moors, Anick; Georgalis, Tina; Armstrong, Gail; Modler, Jeff; Frégeau, Chantal J

    2013-05-01

    A fluorescence-based assay specifically targeting human spermatozoa was tested and optimized for best staining results using a variety of mock sexual assault samples. Swab clippings versus whole swabs were evaluated for best sample preparation and to simplify workflow (direct application versus swab extraction). The practicality and sensitivity of Sperm Hy-Liter™ was compared to our current phase contrast microscopy protocol for searching for the presence of spermatozoa. Sperm Hy-Liter™ was more sensitive than phase contrast microscopy and was able to detect spermatozoa more effectively in actual sexual assault samples (recent [N=240] or 24 years old [N=4]) containing few spermatozoa. Correlations were drawn between the Sperm Hy-Liter™ spermatozoa counts and the AmpFlSTR(®) Profiler(®) Plus male profiles generated from the sperm cell DNA fractions of semen containing swabs and swab clippings. In addition, recovered spermatozoa from Sperm Hy-Liter™-stained slides with greater than 40 spermatozoa produced full STR male profiles in 20.3% of slides tested and partial STR male profiles in 52.8% of slides tested. The adoption of Sperm Hy-Liter™ offers a means to standardize and improve the efficiency of the microscopic screening of sexual assault evidence.

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

    Science.gov (United States)

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

    2003-10-15

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

  3. Effect of antioxidant supplementation on function and fertility of sex-sorted boar spermatozoa.

    Science.gov (United States)

    Xia, Chunmei; Xia, Wei; Yang, Sheng; An, Lei; Li, Xihe; Wu, Zhonghong; Zhang, Jiaxing; Wang, Zhuqing; Tian, Jianhui

    2012-12-01

    The goal of this study was to investigate the effects of antioxidant supplementation on the quality of flow cytometrically-sorted boar spermatozoa. The effects of ascorbic acid-2-glucoside (AA-2G) on the sex-sorting process were evaluated using a variety of concentrations. The effects of different antioxidants (AA-2G, l-glutathione, and vitamin E) on the viability and lifespan of boar spermatozoa were also compared during sorting. Furthermore, the effect of AA-2G on acrosome intactness, the capacitation ability of sorted boar spermatozoa and pregnancy efficiency after artificial insemination (AI) at different sorting-to-insemination intervals were examined. Greater (Psperm head lateral displacement (ALH)) of the sex-sorted boar spermatozoa were greater (Pboar spermatozoa in the AA-2G-supplemented group was less (Pboar spermatozoa for the AA-2G-supplemented group were 59.25%, while the control group remains no sufficient quality semen. This study demonstrates that AA-2G supplementation can improve the quality of flow cytometrically sorted boar spermatozoa and that the optimal concentration of AA-2G for sorting is 0.068 mg/mL.

  4. 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.

  5. Is the distribution of selenium and zinc in the sublocations of spermatozoa regulated?

    Science.gov (United States)

    Bertelsmann, Holger; Sieme, Harald; Behne, Dietrich; Kyriakopoulos, Antonios

    2007-01-01

    In the sperm nuclei, of mammalian species selenium has been found only in the form of sperm nuclei glutathione peroxidase (snGPx) where it is most likely bound to the chromatin of spermatozoa. Over 80% of selenium in sperm is bound to the selenoprotein phospholipid hydroperoxide glutathione peroxidase (PHGPx) in the midpiece of rat sperm. Zinc in sperm is mainly contained in the outer dense fiber (ODF) proteins of the flagella of mammalian spermatozoa. In the sperm nuclei, zinc is predominately located in the chromatin to the protamine proteins. In order to investigate if the insertion of zinc and selenium in sperm chromatin is regulated, the element concentrations were determined in equine spermatozoa and purified sperm nuclei. We found a significant positive correlation between the selenium concentration in equine spermatozoa and sperm nuclei. The same finding was obtained for the zinc concentration in spermatozoa and sperm nuclei. The results assume that the distribution of selenium and zinc in spermatozoa is regulated by cell signaling pathways and in this way determining the selenium and zinc amount in the chromatin of spermatozoa.

  6. In vitro inhibition of superoxide anion production and superoxide dismutase activity by zinc in human spermatozoa.

    Science.gov (United States)

    Gavella, M; Lipovac, V; Vucić, M; Sverko, V

    1999-08-01

    The in vitro effect of zinc on superoxide anion (O2-) generation and on SOD-like activity in spermatozoa of infertile men was investigated. The formation of superoxide anion was stimulated by NADPH and the level of superoxide anion was measured by the reduction of ferricytochrome c. Both Percoll-isolated (n = 14) and washed spermatozoa (n = 14) exposed to 1 mmol/L zinc (60 min, 37 degrees C), released less (p zinc-untreated spermatozoa. These results implicate a possible role for zinc as a scavenger of excessive superoxide anions produced by defective spermatozoa in semen after ejaculation. Additionally, zinc was found to dose-dependently inhibit superoxide dismutase (SOD)-like activity of spermatozoa in vitro. The inhibition of SOD-like activity by an equal concentration of zinc (1 mmol/L) was less pronounced in oligospermic (p zinc to inhibit SOD-like activity may be relevant to the physiological function of spermatozoa in fertilization. The evidence that zinc may elicit an inhibition of both superoxide anion production and SOD-like activity in human spermatozoa, indicate the existence of novel, zinc-related mechanism(s) involved in the oxidative events occurring after ejaculation, with a possible modulatory effect on germ cell function.

  7. Innate immune responses of young bulls to a novel environment.

    Science.gov (United States)

    Razzuoli, Elisabetta; Olzi, Emilio; Calà, Pietro; Cafazzo, Simona; Magnani, Diego; Vitali, Andrea; Lacetera, Nicola; Archetti, Laura; Lazzara, Fabrizio; Ferrari, Angelo; Nanni Costa, Leonardo; Amadori, Massimo

    2016-04-01

    Animal welfare during transportation has been investigated in several studies, as opposed to post-transportation phases. In this study, we evaluated the effect of a novel environment after transportation on 26 Friesian bulls, 242 ± 42 day-old, from ten different dairy farms. Animals were shipped to a breeding center in different seasons, and selected parameters of innate immunity (serum bactericidal activity, hemolytic complement, serum albumin, α, β, and γ-globulins, interleukin-6, TNF-α) were monitored before and after the arrival at days--4/0/4/15/30. Our results showed significant differences of IL-6 and TNF-α protein levels at destination in December (94 ± 1.3 pg/ml) and June (+788 pg/ml), respectively. Moreover, the serum levels of these cytokines increased between days 0 and 15 after the arrival, the modulation of IL-6 being in agreement with established models of physical and/or psychological stress. Concerning the modulation of albumin, alpha and beta-globulins, the highest levels were detected in April, whereas a significant decrease was observed between day 15 and 30 after arrival; on the contrary, γ-globulin levels significantly increased after day 15. The results of this study highlight the occurrence of innate immune responses of young bulls to the combined effects of climate (season) and novel farming conditions.

  8. Sprint swimming performance of wild bull trout (Salvelinus confluentus)

    Science.gov (United States)

    Mesa, M.G.; Phelps, J.; Weiland, L.K.

    2008-01-01

    We conducted laboratory experiments to determine the sprint swimming performance of wild juvenile and adult bull trout Salvelinus confluentus. Sprint swimming speeds were estimated using high-speed digital video analysis. Thirty two bull trout were tested in sizes ranging from about 10 to 31 cm. Of these, 14 fish showed at least one motivated, vigorous sprint. When plotted as a function of time, velocity of fish increased rapidly with the relation linear or slightly curvilinear. Their maximum velocity, or Vmax, ranged from 1.3 to 2.3 m/s, was usually achieved within 0.8 to 1.0 s, and was independent of fish size. Distances covered during these sprints ranged from 1.4 to 2.4 m. Our estimates of the sprint swimming performance are the first reported for this species and may be useful for producing or modifying fish passage structures that allow safe and effective passage of fish without overly exhausting them. ?? 2008 by the Northwest Scientific Association. All rights reserved.

  9. Variance in spermatozoa number among Apis dorsata drones and among Apis mellifera drones

    OpenAIRE

    Koeniger, Gudrun; KOENIGER, Nikolaus; Tingek, Salim; Phiancharoen, Mananya

    2005-01-01

    International audience; Published estimates of the mean spermatozoa numbers for Apis dorsata drones vary from 1.2 × 106 and 2.4 × 106; the number of spermatozoa per individual drone vary from 0.22 × 106 to 2.65 × 106. Counts presented here revealed 1.19 × 106 + 0.25 × 106 spermatozoa in drones sampled near a colony and 1.59 × 106 + 0.18 × 106 in drones sampled at a drone congregation area (DCA) in Sabah, Borneo. The difference between the two sites is significant. Further, the degree of varia...

  10. Binding of free and protein-associated zinc to rat spermatozoa.

    Science.gov (United States)

    Sansone, G; Martino, M; Abrescia, P

    1991-01-01

    1. The zinc content of rat spermatozoa increases, upon ejaculation, from 0.035 to 1.055 micrograms/10(6) cells. 2. The rat seminal plasma holds zinc both as free ion and as protein-bound forms. 3. Zinc-free ions bind in vitro to rat epididymal spermatozoa. 4. Zinc-protein complexes can be isolated, by a chromatographic procedure, from the dorsolateral lobe of rat prostate. 5. The isolated zinc-protein complexes bind in vitro to rat epididymal spermatozoa.

  11. Membrane integrity and fertilizing potential of cryopreserved spermatozoa in European mouflon.

    Science.gov (United States)

    Naitana, S; Ledda, S; Leoni, G; Bogliolo, L; Loi, P; Cappai, P

    1998-08-21

    There is a pressing need to develop and use assisted reproductive techniques in wildlife species living in small and captive groups. We evaluated the effect of freezing on membrane integrity and fertilizing capacity of European mouflon (Ovis gmelini musimon) spermatozoa collected during the breeding season. After thawing, the percentage of live spermatozoa, stained with fluorescein isothiocynate labeled Pisum Sativum agglutinin and propidium iodide, was 47% of which 19% showed intact acrosomal membrane. After culture in TCM 199 + 10% FCS, the number of live spermatozoa was significantly (P European mouflon.

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

    Directory of Open Access Journals (Sweden)

    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.

  13. 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.

  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.

  15. Effects of HeLa and BHK-21 cells on the survival of fowl, bull, ram and boar spermatozoa in vitro

    OpenAIRE

    Ashizawa, Koji; Tokudome, Y; Okauchi, K.; Nishiyama, H

    1982-01-01

    Disclaimer. This is not the definitive version of record of this article. This manuscript has been accepted for publication in Reproduction, but the version presented here has not yet been copy edited, formatted or proofed. Consequently, neither Reproduction, the Society for Reproduction and Fertility nor its publisher, BioScientifica Ltd, accepts responsibility for any errors or omissions it may contain. 1982 Society for Reproduction and Fertility.

  16. Effects of long term storage of semen in liquid nitrogen on the viability, motility and abnormality of frozen thawed Frisian Holstein bull spermatozoa

    Directory of Open Access Journals (Sweden)

    Abdul MALIK

    2015-03-01

    Conclusions: Based on the results in these experiments, it may be concluded that concentration sperm during one year storage in liquid nitrogen resulted in similar concentration storage as long as six years. However, the viability and motility sperm thawed storage in liquid nitrogen during six years was lower than storage on the 1 and 2 years.

  17. Molecular cloning and subcellular localization of Tektin2-binding protein 1 (Ccdc 172) in rat spermatozoa.

    Science.gov (United States)

    Yamaguchi, Airi; Kaneko, Takane; Inai, Tetsuichiro; Iida, Hiroshi

    2014-04-01

    Tektins (TEKTs) are composed of a family of filament-forming proteins localized in cilia and flagella. Five types of mammalian TEKTs have been reported, all of which have been verified to be present in sperm flagella. TEKT2, which is indispensable for sperm structure, mobility, and fertilization, was present at the periphery of the outer dense fiber (ODF) in the sperm flagella. By yeast two-hybrid screening, we intended to isolate flagellar proteins that could interact with TEKT2, which resulted in the isolation of novel two genes from the mouse testis library, referred as a TEKT2-binding protein 1 (TEKT2BP1) and -protein 2 (TEKT2BP2). In this study, we characterized TEKT2BP1, which is registered as a coiled-coil domain-containing protein 172 (Ccdc172) in the latest database. RT-PCR analysis indicated that TEKT2BP1 was predominantly expressed in rat testis and that its expression was increased after 3 weeks of postnatal development. Immunocytochemical studies discovered that TEKT2BP1 localized in the middle piece of rat spermatozoa, predominantly concentrated at the mitochondria sheath of the flagella. We hypothesize that the TEKT2-TEKT2BP1 complex might be involved in the structural linkage between the ODF and mitochondria in the middle piece of the sperm flagella.

  18. Kualitas Spermatozoa Mencit yang Terpapar Radiasi Sinar-X Secara Berulang (SPERMATOZOA QUALITY OF MICE EXPOSED TO X-RAYS RADIATION IN REPEATED

    Directory of Open Access Journals (Sweden)

    Ni Wayan Sudatri

    2015-05-01

    Full Text Available In radiology, X-ray has been used to diagnose disease and therapy. However, behind the technologybenefits provided by the radiation, the negative effects are often debated. The purpose of this study was toinvestigate the effects of repeated radiation on sperm quality mice (Mus musculus L. Thirty- two adultmale mice aged three months were divided into groups P1 (1x 200 rad, P2 (2x200 rad, P3 (3x200 rad andcontrol irradiated with x-rays according to the experimental design . Spermatozoa quality parametersobserved were : number of spermatozoa, motility, viability and morphology of spermatozoa. The results ofthe Post Hoc LSD tests for significant differences (P>0.05 between the control and treatment showed thatthe X-ray radiation exposure to 1x200 rad, 2x200 rad, and 3x200 rad decreases the motility, viability,normal morphology and number spermatozoa produced compared with controls. This is caused by exposureto X-ray radiation causes the formation of free radicals in the body that damage sperm cells mice. Exposureto X-ray radiation repeatedly lowered the quality of spermatozoa of mice.

  19. Surface accumulation of spermatozoa: a fluid dynamic phenomenon

    CERN Document Server

    Smith, David J

    2010-01-01

    Recent mathematical fluid dynamics models have shed light into an outstanding problem in reproductive biology: why do spermatozoa cells show a 'preference' for swimming near to surfaces? In this paper we review quantitative approaches to the problem, originating with the classic paper of Lord Rothschild in 1963. A recent 'boundary integral/slender body theory' mathematical model for the fluid dynamics is described, and we discuss how it gives insight into the mechanisms that may be responsible for the surface accumulation behaviour. We use the simulation model to explore these mechanisms in more detail, and discuss whether simplified models can capture the behaviour of sperm cells. The far-field decay of the fluid flow around the cell is calculated, and compared with a stresslet model. Finally we present some new findings showing how, despite having a relatively small hydrodynamic drag, the sperm cell 'head' has very significant effects on surface accumulation and trajectory.

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

    Science.gov (United States)

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

    2000-08-01

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

  1. 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.

  2. 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.

  3. Evaluation of bull prolificacy on commercial beef cattle ranches using DNA paternity analysis.

    Science.gov (United States)

    Van Eenennaam, A L; Weber, K L; Drake, D J

    2014-06-01

    SNP-based DNA testing was used to assign paternity to 5,052 calves conceived in natural service multisire breeding pastures from 3 commercial ranches in northern California representing 15 calf crops over 3 yr. Bulls present for 60 to 120 d at a 25:1 cow to bull ratio in both fall and spring breeding seasons in ∼40 ha or smaller fenced breeding pastures sired a highly variable (P paternity testing of the subset of those calves born in wk 3 of the calving season was highly predictive of overall bull prolificacy and may offer a reduced-cost DNA-based option for assessing prolificacy. Prolificacy of young bulls in their first breeding season was positively linearly related (P < 0.05) to subsequent breeding seasons, explaining about 20% of the subsequent variation. Prolificacy was also positively linearly related (P < 0.05) to scrotal circumference (SC) EPD for Angus bulls that had SC EPD Beef Improvement Federation accuracies greater than 0.05. Varying prolificacy of herd bulls has implications for the genetic composition of replacement heifers, with the genetics of those bulls siring an increased number of calves being disproportionately represented in the early-born replacement heifer pool.

