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Sample records for boar sperm membranes

  1. Detection of boar sperm plasma membrane protein using Rhodamine 640; implications for cryobiology and physiology

    Science.gov (United States)

    Rhodamine 640 (R640) was used to detect changes in boar sperm plasma membrane protein (PMP) during cryopreservation; a poorly understood phenomenon. The protocol was adapted for boar sperm so that semen samples (n = 17) could be analyzed for PMP (R640 positive) and plasma membrane integrity (PMI; Y...

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

    OpenAIRE

    Laura Parlapan

    2015-01-01

    Preliminary studies have shown that sperm membrane from swine shows high sensitivity to cryopreservation process, causing a dramatic reduction in sperm quality. This has been attributed to the production of reactive oxygen species, that cause lipid peroxidation in sperm membranes. The aim of the present study was to minimize the oxidative attack by adding different concentration of alpha-lipoic acid into the sperm liquid storage at 17ºC for 7 days. Freshly ejaculated boar semen was diluted wi...

  3. Seminal plasma arising from the whole boar sperm-rich fraction increases the stability of sperm membrane after thawing.

    Science.gov (United States)

    Torres, M A; Ravagnani, G M; Leal, D F; Martins, S M M K; Muro, B B D; Meirelles, F V; Papa, F O; Dell'aqua, J A; Alvarenga, M A; Moretti, A S; De Andrade, A F C

    2016-05-01

    Boar spermatozoa arising from the sperm-rich ejaculate fraction are reported to have a more stable plasma membrane and are more resistant to cold shock and premature acrosome reaction than spermatozoa from the whole ejaculate. Furthermore, seminal plasma (SP) can increase the cryotolerance of boar spermatozoa, and in other domestic species, it has the ability to reverse cryopreservation damage. This study aimed to evaluate the effects of boar SP arising from the whole sperm-rich ejaculate fraction (SP-SRF) on the integrity, stability, and peroxidation of sperm membranes after thawing. Each ejaculate ( = 24) was divided among 4 treatments: control (CT), centrifuged and suspended in autologous SP-SRF (CS), centrifuged with withdrawn SP-SRF (CW), and post-thawed SP arising from the whole sperm-rich fraction addition to CW (CWSP). After thawing, all treatments were incubated for 5, 60, and 120 min and were analyzed for membrane integrity, fluidity, and peroxidation by flow cytometer. The absence of SP-SRF increased the lipid disorder ( 0.05) or membrane integrity ( > 0.05). However, the increase in lipid disorder by withdrawal of SP-SRF was reversed by SP-SRF addition ( 0.05) and lipid peroxidation ( > 0.05) were unchanged. In conclusion, despite the centrifugation effects, the addition of SP arising from the whole sperm-rich fraction to post-thawed boar semen decreased sperm lipid disorder without an influence of the sperm membrane integrity and peroxidation. PMID:27285688

  4. Detection of cooling-induced membrane changes in the response of boar sperm to capacitating conditions.

    Science.gov (United States)

    Petrunkina, Anna M; Volker, Gabriele; Weitze, Karl-Fritz; Beyerbach, Martin; Töpfer-Petersen, Edda; Waberski, Dagmar

    2005-05-01

    There is a need for methods of rapid and sensitive sperm function assessment. As spermatozoa are not able to fertilize an oocyte before having undergone a series of complex physiological changes collectively called capacitation, it is logical to assess sperm function under fertilizing conditions in vitro. In this study, the responsiveness of sperm to capacitating conditions in vitro was monitored by changes in sperm response to ionophore and by changes in the amount of intracellular calcium ions in stored boar semen. Boar semen was diluted at 32 and 20 degrees C and stored for 24 and 72 h at 16 and 10 degrees C. Ionophore-induced changes and increased intracellular calcium ion content in boar spermatozoa were recorded by flow cytometry and found to progress as a function of time during incubation under capacitating conditions. All responsiveness parameters (increases in proportions of membrane-defective spermatozoa, acrosome-reacted spermatozoa, and cells with high intracellular calcium levels) were shown to be sensitive to subtle physiological changes occurring at low storage temperatures. The initial levels of sperm with a high calcium content were higher in semen stored at 10 degrees C, but the accumulation of internal calcium was lower than in semen stored at 16 degrees C. The loss of membrane integrity and increase in the proportion of acrosome-reacted cells were higher in semen stored at 10 degrees C. Dilution at 20 degrees C had no negative effect on membrane integrity or responsiveness to capacitating conditions. There was no significant difference between semen stored for 24 and 72 h in terms of membrane integrity, acrosome reaction, and intracellular calcium after capacitation treatment. However, dynamics of cell death and acrosome reaction in response to capacitating conditions were somewhat accelerated after 72 h storage, especially in semen stored at 10 degrees C. It can be concluded that the simultaneous use of the sperm membrane responsiveness and

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

    Directory of Open Access Journals (Sweden)

    Laura Parlapan

    2015-05-01

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

  6. Effect of hyaluronan supplementation on boar sperm motility and membrane lipid architecture status after cryopreservation.

    Science.gov (United States)

    Peña, F J; Johannisson, A; Wallgren, M; Rodriguez-Martinez, H

    2004-01-01

    We investigated the effect of supplementing extended boar semen with different amounts of hyaluronan (HA) prior to freezing on post-thaw sperm characteristics. Using a split sample design, the effect of HA at a final concentration of 500 or 1000 microg/ml semen on post-thaw motility parameters, and membrane lipid architecture status assessed by merocyanine-540/YOPRO-1 and flow cytometry were evaluated. HA-supplementation improved motility parameters (P < 0.05 to P < 0.001) and decreased the percentage of hyperactivated spermatozoa (P < 0.05). HA-supplemented samples had more spermatozoa showing high lipid membrane stability as assessed with merocyanine-540. In conclusion, HA appeared to preserve post-thaw spermatozoa viability in vitro and maintained membrane stability after cryopreservation. PMID:14643862

  7. Factors influencing boar sperm cryosurvival.

    Science.gov (United States)

    Roca, J; Hernández, M; Carvajal, G; Vázquez, J M; Martínez, E A

    2006-10-01

    Optimal sperm cryopreservation is a prerequisite for the sustainable commercial application of frozen-thawed boar semen for AI. Three experiments were performed to identify factors influencing variability of postthaw sperm survival among 464 boar ejaculates. Sperm-rich ejaculate fractions were cryopre-served using a standard freezing-thawing procedure for 0.5-mL plastic straws and computer-controlled freezing equipment. Postthaw sperm motility (assessed with a computer-assisted semen analysis system) and viability (simultaneously probed by flow cytometry analysis after triple-fluorescent stain), evaluated 30 and 150 min postthaw, were used to estimate the success of cryopreservation. In the first experiment, 168 unselected ejaculates (1 ejaculate/boar), from boars of 6 breeds with a wide age range (8 to 48 mo), were cryopreserved over a 12-mo period to evaluate the predictive value of boar (breed and age), semen collection, transport variables (season of ejaculate collection, interval between collections, and ejaculate temperature exposure), initial semen traits, and sperm quality before freezing on sperm survival after freezing-thawing. In Exp. 2, 4 ejaculates from each of 29 boars, preselected according to their initial semen traits and sperm quality before freezing, were collected and frozen over a 6-mo period to evaluate the influence of interboar and intraboar ejaculate variability in the survival of sperm after cryopreservation. In Exp. 3, 12 ejaculates preselected as for Exp. 2, from each of 15 boars with known good sperm cryosurvival, were collected and frozen over a 12-mo period to estimate the sustainability of sperm cryosurvival between ejaculates over time. Boar and semen collection and transport variables were not predictive of sperm cryosurvival among ejaculates. Initial semen traits and sperm quality variables observed before freezing explained 23.2 and 10.9%, respectively, of the variation in postthaw sperm motility and viability. However, more that

  8. Within and between breed differences in freezing tolerance and plasma membrane fatty acid composition of boar sperm.

    Science.gov (United States)

    Waterhouse, K E; Hofmo, P O; Tverdal, A; Miller, R R

    2006-05-01

    The response of sperm to cryopreservation and the fertility of frozen-thawed semen varies between species. Besides species differences in sperm physiology, structure and biochemistry, factors such as sperm transport and female reproductive tract anatomy will affect fertility of frozen-thawed semen. Therefore, studying differences in sperm cryotolerance between breeds and individuals instead of between species may reveal sources of variability in sperm cryotolerance. In the present study, the effect of cooling, re-warming and freezing and thawing on plasma membrane and acrosome integrity of sperm within and between Norwegian Landrace and Duroc breeds was studied. Furthermore, the relation between post-thaw survival rate and fatty acid composition of the sperm plasma membranes was investigated. Flow cytometry assessments of plasma membrane and acrosome integrity revealed no significant differences between breeds; however there were significant male-to-male variations within breeds in post-thaw percentages of live sperm (plasma membrane intact). The most abundant fatty acids in the plasma membranes from both breeds were palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1, n-9), docosapentaenoic acid (22:5, n-6) and docosahexaenoic acid (22:6, n-3). The ratio of sigma operator 22:5, n-6 and 22:6, n-3/ sigma operator all other membrane fatty acids was significantly related to survival rate (plasma membrane integrity) of sperm for both Norwegian Landrace (correlation coefficient (r(s)) = 0.64, P boars. In conclusion, male-to-male differences in sperm survival rate after freezing and thawing may be partly related to the amount of long-chain polyunsaturated fatty acids in the sperm plasma membranes. PMID:16672353

  9. Protective effect of hyaluronic acid on cryopreserved boar sperm.

    Science.gov (United States)

    Qian, Li; Yu, Sijiu; Zhou, Yan

    2016-06-01

    This study aimed to evaluate the effects of supplementing freezing and thawing media with hyaluronic acid (HA) on the quality parameters of frozen-thawed boar spermatozoa. Boar semen samples were collected from seven mature Yorkshire boars once a week using the gloved hand technique; these samples were frozen-thawed in the extender with added HA. Boar sperm was cryopreserved in the extender with HA added at concentrations of 0 (used as control), 4, 6, 8, 8 and 12mg/L, and their effects on the quality of frozen-thawed boar sperm were evaluated. HA addition to the extender significantly improved sperm motility, sperm membrane integrity, mitochondrial activity, acrosomal integrity, superoxide dismutase and catalase activity, but decreased sperm malondialdehyde level (p<0.05). Therefore, HA could be a promising cryoprotectant for boar sperm. PMID:26944660

  10. Effects of in vitro storage time and semen-extender on membrane quality of boar sperm assessed by flow cytometry.

    Science.gov (United States)

    Waterhouse, K E; De Angelis, P M; Haugan, T; Paulenz, H; Hofmo, P O; Farstad, W

    2004-12-01

    The Norwegian AI company Norsvin has used the short-term semen-extender BTS to extend and store boar semen since the late 1980s. Fertility results have been consistent when extended semen has been used for AI within 3 days after collection, however, from a production and economic point of view it is preferable that semen stored for up to 5 days can be used. The aim of this study was to compare membrane quality of sperm stored in BTS for 3 days with sperm stored in the long-term semen-extenders Androstar, Mulberry III and X-cell for 5 days. Using a split-sample design, plasma membrane- and acrosome-integrity were assessed flow cytometrically by use of Yo-Pro-1 and PNA-FITC, and fluidity and phospholipid asymmetry of the membrane were assessed by use of MC540 and Annexin V-FITC. Due to observed sperm fragmentation in Androstar after Day 1, the data for Androstar were excluded from the analyses. After 5 days of storage, the membrane quality of X-cell-stored sperm was not statistically different from that of sperm stored in BTS for 3 days, while membrane quality of sperm stored in Mulberry III was statistically better on Day 5 compared to BTS on Day 3. In conclusion, Mulberry III and X-cell preserve sperm quality, as well as that of BTS on Day 3, for up to 5 days after collection. PMID:15511551

  11. Encapsulation of sex sorted boar semen: sperm membrane status and oocyte penetration parameters.

    Science.gov (United States)

    Spinaci, Marcella; Chlapanidas, Theodora; Bucci, Diego; Vallorani, Claudia; Perteghella, Sara; Lucconi, Giulia; Communod, Ricardo; Vigo, Daniele; Galeati, Giovanna; Faustini, Massimo; Torre, Maria Luisa

    2013-03-01

    Although sorted semen is experimentally used for artificial, intrauterine, and intratubal insemination and in vitro fertilization, its commercial application in swine species is still far from a reality. This is because of the low sort rate and the large number of sperm required for routine artificial insemination in the pig, compared with other production animals, and the greater susceptibility of porcine spermatozoa to stress induced by the different sex sorting steps and the postsorting handling protocols. The encapsulation technology could overcome this limitation in vivo, protecting and allowing the slow release of low-dose sorted semen. The aim of this work was to evaluate the impact of the encapsulation process on viability, acrosome integrity, and on the in vitro fertilizing potential of sorted boar semen. Our results indicate that the encapsulation technique does not damage boar sorted semen; in fact, during a 72-hour storage, no differences were observed between liquid-stored sorted semen and encapsulated sorted semen in terms of plasma membrane (39.98 ± 14.38% vs. 44.32 ± 11.72%, respectively) and acrosome integrity (74.32 ± 12.17% vs. 66.07 ± 10.83%, respectively). Encapsulated sorted spermatozoa presented a lower penetration potential than nonencapsulated ones (47.02% vs. 24.57%, respectively, P 0.05) was observed in terms of total efficiency of fertilization expressed as normospermic oocytes/total oocytes (18.45% vs. 15.43% for sorted diluted and sorted encapsulated semen, respectively). The encapsulation could be an alternative method of storing of pig sex sorted spermatozoa and is potentially a promising technique in order to optimize the use of low dose of sexed spermatozoa in vivo. PMID:23261305

  12. The protective effect of a 17°C holding time on boar sperm plasma membrane fluidity after exposure to 5°C.

    Science.gov (United States)

    Casas, I; Althouse, G C

    2013-02-01

    The holding time (HT) is the period during which an ejaculate, either in a raw or diluted state, is held at 17°C before further processing for cold-storage. In boars, the HT positively influences select sperm quality parameters of semen cooled from 15 to 5°C, a range in temperature during which plasma membrane remodeling occurs. Objective insight into the effect of HT on plasma membrane organization remains unknown. Therefore, the present work sought to elucidate if HT contributes to minimizing alterations in boar sperm plasma membrane fluidity at the initial step of the cooling process in a cryopreservation practice (holding at 5°C) and in relation with select sperm quality parameters. Nineteen ejaculates from five boars were collected and processed according to different treatments: T1) Fresh diluted semen, 0h at 17°C; T2) Fresh diluted semen, 24h at 17°C (HT); T3) Sperm from T1 in a lactose-egg yolk (LEY) extender, 3h at 5°C; T4) Sperm from T2 in LEY, 3h at 5°C; T5) Sperm from T1 in LEY, 24h at 5°C; T6) Sperm from T2 in LEY, 24h at 5°C. Sperm motility was assessed using CASA, and sperm plasma membrane integrity and fluidity were evaluated by flow cytometry with dual labeling (M540/YO-PRO®-1). Results demonstrated that the lack of exposure to a HT (T5) results in reduced sample motility compared to those having a HT (T6), with sperm exposed to HT exhibiting less plasma membrane fluidity. Collectively, these results provide empirical evidence that incorporation of a HT in semen processing protects boar sperm against cold injury through maintenance of lipid architecture of the plasma membrane. PMID:23219919

  13. Novel Flow Cytometry Analyses of Boar Sperm Viability: Can the Addition of Whole Sperm-Rich Fraction Seminal Plasma to Frozen-Thawed Boar Sperm Affect It?

    Science.gov (United States)

    Torres, Mariana Andrade; Díaz, Rommy; Boguen, Rodrigo; Martins, Simone Maria Massami Kitamura; Ravagnani, Gisele Mouro; Leal, Diego Feitosa; Oliveira, Melissa de Lima; Muro, Bruno Bracco Donatelli; Parra, Beatriz Martins; Meirelles, Flávio Vieira; Papa, Frederico Ozanan; Dell'Aqua, José Antônio; Alvarenga, Marco Antônio; Moretti, Aníbal de Sant'Anna; Sepúlveda, Néstor; de Andrade, André Furugen Cesar

    2016-01-01

    Boar semen cryopreservation remains a challenge due to the extension of cold shock damage. Thus, many alternatives have emerged to improve the quality of frozen-thawed boar sperm. Although the use of seminal plasma arising from boar sperm-rich fraction (SP-SRF) has shown good efficacy; however, the majority of actual sperm evaluation techniques include a single or dual sperm parameter analysis, which overrates the real sperm viability. Within this context, this work was performed to introduce a sperm flow cytometry fourfold stain technique for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential. We then used the sperm flow cytometry fourfold stain technique to study the effect of SP-SRF on frozen-thawed boar sperm and further evaluated the effect of this treatment on sperm movement, tyrosine phosphorylation and fertility rate (FR). The sperm fourfold stain technique is accurate (R2 = 0.9356, p > 0.01) for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential (IPIAH cells). Centrifugation pre-cryopreservation was not deleterious (p > 0.05) for any analyzed variables. Addition of SP-SRF after cryopreservation was able to improve total and progressive motility (p boar semen was cryopreserved without SP-SRF; however, it was not able to decrease tyrosine phosphorylation (p > 0.05) or improve IPIAH cells (p > 0.05). FR was not (p > 0.05) statistically increased by the addition of seminal plasma, though females inseminated with frozen-thawed boar semen plus SP-SRF did perform better than those inseminated with sperm lacking seminal plasma. Thus, we conclude that sperm fourfold stain can be used to simultaneously evaluate plasma and acrosomal membrane integrity and mitochondrial membrane potential, and the addition of SP-SRF at thawed boar semen cryopreserved in absence of SP-SRF improve its total and progressive motility. PMID:27529819

  14. The solubilisation of boar sperm membranes by different detergents - a microscopic, MALDI-TOF MS, 31P NMR and PAGE study on membrane lysis, extraction efficiency, lipid and protein composition

    OpenAIRE

    Müller Karin; Braun Beate; Wibbelt Gudrun; Süß Rosmarie; Fuchs Beate; Jakop Ulrike; Schiller Jürgen

    2009-01-01

    Abstract Background Detergents are often used to isolate proteins, lipids as well as "detergent-resistant membrane domains" (DRMs) from cells. Different detergents affect different membrane structures according to their physico-chemical properties. However, the effects of different detergents on membrane lysis of boar spermatozoa and the lipid composition of DRMs prepared from the affected sperm membranes have not been investigated so far. Results Spermatozoa were treated with the selected de...