  4. Glycosidases in the plasma membrane of Ceratitis capitata spermatozoa.

    Science.gov (United States)

    Intra, Jari; De Caro, Daniela; Perotti, Maria-Elisa; Pasini, Maria Enrica

    2011-02-01

    Fruit flies in the family Tephritidae are rated among the world's most destructive agricultural pests. The Mediterranean fruit fly Ceratitis capitata is emerging as a model organism to study the fertilization in Insects. Three integral proteins with glycosidase activity are present in the plasma membrane of spermatozoa. The glycosidases have been purified and characterized. We have demonstrated the presence of three enzymes, a β-N-acetylhexosaminidase, an α-mannosidase and an α-l-fucosidase. The molecular mass of the native enzymes estimated by gel filtration was 160 kDa for β-N-acetylhexosaminidase, 310 kDa for α-mannosidase and 140 kDa for α-l-fucosidase. SDS-PAGE showed that β-N-acetylhexosaminidase is a dimer of a single protein of 73 kDa, α-mannosidase consists of six subunits with different molecular weights and α-l-fucosidase is a dimer made up by two different monomers. Characterization of the purified enzymes included glycosylation pattern, pI, optimal pH, substrate preference, kinetic properties and thermal stability. Soluble forms similar to the sperm associated glycosidases are present. Polyclonal antibodies raised against synthetic peptides designed from the predicted products of the Drosophila melanogaster genes encoding β-N-acetylhexosaminidase and α-l-fucosidase were used. Immunofluorescence labelling of spermatozoa showed that the enzymes are present in the sperm plasma membrane overlying the acrosome and the tail. This work represents the first report on the characterization in C. capitata of sperm proteins that are potentially involved in primary gamete recognition.

  5. Genotoxicity of diuron and glyphosate in oyster spermatozoa and embryos.

    Science.gov (United States)

    Akcha, F; Spagnol, C; Rouxel, J

    2012-01-15

    We investigated the effects of genotoxicant exposure in gametes and embryos to find a possible link between genotoxicity and reproduction/developmental impairment, and explore the impact of chemical genotoxicity on population dynamics. Our study focused on the genotoxic effects of two herbicides on oyster gametes and embryos: glyphosate (both as an active substance and in the Roundup formulation) and diuron. France is Europe's leading consumer of agrochemical substances and as such, contamination of France's coastal waters by pesticides is a major concern. Glyphosate and diuron are among the most frequently detected herbicides in oyster production areas; as oyster is a specie with external reproduction, its gametes and embryos are in direct contact with the surrounding waters and are hence particularly exposed to these potentially dangerous substances. In the course of this study, differences in genotoxic and embryotoxic responses were observed in the various experiments, possibly due to differences in pollutant sensitivity between the tested genitor lots. Glyphosate and Roundup had no effect on oyster development at the concentrations tested, whereas diuron significantly affected embryo-larval development from the lowest tested concentration of 0.05 μg L⁻¹, i.e. an environmentally realistic concentration. Diuron may therefore have a significant impact on oyster recruitment rates in the natural environment. Our spermiotoxicity study revealed none of the tested herbicides to be cytotoxic for oyster spermatozoa. However, the alkaline comet assay showed diuron to have a significant genotoxic effect on oyster spermatozoa at concentrations of 0.05 μg L⁻¹ upwards. Conversely, no effects due to diuron exposure were observed on sperm mitochondrial function or acrosomal membrane integrity. Although our initial results showed no negative effect on sperm function, the possible impact on fertilization rate and the consequences of the transmission of damaged DNA for

  6. Evaluation of protamines 1 and 2 transcript contents in spermatozoa from asthenozoospermic men.

    Directory of Open Access Journals (Sweden)

    Magdalena Depa-Martynow

    2008-04-01

    Full Text Available During mammalian spermatogenesis, the chromatin structure undergoes substantial condensation. The key role in this process is played by protamines 1 and 2 (PRM1, PRM2. We attempted to compare the levels of PRM1 and PRM2 transcripts in mature spermatozoa of normospermic and asthenozoospermic men. Human ejaculates from normozoospermic (n=70 and asthenozoospermic (n=100 donors were purified by centrifugation through discontinuous Percoll density gradient. RNA was isolated from spermatozoa according to the ChomczyĂąski and Sacchi method, treated with DNase I, and reverse-transcribed into cDNA. Using reverse transcription and real-time quantitative polymerase chain reaction analysis, we found a reduction in the levels of PRM1 and PRM2 transcripts in spermatozoa from asthenozoospermic men, as compared to controls (P<0.001. Our findings indicate that a reduction in contents of PRM1 and PRM2 transcripts in spermatozoa may be linked with asthenozoospermia.

  7. KAJI BANDING KUALITAS SPERMATOZOA SAPI SIMMENTAL, LIMOUSIN, DAN FRIESIAN HOLSTEIN TERHADAP PROSES PEMBEKUAN

    Directory of Open Access Journals (Sweden)

    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

  8. The occurrence of spermatozoa in the ovary of the gynogenetic viviparous teleost Poecilia formosa (POECILIIDAE).

    Science.gov (United States)

    Uribe, Mari Carmen; Grier, Harry J; De la Rosa-Cruz, Gabino; Schartl, Manfred

    2016-03-01

    The reproductive mode of the female viviparous teleost Poecilia formosa (Poeciliidae) represents the phenomenon known as gynogenesis; that is, parthenogenetic development is initiated by spermatozoa which are needed for physiological activation of the egg and the initiation of gestation, but spermatozoa are prevented from contributing to the genome of the embryo. For the reason that no previous histological analyses of the ovary of this species during the reproductive cycle has been published the present study has been conducted. This study examined the histology of the ovary of P. formosa during nongestation and gestation phases and identified the presence of spermatozoa inside the ovary. Spermatozoa were observed in folds of the ovarian epithelium of P. formosa during both the nongestation and gestation phases. Sperm storage as documented in this study is a very important trait for the gynogenetic viviparous fish P. formosa contributing to the understanding of this species reproduction.

  9. 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.

  10. 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).

  11. 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).

  12. Impact of maturity rate of daughters on genetic ranking of Holstein bulls.

    Science.gov (United States)

    Norman, H D; Wright, J R; Powell, R L; Vanraden, P M

    2005-09-01

    If genetic evaluations are calculated with a single-trait repeatability model, evaluation changes may be attributed in part to bulls that have daughters that deviate considerably from the typical response to aging. Differences in maturity rate of bull daughters were examined to determine whether they influence change in bull evaluations. Standardized milk records for Holsteins that first calved between 1960 and 1998 were used to calculate 12 tailored predicted transmitting abilities (PTA) for each bull. Predicted transmitting abilities were tailored from combinations of 4 annual cut-off dates and 3 parities. Date screening selected cows first calving before January of 1996, 1997, 1998, or 1999. Parity screening selected milk records from the first 1, 2, or 3 parities. Therefore, 4 evaluations (PTA1) included only first-parity records available for daughters and contemporaries prior to the respective years designated. Four more evaluations (PTA(1,2)) included the records from the first 2 parities for cows first calving prior to those same year cutoffs; likewise, the last 4 evaluations (PTA(1,2,3)) included records from the first 3 parities. Stability of bull evaluations (standard deviations of differences as well as correlations between bull evaluations) across time was compared. Bulls born after 1984 with > or =500 daughters were of interest because of the high precision of evaluations and recent activity. Tailored PTA of those bulls had more uniformity across years in mean records per daughter than did official USDA PTA. Standard deviation of differences in PTA1, PTA(1,2), and PTA(1,2,3) for milk between evaluation years 1996 and 1997 were 28, 28, and 27 kg compared with 63 kg for official evaluations; similarly, between 1996 and 1999, SD were 36, 32, and 32 kg compared with 80 kg. Results suggested that a modification to the current evaluation model to account for maturity rate should reduce fluctuations in individual bull PTA across time and may improve accuracy

  13. 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.

  14. 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.

  15. 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.

  16. 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)

  17. IN VITRO EFFECTS OF COPPER ON THE MOTILITY AND VIABILITY OF SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    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.

  18. Reduced levels of intracellular calcium releasing in spermatozoa from asthenozoospermic patients

    Directory of Open Access Journals (Sweden)

    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

  19. Morphology and ultrastructure of Brachymystax lenok tsinlingensis spermatozoa by scanning and transmission electron microscopy.

    Science.gov (United States)

    Guo, Wei; Shao, Jian; Li, Ping; Wu, Jinming; Wei, Qiwei

    2016-08-01

    This study was conducted to investigate Brachymystax lenok tsinlingensis spermatozoa cell morphology and ultrastructure through scanning and transmission electron microscopy. Findings revealed that the spermatozoa can be differentiated into three major parts: a spherical head without an acrosome, a short mid-piece, and a long, cylindrical flagellum. The mean length of the spermatozoa was 36.11±2.84μm, with a spherical head length of 2.78±0.31μm. The mean anterior and posterior head widths were 2.20±0.42μm and 2.55±0.53μm, respectively. The nuclear fossa was positioned at the base of the nucleus that contained the anterior portion of flagellum and a centriolar complex (proximal and distal centrioles). The short mid-piece was located laterally to the nucleus and possessed just one spherical mitochondrion with a mean diameter of 0.65±0.14μm. The spermatozoa flagellum was long and cylindrical, and could be separated into two parts: a long main-piece and a short end-piece. The main piece of the flagellum had short irregular side-fins. The axoneme composed the typical '9+2' microtubular doublet structure and was enclosed by the cell membran e. This study confirmed that B. lenok tsinlingensis spermatozoa can be categorized as teleostean "Type I" spermatozoa; 'primitive' or 'ect-aquasperm type' spermatozoa. To the best of the authers knowledge, this was the first study conducted on the morphology and ultrastructure of B. lenok tsinlingensis spermatozoa.

  20. AMPK up-activation reduces motility and regulates other functions of boar spermatozoa.

    Science.gov (United States)

    Hurtado de Llera, A; Martin-Hidalgo, D; Gil, M C; Garcia-Marin, L J; Bragado, M J

    2015-01-01

    We recently demonstrated that AMPK inhibition in spermatozoa regulates motility, plasma membrane organization, acrosome integrity and mitochondrial membrane potential. As AMPK activity varies in different energy conditions induced by sperm environment, this work investigates the functional effects of AMPK activation in boar spermatozoa. Spermatozoa were incubated under non-stimulating (TBM) or Ca(2+) and [Formula: see text]-stimulating (TCM) media in the presence/absence of AMPK activator, A769662, for different times. AMPK activity, evaluated as Thr(172) phosphorylation by western blot, is effectively increased by A769662 in spermatozoa. AMPK activation significantly reduces the percentage of motile spermatozoa under Ca(2+) and/or [Formula: see text]-stimulating conditions. Moreover, AMPK activation in spermatozoa incubated in TBM or TCM significantly reduces curvilinear VCL, straight-line VSL and average VAP velocities, which subsequently lead to a significant decrease in the percentage of rapid spermatozoa (VAP > 80 μm/s). The effect of AMPK activation on motility is intensified by the absence of BSA in the incubation medium. AMPK activation for a short time prevents the decline in cell viability and in the sperm population displaying high mitochondrial membrane potential which is induced by Ca(2+) and [Formula: see text]. Sustained (24 h) AMPK activation under TBM or TCM significantly increases both lipid disorganization and phosphatidylserine externalization in the sperm plasma membrane, and diminishes the acrosome membrane integrity. In summary, AMPK activation modifies essential sperm processes such as motility, viability, mitochondrial membrane potential, acrosome membrane integrity, and organization and fluidity of plasma membrane. As these spermatozoa processes are required under different environmental conditions when transiting through the female reproductive tract to achieve fertilization, we conclude that balanced levels of AMPK activity are

  1. PSP-I/PSP-II spermadhesin exert a decapacitation effect on highly extended boar spermatozoa.

    Science.gov (United States)

    Caballero, Ignacio; Vazquez, Juan M; Mayor, Gloria M; Almiñana, Carmen; Calvete, Juan J; Sanz, Libia; Roca, Jordi; Martinez, Emilio A

    2009-10-01

    PSP-I/PSP-II heterodimer is a major protein of boar seminal plasma that is able to preserve, in vitro, the viability, motility and mitochondrial activity of highly-extended boar spermatozoa. However, a relationship between the protective effects of the heterodimer and sperm capacitation is still unclear. The present study investigated the effect of the PSP-I/PSP-II (1.5 mg/mL) on membrane stability, intracellular calcium concentration ([Ca(2+)](I)) and plasma membrane and acrosome integrity of highly extended boar spermatozoa. Boar spermatozoa were diluted to 1 x 10(6) spermatozoa/mL and incubated at 38 degrees C in Phosphate-buffered saline (PBS) for 10, 30, 60, 120 and 300 min or in modified Tris-buffered medium (mTBM) for 10, 20, 30, 60 and 120 min. After each incubation time, the membrane stability (using Merocyanine-540/Yo-Pro-1), elevation of [Ca(2+)](I) (using Fluo-3-AM/PI) and the sperm plasma membrane and acrosome integrity (using SYBR-14/PI/PE-PNA) were evaluated by flow cytometry. As expected, exposure of the spermatozoa to the PSP-I/PSP-II preserved the plasma membrane and acrosome integrity compared to non-exposed spermatozoa in both media PBS and mTBM (p PSP-I/PSP-II compared to controls irrespective of the dilution media. The evaluation of the [Ca(2+)](I) levels showed that while spermatozoa incubated in mTBM and exposed to PSP-I/PSP-II had lower [Ca(2+)](I) than controls (39.08% vs. 47.97%, respectively; p PSP-I/PSP-II. In conclusion, PSP-I/PSP-II exert a non-permanent decapacitation effect on highly extended boar spermatozoa that is related with a delay in the increase of [Ca(2+)](I) levels.

  2. Chronological age and breed-type effects on carcass characteristics and palatability of bull beef.

    Science.gov (United States)

    Riley, R R; Smith, G C; Cross, H R; Savell, J W; Long, C R; Cartwright, T C

    1986-01-01

    Bulls (n = 115) of four slaughter ages (9, 12, 15 or 18 months) and of 15 genotypes were studied. In this analysis, each bullock was assigned to one of four breed groups-British and British crosses, Brahman and Brahman crosses. Jersey and Jersey crosses or Holstein and Holstein crosses. Slaughter age had an (P Brahman-type bulls had higher shear force values (P Brahman-type carcasses were assigned the lowest numerical ratings for juiciness. Breed-type had a greater effect on tenderness of bull beef than did chronological age.

  3. 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.

  4. 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...

  5. The application of image cytometry to viability assessment in dual fluorescence-stained fish spermatozoa.

    Science.gov (United States)

    Flajshans, Martin; Cosson, Jacky; Rodina, Marek; Linhart, Otomar

    2004-01-01

    The viability of spermatozoa has been assessed using SYBR 14 staining for DNA of living cells and propidium iodide staining for DNA of degenerate cells. This dual staining was performed on four fish species (Siberian sturgeon, Acipenser baerii; common carp, Cyprinus carpio; tench, Tinca tinca and wels, Silurus glanis) and the proportions of live and dead spermatozoa were assessed by epifluorescence microscopy and image cytometry. Ten phase contrast and epifluorescent images were recorded per sample, corresponding images were overlaid, and the blended images were evaluated for live and dead spermatozoa, represented by green and red fluorescence signals. Live/dead proportions were assessed, after dual thresholding, by imaging software that counted absolute numbers of objects and computed their frequencies. All sperm heads were found to be labelled, emitting either green or red light. Mean numbers of spermatozoa per image were in the ranges 32-113, 61-105, 48-104 and 29-91 for Siberian sturgeon, common carp, tench and wels, respectively. The corresponding proportions of live spermatozoa were in the ranges 83.56-94.59%, 93.92-97.02%, 76.14-97.76% and 79.45-83.76%. Standard deviations did not exceed 5% of the means. The image cytometric system using dual staining with SYBR 14 and propidium iodide was clearly suitable for assessing the viability of freshwater fish spermatozoa.

  6. Easy sperm processing technique allowing exclusive accumulation and later usage of DNA-strandbreak-free spermatozoa.

    Science.gov (United States)

    Ebner, T; Shebl, O; Moser, M; Mayer, R B; Arzt, W; Tews, G

    2011-01-01

    Sperm DNA fragmentation is increased in poor-quality semen samples and correlates with failed fertilization, impaired preimplantation development and reduced pregnancy outcome. Common sperm preparation techniques may reduce the percentage of strandbreak-positive spermatozoa, but, to date, there is no reliable approach to exclusively accumulate strandbreak-free spermatozoa. To analyse the efficiency of special sperm selection chambers (Zech-selectors made of glass or polyethylene) in terms of strandbreak reduction, 39 subfertile men were recruited and three probes (native, density gradient and Zech-selector) were used to check for strand breaks using the sperm chromatin dispersion test. The mean percentage of affected spermatozoa in the ejaculate was 15.8 ± 7.8% (range 5.0–42.1%). Density gradient did not significantly improve the quality of spermatozoa selected(14.2 ± 7.0%). However, glass chambers completely removed 90% spermatozoa showing strand breaks and polyethylene chambers removed 76%. Both types of Zech-selectors were equivalent in their efficiency, significantly reduced DNA damage (P sperm preparation technique concentrating spermatozoa unaffected in terms of DNA damage. The special chambers most probably select for sperm motility and/or maturity.