  15. Determination of intracellular reactive oxygen species and high mitochondrial membrane potential in Percoll-treated viable boar sperm using fluorescence-activated flow cytometry.

    Science.gov (United States)

    Guthrie, H D; Welch, G R

    2006-08-01

    The use of frozen semen in the swine industry is limited by problems with viability and fertility compared with liquid semen. Part of the reduction in sperm motility and fertility associated with cryopreservation may be due to oxidative damage from excessive or inappropriate formation of reactive oxygen species (ROS). Chemiluminescence measurements of ROS are not possible in live cells and are problematic because of poor specificity. An alternative approach, flow cytometry, was developed to identify viable boar sperm containing ROS utilizing the dyes hydroethidine and 2', 7'-dichlorodihydrofluorescein diacetate as oxidizable substrates and impermeant DNA dyes to exclude dead sperm. The percentage of sperm with high mitochondrial transmembrane potential was determined by flow cytometry using the mitochondrial probe 5, 5', 6, 6'-tetrachloro-1, 1', 3, 3'-tetraethylbenzimidazolylcarbocyanine iodide with propidium iodide staining to exclude nonviable cells. Sperm were incubated with and without ROS generators and free radical scavengers. Basal ROS formation was low (less than 4%) and did not differ (P = 0.26) between viable fresh and frozen-thawed boar sperm. In addition, fresh and frozen-thawed viable sperm were equally susceptible (P = 0.20) to intracellular formation of ROS produced by xanthine/xanthine oxidase (94.4 and 87.9% of sperm, respectively). Menadione increased (P boar sperm, both were quite susceptible to external sources of hydrogen peroxide. PMID:16864869

  16. Dynamics of the induced acrosome reaction in boar sperm evaluated by flow cytometry

    DEFF Research Database (Denmark)

    Birck, Anders; Labouriau, Rodrigo; Christensen, Preben

    2009-01-01

    The present study investigated the dynamics of the in vitro induced acrosome reaction (AR) in boar sperm in response to medium composition, incubation time and ionophore concentration. The AR is a prerequisite for normal sperm fertilizing capability and can be studied in vitro following induction...... information on sperm viability and acrosomal status. The ionophore induced AR was dependent on extracellular Ca2+, but could be easily induced in boar sperm without capacitation. Capacitation-associated plasma membrane phospholipid scrambling was assessed and a medium specific ability to induce these membrane...... changes was observed. Both sperm viability and the induced AR were significantly affected by sperm capacitation, incubation time and ionophore concentration. The results lead to suggestions for an optimized AR induction protocol that takes both sperm viability and the effectiveness of AR induction...

  17. Protein tyrosine phosphorylation in boar sperm

    Czech Academy of Sciences Publication Activity Database

    Pěknicová, Jana; Geussová, Gizela; Kaláb, P.

    Česká republika: XXX , 2002. s. X-X. [Symposium českých reprodukčních imunologů s mezinárodní účastí/8./. 16.05.2002-19.05.2002, Žďár nad Sázavou] R&D Projects: GA ČR GA303/00/1651; GA ČR GA204/02/1373; GA MZd NJ5851 Keywords : boar sperm * phosphorylation * VCP Subject RIV: EB - Genetics ; Molecular Biology

  18. Effect of different monosaccharides and disaccharides on boar sperm quality after cryopreservation.

    Science.gov (United States)

    Gómez-Fernández, José; Gómez-Izquierdo, Emilio; Tomás, Cristina; Mocé, Eva; de Mercado, Eduardo

    2012-07-01

    The aim of the present study was to evaluate the cryoprotectant effect of different non-permeating sugars for boar sperm. Pooled semen from three boars was used for the experiments. In the first experiment, the sperm quality of boar sperm cryopreserved with an egg-yolk based extender supplemented with different monosaccharides (glucose, galactose or fructose) was compared to a control cryopreserved in lactose-egg yolk extender. In the second experiment, the effect of five disaccharides (lactose, sucrose, lactulose, trehalose or melibiose) on boar sperm cryosurvival was studied. Several sperm quality parameters were assessed by flow cytometry in samples incubated for 30 and 150 min at 37°C after thawing: percentages of sperm with intact plasma membrane (SIPM), sperm presenting high plasma membrane fluidity (HPMF), sperm with intracellular reactive oxygen substances production (IROSP) and apoptotic sperm (AS). In addition, the percentages of total motile (TMS) and progressively motile sperm (PMS) were assessed at the same incubation times with a computer-assisted sperm analysis system. Freezing extenders supplemented with each of the monosaccharide presented smaller cryoprotective effect than the control extender supplemented with lactose (P<0.05). However, from the three monosaccharides tested, glucose provided the best sperm quality after freezing-thawing. With respect to the disaccharides studied, samples frozen with the extender supplemented with lactulose exhibited in general the lowest sperm quality, except for the percentage of capacitated sperm, which was highest (P<0.05) in the samples cryopreserved with the trehalose extender. Our results suggest that disaccharides have higher cryoprotective effect than monosaccharides, although the monosaccharide composition of the disaccharides is also important, since the best results were obtained with those disaccharides presenting glucose in their composition. PMID:22771077

  19. Seminal plasma applied post-thawing affects boar sperm physiology: a flow cytometry study.

    Science.gov (United States)

    Fernández-Gago, Rocío; Domínguez, Juan Carlos; Martínez-Pastor, Felipe

    2013-09-01

    Cryopreservation induces extensive biophysical and biochemical changes in the sperm. In the present study, we used flow cytometry to assess the capacitation-like status of frozen-thawed boar spermatozoa and its relationship with intracellular calcium, assessment of membrane fluidity, modification of thiol groups in plasma membrane proteins, reactive oxygen species (ROS) levels, viability, acrosomal status, and mitochondrial activity. This experiment was performed to verify the effect of adding seminal plasma on post-thaw sperm functions. To determine these effects after cryopreservation, frozen-thawed semen from seven boars was examined after supplementation with different concentrations of pooled seminal plasma (0%, 10%, and 50%) at various times of incubation from 0 to 4 hours. Incubation caused a decrease in membrane integrity and an increase in acrosomal damage, with small changes in other parameters (P > 0.05). Although 10% seminal plasma showed few differences with 0% (ROS increase at 4 hours, P boar spermatozoa, possibly through membrane changes and ROS increase. Although some effects were detrimental, the stimulatory effect of 50% seminal plasma could favor the performance of post-thawed boar semen, as showed in the field (García JC, Domínguez JC, Peña FJ, Alegre B, Gonzalez R, Castro MJ, Habing GG, Kirkwood RN. Thawing boar semen in the presence of seminal plasma: effects on sperm quality and fertility. Anim Reprod Sci 2010;119:160-5). PMID:23756043

  20. Deep freezing of concentrated boar semen for intra-uterine insemination: effects on sperm viability.

    Science.gov (United States)

    Saravia, Fernando; Wallgren, Margareta; Nagy, Szabolcs; Johannisson, Anders; Rodríguez-Martínez, Heriberto

    2005-03-15

    The use of deep-frozen boar semen for artificial insemination (AI) is constrained by the need for high sperm numbers per dose, yielding few doses per ejaculate. With the advancement of new, intra-uterine insemination strategies, there is an opportunity for freezing small volumes containing high sperm numbers, provided the spermatozoa properly sustain cryopreservation. The present study aimed to concentrate (2 x 10(9) spz/mL) and freeze boar spermatozoa packed in a 0.5 mL volume plastic medium straw (MS) or a multiple FlatPack (MFP) (four 0.7 mL volume segments of a single FlatPack [SFP]) intended as AI doses for intra-uterine AI. A single freezing protocol was used, with a conventional FlatPack (SFP, 5 x 10(9) spz/5 mL volume) as control. Sperm viability post-thaw was monitored as sperm motility (measured by computer-assisted sperm analysis, CASA), as plasma membrane integrity (PMI, assessed either by SYBR-14/PI, combined with flow cytometry, or a rapid hypo-osmotic swelling test [sHOST]). Sperm motility did not differ statistically (NS) between test-packages and control, neither in terms of overall sperm motility (range of means: 37-46%) nor sperm velocity. The percentages of linearly motile spermatozoa were, however, significantly higher in controls (SFP) than in the test packages. Spermatozoa frozen in the SFP (control) and MFP depicted the highest PMI (54 and 49%, respectively) compared to MS (38%, P flow cytometry. In absolute numbers, more viable spermatozoa post-thaw were present in the MFP dose than in the MS (P boar variation was present, albeit only significant for MS (sperm motility) and SFP (PMI). In conclusion, the results indicate that boar spermatozoa can be successfully frozen when concentrated in a small volume. PMID:15725440

  1. Microscopic analysis of MTT stained boar sperm cells

    Directory of Open Access Journals (Sweden)

    B.M. van den Berg

    2015-06-01

    Full Text Available The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few min in a diffuse staining of the midpiece of the sperm cells. Upon further incubation the staining of the midpiece became more intense, and gradually the formation of packed formazan granules became more visible. At the same time, a small formazan stained granule appeared medially on the sperm head, which increased in size during further incubation. After incubation for about 1 h the midpiece granules were intensely stained and more clearly distinct as granules, while aggregation of sperm cells occurred. Around 90% of the sperm cells showed these staining events. At the end of the staining the formazan granules have disappeared from both the sperm cells and medium, whereas formazan crystals appeared as thin crystal threads, that became heavily aggregated in the incubation medium. It was concluded that formazan is taken up by lipid droplets in the cytoplasm. Further, the use of the MTT assay to test for sperm viability should be regarded as a qualitative assay, whereas its practical use at artificial insemination (AI Stations is limited.

  2. Evaluation of sperm chromatin structure in boar semen

    Directory of Open Access Journals (Sweden)

    Banaszewska Dorota

    2015-06-01

    Full Text Available This study was an attempt to evaluate sperm chromatin structure in the semen of insemination boars. Preparations of semen were stained with acridine orange, aniline blue, and chromomycin A3. Abnormal protamination occurred more frequently in young individuals whose sexual development was not yet complete, but may also be an individual trait. This possibility is important to factor into the decision regarding further exploitation of insemination boars. Thus a precise assessment of abnormalities in the protamination process would seem to be expedient as a tool supplementing morphological and molecular evaluation of semen. Disruptions in nucleoprotein structure can be treated as indicators of the biological value of sperm cells.

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

    Science.gov (United States)

    Kuster, C E; Althouse, G C

    2016-01-01

    Bacteriospermia is a documented risk to reproductive performance when using extended boar semen for artificial insemination. A substantial list of bacteria have been recovered from boar semen attributed to fecal, preputial, skin, and hair microorganisms, with these and other environmental bacteria from processing areas identified in doses prepared for artificial insemination. Gram-negative bacteria are most commonly recovered from extended doses, including both Enterobacteriaceae and environmental contaminants, such as those that inhabit water purification systems. The method of processing, distributing, and storing fresh liquid boar semen before insemination plays a role in bacterial growth dynamics and the degree to which the bacteria may damage the sperm or affect the sow. Not all bacterial isolates or contamination levels have the same impact on sperm, with multiple factors governing if and when storage longevity will be reduced through sperm-to-sperm agglutination, impaired motility, acrosome disruption, or loss of membrane viability. Suboptimal reproductive performance can occur because of reduced fertilizing capacity of the sperm or induction of a uterine environment hostile to sperm and/or embryonic survival. Effective bacterial control strategies are necessary to minimize the risk of bacteria contaminating extended semen doses, including monitoring programs designed for quick detection and intervention, should the need arise. PMID:26525397

  4. Relationship of flow cytometric sperm integrity assessments with boar fertility performance under optimized field conditions.

    Science.gov (United States)

    Broekhuijse, M L W J; Šoštarić, E; Feitsma, H; Gadella, B M

    2012-12-01

    The number of intact and functional spermatozoa in semen can be assessed with flow cytometry and is believed to relate to male fertility. The aim of this study was to examine whether currently used sperm integrity assessments with flow cytometry correlate with field fertility data obtained for boar semen. For this purpose, 20 boars were followed for a 20-wk period (with a total average production of 33 ejaculates per boar) and the obtained fertility results (farrowing rate and number of piglets born) of commercial artificial insemination doses made from these ejaculates were recorded. Fertility results were corrected for farm, sow, boar, and semen-related parameters. From the same semen samples, sperm cell integrity was assessed with respect to DNA and to membrane integrity, acrosome intactness and responsiveness, and mitochondrial potential using established flow cytometric assays. This was done on freshly produced semen and on semen stored for up to 15 d. Remarkably, none of the individual membrane integrity variables was significantly related to fertility results. In contrast, the amount of DNA damage as assessed at 7 to 10 d and at 14 to 15 d of semen storage related to farrowing rate (P = 0.0400) and total number of piglets born (P = 0.0310), respectively. Therefore, the degree of DNA damage in stored boar semen samples may be a useful factor to evaluate semen as an indicator for litter size and farrowing rate. PMID:23255815

  5. Post-thaw motility of frozen boar sperm does not predict success with in vitro fertilization

    Science.gov (United States)

    Using cryopreserved boar sperm rather than liquid semen for in vitro fertilization (IVF) allows improved IVF consistency. However, cryopreservation of boar sperm results in reduced post-thaw motility, fertilization and embryo development. Boars are often screened on an individual basis prior to use ...

  6. High resolution DNA flow cytometry of boar sperm cells in identification of boars carrying cytogenetic aberrations

    DEFF Research Database (Denmark)

    Larsen, Jacob; Christensen, Knud; Larsen, Jørgen K;

    2004-01-01

    The cytogenetic quality of boars used for breeding determines the litter outcome and thus has large economical consequences. Traditionally, quality controls based on the examination of simple karyograms are time consuming and sometimes give uncertain results. As an alternative, the use of high......-resolution DNA flow cytometry on DAPI-stained sperm cell nuclei (CV...

  7. Fluorescent analysis of boar sperm capacitation process in vitro

    Czech Academy of Sciences Publication Activity Database

    Děd, Lukáš; Dostálová, Pavla; Dorosh, Andriy; Pěknicová, Jana

    Portland : Society for the Study of Reproduction, 2011 - ( Perreault Darney, S.; Robaire, B.; Murphy, B.). s. 10-10 [44th Annual Meeting Society for the Study of Reproduction : Reproduction and the World's Future.. 31.07.2011-04.08.2011, Portland] R&D Projects: GA MŠk(CZ) 1M06011 Institutional research plan: CEZ:AV0Z50520701 Keywords : capacitation * boar sperm * fluorescent microscopy * flow cytometry * acrosomal reaction Subject RIV: EI - Biotechnology ; Bionics

  8. Specific LED-based red light photo-stimulation procedures improve overall sperm function and reproductive performance of boar ejaculates.

    Science.gov (United States)

    Yeste, Marc; Codony, Francesc; Estrada, Efrén; Lleonart, Miquel; Balasch, Sam; Peña, Alejandro; Bonet, Sergi; Rodríguez-Gil, Joan E

    2016-01-01

    The present study evaluated the effects of exposing liquid-stored boar semen to different red light LED regimens on sperm quality and reproductive performance. Of all of the tested photo-stimulation procedures, the best pattern consisted of 10 min light, 10 min rest and 10 min of further light (10-10-10 pattern). This pattern induced an intense and transient increase in the majority of motility parameters, without modifying sperm viability and acrosome integrity. While incubating non-photo-stimulated sperm at 37 °C for 90 min decreased all sperm quality parameters, this reduction was prevented when the previously-described light procedure was applied. This effect was concomitant with an increase in the percentage of sperm with high mitochondrial membrane potential. When sperm were subjected to 'in vitro' capacitation, photo-stimulation also increased the percentage of sperm with capacitation-like changes in membrane structure. On the other hand, treating commercial semen doses intended for artificial insemination with the 10-10-10 photo-stimulation pattern significantly increased farrowing rates and the number of both total and live-born piglets for parturition. Therefore, our results indicate that a precise photo-stimulation procedure is able to increase the fertilising ability of boar sperm via a mechanism that could be related to mitochondrial function. PMID:26931070

  9. Specific LED-based red light photo-stimulation procedures improve overall sperm function and reproductive performance of boar ejaculates

    Science.gov (United States)

    Yeste, Marc; Codony, Francesc; Estrada, Efrén; Lleonart, Miquel; Balasch, Sam; Peña, Alejandro; Bonet, Sergi; Rodríguez-Gil, Joan E.