  7. A comparison of ejaculated and testicular spermatozoa aneuploidy rates in patients with high sperm DNA damage.

    Science.gov (United States)

    Moskovtsev, Sergey I; Alladin, Naazish; Lo, Kirk C; Jarvi, Keith; Mullen, J Brendan M; Librach, Clifford L

    2012-06-01

    Testicular spermatozoa are utilized to achieve pregnancy in couples with severe male factor infertility. Several studies suggest that aneuploidy rates in spermatozoa are elevated at the testicular level in infertile patients compared to ejaculates of normal controls. However, essential data regarding aneuploidy rates between ejaculated and testicular spermatozoa in the same individuals is lacking. The purpose of our study was to compare aneuploidy rates at the testicular and post-testicular level from the same patients with persistently high sperm DNA damage. Ejaculates and testicular biopsies were obtained from eight patients with persistently high DNA damage (>30%). Both ejaculated and testicular samples were analyzed for sperm DNA damage and sperm aneuploidy for chromosomes 13, 18, 21, X, and Y. In addition, semen samples from ten normozoospermic men presenting for fertility evaluation served as a control group. A strong correlation between the alteration of spermatogenesis and chromatin deterioration was observed in our study. In the same individuals, testicular samples showed a significantly lower DNA damage compared to ejaculated spermatozoa (14.9% ± 5.0 vs. 40.6% ± 14.8, P<0.05), but significantly higher aneuploidy rates for the five analyzed chromosomes (12.41% ± 3.7 vs. 5.77% ± 1.2, P<0.05). While testicular spermatozoa appear favourable for ICSI in terms of lower DNA damage, this potential advantage could be offset by the higher aneuploidy rates in testicular spermatozoa.

  8. Quality of human spermatozoa: relationship between high-magnification sperm morphology and DNA integrity.

    Science.gov (United States)

    Maettner, R; Sterzik, K; Isachenko, V; Strehler, E; Rahimi, G; Alabart, J L; Sánchez, R; Mallmann, P; Isachenko, E

    2014-06-01

    The aim of this work is to establish the relationship between the morphology of Intracytoplasmic Morphologically Selected Sperm Injection (IMSI)-selected spermatozoa and their DNA integrity. The 45 ejaculates were randomly distributed into three treatment groups: normozoospermic, oligoasthenozoospermic and oligoasthenotheratozoospermic samples. The evaluation of DNA integrity was performed using the sperm chromatin dispersion test. It was established that DNA integrity of spermatozoa is strongly dependent on ejaculate quality (P 0.1). With decreased ejaculate quality, the percentage of spermatozoa with nonfragmented DNA decreased significantly (P < 0.05) independent from morphological class. Nevertheless, the rate of IMSI-selected spermatozoa with fragmented DNA within of Class 1 in normozoospermic (Group 1), in oligoasthenozoospermic (Group 2) and in oligoasthenotheratozoospermic (Group 3) samples was 21.1%, 31.8% and 54.1%, respectively. In conclusion, there is a direct relationship between morphological parameters of spermatozoa and their DNA integrity. However, the IMSI technique alone is not enough for the selection of spermatozoa with intact nuclei.

  9. Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

    Science.gov (United States)

    Nguyen, Quynh Thu; Wallner, Ulrike; Schmicke, Marion; Waberski, Dagmar

    2016-01-01

    ABSTRACT 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. PMID:27612509

  10. 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.

  11. Kruger strict morphology and post-thaw progressive motility in cryopreserved human spermatozoa.

    Science.gov (United States)

    Lee, C-Y; Lee, C-T; Wu, C-H; Hsu, C-S; Hsu, M-I

    2012-05-01

    The purpose of this prospective study was to evaluate Kruger strict morphology and conventional semen analysis in predicting cryosurvival and the progressive motility recovery rate of frozen spermatozoa. Our study included 56 semen samples with >10 million spermatozoa per ejaculate. The main outcome measures were conventional semen analysis, strict morphology analysis by the Kruger method, cryosurvival rate and post-thaw sperm motility. A significant reduction in sperm motility after cryopreservation was demonstrated. The freeze-thawing process caused a 66% reduction in rapid progressive motile spermatozoa, a 45% reduction in slow progressive motile spermatozoa and a 2% reduction in nonprogressive motile spermatozoa. The cryosurvival and progressive motility recovery rates were not correlated with parameters of conventional semen analysis, such as sperm concentration, motility, WHO morphology and total motile count, but the progressive motility recovery rate was significantly correlated with the percentage of spermatozoa exhibiting Kruger normal morphology (P = 0.028). The recovery rate of rapidly progressive motility was profoundly decreased compared with slow progressive motility following the frozen-thaw procedure of semen. Kruger strict morphology assessment was a better predictor of the progressive motility recovery rate following the freezing-thaw procedure than parameters of conventional semen analysis.

  12. Selenium in blood, semen, seminal plasma and spermatozoa of stallions and its relationship to sperm quality.

    Science.gov (United States)

    Bertelsmann, H; Keppler, S; Höltershinken, M; Bollwein, H; Behne, D; Alber, D; Bukalis, G; Kyriakopoulos, A; Sieme, H

    2010-01-01

    The essential trace element selenium is indispensable for male fertility in mammals. Until now, little data existed regarding the relationship between selenium and sperm quality in the stallion. Selenium, or selenium-dependent glutathione peroxidase activity, was determined in red blood cells, semen, seminal plasma and spermatozoa, and the percentages of spermatozoa with progressive motility (PMS), intact membranes (PMI), altered (positive) acrosomal status (PAS) and detectable DNA damage, determined by the sperm chromatin structure assay, were evaluated in 41 healthy stallions (three samples each). The pregnancy rate per oestrus cycle (PRC) served as an estimation of fertility. An adverse effect on stallion fertility caused by low dietary selenium intake was excluded, as all stallions had sufficient selenium levels in their blood. Interestingly, no significant correlations (P > 0.05) between the selenium level in blood and the selenium level in seminal plasma or spermatozoa were found, suggesting that the selenium level in blood is no indicator of an adequate selenium supply for spermatogenesis. The selenium level in spermatozoa (nmol billion(-1)) was correlated with PMI, PMS and PAS (r = 0.40, r = 0.31 and r = -0.42, respectively; P selenium concentration in spermatozoa (nmol g(-1)) was correlated with PRC (r = 0.40, P selenium status for the male equine reproduction requires the analysis of selenium in spermatozoa. Furthermore, selenium is associated with improved sperm quality and fertility in the stallion.

  13. Concentration of trace elements in human semen and relation to spermatozoa quality.

    Science.gov (United States)

    Slivkova, Jana; Popelkova, Miroslava; Massanyi, Peter; Toporcerova, Silvia; Stawarz, Robert; Formicki, Grzegorz; Lukac, Norbert; Putała, Aldona; Guzik, Marek

    2009-03-01

    In this study the concentrations of trace elements such as lead, cadmium, iron, nickel, copper and zinc in the human semen (n = 47), occurrence of pathological spermatozoa, and correlations of these elements to pathological forms were investigated. For each sample of human spermatozoa at least 500 spermatozoa were evaluated. Metal contents were determined by the voltametric method and flame absorption spectrophotometry method. The concentrations of trace elements in human semen were: lead 1.49 +/- 0.40 mg x kg(- 1), cadmium 0.13 +/- 0.15 mg x kg(- 1), iron 2.59 +/- 0.21 mg x kg(- 1), nickel 0.40 +/- 0.07 mg x kg(- 1), copper 0.28 +/- 0.06 mg x kg(- 1), and zinc 153.93 +/- 67.08 mg x kg(- 1), respectively. The total percentage of pathological spermatozoa was 41.61 +/- 9.80% with predominancy of broken flagellum, flagellum torso and separated flagellum. In relation to trace elements the analysis showed correlation between copper and lead (r = -0.47), nickel and iron (r = 0.36), lead and flagellum ball (r = -0.39), cadmium and large heads (r = 0.37) and between iron and other forms of pathological spermatozoa (r = -0.32). Results of this study describe possible effects of trace elements on the spermatozoa quality in normal human sperm.

  14. Effect of NGF on the motility and acrosome reaction of golden hamster spermatozoa in vitro.

    Science.gov (United States)

    Jin, WanZhu; Tanaka, Akane; Watanabe, Gen; Matsuda, Hiroshi; Taya, Kazuyoshi

    2010-08-01

    Motility and fertilizing ability are known to be two important physiological attributes of a mature sperm, yet the mechanism by which spermatozoa mature and become motile remains largely unknown. It has been shown that nerve growth factor (NGF) is a protein essential for the development, maintenance and survival of the peripheral and central nervous systems. However, the presence of high levels of NGF protein and mRNA do not correlate with the innervations by NGF sensitive fibers in tissues such as the testis, prostate and seminal vesicles. These observations have shifted the attention of research to the role of NGF outside of the nervous system. Here, we demonstrate that NGF and its receptors TrkA and p75 are widely expressed in the testis, accessory reproductive organ, and the epididymal sperms. We also show that NGF stimulates two important aspects of sperm functions, motility and the acrosome reaction, in a time- and dose-dependent manner. NGF activated the sperm cell acrosome reaction, while addition of inhibitors specific for MAPK kinase significantly blocked the sperm acrosome reaction. Taken together, our findings suggest that NGF plays an integral role in sperm motility and the acrosome reaction through, at least in part, the MAPK signalling pathway.

  15. 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.

  16. Promoter region of the bovine growth hormone receptor gene: single nucleotide polymorphism discovery in cattle and association with performance in Brangus bulls.

    Science.gov (United States)

    Garrett, A J; Rincon, G; Medrano, J F; Elzo, M A; Silver, G A; Thomas, M G

    2008-12-01

    Expression of the GH receptor (GHR) gene and its binding with GH is essential for growth and fat metabolism. A GT microsatellite exists in the promoter of bovine GHR segregating short (11 bp) and long (16 to 20 bp) allele sequences. To detect SNP and complete an association study of genotype to phenotype, we resequenced a 1,195-bp fragment of DNA including the GT microsatellite and exon 1A. Resequencing was completed in 48 familialy unrelated Holstein, Jersey, Brown Swiss, Simmental, Angus, Brahman, and Brangus cattle. Nine SNP were identified. Phylogeny analyses revealed minor distance (i.e., Brahman cattle averaged 27.4 +/- 0.07% divergence from the Bos taurus breeds, whereas divergence of Brangus was intermediate. An association study of genotype to phenotype was completed with data from growing Brangus bulls (n = 553 from 96 sires) and data from 4 of the SNP flanking the GT microsatellite. These SNP were found to be in Hardy-Weinberg equilibrium and in phase based on linkage disequilibrium analyses (r(2) = 0.84 and D'= 0.92). An A/G tag SNP was identified (ss86273136) and was located in exon 1A, which began 88 bp downstream from the GT microsatellite. Minor allele frequency of the tag SNP was greater than 10%, and Mendelian segregation was verified in 3 generation pedigrees. The A allele was derived from Brahman, and the G allele was derived from Angus. This tag SNP genotype was a significant effect in analyses of rib fat data collected with ultrasound when bulls were ~365 d of age. Specifically, bulls of the GG genotype had 6.1% more (P = 0.0204) rib fat than bulls of the AA and AG genotypes, respectively. Tag SNP (ss86273136), located in the promoter of GHR, appears to be associated with a measure of corporal fat in Bos taurus x Bos indicus composite cattle.

  17. USFWS Land Interests Associated with Bull Mountain Game Range [Land Status Map

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This map was produced by the Division of Realty to depict landownership at Bull Mountain Game Range. It was generated from rectified aerial photography, cadastral...

  18. Longitudinal strain bull's eye plot patterns in patients with cardiomyopathy and concentric left ventricular hypertrophy.

    Science.gov (United States)

    Liu, Dan; Hu, Kai; Nordbeck, Peter; Ertl, Georg; Störk, Stefan; Weidemann, Frank

    2016-05-10

    Despite substantial advances in the imaging techniques and pathophysiological understanding over the last decades, identification of the underlying causes of left ventricular hypertrophy by means of echocardiographic examination remains a challenge in current clinical practice. The longitudinal strain bull's eye plot derived from 2D speckle tracking imaging offers an intuitive visual overview of the global and regional left ventricular myocardial function in a single diagram. The bull's eye mapping is clinically feasible and the plot patterns could provide clues to the etiology of cardiomyopathies. The present review summarizes the longitudinal strain, bull's eye plot features in patients with various cardiomyopathies and concentric left ventricular hypertrophy and the bull's eye plot features might serve as one of the cardiac workup steps on evaluating patients with left ventricular hypertrophy.

  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. 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.

  1. Pemisahan Spermatozoa Berkromosom X dan Y Kambing Boer dan Aplikasinya Melalui Inseminasi Buatan Untuk Mendapatkan Jenis Kelamin Anak Sesuai Harapan

    Directory of Open Access Journals (Sweden)

    Dasrul Dasrul

    2013-04-01

    Full Text Available Separation of spermatozoa with x and y chromosome at boer goat and its application by artificial insemination for kid sex purpose ABSTRACT. The purposes of this experiment are to investigate the separation of X and Y spermatozoa by measuring the spermatozoa quality, sex ratio between X and Y, capacity of fertility indicated by conception rate and sex ratio of goat boar kids. Samples, used in this experiment, are fresh semen from Boer goat with high quality consists of 4 group treatments with 6 replications 1 group of spermatozoa without separation (control, 2 group of spermatozoa separated by percoll gradient density centrifugation 3 levels (P1, 5 levels (P2 and swine up (P3. The observed parameters are spermatozoa quality, X and Y spermatozoa ratio, fertility’s capacity and sex ratio on the birth. Quality examination of spermatozoa and identify X and Y spermatozoa is based on the standard method of WHO. The conception rate was based on the ratio of pregnant goat after the first insemination. Data of spermatozoa quality and spermatozoa ratio were analyzed by using analisis of variance (ANOVA and further analysis by LSD if there were differences between treatments. The results of this experiment showed that spermatozoa quality Boer goat significantly reduced (p<0,05 after separation with percoll gradient density centrifugation  and swim up. Percentage spermatozoa X after percoll gradient density centrifugation was significantly higher (P<0,05 compared to control and swim up. Meanwhile, the Y spermatozoa population was significantly higher (P<0,05 after swim up treatment compared to percoll gradient density centrifugation and control. The percentage of sex ratio (male: female after insemination from percoll gradient density centrifugation produced more female than male. On the other hand, insemination from swim up produced more male than female. Sex ratio produced from separation of percoll gradient density centrifugation, swim up was

  2. 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 <0.005). Additional mental stress further increased both systolic BP and diastolic BP (+3 mm Hg, p <0.05) and heart rate (+13 beats/min, p <0.005) in response to 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.

  3. Ruptured urinary bladder attributable to urethral compression by a haematoma after vertebral fracture in a bull

    OpenAIRE

    Braun, U.; Trösch, L; Sydler, T.

    2014-01-01

    BACKGROUND: In male cattle, rupture of the urinary bladder is usually associated with urethral obstruction by uroliths. Less common causes include urethral compression or stricture. This case report describes the findings in a young Limousion breeding bull with rupture of the urinary bladder because of urethral compression by a haematoma after coccygeal fracture. CASE PRESENTATION: The bull had been introduced into a 40-head Red-Holstein herd one week before being injured. One week after intr...

  4. Population Structure Analysis of Bull Genomes of European and Western Ancestry

    OpenAIRE

    Neo Christopher Chung; Joanna Szyda; Magdalena Frąszczak

    2017-01-01

    Since domestication, population bottlenecks, breed formation, and selective breeding have radically shaped the genealogy and genetics of Bos taurus. In turn, characterization of population structure among diverse bull (males of Bos taurus) genomes enables detailed assessment of genetic resources and origins. By analyzing 432 unrelated bull genomes from 13 breeds and 16 countries, we demonstrate genetic diversity and structural complexity among the European/Western cattle population. Important...