    2016-01-01

    The present study evaluated the effects of exposing liquid-stored boar semen to different red light LED regimens on sperm quality and reproductive performance. Of all of the tested photo-stimulation procedures, the best pattern consisted of 10 min light, 10 min rest and 10 min of further light (10-10-10 pattern). This pattern induced an intense and transient increase in the majority of motility parameters, without modifying sperm viability and acrosome integrity. While incubating non-photo-stimulated sperm at 37 °C for 90 min decreased all sperm quality parameters, this reduction was prevented when the previously-described light procedure was applied. This effect was concomitant with an increase in the percentage of sperm with high mitochondrial membrane potential. When sperm were subjected to ‘in vitro’ capacitation, photo-stimulation also increased the percentage of sperm with capacitation-like changes in membrane structure. On the other hand, treating commercial semen doses intended for artificial insemination with the 10-10-10 photo-stimulation pattern significantly increased farrowing rates and the number of both total and live-born piglets for parturition. Therefore, our results indicate that a precise photo-stimulation procedure is able to increase the fertilising ability of boar sperm via a mechanism that could be related to mitochondrial function. PMID:26931070

  10. Current knowledge on boar sperm metabolism: Comparison with other mammalian species.

    Science.gov (United States)

    Rodríguez-Gil, Joan E; Bonet, Sergi

    2016-01-01

    A practical consequence of the specific pig reproductive cycle is that the main functional features that distinguish boar spermatozoa cannot be extrapolated to other species. This prevents an overall picture that explains mammalian sperm function from being assumed. Furthermore, the extraordinary complexity of the molecular mechanisms implied in the control and modulation of mature boar sperm functions makes it impossible to provide a complete description of these mechanisms in the limited space of this chapter. Taking this into account, this chapter centers on the description of three highly important specific aspects of boar sperm function. The first aspect is the mechanisms by which boar sperm cells uptake extracellular energy sources. The second aspect is the necessity of mammalian sperm to use other hexoses than glucose as feasible energy sources. The third aspect would be an analysis of the roles that mitochondria could play in the regulation of the overall boar sperm function. As a whole, this revision intends to be an overall picture of regulatory mechanisms involved in the maintenance of proper energy levels of boar sperm and their relationship with the control of the overall boar sperm function. PMID:26094247

  11. Effect of alpha-tocopherol supplementation during boar semen cryopreservation on sperm characteristics and expression of apoptosis related genes.

    Science.gov (United States)

    Jeong, Yeon-Ji; Kim, Mi-Kyeong; Song, Hye-Jin; Kang, Eun-Ju; Ock, Sun-A; Kumar, B Mohana; Balasubramanian, S; Rho, Gyu-Jin

    2009-04-01

    Boar semen is extremely vulnerable to cold shock and sensitive to peroxidative damage due to high content of unsaturated fatty acids in the phospholipids of the plasma membrane and the relatively low antioxidant capacity of seminal plasma. The present study evaluated the influence of alpha-tocopherol supplementation at various concentrations in the boar semen extender during cryopreservation on post-thawed sperm motility characteristics (total sperm motility, MOT; local motility, LCM; curvilinear velocity, VCL; straight linear velocity, VSL; and average path velocity, VAP), sperm qualities (viability, acrosomal integrity and apoptosis), expression of stress protein (HSP70), and the expression of pro-apoptotic (Bax and Bak) and anti-apoptotic (Bcl-2l and Bcl-xl) genes. Semen collected from 10 Duroc boars was cryopreserved in lactose-egg yolk buffer supplemented with various concentrations of alpha-tocopherol (0, 100, 200, 400, 600 and 800 microM) using the straw-freezing procedure and stored at -196 degrees C for a minimum period of one month. In frozen-thawed groups, sperm motility was significantly (Psperm. In fresh sperm, HSP70 immunoreactivity expression was observed in the equatorial region, but in frozen-thawed groups, expressions were mostly observed in the sperm head. Higher apoptosis rates were observed in 600 and 800 microM alpha-tocopherol supplemented frozen-thawed groups. In alpha-tocopherol supplemented frozen-thawed groups immediately after thawing, the expression was similar to that of fresh group. But after incubation at 37 degrees C for 3h, the expression in 200 and 800 microM alpha-tocopherol supplemented groups was higher than that of others. Expression of pro-apoptotic genes was significantly higher and anti-apoptotic genes was significantly (Psperm group. In conclusion, alpha-tocopherol, supplemented at 200 microM concentration in boar semen extender during cryopreservation had a positive effect on post-thawed sperm survivability. PMID:19141297

  12. Reduced curvilinear velocity of boar sperm on substrates with increased hydrophobicity

    OpenAIRE

    Mears, M.; Kennelly, T.M.; Geoghegan, M; Howse, J.R.; Tarmey, D.S.; Pacey, A. A.

    2014-01-01

    The curvilinear velocity (VCL) of boar spermatozoa between standard microscopy glassware decreases when the slides are coated with the hydrophobic polymer polystyrene (PS) compared with the less hydrophobic poly(methyl methacrylate) (PMMA) coating. Sperm from three boars were observed and analyzed using particle tracking software. The VCL did not differ significantly between coatings of different thickness, indicating no penetration of the sperm into the coating and that only the surface laye...

  13. Antioxidative effects of magnetized extender containing bovine serum albumin on sperm oxidative stress during long-term liquid preservation of boar semen.

    Science.gov (United States)

    Lee, Sang-Hee; Park, Choon-Keun

    2015-08-21

    Magnetized water is defined as water that has passed through a magnet and shows increased permeability into cells and electron-donating characteristics. These attributes can protect against membrane damage and remove reactive oxygen species (ROS) in mammalian cells. We explored the effects of improved magnetized semen extenders containing bovine serum albumin (BSA) as antioxidants on apoptosis in boar sperm. Ejaculated semen was diluted in magnetized extender (0G and 6000G) with or without BSA (0G + BSA and 6000G + BSA), and sperm were analyzed based on viability, acrosome reaction, and H2O2 level of live sperm using flow cytometry. Sperm were then preserved for 11 days at 18 °C. We found that viability was significantly higher in 6000G + BSA than under the other treatments (P boar sperm. PMID:26143531

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

    Science.gov (United States)

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

    2015-08-01

    This study was undertaken to examine the effect of cholesterol-loaded-cyclodextrin (CLC) on boar sperm viability and spermatozoa cryosurvival during boar semen cryopreservation, and methyl-β-cyclodextrin (MBCD) was treated for comparing with CLC. Boar semen treated with CLC and MBCD before freezing process to monitor the effect on survival and capacitation status by flow cytometry with appropriate fluorescent probes. Sperm viability was higher in 1.5mg CLC-treated sperm (76.9±1.01%, Psemen, in which CLC treatment prior to freezing and thawing increased the development of oocytes to blastocyst stage in vitro. In conclusion, CLC could protect the viability of spermatozoa from cryodamage prior to cryopreservation in boar semen. PMID:26091957

  15. The Fertility of Frozen Boar Sperm When used for Artificial Insemination.

    Science.gov (United States)

    Knox, R V

    2015-07-01

    One of the limits to practical use of frozen boar sperm involves the lowered fertility when used for artificial insemination. Years of studies have shown that 5-6 billion sperm (approximately 3 billion viable) used in single or multiple inseminations results in pregnancy rates most often between 60 and 70% and with litter sizes between nine and 10 pigs. Yet today, it is not uncommon for studies to report pregnancy rates from 70 to 85% and litter sizes with 11-12 pigs. While global statements about the incidence and reasons for higher fertility are not conclusive, incremental fertility improvements appear independently associated with use of a minimum number of viable sperm (1-2 billion), insemination timing that increases the probability that sperm will be present close to ovulation for groups of females, selection for boar sperm survival following cryopreservation, and modification of the freeze and thaw conditions using additives to protect sperm from oxidative damage. Studies show that techniques such as intrauterine and deep uterine insemination can provide an opportunity to reduce sperm numbers and that control of time of ovulation in groups of females can reduce the need for multiple inseminations and improve the chance for AI close to ovulation. However, optimal and consistent fertility with cryopreserved boar sperm may require a multifaceted approach that includes boar selection and screening, strategic use of additives during the freezing and thawing process, post-thaw evaluation of sperm and adjustments in sperm numbers for AI, assessment of female fertility and ovulation induction for single insemination. These sequenced procedures should be developed and incorporated into a quality control system for improved fertility when using minimal numbers of cryopreserved boar sperm. PMID:26174925

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

    Science.gov (United States)

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

  17. Validation of the FACSCount AF system for determination of sperm concentration in boar semen

    DEFF Research Database (Denmark)

    Hansen, C.; Christensen, P.; Stryhn, H.;

    2002-01-01

    Biosciences) was compared with microscopic counting using a Burker-Turk haemocytometer. In addition, sperm concentration was determined using the Corning 254 spectrophotometer which is used routinely by Danish artificial insemination stations for boars. The results show that the agreement between flow...... with the spectrophotometric method ( CV = 6.3%). These results indicate that the FACSCount AF System is a valuable tool for precise and accurate assessment of sperm concentration in boar semen and that use of this system may lead to production of more uniform insemination doses containing a specific number of sperm per dose....