  5. Traumatic Brain Injury Due to Bull Assault in a Girl: a Case Report

    Science.gov (United States)

    ALVIS-MIRANDA, Hernando Raphael; CASTELLAR-LEONES, Sandra Milena; VELÁSQUEZ-LOPERENA, Dufays Danith; VILLA-DELGADO, Rosmery; ALCALA-CERRA, Gabriel; MOSCOTE-SALAZAR, Luis Rafael

    2013-01-01

    ABSTRACT Traumatic brain injury is a common condition in the emergency services, affecting the pediatric and adult population significantly. Patterns of head injury as well as management principles in children are important differences compared to adults. Traumatic brain injury by bull rush is usually seen in adults but has not been described in children-report a pediatric cranial trauma present bull rush, which to our knowledge is the first report in the literature of this nature. PMID:24790672

  6. TINJAUAN PENAMBAHAN MINERAL ZN DALAM PAKAN TERHADAP KUALITAS SPERMATOZOA PADA SAPI FRISIAN HOLSTEIN JANTAN

    Directory of Open Access Journals (Sweden)

    Sus Derthi Widhyari

    2015-04-01

    Full Text Available Bulls are expected to be able to produce a good quality and quantity of sperm. The objective of this experiment was to study the effect of Zn supplementation on the sperm quality, in Frisian holstein bulls. Ten bulls, 1618 months of age were used in this experiment. The experimental bulls were divided into two groups, i.e.,group without Zn supplementation (control and group with 60 ppm of Zn supplementation. Zn supplementations were given everyday for a period of four months. Semen was collected by using artificial vaginaat the end of the experiment. Semen quality was evaluated macroscopically and microscopically. The variables measured were semen volume, semen pH, sperm motility, sperm concentration, sperm viability, and sperm abnormality. The results showed that Zn supplementation significantly increased sperm motility and sperm concentration (P<0.05, whereas there was no significant difference in other parameters.

  7. 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

  8. 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.

    2016-01-01

    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.

  9. 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.

  10. Growth performance, carcass characteristics and meat quality of finishing bulls fed crude glycerin-supplemented diets

    Directory of Open Access Journals (Sweden)

    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.

  11. Reaction norms in weights at 365 days old in nellore bulls in northern Brazil

    Directory of Open Access Journals (Sweden)

    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. 

  12. Binding of semenogelin I to intact human spermatozoa studied by flow cytometry and surface plasmon resonance.

    Science.gov (United States)

    Jonsson, Magnus; Frohm, Birgitta; Malm, Johan

    2010-01-01

    Approximately 1 in 10 couples is infertile. No definite cause can be found in about 25% of those cases. Studies have indicated that seminal vesicle secretion functions as an optimizer of fertilization. The Zn(2+) binding protein semenogelin I (SgI) represents a major fraction of the proteins present in seminal vesicle fluid, and it serves as a structural component of the coagulum that is formed after ejaculation. Cleavage of SgI by prostate-specific antigen results in liquefaction of the coagulum. Fragmented SgI has antibacterial effects and inhibits spermatozoa mobility. SgI has also been found complexed to eppin on spermatozoa, and this complex has been suggested to be of importance for fertility. Here, we used flow cytometry and surface plasmon resonance to study SgI regarding its association with spermatozoa and the interaction dependency on Zn(2+). The concentration of Zn(2+) in seminal plasma is approximately 100 times higher than in blood plasma, and the metal ion is known to change the structure of SgI. We found that SgI binds to spermatozoa in a concentration-dependent and saturable manner. In solution, SgI bound to spermatozoa in a non-Zn(2+)-dependent way, whereas immobilized SgI interacts with spermatozoa only in the presence of Zn(2+). It indicates that SgI must exhibit a specific structure or free flexibility to be able to interact with that ligand. Our results indicate that the association of SgI to spermatozoa is conformation dependent and specific. These findings could constitute a basis for the development of a male contraceptive.

  13. Resistance of human spermatozoa to cryoinjury in repeated cycles of thaw-refreezing

    Directory of Open Access Journals (Sweden)

    Sidney Verza Jr.

    2009-10-01

    Full Text Available Objective: To study the resistance of human spermatozoa to cryoinjury in repeated cycles of thaw-refreezing by using the fast liquid nitrogen vapor method. Material and Methods: Semen specimens were obtained from sixteen normal and oligozoospermic individuals who required disposal at the sperm bank. Five of them had testicular cancer. Specimens were thawed and an aliquot was removed for analysis. The remaining specimens were refrozen without removing the cryomedia. Repeated freeze-thaw cycles were performed until no motile sperm were observed. Sperm motility, number of motile spermatozoa and viability were determined after thawing. Resistance to cryoinjury was compared between groups and also after each refreezing cycle within groups. Results: Motile spermatozoa were recovered after five and two refreeze-thawing cycles in normozoospermic and oligozoospermic specimens, respectively. There were no significant differences in the recovery of motile spermatozoa between thaws within each group of normal and oligozoospermic specimens, but percentage motility and total number of motile spermatozoa were significantly lower in the oligozoospermic one. Specimens from men with cancer were exposed to six refreeze-thawing cycles. Although recovery of motile spermatozoa was significantly impaired after each thawing, there were no significant differences in the recovery of motile sperm between thaws in cancer and non-cancer groups. Conclusions: Human spermatozoa resist repeated cryopreservation using the fast liquid nitrogen vapor method. Normozoospermic specimens withstand refreezing for an average two cycles longer than oligozoospermic ones. Specimens from cancer patients seem to resist repeated cryoinjury similarly to non-cancer counterparts. Resistance to repeated cryoinjury was related to the initial semen quality.

  14. Effects of the seminal plasma zinc content and catalase activity on the semen quality of water buffalo (Bubalus bubalis) bulls.

    Science.gov (United States)

    Alavi-Shoushtari, S M; Rezai, S Asri; Ansari, M H Kh; Khaki, A

    2009-01-15

    In order to determine zinc and catalase content of seminal plasma in the buffalo and to study their associations with the semen characteristics, 54 semen samples were collected from 10 buffalo bulls; semen volume and sperm concentration, gross and progressive motility and viability were evaluated, seminal plasma was then harvested by centrifugation and its zinc content was estimated by atomic absorption spectrophotometer and its catalase activity determined by using a commercial kit. The zinc content of the seminal plasma (Mean +/- SEM) was recorded as 154.40 +/- 1.74 mg L(-1), while, the mean catalase value was 32.00 +/- 0.42 U mL(-1). The mean zinc values was highly correlated with sperm progressive motility and viability and with catalase values (p = 0.000 for all) and also was associated with gross motility (p = 0.020) and negatively with abnormal morphology (p = 0.049). The catalase values were highly associated with sperm progressive motility, viability and zinc content (p = 0.000 for all) and was associated with sperm gross motility (p = 0.024). For further clarification of these correlations, the samples were categorized in three groups of excellent (Ex, >90% motile, n = 33), good (Go, 80-89% motile, n = 15) and moderate (Mo, zinc and catalase values were recorded as 161.07 +/- 1.63 mg L(-1) and 33.41 +/- 0.34 U mL(-1) in Ex, 146.70 +/- 1.91 mg L(-1) and 31.01 +/- 0.67 in Go and 136.42 +/- 4.97 mg L(-1) and 26.51 +/- 0.87 U mL(-1) in Mo groups. The mean zinc value in Ex group was highly associated with sperm motility, viability and catalase values, in Go group was associated with catalase values and highly associated with sperm abnormal morphology and in Mo group it was highly associations with catalase values only. The mean catalase value in Ex group, was highly associated with sperm motility and viability, in Go group was associated with zinc content and in Mo groups was highly associated with the zinc content. These results show that seminal plasma zinc

  15. Use of Bayesian Inference to Correlate In Vitro Embryo Production and In Vivo Fertility in Zebu Bulls

    Directory of Open Access Journals (Sweden)

    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.

  16. DAG defect in Murrah buffalo bulls in Brazil

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    W.G. Vale

    2010-02-01

    Full Text Available Dag defect sperms have an abnormal tail coiled morphology are totally or almost totally immotile and have severe abnormalities on the fibers in the axial filament. This defect in bovine was shown to be due an autosomal recessive gene. In the present study 23 buffalo bulls age 18-34 months examined for BSE (Breeding Soundness Examination showed sperm motility varied between 0 to 70% accomplished with 10 a 65% of sperm pathology with or not a predominance of strong coiled tails with or without persistence of droplet. In Brazil Murrah breed have had only eight female PO (pure of origin - India gave origin of the herd and not more than four males where also imported from India, although in the last two decades some quantity of frozen semen were introduced in the country imported from India. As in the other domestic species inbreed seems to be the main cause of this abnormalities in different breeds. The knowledge and further study of such abnormalities is the only way to overcome a problem that if not controlled will tend to spread out without control causing in the future more problem for development of some herds.

  17. Behaviour and meat quality of Podolian young bulls

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    Fabio Napolitano

    2010-01-01

    Full Text Available From April to August 2008, twelve Podolian subjects, aged about 11 months at the beginning of the experimental period, were used to evaluate the effect of rearing system (Confined vs. Freerange and season (spring vs. summer on their behaviour and meat quality. Nine sessions of behavioural observations were performed. During a 6-h period, the behaviour of a focal animal, was continuously monitored. In each session a different animal was chosen. All the animals were slaughtered at 18 months of age. Walking (P<0.001 and standing (P<0.05 were lower in summer, whereas inactivity was higher (P<0.05. Free-range bulls spent more time walking (P<0.05, feeding (P<0.001 and standing (P<0.01 and showed a lower number of agonistic (P<0.05 and non-agonistic social interaction than confined animals (P<0.01. Self- and allo-grooming were not affected by rearing system, whereas season influenced self-grooming with higher values in spring (P<0.05. Confined animals showed higher final weights (P<0.05 and a lighter meat (P<0.05, whereas no differences between groups were observed for average daily gains, carcass yield, water holding capacity and a* and b* indexes. Confinement markedly affected the behaviour of the animals, whereas free-ranging had only minor negative effects on meat lightness.

  18. Molecular Cloning of Spergen-4, Encoding a Spermatogenic Cell-Specific Protein Associated with Sperm Flagella and the Acrosome Region in Rat Spermatozoa.

    Science.gov (United States)

    Howida, Ali; Salaheldeen, Elsaid; Iida, Hiroshi

    2016-04-01

    We used a differential display in combination with complementary DNA (cDNA) cloning approach to isolate a novel rat gene LOC690919 with an open reading frame of 1227-length nucleotides encoding a protein of 409 amino acids. This gene was designated as Spergen-4 (a spermatogenic cell-specific gene-4). Spergen-4 mRNA was highly expressed in testis, and its expression was detected in rat testis starting at three weeks of postnatal development and persisting up to adulthood. Mouse and human orthologs, which lack N-terminal 77 amino acid residues of rat Spegen-4, were found in the database. Immunofluorescence microscopy and immunoblot analysis demonstrated that Spergen-4 was not expressed in spermatogonia, spermatocytes, and round spermatids, but was restrictedly detected at sperm head, cytoplasm, and developing flagella of elongated spermatids in rat testis. In mature spermatozoa, Spergen-4 was detected at the acrosome region as well as the principal piece of flagella. Spergen-4 immunosignal disappeared from sperm heads on acrosome reaction induced by progesterone. These data suggest that Spergen-4 integrated into elongated spermatids during spermiogenesis serves as a constituent for acrosome region and flagella of rat spermatozoa.

  19. 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 a

  20. Bull Trout Distribution and Abundance in the Waters on and Bordering the Warm Springs Indian Reservation, 2000 Annual Report.

    Energy Technology Data Exchange (ETDEWEB)

    Brun, Christopher

    2000-01-01

    The range of bull trout (Salvelinus confluentus) in the Deschutes River basin has decreased from historic levels due to many factors including dam construction, habitat degradation, brook trout introduction and eradication efforts. While the bull trout population appears to be stable in the Metolius River-Lake Billy Chinook system they have been largely extirpated from the upper Deschutes River (Buchanan et al. 1997). Little was known about bull trout in the lower Deschutes basin until BPA funded project No.9405400 began during 1998. In this progress report we describe the findings from the third year (2000) of the multi-year study aimed at determining the life history, genetics, habitat needs and limiting factors of bull trout in the lower Deschutes subbasin. Juvenile bull trout and brook trout (Salvelinus fontinalis) relative abundance was assessed in the Warm Springs River and Shitike Creek by night snorkeling. In the Warm Springs R. juvenile bull trout were slightly more numerous than brook trout, however, both were found in low densities. Relative densities of both species declined from 1999 observations. Juvenile bull trout vastly out numbered brook trout in Shitike Cr. Relative densities of juvenile bull trout increased while brook trout abundance was similar to 1999 observations in eight index reaches. The utility of using index reaches to monitor trends in juvenile bull trout and brook trout relative abundance was assessed in the Warm Springs R. for the second year. Mean relative densities of both species, within the index reaches was slightly higher than what was observed in a 2.4 km control reach. Mill Creek was surveyed for the presence of juvenile bull trout. The American Fisheries Society ''Interim protocol for determining bull trout presence'' methodology was field tested. No bull trout were found in the 2 km survey area.

  1. Identifying factors affecting age at first semen freezing and age at first semen use in Sahiwal bulls

    OpenAIRE

    Naha, B. C.; Chakravarty, A. K.; Mir, M. A.; V. Jamuna; Singh, A. P.; Maher, D

    2015-01-01

    Aim: The objective of the study was to evaluate the effects of non-genetic factors on reproduction traits viz. age at first semen freezing and age at first semen use of breeding bulls in Sahiwal bulls by fitting least-squares analysis. Materials and Methods: The information on reproduction traits of 43 Sahiwal breeding bulls belonging to 8 sets of Sahiwal breeding program at Indian Council of Agricultural Research-National Dairy Research Institute (ICAR-NDRI), Karnal (Haryana), India durin...

  2. Osmotic stress stimulates generation of superoxide anion by spermatozoa in horses.

    Science.gov (United States)

    Burnaugh, L; Ball, B A; Sabeur, K; Thomas, A D; Meyers, S A

    2010-02-01

    The objective of this study was to examine the interplay between osmotic and oxidative stress as well as to determine mechanisms by which osmotic stress increases superoxide generation in spermatozoa of horses. Superoxide production, as measured by dihydroethidium (DHE), increased when spermatozoa of horses were incubated under either hyperosmotic or hyposmotic conditions. This increase in superoxide production was inhibited by the MAP kinase p38 inhibitor, SB203580, and by the superoxide scavenger, tiron. Incubation of spermatozoa under hyperosmotic conditions increased overall protein tyrosine phosphorylation as measured by western blotting techniques; however, a similar increase was not detected when spermatozoa were incubated under hyposmotic conditions. The general protein kinase C (PKC) and protein tyrosine kinase (PTK) inhibitor staurosporine inhibited (P<0.05) tyrosine phosphorylation in samples from cells under hyperosmotic conditions. In addition, the NADPH oxidase inhibitor diphenyleneiodonium (DPI) also inhibited (P<0.05) protein tyrosine phosphorylation in cells under hyperosmotic conditions. In summary, these data indicate that incubation of equine spermatozoa under both hyposmotic and hyperosmotic conditions can increase superoxide anion generation. Under hyperosmotic conditions, this increased generation of superoxide anion was accompanied by increased protein tyrosine phosphorylation.

  3. Increased chromatin fragmentation and reduced acrosome integrity in spermatozoa of red deer from lead polluted sites.

    Science.gov (United States)

    Castellanos, Pilar; del Olmo, Enrique; Fernández-Santos, M Rocío; Rodríguez-Estival, Jaime; Garde, J Julián; Mateo, Rafael

    2015-02-01

    Vertebrates are constantly exposed to a diffuse pollution of heavy metals existing in the environment, but in some cases, the proximity to emission sources like mining activity increases the risk of developing adverse effects of these pollutants. Here we have studied lead (Pb) levels in spermatozoa and testis, and chromatin damage and levels of endogenous antioxidant activity in spermatozoa of red deer (Cervus elaphus) from a Pb mining area (n=37) and a control area (n=26). Deer from the Pb-polluted area showed higher Pb levels in testis parenchyma, epididymal cauda and spermatozoa, lower values of acrosome integrity, higher activity of glutathione peroxidase (GPx) and higher values of DNA fragmentation (X-DFI) and stainability (HDS) in sperm than in the control area. These results indicate that mining pollution can produce damage on chromatin and membrane spermatozoa in wildlife. The study of chromatin fragmentation has not been studied before in spermatozoa of wildlife species, and the sperm chromatin structure assay (SCSA) has been revealed as a successful tool for this purpose in species in which the amount of sperm that can be collected is very limited.

  4. 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.

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

    Directory of Open Access Journals (Sweden)

    Kondracki S.

    2013-01-01

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

  6. Effects of No on spontaneous acrosome reaction in AsAb positive rat spermatozoa

    Institute of Scientific and Technical Information of China (English)

    ZhanW; NiJ

    2002-01-01

    Objective:To explore the effect of NO on the spontaneous acrosome reaction in antisperm antibody (AsAb) positive rat spermatozoa.Methods:The rat model of AsAb was set up by artificial immunization.The level of AsAb in blood serum was determined by TAT and ELISA.Rat spermatozoa was visualized by staining the acrosome with Coomassie brilliant blue.The NO concentration in rat spermatozoa was assayed by HPLC.Results:The percentage of acrosome reaction,NO concentration,superoxide dismutase(SOD) and Na+-K+ ATPase activity in AsAb positive rat spermatozoa were significantly decreased compared with the control group.Low dose of NO(SNP 10-9-10-8mol/L) increased the percentage of acrosome reaction and SOD activity,but had no effect on Na+-K+ATPase activity.High dose of NO(SNP 10-6-10-4mol/L) decreased the three items.Conclusion:The decrease in acrosome reaction in positive AsAb rat spermatozoa might be related to a decrease in NO and increase in O2-.(the SOD activity was decreased)in sperm.A low dose of NO might increase the percentage of acrosome reaction in AsAb positive rat by inhibiting the superoxide,while a high dose of NO was harmful to sperm function.

  7. Evaluation of chromatin condensation in human spermatozoa: a flow cytometric assay using acridine orange staining.

    Science.gov (United States)

    Golan, R; Shochat, L; Weissenberg, R; Soffer, Y; Marcus, Z; Oschry, Y; Lewin, L M

    1997-01-01

    The quality of sperm chromatin is an important factor in fertilization and is especially critical where one spermatozoon is artificially selected for fertilizing an egg (as in intracytoplasmic sperm injection). In this study, flow cytometry after staining of human spermatozoa with Acridine Orange was used to study chromatin structure. A method is described for estimating the percentage of cells in a human sperm sample that have completed epididymal maturation in regard to chromatin condensation. Of the 121 samples of the semen that were examined, nine contained a higher percentage of hypocondensed spermatozoa and six samples contained elevated amounts of hypercondensed spermatozoa. In addition to aberrancies in chromatin condensation other defects showed up as satellite populations of spermatozoa with higher than normal ratios of red/green fluorescence after Acridine Orange staining. Such defects were found in 15 semen samples. The use of swim-up and Percoll gradient centrifugation methods was shown to improve the percentage of spermatozoa with normal chromatin structure in some samples with poor initial quality.

  8. Effect of dietary selenium deficiency on the in vitro fertilizing ability of mice spermatozoa.

    Science.gov (United States)

    Sánchez-Gutiérrez, M; García-Montalvo, E A; Izquierdo-Vega, J A; Del Razo, L M

    2008-08-01

    Selenium is an essential micronutrient for mammals, being integral part of antioxidant system. The aim of the study was to evaluate the effect of selenium deficiency on in vitro fertilization (IVF) capacity of spermatozoa and on oxidative stress in these cells. Male C57BL/6N mice were maintained on selenium-deficient or selenium-sufficient diets (0.02 or 0.2 ppm of selenium as selenomethionine, respectively) for 4 months. Liver glutathione peroxidase activity measurements were used to confirm selenium deficiency. Sperm quality and IVF capability among both groups were evaluated. To assess oxidative damage, lipid peroxidation as malondialdehyde production was determined in spermatozoa as well as the testes. Ultrastructural analyses of spermatozoa nuclei using transmission electron microscopy were also performed. The percentage of eggs fertilized with sperm from selenium-deficient mice was significantly decreased by approximately 67%. This reduced fertilization capacity was accompanied by increased levels of lipid peroxidation in both the testes and sperm, indicating that selenium deficiency induced oxidative stress. Consistent with this finding, spermatozoa from selenium-deficient animals exhibited altered chromatin condensation. Deficiency in dietary selenium decreases the reproductive potential of male mice and is associated with oxidative damage in spermatozoa.

  9. Seminal concentration of trace elements in fox and relationships to spermatozoa quality.

    Science.gov (United States)

    Massányi, Peter; Trandzik, Jozef; Nad, Pavol; Skalická, Magdalena; Koréneková, Beata; Lukac, Norbert; Fabis, Marian; Toman, Robert

    2005-01-01

    Concentrations of copper, zinc, iron, cadmium, lead, and nickel in the semen of foxes (Vulpes vulpes, n = 10), microscopic analysis of occurrence of pathological spermatozoa, and correlations of these elements with pathological forms were studied. Samples were analyzed by using an atomic absorption spectrophotometer. For analysis of pathological spermatozoa semen samples fixed with Hancock's solution and stained with Giemsa were prepared. For each fox at least 1000 spermatozoa were evaluated. The concentrations of copper, zinc, and iron in semen of foxes were found to be 2.16+/-0.53 mg/kg, 13.09+/-5.22 mg/kg, and 33.16+/-24.36 mg/kg, respectively, on wet weight basis. Concentration of cadmium was low (0.07+/-0.05 mg/kg). The levels of lead and nickel in the semen of foxes were 0.08+/-0.06 mg/kg and 0.35+/-0.24 mg/kg, respectively. The total percentage of pathological spermatozoa was 7.76+/-1.33% with predominancy of knob twisted flagellum, separated flagellum, and broken flagellum. In relation to trace elements the analysis showed significant (p spermatozoa (r = -0.72), zinc and broken flagellum (r = -0.69), iron and retention of cytoplasmic drop (r = 0.87), cadmium and separated flagellum (r = -0.68), and between cadmium retention of cytoplasmic drop (r = 0.87).

  10. Effects of alginate on frozen-thawed boar spermatozoa quality, lipid peroxidation and antioxidant enzymes activities.

    Science.gov (United States)

    Hu, Jinghua; Geng, Guoxia; Li, Qingwang; Sun, Xiuzhu; Cao, Hualin; Liu, Yawei

    2014-06-30

    Although alginate was reported to play an important role as free radical scavengers in vitro and could be used as sources of natural antioxidants, there was no study about the cryoprotective effects of alginate on boar spermatozoa freezing. The objective of this research was to evaluate the effects of different concentrations of alginate added to the freezing extenders on boar spermatozoa motility, plasma membrane integrity, acrosomal integrity, mitochondrial activities, lipid peroxidation and antioxidative enzymes activities (SOD and GSH-Px) after thawing. Alginate was added to the TCG extender to yield six different final concentrations: 0, 0.2, 0.4, 0.6, 0.8, and 1.0mg/mL. The semen extender supplemented with various doses of alginate increased (Palginate also provided significantly positive effect on post-thaw boar spermatozoa acrosomal integrity at concentrations of 0.6, 0.8, 1.0mg/mL, compared with that of the control (Palginate led to higher SOD and GSH-Px activities and lower MDA levels, in comparison to the control (Palginate exhibited a dose-related response on frozen-thawed boar spermatozoa motility, functional integrity and antioxidative capacity at appropriate concentrations. Therefore alginate could be employed as an effective cryoprotectant in boar spermatozoa cryopreservation.

  11. Breeding soundness evaluation of bulls by semen analysis, testicular fine needle aspiration cytology and trans-scrotal ultrasonography

    Directory of Open Access Journals (Sweden)

    Chapwanya A

    2008-05-01

    Full Text Available Abstract The aim of this study was to evaluate the usefulness of trans-scrotal ultrasonography and testicular fine needle aspiration cytology in assessing bulls for breeding suitability. These two techniques were also compared with semen analysis. Bulls presented for breeding soundness evaluation were assessed using all three techniques. The findings of each technique were compared. There was agreement in classification of fertile bulls using all three techniques, suggesting that the combined use of these techniques enhances routine breeding soundness examination. Use of the three techniques also enhances detailed investigation of suspected sub-fertile bulls while accurately identifying testicular cause(s of sire sub-fertility.

  12. The Effect of Production System on Fatty Acid Composition in Beef Meat of Cika Young Bulls

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    Mojca Simčič

    2013-09-01

    Full Text Available The aim of the study was to determine chemical and fatty acid composition in meat of Cika bulls fattened in two production systems. Eight bulls were semi-intensively fattened with a total mix ratio (TMR, while ten were grazed without cereals supplementation. Housed bulls were slaughtered at the age of 20.0 months while grazed bulls were slaughtered at the age of 23.5 months. Samples of M. longissimus dorsi located between the 7th and 8th rib were used to determine the chemical and fatty acid composition. Data were analysed using the GLM procedure of the SAS/ STAT. Model 1 was used to test the effect of production system on the chemical composition of meat. Model 2 was used to test the effects of production system and the intramuscular fat content nested within the production system as linear regression on the fatty acid composition. Significant differences were determined in intramuscular fat (14.54 vs. 8.57 g/kg, P = 0.015 and in dry matter content (231.85 vs. 239.49 g/kg, P = 0.032 in beef meat of TMR fed and in grazed bulls, respectively. Beef meat of grazed bulls contained significantly lower palmitic acid (19.87 vs. 21.03 wt.%, P = 0.002, oleic acid (26.18 vs. 30.01 wt.%, P = 0.007 and MUFA (29.88 vs. 33.80 wt.%, P = 0.010 and higher α-linolenic (3.33 vs. 1.45 wt.%, P = 0.014, EPA (1.40 vs. 0.57 wt.%, P = 0.002, DPA (1.84 vs. 1.06 wt.%, P = 0.001, PUFA (21.42 vs. 19.43 wt.%, P = 0.047 and n-3PUFA (6.80 vs. 3.24 wt.%, P = 0.001 compared to TMR fed bulls.

  13. Biochemical and organoleptic characteristics of muscle from early and late maturing bulls in different production systems.

    Science.gov (United States)

    Mezgebo, G B; Monahan, F J; McGee, M; O'Riordan, E G; Picard, B; Richardson, R I; Moloney, A P

    2016-12-28

    In grass-based beef production systems (PS), early maturing (EM) breed types may be preferable to late maturing (LM) breed types in achieving adequate carcass fat cover. Biochemical and organoleptic characteristics of muscle from suckler bulls were investigated in EM and LM (n=28/breed) assigned to one of two PS (ad libitum concentrates and grass silage to slaughter (C) or ad libitum silage plus 2 kg concentrate daily during winter followed by 99 days at pasture and then an indoor finishing period on C (GSPC)) in a 2 breed type×2 PS factorial arrangement of treatments. Bulls were managed to have a common target carcass weight of 380 kg. Intramuscular fat (IMF) content was higher (P<0.05) for EM than LM, and for C than GSPC bulls. Collagen solubility was higher (P<0.05) for C than GSPC bulls. Lactate dehydrogenase (LDH) and phosphofructokinase activities were higher (P<0.05) for LM than EM. Isocitrate dehydrogenase activity and the Type I myosin heavy chain (MyHC) proportion were higher (P<0.05) for EM than LM. The LDH activity and the Type IIX MyHC proportion were higher (P<0.05) for C than GSPC bulls. Sensory ratings for tenderness and juiciness were higher (P<0.01) for beef from EM than LM while sensory ratings for tenderness, flavour liking and overall liking were higher (P<0.001) for C than for GSPC bulls. Differences in sensory quality were largely eliminated when adjusted for IMF. Overall, carcass fat scores, IMF and sensory scores were higher in EM than LM and in C than GSPC bulls but most differences in sensory quality could be attributed to differences in IMF.

  14. Physical, biotic, and sampling influences on diel habitat use by stream-dwelling bull trout

    Science.gov (United States)

    Banish, N.P.; Peterson, J.T.; Thurow, R.F.

    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 benthic and solitary; most (88%) were observed at night. We developed a conditional logistic regression model to account for the effect of fish movement in response to snorkeling, and we fitted 18 candidate models to evaluate the relative influences of biotic and abiotic factors on habitat use. The candidate models were also fitted with a naive logistic regression (i.e., no movement) to evaluate the effects of movement on inferences of microhabitat use. The most plausible model describing bull trout habitat use was the same for the conditional and nai??ve regressions and included depth, velocity, percent rubble substratum, and the day X depth, body size X depth, and body size X day X depth interactions. The presence of brook trout S. fontinalis and the abundance of conspecifics did not strongly influence microhabitat use by bull trout. The relative rankings of the remaining models differed substantially between the conditional and nai??ve models. Relative to the conditional models, the naive models overestimated the importance of diurnal differences in habitat use and overestimated the use of deepwater habitats, particularly during the day. Both model types suggested that all sizes of bull trout were generally found in deeper, low-velocity habitat at night, whereas small bull trout (70-90 mm total length) were found in shallower habitats during the day. We recommend lhat biologists account for fish movement in response to sampling to avoid biasing modeled habitat use patterns by bull trout. ?? Copyright by the American Fisheries Society 2008.

  15. Natural or artificial? Habitat-use by the bull shark, Carcharhinus leucas.

    Directory of Open Access Journals (Sweden)

    Jonathan M Werry

    Full Text Available BACKGROUND: Despite accelerated global population declines due to targeted and illegal fishing pressure for many top-level shark species, the impacts of coastal habitat modification have been largely overlooked. We present the first direct comparison of the use of natural versus artificial habitats for the bull shark, Carcharhinus leucas, an IUCN 'Near-threatened' species--one of the few truly euryhaline sharks that utilises natural rivers and estuaries as nursery grounds before migrating offshore as adults. Understanding the value of alternate artificial coastal habitats to the lifecycle of the bull shark is crucial for determining the impact of coastal development on this threatened but potentially dangerous species. METHODOLOGY/FINDINGS: We used longline surveys and long-term passive acoustic tracking of neonate and juvenile bull sharks to determine the ontogenetic value of natural and artificial habitats to bull sharks associated with the Nerang River and adjoining canals on the Gold Coast, Australia. Long-term movements of tagged sharks suggested a preference for the natural river over artificial habitat (canals. Neonates and juveniles spent the majority of their time in the upper tidal reaches of the Nerang River and undertook excursions into adjoining canals. Larger bull sharks ranged further and frequented the canals closer to the river mouth. CONCLUSIONS/SIGNIFICANCE: Our work suggests with increased destruction of natural habitats, artificial coastal habitat may become increasingly important to large juvenile bull sharks with associated risk of attack on humans. In this system, neonate and juvenile bull sharks utilised the natural and artificial habitats, but the latter was not the preferred habitat of neonates. The upper reaches of tidal rivers, often under significant modification pressure, serve as nursery sites for neonates. Analogous studies are needed in similar systems elsewhere to assess the spatial and temporal generality of

  16. Effect of dialysis on the proacrosin/acrosin system and motility of turkey (Meleagris gallopavo) spermatozoa during liquid storage.

    Science.gov (United States)

    Słowińska, M; Dietrich, G J; Liszewska, E; Kozłowski, K; Jankowski, J; Ciereszko, A

    2013-01-01

    1. The effect of dialysis on the proacrosin/acrosin system and motility of turkey spermatozoa were examined after 24 and 48 h of liquid storage at 4°C. 2. Fifteen pools of semen diluted in extender were dialysed against Clemson Turkey Semen Diluent (dialysed semen) or stored in aerobic conditions (undialysed semen). Semen quality was assessed by measuring spermatozoa motility, amidase activity of spermatozoa suspension, spermatozoa extract and seminal plasma and anti-trypsin activity of seminal plasma. 3. Extracted amidase activity of dialysed semen was lower than undialysed by 28%. Higher values for speed parameters of spermatozoa were found in dialysed semen in comparison to undialysed, for example, 81.6 µm/s versus 75.0 µm/s for straight-line velocity (VSL), 114.7 µm/s versus 110.3 µm/s for curvilinear velocity (VCL) and 86.6 µm/s versus 79.8 µm/s for average path velocity (VAP). 4. It was concluded that dialysis caused lower amidase activity of spermatozoa and increased speed parameters of progressively motile turkey spermatozoa during storage. Lower extracted amidase activity of dialysed semen reflected better membrane integrity of dialysed semen and suggests that the proacrosin/acrosin system of dialysed spermatozoa is less susceptible to activation compared to undialysed semen.

  17. Cryopreservation-induced alterations in protein tyrosine phosphorylation of spermatozoa from different portions of the boar ejaculate.

    Science.gov (United States)

    Kumaresan, A; Siqueira, A P; Hossain, M S; Bergqvist, A S

    2011-12-01

    Previous studies have shown that boar sperm quality after cryopreservation differs depending on the ejaculate fraction used and that spermatozoa contained in the first 10mL (P1) of the sperm-rich fraction (SRF) show better cryosurvival than those in the SRF-P1. Since protein tyrosine phosphorylation (PTP) in spermatozoa is related with the tolerance of spermatozoa to frozen storage and cryocapacitation, we assessed the dynamics of cryopreservation-induced PTP and intracellular calcium ([Ca(2+)]i) in spermatozoa, using flow cytometry, from P1 and SRF-P1 of the boar ejaculate at different stages of cryopreservation. Sperm kinetics, assessed using a computer-assisted semen analyzer, did not differ between P1 and SRF-P1 during cryopreservation but the decrease in sperm velocity during cryopreservation was significant (Psperm PTP. The proportion of spermatozoa with PTP did not differ significantly between portions of the boar ejaculate. However at any given step during cryopreservation the percentage of spermatozoa with PTP was comparatively higher in SRF-P1 than P1. A 32kDa tyrosine phosphorylated protein, associated with capacitation, appeared after cooling suggesting that cooling induces capacitation-like changes in boar spermatozoa. In conclusion, the study has shown that the cryopreservation process induced PTP in spermatozoa and their proportions were similar between portions of SRF.