  18. Effect of homeopathic treatment used in commercial boar semen diluent on sperm viability

    Directory of Open Access Journals (Sweden)

    Mayra Assunpção

    2012-09-01

    Full Text Available Background: It has been speculated that the homeopathic treatment of sperm cells in order to improve semen quality could be promising. However, few data is available and its use in spermatozoa requires investigation. It is well established that mitochondrial membrane potential is an important viability parameter of spermatozoa and it is intimately related to reproductive efficiency. In this manner, new technologies in order to improve the activity of sperm cells and, finally, the fecundity of swine herds are of extremely importance. Due to the lack of knowledge of homeopathic treatment effect on spermatozoa, the aim of the present study was to verify the effect of three different homeopathic treatments on viability of boar sperm cells. Methods: semen samples were obtained from two sexually mature boars (18 mo of age. The boars were cross bred, with similar genetics of Pietrain versus Duroc, BP 450 progeny from a supplier company of similar reproductive performance animals. The animals were maintained in individual stalls, study conducted in Sao Paulo - Brazil. Three homeopathic treatments: Pulsatilla 6CH, Avena 6 CH or both, compared to placebo treatment (sucrose, the homeopathic medicaments or the control were administrated as globules manipulated according Brazilian Homeopathic Pharmacology. Each globule weighted 30 mg and contained sucrose as vehicle. One dose of two globules was added per 100 mL of diluted boar semen, which were chilled for 24 or 48 hours. All samples were labeled in codes in order to allow all laboratory analysis and evaluations being performed as a blind test. Data were tested for normality of residues and homogeneity of variances using the Guided Data Analysis software. Variables and interactions were analyzed by the PROC MIXED of the SAS package (SAS Institute Ins. Cary, NC. Adjusted least squares means (LSMEANS of treatments were compared using the Tukey Test. Results: The different treatments contributed to

  19. Increasing storage time of extended boar semen reduces sperm DNA integrity.

    Science.gov (United States)

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

    2005-04-15

    There is an extensive use of artificial insemination (AI) in the pig industry. Extended liquid boar semen may be used for insemination for up to 5 days after collection. The objective of this study was to determine the changes in sperm quality, when boar semen was extended and stored at 18 degrees C for up to 72 h post-collection. The study included three ejaculates from five boars, for each of the four breeds: Duroc, Hampshire, Landrace and Danish Large White (n=60 ejaculates). The sperm chromatin structure assay (SCSA) showed an increase in DNA fragmentation index (DFI) after 72 h of incubation (Pboars, all ejaculates had a large increase in DFI after 24 h of incubation. The standard deviation of DFI (SD-DFI) differed between breeds, with the SD-DFI for Hampshire being significantly greater than for the other breeds. The SD-DFI did not change during the 72 h of storage. Sperm viability was determined using SYBR-14 and propidium iodide in combination with flow cytometry. The sperm viability did not differ between breeds (P=0.21), but a difference in viability during storage (Pboars and storage of extended boar semen at 18 degrees C for 72 h significantly decreased the integrity of sperm DNA. PMID:15823356

  20. Enhanced fertility prediction of cryopreserved boar spermatozoa using novel sperm function assessment

    Science.gov (United States)

    Cryopreserved semen is seldom used for commercial porcine artificial insemination (AI) despite many advantages that cryopreservation provides. Compared to fresh semen, the fertility of frozen-thawed boar sperm is more variable but usually less. Predicting the fertility of individual ejaculates for s...

  1. In search of epigenetic marks in testes and sperm cells of differentially fed boars.

    Directory of Open Access Journals (Sweden)

    Rémy Bruggmann

    Full Text Available In search of transmittable epigenetic marks we investigated gene expression in testes and sperm cells of differentially fed F0 boars from a three generation pig feeding experiment that showed phenotypic differences in the F2 generation. RNA samples from 8 testes of boars that received either a diet enriched in methylating micronutrients or a control diet were analyzed by microarray analysis. We found moderate differential expression between testes of differentially fed boars with a high FDR of 0.82 indicating that most of the differentially expressed genes were false positives. Nevertheless, we performed a pathway analysis and found disparate pathway maps of development_A2B receptor: action via G-protein alpha s, cell adhesion_Tight junctions and cell adhesion_Endothelial cell contacts by junctional mechanisms which show inconclusive relation to epigenetic inheritance. Four RNA samples from sperm cells of these differentially fed boars were analyzed by RNA-Seq methodology. We found no differential gene expression in sperm cells of the two groups (adjusted P-value>0.05. Nevertheless, we also explored gene expression in sperm by a pathway analysis showing that genes were enriched for the pathway maps of bacterial infections in cystic fibrosis (CF airways, glycolysis and gluconeogenesis p.3 and cell cycle_Initiation of mitosis. Again, these pathway maps are miscellaneous without an obvious relationship to epigenetic inheritance. It is concluded that the methylating micronutrients moderately if at all affects RNA expression in testes of differentially fed boars. Furthermore, gene expression in sperm cells is not significantly affected by extensive supplementation of methylating micronutrients and thus RNA molecules could not be established as the epigenetic mark in this feeding experiment.

  2. Uterine activity, sperm transport, and the role of boar stimuli around insemination in sows.

    Science.gov (United States)

    Langendijk, P; Soede, N M; Kemp, B

    2005-01-15

    This paper describes changes in spontaneous myometrial activity around estrus, factors that affect myometrial activity, and the possible role of uterine contractions in the process of (artificial) insemination, sperm transport and fertilization. Myometrial activity in the sow increases during estrus. The activity is myogenic in origin, but several factors have been shown to affect myometrial activity. Natural mating stimulates uterine contractions through several mechanisms. The presence of a boar, rather than the act of mating, induces central oxytocin release in the sow and thus increases uterine activity. Estrogens in the ejaculate of a boar can trigger prostaglandin release by the endometrium and thus increase uterine activity. Tactile stimulation of the genital tract (cervix) or tactile stimulation of the back and flanks of the sow during artificial insemination does not cause a release of oxytocin. There is hardly any evidence for the effects of these latter stimuli on uterine activity, and if they are present at all, the effects are very small. Evidence for the effects of synthetic boar odor on oxytocin release and/or uterine activity is inconsistent. The mere presence of a boar during insemination, in contrast, clearly stimulates uterine activity through the release of oxytocin. Hormonal stimulation (intrauterine) of uterine activity with estrogens, prostaglandins, or oxytocins before, during or after insemination generally improves fertilization rate, especially in situations with reduced fertility. Therefore, uterine contractions are believed to play an important role in the transport of sperm cells to the oviducts after insemination. Whether uterine contractions are absolutely necessary for sperm transport through the uterine horns, however, is not clear. Intensive stimulation of uterine contractions using hormones can also reduce the fertilization rate, probably by increasing the reflux of sperm cells during insemination. In this respect, the presence

  3. MicroRNA in sperm from Duroc, Landrace and Yorkshire boars

    Science.gov (United States)

    Kasimanickam, Vanmathy; Kastelic, John

    2016-01-01

    Sperm contain microRNAs (miRNAs), which may have roles in epigenetic control. Regarding phylogenetic relationships among various swine breeds, Yorkshire and Landrace, are considered phenotypically and genetically very similar, but distinctly different from Duroc. The objective of the present study was to compare abundance of boar sperm miRNAs in these three breeds. Overall, 252 prioritized miRNAs were investigated using real-time PCR; relative expression of miRNAs in sperm was similar in Yorkshire and Landrace boars, but significantly different compared to Duroc. Seventeen miRNAs (hsa-miR-196a-5p, hsa-miR-514a-3p, hsa-miR-938, hsa-miR-372-3p, hsa-miR-558, hsa-miR-579-3p, hsa-miR-595, hsa-miR-648, hsa-miR-524-3p, hsa-miR-512-3p, hsa-miR-429, hsa-miR-639, hsa-miR-551a, hsa-miR-624-5p, hsa-miR-585-3p, hsa-miR-508-3p and hsa-miR-626) were down-regulated (P < 0.05; fold regulation ≤−2) in Yorkshire and Landrace sperm, compared to Duroc sperm. Furthermore, three miRNAs (hsa-miR-9-5p, hsa-miR-150-5p, and hsa-miR-99a-5p) were significantly up-regulated in Yorkshire and Landrace sperm compared to Duroc sperm, However, 240 miRNAs were not significantly different (within + 2 fold) between Yorkshire and Landrace sperm. We concluded that miRNAs in sperm were not significantly different between Yorkshire and Landrace boars, but there were significant differences between those two breeds and Duroc boars. Furthermore, integrated target genes for selected down-regulated miRNAs (identified via an in-silico method) appeared to participate in spermatogenesis and sperm functions. PMID:27597569

  4. Boar sperm quality in lines of pigs selected for either ovulation rate or uterine capacity.

    Science.gov (United States)