  18. Effects of storage in different semen extenders on the pre-freezing and post-thawing quality of boar spermatozoa.

    Science.gov (United States)

    Dziekońska, A; Zasiadczyk, Ł; Lecewicz, M; Strzeżek, R; Koziorowska-Gilun, M; Fraser, L; Mogielnicka-Brzozowska, M; Kordan, W

    2015-01-01

    The aim of this study was to investigate the effects of storage of semen in different commercial extenders on the pre-freezing and post-thawing quality of boar spermatozoa. Semen was diluted in BTS, Androhep (AH) and Gedil (GD), stored for 24 h at 17°C, and then frozen in accordance with the cryopreservation protocol. Analyses of the quality of spermatozoa included: motility, normal apical ridge (NAR) acrosome, plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), measurements of ATP content and activity of superoxidase dismutase (SOD) and glutathione peroxidase (GPx). Prior to the freezing process, no significant effect of the extender on the sperm quality parameters was noted. After thawing the spermatozoa it was demonstrated that the type of extender used influenced PMI, MMP, ATP content and activity of GPx. In the AH extender the percentage of spermatozoa with PMI and ATP content in spermatozoa was significantly higher (Pboar spermatozoa stored for 24 hours in liquid state can be used. However, the type of extender used prior to freezing may have a significant effect on the post-thawing quality of the spermatozoa. The AH extender better secured the quality of thawed boar spermatozoa as compared with the BTS or GD.

  19. Inhibition of human spermatozoa-zona pellucida binding by a combinatorially derived peptide from a synthetic target.

    Science.gov (United States)

    Pieczenik, George; Garrisi, John; Cohen, Jacques

    2006-09-01

    Intact zona-free human oocytes were screened using a combinatorial peptide library selection protocol. Pieczenik Peptide Sequence 1 (PPS1) HEHRKRG binds human spermatozoa. A complementary and unique binding sequence HNSSLSPLATPA (PPS2) was developed from the first PPS1 ligand that binds to the human zona pellucida or oolemma. Cytoplasm-free zonae from unfertilized eggs were obtained and used as an assay system to test the effects of exposure to these two ligands. Spermatozoa were inserted into evacuated zonae and their behaviour and binding activity were assessed at regular intervals. The behaviour of spermatozoa exposed to PPS1 and unlabelled spermatozoa injected into unexposed zonae was similar as far as binding was concerned (50 and 54% binding), but PPS1 exposed spermatozoa had higher motility and displacement, marked by their escape from the zona pellucida. Zonae exposed to PPS2 inhibited the interaction between injected spermatozoa and the inside of the zona when compared with controls (8.3 and 53.8% attached respectively, P movie sequence taken approximately 30 min after injection of spermatozoa into empty human zonae pellucidae shows behaviour of non-manipulated spermatozoa into zonae not exposed or exposed to ligand. This may be purchased for viewing on the Internet at www.rbmonline.com/Article/2159 (free to web subscribers).

  20. 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 boar

  1. Nucléation, ascension et éclatement d'une bulle de champagne

    Science.gov (United States)

    Liger-Belair, G.

    2006-03-01

    People have long been fascinated by bubbles and foams dynamics, and since the pioneering work of Leonardo da Vinci in the early 16th century, this subject has generated a huge bibliography. However, only quite recently, much interest was devoted to bubbles in Champagne wines and carbonated beverages. Since the time of the benedictine monk dom Pierre Perignon (1638-1715), champagne is the wine of celebration. This fame is largely linked to the elegance of its effervescence and foaming properties. In this book, the latest results about the chemical physics behind the bubbling properties of Champagne and sparkling wines are collected and fully illustrated. The first chapter is devoted to the history of champagne and to a presentation of the tools of the physical chemistry of interfaces needed for a whole comprehension of the book. Then, the three main steps of a fleeting champagne bubble's life are presented in chronological order, that is, the bubble nucleation on the glass wall (Chap.2), the bubble ascent and growth through the liquid matrix (Chap.3), and the bursting of bubbles at the liquid surface (Chap.4), which constitutes the most intriguing, functional, and visually appealing step. L'objectif général de ce travail consacré à l'étude des processus physicochimiques liés à l'effervescence des vins de Champagne était de décortiquer les différentes étapes de la vie d'une bulle de champagne en conditions réelles de consommation, dans une flûte. Nous résumons ci-après les principaux résultats obtenus pour chacune des étapes de la vie de la bulle, depuis sa naissance sur les parois d'une flûte, jusqu'à son éclatement en surface. Nucléation À l'aide d'une caméra rapide munie d'un objectif de microscope, nous avons pu mettre à mal une idée largement répandue. Ce ne sont pas les anfractuosités de la surface du verre ou de la flûte qui sont responsable de la nucléation hétérogène des bulles, mais des particules adsorbées sur les parois du

  2. 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

  3. Evaluation of cheetah and leopard spermatozoa developmental capability after interspecific ICSI with domestic cat oocytes.

    Science.gov (United States)

    Moro, L N; Sestelo, A J; Salamone, D F

    2014-08-01

    The ICSI procedure is potentially of great value for felids, and it has not been extensively studied in these species. The objectives of this work were to determine the best conditions for ICSI in the domestic cat (DC) to generate interspecific embryos by injecting cheetah (Ch) and leopard (Leo) spermatozoa. Firstly, DC oocytes were matured with insulin-transferrin-selenium (ITS) or without it (MM) and cultured using atmospheric (21%) or low (5%) oxygen tension after ICSI. The group ITS-5%O2 showed the highest blastocyst rate (p spermatozoa were able to generate blastocysts without artificial activation, which suggests that developmental capacity of wild felid spermatozoa can be evaluated by interspecific ICSI. This technique should be used to assist wild felid reproduction.

  4. Severe tail defects in the spermatozoa ejaculated by an English bulldog.

    Science.gov (United States)

    Rota, Ada; Manuali, Elisabetta; Caire, Simona; Appino, Simonetta

    2008-01-01

    This paper describes the case of a 2-year-old English Bulldog with severe teratozoospermia that consisted mainly of "Dag-like" defects, which is strong coiling of the tail. Although libido, semen volume and sperm concentration were normal, sperm motility was 5.0%, and 93.3% of spermatozoa exhibited morphological abnormalities affecting the tail. Transmission electron microscopic examination of the spermatozoa revealed strong folding, coiling and fracture of sperm midpieces and tails, axonemal defects and the presence of swollen and unevenly distributed mitochondria. Taking into account the dog's history and examinations and the constantly high percentage of abnormal spermatozoa over time, the defect was considered to be genetic in origin.

  5. IN VITRO EFFECT OF SILVER ENGINEERED NANOPARTICLES ON HUMAN SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    G. Terzuoli

    2012-08-01

    Full Text Available Silver (Ag NPs are among the most commercialized NPs due to their antimicrobial potential. They are highly at- tractive for possible applications in manufacture of medical device. However there is a serious lack of information concerning their impact on the human health and the environment. Moreover studies on the effects of NPs on ejaculated sperm are rather limited. For these reasons our study explored the in vitro effects of Ag NPs on human ejaculated spermatozoa. Ag NPs have been produced, characterized, and furnished by Colorobbia Industry, Sovigliana (Vinci, Florence, Italy. Aliquots of total semen were incubated at 37°C for 60 minutes (min and 120 min at the concentration of 125 μM, 250 μM, and 500 μM of engineered Ag NPs. The control was represented by specimens of semen samples treated with the same procedure without NPs. After the incubations, sperm motility was evaluated following WHO guidelines and sperm viability was eval- uated by Eosin Y test. At the end of incubation with Ag NPs the samples were processed by a Field Emission Gun-based Scanning Transmition Electron Microscope/ Energy Dispertion Spectrometry (STEM/EDS. We observed that sperm motility percentage was significantly reduced in semen samples treated with 125 μM, 250 μM and 500 μM of Ag NPs after 60 min and 120 min of incubation respect to controls (P<0.001; P<0.01, 125 μM at 60 min. Sperm viability percentage significantly decreased in a progressive manner after 125 μM (P<0.05, 250 μM (P<0.05 and 500 μM (P<0.001 Ag NPs incubation at 60 min and 120 min. We did not find any significant difference between the values assessed after 60 min of NPs incubation and those estimated after 120 min of incubation. In the control samples, the sperm motility and the sperm viability percentages significantly decreased after 120 min of incubation (P<0.001 respect to the basal values. Ag NPs were undetectable in all treated samples by STEM/EDS. These in vitro results show a

  6. Preliminary studies on the cryopreservation of spermatozoa in the fresh water fish common carp (Cyprinus carpio L.)

    Institute of Scientific and Technical Information of China (English)

    Kuppusamy Umaa Rani; Natesan Munuswamy

    2014-01-01

    Objective: To investigate the effects of various extenders containing different cryoprotectants on post-thaw viability, motility and scanning electron microscopic study of frozen spermatozoa from carp. Methods:cryopreservation has been achieved using extender and cryoprotectants like dimethylacetamide After screening a variety of cryoprotectants and extenders, a protocol for the 5%-20% at appropriate dilution ratio (1:10). For all experimental tests, the motility and viability percentage of spermatozoa were examined after storage of 5 d at 4 °C. Results: The maximum motility of 75% has been observed with 10% dimethyl sulfoxide. Scanning electron microscopic studies on normal and cryopreserved spermatozoa showed prominent head, middle piece and different segments of flagellum. There were no significant deformities noticed on the surface topography of cryopreserved spermatozoa. Conclusions:Thus the results clearly documented that cryoprotectant 10% dimethyl sulfoxide affords better cryopreservation at 4 °C for the spermatozoa of Cyprinus carpio.

  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.

    Science.gov (United States)

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

    2014-01-01

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

  9. Wigwam River Juvenile Bull Trout and Fish Habitat Monitoring Program : 2001 Data Report.

    Energy Technology Data Exchange (ETDEWEB)

    Cope, R.S.; Morris, K.J.; Bisset, J.E.

    2002-03-01

    The Wigwam River juvenile bull trout 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. The Wigwam River has been characterized as the single most important bull trout spawning stream in the Kootenay Region. This report provides a summary of results obtained during the second year (2001) of the juvenile bull trout enumeration and fish habitat assessment program. This project was commissioned in planning for fish habitat protection and forest development within the upper Wigwam River valley. 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 in the upper Wigwam River, especially as they relate to spawning and rearing habitat quality. Five permanent sampling sites were established August 2000 in the Wigwam river drainage (one site on Bighorn Creek and four sites on the mainstem Wigwam River). At each site, juvenile (0{sup +}, 1{sup +} and 2{sup +} age classes) fish densities and stream habitat conditions were measured over two stream meander wavelengths. Bull trout represented 95.1% of the catch and the mean density of juvenile bull trout was estimated to be 20.7 fish/100m{sup 2} (range 0.9 to 24.0 fish/100m{sup 2}). This compares to 17.2 fish/100m{sup 2} (+20%) for the previous year. Fry (0{sup +}) dominated the catch and this was a direct result of juvenile bull trout ecology and habitat partitioning among life history stages. Site selection was biased towards sample sites which favored high bull trout fry capture success. Comparison of fry density estimates replicated across both the preliminary survey (1997) and the current study (Cope and Morris 2001) illustrate the stable nature of these high densities. Bull trout populations have been shown to be extremely susceptible to habitat degradation and over-harvest and are

  10. Induction of gynogenesis in red crucian carp using spermatozoa of blunt snout bream

    Institute of Scientific and Technical Information of China (English)

    SUN Yuandong; LIU Yun; TAO Min; LIU Shaojun; ZHANG Chun; DUAN Wei; SHEN Jiamin; WANG Jing; ZENG Chen; LONG Yu

    2007-01-01

    Gynogenetic diploid was induced in red crucian carp (RCC) ( Carassius auratus Red Variety) eggs using UV-irradiated spermatozoa from blunt snout bream (B) (Megalobrama amblycephala ) or from mirror carp (C) (Cyprinus carpio. L). Spermatozoa were genetically inactivated by an appropriate UV dosage, and then the maternal DNA was duplicated with cold shock at 0-4 ℃. When using the spermatozoa of B, the fertilization rate, hatching rate and survival at first feeding were 52.6 ± 3.0 %, 23.6 ± 4.1 % and 15.7 ± 3.4 %, respectively, and the survival at first feeding was significantly higher than that ( 11.3 ± 2.2 % ) when using the spermatozoa of C (Cyprinus carpio. L). According to the morphological characteristics, the chromosome number and the degree of gonadal development, gynogenetic RCC could be distinguished from the control hybrids of RCC♀ × B ♂ . The individuals with red body color, 100 chromosomes and normal gonadal development were successful gynogenetic RCC, while the individuals with 124 or 148 chromosomes and delayed gonadal development were hybrids of (RCC × B). The triploid hybrids (RCC × B) (2 years old) were sterile, but the tetraploid hybrids (RCC × B) were sexually mature age of two. In the present study, compared to the spermatozoa of C, the advantages of spermatozoa of B as the activation source were that could increase the survival at first feeding of gynogenetic individuals and simplify the confirmation of gynogenetic status, which suggested that the spermof B was an effective activation source for inducing gynogenesis in crucian carp.

  11. Fusion of artificial membranes with mammalian spermatozoa. Specific involvement of the equatorial segment after acrosome reaction.

    Science.gov (United States)

    Arts, E G; Kuiken, J; Jager, S; Hoekstra, D

    1993-11-01

    The fusogenic properties of bovine and human spermatozoa membranes were investigated, using phospholipid bilayers (liposomes) as target membranes. Fusion was monitored by following lipid mixing, as revealed by an assay based on resonance-energy transfer. In addition, fusion was visualized by fluorescence microscopy, using fluorescent lipid vesicles. Cryopreserved bovine sperm fused with liposomes before induction of the acrosome reaction, fluorescence being located in essentially all spermatozoa membrane domains. Fresh bovine and human spermatozoa fused with liposomes only after the induction of the acrosome reaction, as triggered by calcium ionophore A23187 or zonae pellucidae (proteins), while the fluorescence distribution was mainly restricted to the equatorial segment (ES). However, with spermatozoa that had undergone a freeze/thawing cycle, domains other than ES also became labeled. Hence, the redistribution of the lipid probes over the entire membrane occurring during lipid mixing with cryopreserved bovine sperm is probably related to membrane perturbations caused by long-term cryopreservation. Fusion with liposomes was governed by spermatozoa factors and required the presence of acidic phospholipids like cardiolipin and phosphatidylserine in the liposomal bilayer. Incorporation of the zwitterionic lipid phosphatidylcholine in the vesicles inhibited the fusion reaction. Fusion was pH dependent. The results indicate that the ES is the primary domain of spermatozoa membranes that harbours the fusogenic capacity of sperm. Liposomes appear a valuable tool in further characterizing the properties of this domain, which has been claimed [Yanagimachi, R. (1988) in The physiology of reproduction (Knobil, E. & Neill, J., eds) pp. 135-185, Raven Press, New York] to represent the putative, initial fusion site for the oocyte.

  12. 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.

  13. Application of quantum dot nanoparticles for potential non-invasive bio-imaging of mammalian spermatozoa

    Directory of Open Access Journals (Sweden)

    Feugang Jean M

    2012-12-01

    Full Text Available Abstract Background Various obstacles are encountered by mammalian spermatozoa during their journey through the female genital tract, and only few or none will reach the site of fertilization. Currently, there are limited technical approaches for non-invasive investigation of spermatozoa migration after insemination. As the knowledge surrounding sperm behavior throughout the female genital tract still remains elusive, the recent development of self-illuminating quantum dot nanoparticles may present a potential means for real-time in vitro and in vivo monitoring of spermatozoa. Results Here, we show the ability of boar spermatozoa to harmlessly interact and incorporate bioluminescent resonance energy transfer-conjugated quantum dot (BRET-QD nanoparticles. The confocal microscope revealed in situ fluorescence of BRET-QD in the entire spermatozoon, while the ultra-structural analysis using the transmission electron microscope indicated BRET-QD localization on the sperm plasma membrane and intracellular compartment. In controlled-in vitro assays, bioluminescent imaging demonstrated that spermatozoa incubated with BRET-QD and luciferase substrate (coelenterazine emit light (photons/sec above the background, which confirmed the in situ fluorescence imaging. Most importantly, sperm motility, viability, and fertilizing potential were not affected by the BRET-QD incorporation when used at an appropriated ratio. Conclusions Our results demonstrate that pig spermatozoa can incorporate BRET-QD nanoparticles without affecting their motility and capacity to interact with the oocyte when used at an appropriated balance. We anticipate that our study will enable in-depth exploration of the male components of in vivo migration, fertilization, and embryonic development at the molecular level using this novel approach.