    Freking, B A; Purdy, P H; Spiller, S F; Welsh, C S; Blackburn, H D

    2012-08-01

    Selection for 11 generations in swine for ovulation rate (OR) or uterine capacity (UC) resulted in significant changes in component traits of litter size. Our objective was to conserve the unique germplasm for the future and to characterize sperm quality as a correlated response to the selection criterion imposed compared with an unselected control line (CO). Boars representing genetic diversity available in all 3 lines were produced in 2 farrowing seasons. Season 1 was born in September 2005 and was sampled for semen characteristics in October 2006. Season 2 was born in March 2006 and was sampled for semen characteristics in February and March 2007. Each boar (n = 60) was collected twice. The sperm-rich fraction was obtained, and volume and concentration of sperm cells were measured to estimate total sperm production. Each ejaculate was extended 1:3 (vol/vol) with Androhep Plus (Minitube, Verona, WI) and was packed for shipping to the National Animal Germplasm Program laboratory for processing into frozen straws. Semen quality was measured by computer-assisted semen analysis at 3 semen processing points: fresh (FR), 24 h after extender added (E), and postthaw (PT). A mixed model ANOVA was applied to the data. Fixed effects of farrowing season, line, and 2-way interactions were fitted. The random effect of boar (n = 60) within farrowing season and line was used to test line differences. Sperm concentration was not different (P = 0.18) among the lines (0.594 × 10(9), 0.691 × 10(9), and 0.676 × 10(9) cells/mL for CO, OR, and UC lines, respectively). However, significance (P = 0.04) was detected for the volume of the sperm-rich fraction, greatest for OR (86.4 mL), intermediate for UC (75.5 mL), and least for CO (70.2 mL). Line differences were thus detected (P = 0.02) for total sperm production per ejaculate, greatest for OR (54.9 × 10(9)), intermediate for UC (48.7 × 10(9)), and least for CO (40.5 × 10(9)). A larger percentage of progressively motile sperm and

  5. Interaction of milk proteins and Binder of Sperm (BSP) proteins from boar, stallion and ram semen

    OpenAIRE

    Plante, Geneviève; Lusignan, Marie-France; Lafleur, Michel; Manjunath, Puttaswamy

    2015-01-01

    Background Mammalian semen contains a family of closely related proteins known as Binder of SPerm (BSP proteins) that are added to sperm at ejaculation. BSP proteins extract lipids from the sperm membrane thereby extensively modifying its composition. These changes can ultimately be detrimental to sperm storage. We have demonstrated that bovine BSP proteins interact with major milk proteins and proposed that this interaction could be the basis of sperm protection by milk extenders. In the pre...

  6. Antioxidative effects of magnetized extender containing bovine serum albumin on sperm oxidative stress during long-term liquid preservation of boar semen

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Sang-Hee; Park, Choon-Keun, E-mail: parkck@kangwon.ac.kr

    2015-08-21

    Magnetized water is defined as water that has passed through a magnet and shows increased permeability into cells and electron-donating characteristics. These attributes can protect against membrane damage and remove reactive oxygen species (ROS) in mammalian cells. We explored the effects of improved magnetized semen extenders containing bovine serum albumin (BSA) as antioxidants on apoptosis in boar sperm. Ejaculated semen was diluted in magnetized extender (0G and 6000G) with or without BSA (0G + BSA and 6000G + BSA), and sperm were analyzed based on viability, acrosome reaction, and H{sub 2}O{sub 2} level of live sperm using flow cytometry. Sperm were then preserved for 11 days at 18 °C. We found that viability was significantly higher in 6000G + BSA than under the other treatments (P < 0.05). The acrosome reaction was significantly lower in the 6000G + BSA group compared with the other treatments (P < 0.05). Live sperm with high intracellular H{sub 2}O{sub 2} level were significantly lower in the 6000G + BSA group than under other treatments (P < 0.05). Based on our results, magnetized extenders have antioxidative effects on the liquid preservation of boar sperm. - Highlights: • Magnetized water is water that has been passed through a magnetic field. • Magnetized extender improve viability and decrease oxidative stress of boar sperm for preservation. • Ejaculated semen diluted with magnetized extender can improve liquid preservation period.

  7. Antioxidative effects of magnetized extender containing bovine serum albumin on sperm oxidative stress during long-term liquid preservation of boar semen

    International Nuclear Information System (INIS)

    Magnetized water is defined as water that has passed through a magnet and shows increased permeability into cells and electron-donating characteristics. These attributes can protect against membrane damage and remove reactive oxygen species (ROS) in mammalian cells. We explored the effects of improved magnetized semen extenders containing bovine serum albumin (BSA) as antioxidants on apoptosis in boar sperm. Ejaculated semen was diluted in magnetized extender (0G and 6000G) with or without BSA (0G + BSA and 6000G + BSA), and sperm were analyzed based on viability, acrosome reaction, and H2O2 level of live sperm using flow cytometry. Sperm were then preserved for 11 days at 18 °C. We found that viability was significantly higher in 6000G + BSA than under the other treatments (P < 0.05). The acrosome reaction was significantly lower in the 6000G + BSA group compared with the other treatments (P < 0.05). Live sperm with high intracellular H2O2 level were significantly lower in the 6000G + BSA group than under other treatments (P < 0.05). Based on our results, magnetized extenders have antioxidative effects on the liquid preservation of boar sperm. - Highlights: • Magnetized water is water that has been passed through a magnetic field. • Magnetized extender improve viability and decrease oxidative stress of boar sperm for preservation. • Ejaculated semen diluted with magnetized extender can improve liquid preservation period

  8. Effects of straw volume and Equex-STM on boar sperm quality after cryopreservation.

    Science.gov (United States)

    Buranaamnuay, K; Tummaruk, P; Singlor, J; Rodriguez-Martinez, H; Techakumphu, M

    2009-02-01

    The present experiments were designed to study the effect of adding the detergent Equex-STM to freezing extender, and of straw volume (0.25 ml vs 0.5 ml), on boar sperm quality after cryopreservation. Three ejaculates from each of four purebred boars (three Landrace and one Yorkshire) were collected and frozen with a lactose-egg yolk extender containing glycerol with or without 1.5% Equex-STM. The extended semen was loaded into either 0.25- or 0.5-ml straws. The straws were placed in liquid nitrogen (LN(2)) vapour approximately 3 cm above the level of LN(2) for 20 min and then were plunged into LN(2). Thawing was achieved in warm water at 50 degrees C for 12 s and then was incubated in a 38 degrees C water-bath for 30 min before evaluating sperm quality. Results showed that the individual motility, viability and acrosomal normal apical ridge (NAR) were improved (p 0.05). The results of these investigations suggested that Equex-STM exerts a beneficial effect on the quality of cryopreserved boar semen and this cryopreservation protocol was favourable for a 0.5-ml straw. PMID:18484955

  9. Effects of exposure of pre-pubertal boars to di(2-ethylhexyl) phthalate on their frozen-thawed sperm viability post-puberty.

    Science.gov (United States)

    Spjuth, L; Saravia, F; Johannisson, A; Lundeheim, N; Rodríguez-Martínez, H

    2006-10-01

    Late effects of pre-pubertal oral exposure to di(2-ethylhexyl) phthalate (DEHP), a plastic softener used in, for example, polyvinyl chloride-products, on semen quality in young boars have not been clear-cut. The aim of this study was to determine whether stress imposed on spermatozoa would reveal such effects. Semen was collected from post-pubertal boars (8-9 months of age), which had been exposed to 300 mg kg(-1) body weight of DEHP per os three times a week from 3 to 7 weeks of age and from control siblings given placebo (water). The semen was cryopreserved and examined for plasma membrane integrity post-thaw using the short hypo-osmotic swelling test and flow cytometry (propidium iodide /SYBR-14). Sperm motility was assessed by computer-assisted sperm analysis. No significant difference in plasma membrane integrity could be found between the groups. The DEHP-exposed group had a significantly lower percentage of linearly motile spermatozoa at 30 min (P boars pre-pubertally exposed to low doses of DEHP, showed kinematic deviations post-thaw that could be related to DEHP exposure. PMID:16961572

  10. Variability of ejaculate volume and sperm motility depending on the age and intensity of utilization of boars

    OpenAIRE

    Savić R.; Petrović M.; Radojković D.; Radović Č.; Parunović N.; Pušić M.; Radišić R.

    2013-01-01

    The main objective of this study was to evaluate the effect of age (A) and the intensity of the boars' utilization (s) on the phenotypic variability of ejaculate volume (VOL) and sperm motility (MO). The study included 274 ejaculates of Large White boars (LW). Boars were divided into six classes according the age when the ejaculate was taken (10-13, 14-17, 18-21, 22-25, 26-29 and ≥30 months). Semen samples were analyzed during four seasons (spring, summer, ...

  11. Sperm treatment affects capacitation parameters and penetration ability of ejaculated and epididymal boar spermatozoa.