  14. Boar sperm quality in relation to presence of sp32-like protein in spermatozoa - preliminary studies

    Directory of Open Access Journals (Sweden)

    Orzołek Aleksandra

    2015-06-01

    Full Text Available The aim of the study was to analyse sperm proteomes of ejaculates from Polish Large White (PLW and Polish Landrace (PL boars and to identify differences which putatively influence semen quality. Spermatozoa protein profiles were analysed by electrophoretic methods followed by selected techniques to evaluate semen quality on the following factors: sperm motility, lipid peroxidation levels (MDA production, ATP content, activities of superoxide dismutase (SOD and catalase (CAT, total antioxidant status (TAS, and total oxidant status (TOS of seminal plasma. A protein with an estimated molecular weight of 30 kDa was found in spermatozoa of selected ejaculates. Mass spectrometry demonstrated that this polypeptide is most similar to proacrosin binding protein (sp32. The presence of the protein was more frequently observed in sperm extracts obtained in spring-summer period. Ejaculates containing sp32-like protein demonstrated significantly higher spermatozoa motility, lower inhibition of MDA production by seminal plasma, and higher SOD activity in seminal plasma. Boar semen which included sp32-like protein also demonstrated lower ATP levels in spermatozoa as well as higher TAS and lower TOS of seminal plasma, though the differences were not statistically significant. Ejaculates from PLW boars, with sp32-like protein present in sperm, were characterised by significantly higher sperm motility, lower ATP content in spermatozoa, and higher TAS of seminal plasma. The diminished parameters of semen quality were observed in ejaculates from PL boars that also contained the discussed protein, but the differences were not statistically significant. These findings suggest that the presence of sp32-like protein in boar spermatozoa could influence semen quality

  15. Methyl-parathion decreases sperm function and fertilization capacity after targeting spermatocytes and maturing spermatozoa.

    Science.gov (United States)

    Piña-Guzmán, B; Sánchez-Gutiérrez, M; Marchetti, F; Hernández-Ochoa, I; Solís-Heredia, M J; Quintanilla-Vega, B

    2009-07-15

    Paternal germline exposure to organophosphorous pesticides (OP) has been associated with reproductive failures and adverse effects in the offspring. Methyl-parathion (Me-Pa), a worldwide-used OP, has reproductive adverse effects and is genotoxic to sperm, possibly via oxidative damage. This study investigated the stages of spermatogenesis susceptible to be targeted by Me-Pa exposure that impact on spermatozoa function and their ability to fertilize. Male mice were exposed to Me-Pa (20 mg/kg bw, i.p.) and spermatozoa from epididymis-vas deferens were collected at 7 or 28 days post-treatment (dpt) to assess the effects on maturing spermatozoa and spermatocytes, respectively. Spermatozoa were examined for DNA damage by nick translation (NT-positive cells) and SCSA (%DFI), lipoperoxidation (LPO) by malondialdehyde production, sperm function by spontaneous- and induced-acrosome reactions (AR), mitochondrial membrane potential (MMP) by using the JC-1 fluorochrome, and fertilization ability by an in vitro assay and in vivo mating. Alterations on DNA integrity (%DFI and NT-positive cells) in spermatozoa collected at 7 and 28 dpt, and decreases in sperm quality and induced-AR were observed; reduced MMP and LPO were observed at 7 dpt only. Negative correlations between LPO and sperm alterations were found. Altered sperm functional parameters evaluated either in vitro or in vivo were associated with reduced fertilization rates at both times. These results show that Me-Pa exposure of maturing spermatozoa and spermatocytes affects many sperm functional parameters that result in a decreased fertilizing capacity. Oxidative stress seems to be a likely mechanism of the detrimental effects of Me-Pa exposure in male germ cells.

  16. Zona Pellucida Induces Activation of Phospholipase A2 During Acrosomal Exocytosis in Guinea PigSpermatozoa

    Institute of Scientific and Technical Information of China (English)

    袁玉英; 陈文颖; 石其贤; 毛丽珍; 俞淑清; 方昕; E.R.S.Roldan

    2005-01-01

    Objective To investigate whether PLA2 is involved in ZP-stimulated acrosomal exocytosis,if Ca2+ is required for activation of PLA2,and sigal transduction pathways modulating PLA2. Methods Guinea-pig spermatozoa were capacitated and labeled in low Ca2+ medium with [14C]choline chloride or [14C] arachidonic acid,and were then exposed to millimolar Ca2+ and various reagents and stimulated with ZP. Results Precapacitated spermatozoa exposed to millimolar Ca2+ and stimulated with ZP experienced increases in arachidonic acid(AA)and lysophosphatidylcholine (lysoPC) and a parallel decrease in phosphatidylcholine (PC);these chages are indicative of PLA2 activation.Simulation with ZP also led to acrosomal exocytosis in a high proportion of spermatozoa. Lipid changes and exocytosis were prevented if spermatozoa were exposed to aristolochic acid,a PLA2 inhibitor, before treatment with ZP.Stimulation with ZP in medium without added Ca2+,or in medium with millimolar Ca2+ and EGTA or La3+ resulted in no lipid changes or exocytosis. Pretreatment with pertussis toxin,a G1 protein inhibitor, before stimulation with ZP,blocked the release of AA and lysoPC and acrosomal exocytosis.Exposure of spermatozoa to the DAG kinase inhibitor R59022 before ZP stimulation led to a significant increase in the generation of lysoPC and exocytosis.Conclusion These results indicate very strongly that PLA2 plays an essential role in ZP-induced exocytosis in spermatozoa, that PLA2 activation requires Ca2+ internalization,and that it is regulated by signal transduction pathways involving G proteins and DAG.

  17. Sodium Hydrosulfide Exerts a Transitional Attenuating Effect on Spermatozoa Migration in Vitro.

    Science.gov (United States)

    Wiliński, Bogdan; Wiliński, Jerzy; Gajda, Mariusz; Jasek, Ewa; Somogyi, Eugeniusz; Głowacki, Mikołaj; Śliwa, Leopold

    2015-01-01

    Hydrogen sulfide (H2S) has been shown to have a prominent role in the regulation of reproductive system function and fertility. The aim of the study was to assess the effect of a H2S donor, sodium hydrosulfide (NaHS), on mouse sperm migration in vitro. Special plates with 4 corner wells filled with balanced salt solution (control) and various NaHS solutions in concentrations of 2.5 mmol/l, 5 mmol/l or 10 mmol/l were applied. Spermatozoa from each male mouse were injected (the experiment was repeated with ten BALB/c 5-month old males) into the central pocket, connected with the wells with ducts. After 1 h, 2 h and 4 h of incubation, the number of spermatozoa in each well was determined using Bürker's counting chambers. The number ofspermatozoa in all corner wells were summed and the number of the cells in each well was treated as the percentage share of all the migrated spermatozoa. At the time points of 1 hour and 4 hours, no differences regarding chemotactic features of spermatozoa to the utilized solutions were observed. After two hours of incubation the attenuating effect of NaHS medium and high level solutions on spermatozoa migration was observed, but not for the low concentration mixture: H(3, N = 40) = 9.65, P = 0.022; control group vs 5 mmol/l NaHS solution: 36.0% vs 18.5%, P = 0.023; control group vs 10 mmol/l NaHS solution group: 36.0% vs 17.0%, P 0.011. In conclusion, NaHS has a transitional attenuating effect on spermatozoa migration in vitro.

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

    Science.gov (United States)

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

    2015-08-01

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

  19. Changes in mean scrotal circumference in performance tested Swedish beef bulls over time

    Directory of Open Access Journals (Sweden)

    Eriksson Pernilla

    2012-12-01

    Full Text Available Abstract Background There is a growing interest in beef cattle breeding in Sweden. The majority of the females are bred naturally, which is why it is important to choose healthy fertile bulls to obtain good reproduction and profitability. The breeding soundness evaluation includes measurement of scrotal circumference (SC. Our aim was to analyze if the SC of performance tested beef bulls has changed over the years. In total, 1332 bulls (Angus, Charolais, Hereford and Simmental from 13 batches (1997-2010 were included in the study. Case book entries from final evaluation of the bulls, 11-13 months old, were compiled and analyzed. Results An overall mean SC of 34.7 cm independent of breed and age was found which is above the set minimum level. Only eleven bulls did not reach the minimum level. An increase in SC of 0.06-0.07 cm/year was shown for all breeds. In all (1997-2010, the increase of the average SC (independent of breed and age was approximately 1 cm. The positive trend was apparent for all breeds but only statistically significant for the Charolais breed. Conclusions In conclusion, our results indicated an increase in the SC over time, which improves the possibilities to obtain performance tested beef sires in Sweden with the potential for achieving better fertility results.

  20. Marine Habitat Use by Anadromous Bull Trout from the Skagit River, Washington

    Science.gov (United States)

    Hayes, Michael C.; Rubin, Steve P.; Reisenbichler, Reginald; Goetz, Fred A.; Jeanes, Eric; McBride, Aundrea

    2011-01-01

    Acoustic telemetry was used to describe fish positions and marine habitat use by tagged bull trout Salvelinus confluentus from the Skagit River, Washington. In March and April 2006, 20 fish were captured and tagged in the lower Skagit River, while 15 fish from the Swinomish Channel were tagged during May and June. Sixteen fish tagged in 2004 and 2005 were also detected during the study. Fish entered Skagit Bay from March to May and returned to the river from May to August. The saltwater residency for the 13 fish detected during the out-migration and return migration ranged from 36 to 133 d (mean ± SD, 75 ± 22 d). Most bull trout were detected less than 14 km (8.5 ± 4.4 km) from the Skagit River, and several bay residents used the Swinomish Channel while migrating. The bull trout detected in the bay were associated with the shoreline (distance from shore, 0.32 ± 0.27 km) and occupied shallow-water habitats (mean water column depth, Zostera sp.) vegetation classes made up more than 70% of the area used by bull trout. Our results will help managers identify specific nearshore areas that may require further protection to sustain the unique anadromous life history of bull trout.

  1. Collection of preputial material by scraping and aspiration for the diagnosis of Tritrichomonas foetus in bulls

    Directory of Open Access Journals (Sweden)

    P.C. Irons

    2002-07-01

    Full Text Available Two trials were carried out to assess the diagnostic sensitivity and practicability of preputial scraping as a method of collecting preputial material from bulls infected with Tritrichomonas foetus. In the 1st trial, preputial material was collected by simultaneous scraping and aspiration from 3 infected and 1 uninfected bull 10 times over a 5-week period. In the 2nd trial, samples from 5 infected bulls were collected by both sheath washing and scraping on 6 occasions, while 8 uninfected animals were sampled 3 times. Samples were cultured using a modified Trichomonas culture medium (Oxoid. In the first trial, 29 of 30 samples from infected bulls were found to be positive. In the second trial, 83 % of samples collected by both methods tested positive. In neither trial were any samples from the control bulls found to be positive. Scraping was found to be quick and safe, and offered advantages over preputial washing in that urine contamination was easily avoided, samples were smaller and more concentrated and contamination was reduced. It may, however, be subject to greater operator variability than sheath washing. It is concluded that preputial scraping is as effective as washing and represents a suitable alternative for the collection of material for direct examination and culture of Tritrichomonas foetus.

  2. Collection of preputial material by scraping and aspiration for the diagnosis of Tritrichomonas foetus in bulls.

    Science.gov (United States)

    Irons, P C; Henton, M M; Bertschinger, H J

    2002-06-01

    Two trials were carried out to assess the diagnostic sensitivity and practicability of preputial scraping as a method of collecting preputial material from bulls infected with Tritrichomonas foetus. In the 1st trial, preputial material was collected by simultaneous scraping and aspiration from 3 infected and 1 uninfected bull 10 times over a 5-week period. In the 2nd trial, samples from 5 infected bulls were collected by both sheath washing and scraping on 6 occasions, while 8 uninfected animals were sampled 3 times. Samples were cultured using a modified Trichomonas culture medium (Oxoid). In the first trial, 29 of 30 samples from infected bulls were found to be positive. In the second trial, 83 % of samples collected by both methods tested positive. In neither trial were any samples from the control bulls found to be positive. Scraping was found to be quick and safe, and offered advantages over preputial washing in that urine contamination was easily avoided, samples were smaller and more concentrated and contamination was reduced. It may, however, be subject to greater operator variability than sheath washing. It is concluded that preputial scraping is as effective as washing and represents a suitable alternative for the collection of material for direct examination and culture of Tritrichomonas foetus.

  3. Serum gonadotropins after gonadotropin-releasing hormone injection in bulls subjected to spacial restriction.

    Science.gov (United States)

    Gwazdauskas, F C; Bindas, E M; Anderson, G W; McGilliard, M L

    1984-12-01

    Response patterns of luteinizing hormone, follicle-stimulating hormone, and testosterone after injection of gonadotropin-releasing hormone were investigated in bulls grouped by weight (250 to 459 kg body weight) and confined five per pen in 9.2 or 6.4 m2 space per bull in two replicates. Blood samples were collected for 1 h prior to injection of 100 micrograms gonadotropin releasing hormone and 5 h after injection at 15-min intervals. Overall mean luteinizing hormone concentrations were not affected by spacial restriction or replicate. Interaction of treatment by time revealed that luteinizing hormone response curves were not similar. Restricted bulls had a higher response of luteinizing hormone to gonadotropin-releasing hormone. Follicle-stimulating hormone increased in all groups within 15 min and peaked at 219.4 ng/ml at 45 min. Both gonadotropin responses returned to preinjection concentrations by 4 h. Testosterone was affected by treatment, replicate, and time of sampling. Testosterone was higher in restricted bulls and higher in replicate 2. Mean testosterone peak following gonadotropin-releasing hormone was 3.86 ng/ml and occurred between 105 and 120 min which was approximately 90 min after the gonadotropin peaks. It appears that hormone responses to gonadotropin-releasing hormone were not depressed by spacial restriction, and additional spacial restriction of young bulls could be used commercially.

  4. Effect of different disaccharides on the integrity and fertilising ability of freeze-dried boar spermatozoa: a preliminary study.

    Science.gov (United States)

    Garcia, A; Gil, L; Malo, C; Martinez, F; Kershaw-Young, C; de Blas, I

    2014-01-01

    Freeze-drying spermatozoa is a developing technique that facilitates semen storage and transport. However, freeze-dried sperm exhibits impaired DNA integrity, which is associated with reduced fertilizing ability. Boar spermatozoa were freeze-dried in three different freeze-drying EDTA buffers with trehalose (75mM) and lactose (75mM) (EDTA-TL), (2) with sucrose (75mM) and lactose (75mM) (EDTA-SL) or just lactose (150mM) (EDTA-LL) using two freeze-drying protocols. In experiment 1 a one-step protocol was used and in experiment 2 a two-steps protocol was used. Spermatozoa were stored in1.5 mL cryo-tubes and 1.5 mL glass ampules at both 16 degree C and 25 degree C for 1 month. Successfully freeze-dried spermatozoa were stained with acridine-orange to assess chromatin stability. Freeze-drying was most successful when the 2-step protocol was used (experiment 2). Chromatin stability was greater in samples stored in glass ampules compared to cryo tubes. Chromatin stability was also greater in samples freeze-dried in EDTA-LL compared to EDTA-SL or EDTA-TL buffers. Spermatozoa freeze-dried in EDTA-LL and stored for 14 and 28 days at either 16 degree C or 25 degree C were utilized for ICSI. Two pronuclear formation wasgreatest using spermatozoa stored at 25 degree C (69.23%) and for 28 days (50%). Although 16 degree C spermatozoa samples had better stable chromatin, 25 degree C spermatozoa samples offered better two pronuclear formation results. In conclusion, boar spermatozoa freeze-dried using media containing disaccharides exhibit high chromatin stability and are able to fertilise oocytes following ICSI. Disaccharides may therefore advance the development of freeze-drying techniques for spermatozoa enabling ease of sperm storage and transportation.

  5. 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...... 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...

  6. Mosaic male honey bees produced by queens inseminated with frozen spermatozoa.

    Science.gov (United States)

    Harbo, J R

    1980-01-01

    Mosaic male honey bees were found as the progeny of queens that had been inseminated with spermatozoa stored in liquid nitrogen. The origins of these mosaics and the genotype of their gametes were determined by using mutant markers. The mosaics probably developed from an egg pronucleus and a sperm pronucleus that did not unite after the latter had entered the egg. Instead, both pronuclei produced haploid tissue independently. The three mosaics that were mated to queens all had mosaic testes. Therefore, these were situations in which a male honey bee produced two types of spermatozoa.

  7. Quantitative evaluation of p53 as a new indicator of DNA damage in human spermatozoa

    Directory of Open Access Journals (Sweden)

    Salvatore Raimondo

    2014-01-01

    The aim of this study was to assess if a p53 ELISA assay could be a new indicator of DNA damage in human spermatozoa. Materials and Methods: 103 human semen samples were evaluated using both Acridine Orange test and p53 ELISA and results were compared. Results: A clear correlation between the values measured by two methods was obtained. Conclusions: If this hypothesis will be confirmed by further studies, the p53 ELISA assay could become a new and more precise indicator of DNA damage in human spermatozoa.

  8. A radiomicroassay for cytotoxic antibody to human spermatozoa. Quantification by tritiated actinomycin d.

    Science.gov (United States)

    Sung, J S; Shizuya, H; Black, D D; Mumford, D M

    1977-03-01

    A radiomicroassay for titration of spermocytotoxic antibody is described. The assay used [3H]AACTINOMYCIN D ([3H]Act D) to label damaged spermatozoa in a fashion analogous to penetration by vital dye. Optimal conditions for and some kinetics of the assay are presented. The assay is sensitive, reliable, simple to perform and uses only small amounts of serum and spermatozoa. Applied to sperm antibody positive human postvasectomy sera, the assay compared favourably in sensitivity eith vital dye microscopic observations and with parallel titration by the Isojima's immobilization tests.

  9. Expression of NDUFA13 in asthenozoospermia and possible pathogenesis.

    Science.gov (United States)

    Yang, Yang; Cheng, Laiyang; Wang, Ying; Han, Yilong; Liu, Jin; Deng, Xiaohui; Chao, Lan

    2017-01-01

    Asthenozoospermia is a common cause of male infertility, which is characterized by reduced forward motility of spermatozoa. The cause and pathogenesis of asthenozoospermia are not fully understood. The purpose of this study was to investigate the expression of nicotinamide adenine dinucleotide (NADH) dehydrogenase (ubiquinone) 1 alpha subcomplex, 13 (NDUFA13) in the spermatozoa of men with asthenozoospermia and its possible pathogenesis. Protein content of NDUFA13 in spermatozoa was measured by Western blot analysis. The results showed that NDUFA13 expression in spermatozoa was significantly lower in men with asthenozoospermic than in men with normozoospermia (P asthenozoospermia. NDUFA13-specific small interfering RNA was used in the mouse spermatocyte GC2-spd cell line to down-regulate the expression of NDUFA13. The knockdown of NDUFA13 in the GC2-spd cells caused a collapse of mitochondrial membrane potential, an increase in ROS level and more apoptotic cells. Our study showed that NDUFA13 deficiency may be associated with asthenozoospermia through the disturbance of spermatozoa mitochondrial membrane potential and by increasing apoptosis and intracellular ROS.

  10. SEASONAL INFLUENCE ON SEMINAL PATTERN AND FREEZABILITY OF PANTANEIRO BULL SEMEN

    Directory of Open Access Journals (Sweden)

    Nathalia Hack Moreira

    2016-01-01

    Full Text Available The purpose of this paper was to investigate the effect of the season of the year on testicular morphology, fresh and frozen/thawed semen quality from Pantaneiro bulls bred in Brasilia, DF. Six bulls were submitted, once a month for one year, to evaluation of testicular measurements, semen collection using an artificial vagina and cryopreserved with medium containing Tris-based, egg  yolk and glycerol. The testicular length and volume were greater (P 0.05. In conclusion, Pantaneiro bulls, bred in the Midwest region of Brazil, can be used in natural mating programs in all seasons. Aiming to improve the Brazilian Animal Germplasm Bank with a higher number of samples collected, we suggest that the freezing of the semen is done between June and August, although there will be no losses of semen quality at any time. Keywords: bovine; conservation; cryopreservation; reproduction; seasonality.

  11. Polarization-independent split bull's eye antennas for infrared nano-photodetectors.

    Science.gov (United States)

    Yang, Meng; Ren, Fang-Fang; Pu, Lin; Xiao, Long; Sheng, Yun; Wang, Junzhuan; Lu, Hai; Zheng, Youdou; Shi, Yi

    2016-12-19

    Split bull's eye (SBE) antennas exhibit much larger extraordinary optical transmission and strong polarization dependence rather than bull's eye (BE) antennas in the infrared range due to the introduced sub-wavelength slit. Here, we demonstrate a dual-split bull's eye (DSBE) antenna, which consists of two sub-wavelength slits crossing through the center of the BE antenna with an intersection angle θ. The polarization dependence in transmission can be flexibly tailored by adjusting the intersection angle, following a cos(2) (Φ + θ/2) angular dependence on polarization angle Φ. When θ = 90°, the DSBE antenna yields high and polarization-independent transmission enhancement over the entire infrared spectrum. It presents highly promising applications for polarization-insensitive photodetectors and other optoelectronic devices.

  12. Evaluate Bull Trout Movements in the Tucannon and Lower Snake Rivers, 2002-2003 Annual Report.

    Energy Technology Data Exchange (ETDEWEB)

    Faler, Michael P. (US Fish and Wildlife Service, Idaho Fishery Resource Office, Ahsahka, ID); Mendel, Glen W.; Fulton, Carl (Washington Department of Fish and Wildlife, Fish Management Division, Dayton, WA)

    2004-04-01

    We collected 279 adult bull trout (Salvelinus confluentus) in the Tucannon River during the Spring and Fall of 2003. Passive Integrated Transponder (PIT) tags were inserted in 191 of them, and we detected existing PIT tags in an additional 31bull trout. Thirty five of these were also surgically implanted with radio-tags, and we monitored the movements of these fish throughout the year. Fourteen radio-tags were recovered shortly after tagging, and as a result, 21 remained in the river through December 31, 2003. Four bull trout that were radio-tagged in spring 2002 were known to survive and carry their tags through the spring and/or summer of 2003. One of these fish spent the winter near river mile (RM) 13.0; the other 3 over-wintered in the vicinity of the Tucannon Hatchery between RM 34 and 36. Twenty-one radio tags from bull trout tagged in 2002 were recovered during the spring and summer, 2003. These tags became stationary the winter of 2002/2003, and were recovered between RM 11 and 55. We were unable to recover the remaining 15 tags from 2002. During the month of July, radio-tagged bull trout exhibited a general upstream movement into the upper reaches of the Tucannon subbasin. We observed some downstream movements of radio-tagged bull trout in mid to late September and throughout October. By late November and early December, radio tagged bull trout were relatively stationary, and were distributed from the headwaters downstream to river mile 6.4, near Lower Monumental Pool. As in 2002, we did not conduct work associated with objectives 2, 3, or 4 of this study, because we were unable to monitor migratory movement of radio-tagged bull trout into the Federal hydropower system on the mainstem Snake River. Transmission tests of submerged ATS model F1830 radio-tags in Lower Granite Pool showed that audible detection and individual tag identification was possible at depths of 20 and 30 ft. Tests were conducted using an ATS R-4000 Receiver equipped with an &apos

  13. Production and quality of beef from young bulls fed diets supplemented with peanut cake.

    Science.gov (United States)

    Correia, B R; Carvalho, G G P; Oliveira, R L; Pires, A J V; Ribeiro, O L; Silva, R R; Leão, A G; Simionato, J I; Carvalho, B M A

    2016-08-01

    Peanut cake is a biodiesel byproduct that has been tested as an alternative feed additive for use in cattle production. This study aimed to assess the importance of dietary peanut cake inclusion for young bull growth rate, beef production, and beef quality. In total, 32 Nellore young bulls individually housed in stalls with a mean initial body weight of 390±43.5kg were distributed in a completely randomized design for the experiment. The animals were fed Tifton 85 hay and one of four concentrate mixtures with 0, 33, 66 or 100% peanut cake instead of soybean meal. There was a linear reduction (Pcake at levels up to 100% in the diet of feedlot-finished young bulls promotes a beneficial increase in the levels of PUFAs and the following nutraceutical compounds: conjugated linoleic acid (CLA) and Ω3 and Ω6 fatty acids.

  14. THE SPECTACLE OF REDEMPTION: GUILT AND VIOLENCE IN MARTIN SCORSESE’S RAGING BULL

    Directory of Open Access Journals (Sweden)

    Arturo Serrano

    2015-06-01

    Full Text Available Of all the characters that undertake a search for redemption in Martin Scorsese’s films, perhaps it is the story of Jake La Motta in Raging Bull that for many reasons presents the greatest challenge to understanding redemption’s role in the narratives of his films. Is Jake La Motta a redeemed character at the end of Raging Bull? I argue that Scorsese uses Raging Bull to criticize a ritualistic view of redemption by portraying the beginning of Jake’s search as a futile attempt to submit himself to a public spectacle of ritual violence in the boxing ring while visually relating this to the Catholic sacraments and the crucifixion. It will only be later—in the loneliness of a jail cell, estranged from his family and without having to have had gone through a rite—that Jake achieves the self-awareness redemption requires.

  15. Use of cover habitat by bull trout Salvelinus confluentus and lake trout Salvelinus namaycush in a laboratory environment

    Science.gov (United States)

    Meeuwig, Michael H.; Guy, Christopher S.; Fredenberg, Wade A.

    2011-01-01

    Lacustrine-adfluvial bull trout, Salvelinus confluentus, migrate from spawning and rearing streams to lacustrine environments as early as age 0. Within lacustrine environments, cover habitat pro- vides refuge from potential predators and is a resource that is competed for if limiting. Competitive inter- actions between bull trout and other species could result in bull trout being displaced from cover habitat, and bull trout may lack evolutionary adaptations to compete with introduced species, such as lake trout, Salvelinus namaycush. A laboratory experiment was performed to examine habitat use and interactions for cover by juvenile (i.e., bull trout and lake trout. Differences were observed between bull trout and lake trout in the proportion of time using cover (F1,22.6=20.08, Pbull trout using cover and bottom habitats more than lake trout. Habitat selection ratios indicated that bull trout avoided water column habitat in the presence of lake trout and that lake trout avoided bottom habitat. Intraspecific and interspecific agonistic interactions were infrequent, but approximately 10 times greater for intraspecific inter- actions between lake trout. Results from this study provide little evidence that juvenile bull trout and lake trout compete for cover, and that species-specific differences in habitat use and selection likely result in habitat partitioning between these species.

  16. Effects of slaughter weight and carcass electrical stimulation on the quality and palatability of beef from young purebred bulls.

    Science.gov (United States)

    Solomon, M B; West, R L; Hentges, J F

    1986-12-01

    Seventy-eight purebred Angus and Brahman bulls (10 to 18 mo at slaughter) were used to determine the effects of slaughter weight (60, 80, 90 or 100% of the average mature-cow weight for the respective breed) and carcass electrical stimulation (500 V, 20 2-s impulses on the right side) on carcass and meat characteristics. Angus bulls grazed summer forage (millet) after weaning, while Brahman bulls were fed to stimulate gains achieved on forage by Angus bulls. Bulls were then placed in the feedlot for finishing to their designated slaughter weight (293, 381, 412 and 463 kg for Angus and 316, 420, 463 and 516 kg for Brahman). Coarse dark band formation and lean color problems normally associated with bullock carcasses were either eliminated or reduced by stimulation. Increasing slaughter weight from 60 to 90% was associated with an increase in panel tenderness scores for loin steaks. However, increasing slaughter weight from 90 to 100% little change was detected. On the other hand, shear values for loin steaks decreased with increasing slaughter weight. Stimulation increased the tenderness of loin steaks as determined by both panel scores and shear values, and of bottom round steaks (shear-force values). The data from this study suggest that meat from the bulls slaughtered at lighter weights was generally tough, but was improved when bulls were fed to heavier, yet acceptable weights. This research encourages implementation of electrical stimulation on carcasses from young bulls to improve quality-indicating factors.

  17. 78 FR 20133 - Notice of Intent To Prepare an Environmental Impact Statement for the Proposed Bull Mountain Unit...

    Science.gov (United States)

    2013-04-03

    ... Environmental Impact Statement (EIS) to analyze a Master Development Plan (MDP) that proposes to drill up to 150... preparing an Environmental Assessment (EA) for the 150-well Bull Mountain Unit MDP from September 21 to... April 23, 2012. ADDRESSES: You may submit comments related to the proposed Bull Mountain Unit MDP by...

  18. Histopathological and histomorphometric testicular characteristics associated to reproductive condition in Bos indicus (Nellore bulls

    Directory of Open Access Journals (Sweden)

    Kethleen Mesquita da Silva

    2015-07-01

    Full Text Available The aim of this study was to evaluate morphological, morphometric and functional aspects of spermatogenesis based on reproductive conditions of Nellore bulls. The study used 25 bulls Bos indicus (Nellore, which were classified as satisfactory (n=10 and unsatisfactory (n=15 for reproduction. After orchiectomy, fragments of the right testis of each animal were processed routinely and stained with hematoxylin and eosin (HE and periodic acid-Schiff (PAS. The slides were analyzed in bright field microscopy at different magnifications. The tubular segments were classified into four levels (normal, mild and moderate degeneration, severe degeneration, and tubular hypoplasia based on the organization and architecture of the tubular tissue. The average height of the seminiferous epithelium, the average thickness of propria tunic and the average tubular diameter were obtained and the proportion of seminiferous tubules, interstitial tissue, lymphatic vessel and blood vessel of each bull testicle were recorded. Furthermore, the frequency of the three classifications of the spermatogenic cycle A, B and C (A = group of stages I, II, and III which comprise the initial phases of the cycle and mitotic proliferation, B = group of stages IV and V, which comprise intermediate phases and intensive mitosis, C = group stages VI, VII and VIII which comprise the final phases and post-meiosis was compared. The results showed that the degenerative changes in germ cells are associated with histomorphometric variations, including those in the density of the structures that comprise the testicular parenchyma, and are more representative in bulls with poor semen quality. In addition, the unsatisfactory bulls for reproduction showed reduced meiotic potential compared to that of satisfactory reproductive bulls.

  19. Incorporating movement patterns to improve survival estimates for juvenile bull trout

    Science.gov (United States)

    Bowerman, Tracy; Budy, Phaedra

    2012-01-01

    Populations of many fish species are sensitive to changes in vital rates during early life stages, but our understanding of the factors affecting growth, survival, and movement patterns is often extremely limited for juvenile fish. These critical information gaps are particularly evident for bull trout Salvelinus confluentus, a threatened Pacific Northwest char. We combined several active and passive mark–recapture and resight techniques to assess migration rates and estimate survival for juvenile bull trout (70–170 mm total length). We evaluated the relative performance of multiple survival estimation techniques by comparing results from a common Cormack–Jolly–Seber (CJS) model, the less widely used Barker model, and a simple return rate (an index of survival). Juvenile bull trout of all sizes emigrated from their natal habitat throughout the year, and thereafter migrated up to 50 km downstream. With the CJS model, high emigration rates led to an extreme underestimate of apparent survival, a combined estimate of site fidelity and survival. In contrast, the Barker model, which allows survival and emigration to be modeled as separate parameters, produced estimates of survival that were much less biased than the return rate. Estimates of age-class-specific annual survival from the Barker model based on all available data were 0.218±0.028 (estimate±SE) for age-1 bull trout and 0.231±0.065 for age-2 bull trout. This research demonstrates the importance of incorporating movement patterns into survival analyses, and we provide one of the first field-based estimates of juvenile bull trout annual survival in relatively pristine rearing conditions. These estimates can provide a baseline for comparison with future studies in more impacted systems and will help managers develop reliable stage-structured population models to evaluate future recovery strategies.

  20. Effect of preputial washing on bacterial load and preservability of semen in Murrah buffalo bulls

    Directory of Open Access Journals (Sweden)

    G. S. Meena

    2015-06-01

    Full Text Available Aim: To study the effect of preputial washing on bacterial load, preservability and semen quality in Murrah buffalo bulls Materials and Methods: A total of 36 collections of three Murrah buffalo bulls maintained at Artificial Breeding Research Centre, ICAR-National Dairy Research Institute, Karnal, were collected at weekly intervals from each bull without preputial washing and latter ejaculates from same bull with preputial washing by infusing normal saline (0.85%, KMnO4 (0.02% and savlon (2.0% to first, second and third bull, respectively. The microbial load and semen quality were evaluated during different hours of storage at refrigerated temperature (0, 24 and 48 h and after thrawing of cryopreserved (at −196°C semen. Results: The results of preservation of semen at refrigerated temperature showed that bacterial load was markedly lower in ejaculates of bulls subjected to preputial washing. Semen preserved at refrigerator temperature and cryopreserved, the effect of washing solution was significant for individual motility (IM, non-eosiniphilic count, hypo-osmotic swelling reactivity (HOST, total plate count (TPC and ac