    Science.gov (United States)

    Matás, C; Sansegundo, M; Ruiz, S; García-Vázquez, F A; Gadea, J; Romar, R; Coy, P

    2010-11-01

    This work was designed to study how this ability is affected by different sperm treatments routinely used for in vitro fertilization (IVF) assay. In this study, boar sperm samples from epididymal or ejaculated origin were processed by three different methods: left unwashed (NW group), washed in Dulbecco's phosphate-buffered saline supplemented with 0.1% BSA (BSA group), and washed on a Percoll(®) gradient (PERCOLL group). After preparation of semen samples, changes in motility patterns were studied by CASA, calcium uptake by spectrofluorimetry, and ROS generation, spontaneous acrosome reaction, and lipid disorder by means of flow cytometry. Finally IVF assays were also performed with the different semen samples and penetrability results evaluated at 2 and 4 h post insemination (hpi). Independently of the sperm treatment, epididymal spermatozoa showed higher values of progressive motility, percentage of live cells with low lipid disorder, and penetration ability at 4 hpi than the corresponding ejaculated spermatozoa. Ejaculated spermatozoa showed higher levels of calcium uptake, ROS generation and percentage of spontaneous acrosome reaction than epididymal sperm. Regarding sperm treatments, PERCOLL group showed the highest values for some motility parameters (linearity of the curvilinear trajectory, straightness, and average path velocity/curvilinear velocity), ROS generation and penetration ability at 2 and 4 hpi; however this same group showed the lowest values for sperm curvilinear velocity and lateral head displacement. From all experimental groups, ejaculated-PERCOLL-treated spermatozoa showed the highest fertilization ability after 2 hpi. Results suggest that capacitation pathways can be regulated by suitable treatments making the ejaculated sperm able to reach capacitation and fertilize oocytes in similar levels than epididymal spermatozoa, although most of the studied capacitation-associated changes do not correlate with this ability. PMID:20688369

  12. Identification and isolation of boar sperm specific antigens with potential role in sperm-egg interaction

    Czech Academy of Sciences Publication Activity Database

    Baracova, V.; Mollova, M.; Stamenova, M.; Ivanova, M.; Pěknicová, Jana

    2004-01-01

    Roč. 64, 1-2 (2004), s. 91-106. ISSN 0165-0378 R&D Projects: GA ČR(CZ) GA204/02/1373; GA MZd(CZ) NJ7463 Institutional research plan: CEZ:AV0Z5052915 Keywords : spermatozoa * sperm surface antigens * sperm-egg fusion Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.726, year: 2004

  13. Effect of homeopathic treatment used in commercial boar semen diluent on sperm viability

    OpenAIRE

    Mayra Assunpção; Marcelo Goissis; Nilson Roberti Benites; Flavia Regina Barros; Sergio Azevedo; Leoni Villano Bonamin; Erlete Rosalina Vuaden; Cidéli de Paula Coelho; Francisco Rafael Soto; José Antonio Visitin; Mariana Grocke Marques

    2012-01-01

    Background: It has been speculated that the homeopathic treatment of sperm cells in order to improve semen quality could be promising. However, few data is available and its use in spermatozoa requires investigation. It is well established that mitochondrial membrane potential is an important viability parameter of spermatozoa and it is intimately related to reproductive efficiency. In this manner, new technologies in order to improve the activity of sperm cells and, finally, the fecundity of...

  14. Effect of storage in short--and long-term commercial semen extenders on the motility, plasma membrane and chromatin integrity of boar spermatozoa.

    Science.gov (United States)

    De Ambrogi, Marco; Ballester, Juan; Saravia, Fernando; Caballero, Ignacio; Johannisson, Anders; Wallgren, Margareta; Andersson, Magnus; Rodriguez-Martinez, Heriberto

    2006-10-01

    For artificial insemination (AI) in pigs, preservation of liquid boar semen at 16-20 degrees C is still common practice as sperm cryopreservation remains suboptimal in this species. To meet the different needs of the swine industry, several extenders have been developed to preserve semen in liquid form for short--and long-term storage. In the present study, three different commercial extenders devised for short-term (BTS+) or long-term preservation (MR-A and X-Cell), were used to test whether storage of semen from four mature, fertile boars at 17 degrees C for 96 h would affect sperm characteristics relevant for fertility, such as motility, membrane integrity and chromatin stability. Computer-assisted sperm analysis, and stainings with the acylated membrane dye SYBR-14/propidium iodide, and acridine orange in connection with flow cytometry were used to evaluate these variables. Percentages of total motile spermatozoa decreased slightly, but significantly, after 72-96 h. While membrane integrity values varied during the period of study, no significant changes in either membrane integrity or chromatin stability were, however, registered. This suggests a customary 96-day storage at 17 degrees C in these extenders was too short an interval to cause losses of integrity in nuclear DNA in the boar population studied. PMID:16573706

  15. The relationship between seminal plasma aspartate aminotransferase activity, sperm osmotic resistance test value, and semen quality in boars

    Directory of Open Access Journals (Sweden)

    Jacyno Eugenia

    2013-01-01

    Full Text Available The relationship between the activity of aspartate aminotransferase (AspAT in seminal plasma and the values of the osmotic resistance test (ORT of acrosomal membranes and semen traits was examined on 120 young hybrid Pietrain and Duroc boars. The following semen quality traits were determined: the volume of the ejaculate, the percentage of spermatozoa with progressive motility, sperm concentration and the total number of spermatozoa in the ejaculate, percentage of spermatozoa with normal acrosome, the percentage of spermatozoa with major and minor morphological defects, ORT, and the activity of AspAT in seminal plasma. The activity of AspAT in seminal plasma was negatively correlated (p_0.01 with the spermatozoa concentration and total number per ejaculate, percentage of spermatozoa with progressive motility and percentage of spermatozoa with a normal acrosome, while positively with the percentage of spermatozoa with major (p≤0.001 and minor (p≤0.01 morphological defects. The ORT values negatively correlated with the percentage of spermatozoa with major (p≤0.05 and minor (p≤0.01 morphological defects, while positively (p≤0.001 with the percentage of spermatozoa with a normal acrosome.

  16. Using fluorescence-activated flow cytometry to determine reactive oxygen species formation and membrane lipid peroxidation in viable boar spermatozoa.

    Science.gov (United States)

    Guthrie, H David; Welch, Glenn R

    2010-01-01

    Fluorescence-activated flow cytometry analyses were developed for determination of reactive oxygen species (ROS) formation and membrane lipid peroxidation in live spermatozoa loaded with, respectively, hydroethidine (HE) or the lipophilic probe 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid, C(11)BODIPY(581/591) (BODIPY). ROS was detected by red fluorescence emission from oxidization of HE and membrane lipid peroxidation was detected by green fluorescence emission from oxidation of BODIPY in individual live sperm. Of the reactive oxygen species generators tested, BODIPY oxidation was specific for FeSo4/ascorbate (FeAc), because menadione and H(2)O(2) had little or no effect. The oxidization of hydroethidine to ethidium was specific for menadione and H(2)O(2); FeAc had no effect. The incidence of basal or spontaneous ROS formation and membrane lipid peroxidation were low in boar sperm (semen or after low temperature storage; however the sperm were quite susceptible to treatment-induced ROS formation and membrane lipid peroxidation. PMID:20072917

  17. Relationship of sperm small heat-shock protein 10 and voltage-dependent anion channel 2 with semen freezability in boars.

    Science.gov (United States)

    Vilagran, Ingrid; Yeste, Marc; Sancho, Sílvia; Casas, Isabel; Rivera del Álamo, Maria M; Bonet, Sergi

    2014-08-01

    Freezability differences between boar ejaculates exist, but there is no useful method to predict the ejaculate freezability before sperm cryopreservation takes place. In this context, the present study sought to determine whether the amounts of small heat-shock protein 10 (also known as outer dense fiber protein 1) (ODF1/HSPB10) and voltage-dependent anion channel 2 (VDAC2) may be used as boar sperm freezability markers. With this aim, 26 boar ejaculates were split into two fractions: one for protein extraction and the other for cryopreservation purposes. Ejaculates were subsequently classified into two groups (good freezability ejaculates [GFE] and poor freezability ejaculates [PFE]) based on viability and sperm motility assessments after 30 and 240 minutes of after thawing. Although the VDAC2 amounts, analyzed through Western blot, were significantly higher (P cryopreservation procedures. PMID:24933094

  18. Substantial decrease of heat-shock protein 90 precedes the decline of sperm motility during cooling of boar spermatozoa.

    Science.gov (United States)

    Huang, S Y; Kuo, Y H; Lee, W C; Tsou, H L; Lee, Y P; Chang, H L; Wu, J J; Yang, P C

    1999-04-01

    The decline in boar semen quality after cryopreservation may be attributed to changes in intracellular proteins. Thus, the aim of the present study was to evaluate the change of protein profiles in boar spermatozoa during the process of cooling and after cryopreservation. A total of 9 sexually mature boars (mean age = 25.5+/-12.3 mo) was used. Samples for protein analysis were collected before chilling, after cooling to 15 degrees C, after cooling to 5 degrees C, following thawing after freezing to -100 degrees C, and following thawing after 1 wk of cryopreservation at -196 degrees C. Semen characteristics evaluated included progressive motility and the percentage of morphologically normal spermatozoa. Total proteins from 5x10(6) spermatozoa were separated and analyzed by SDS-PAGE. The results revealed that there was a substantial decrease of a 90 kDa protein in the frozen-thawed spermatozoa. Western blot analysis demonstrated that this protein was 90 kDa heat-shock protein (HSP90). Time course study showed that the decrease of HSP90 in spermatozoa initially occurred in the first hour during cooling to 5 degrees C. When compared with the fresh spermatozoa before chilling, there was a 64% decrease of HSP90 in spermatozoa after cooling to 5 degrees C. However, the motility and percentage of normal spermatozoa did not significantly decrease during this period of treatment. Both declined substantially as the semen was thawed after freezing from -100 degrees C. The results indicated that the decrease of HSP90 precedes the decline of semen characteristics. The length of time between a decrease of HSP90 and the decline in sperm motility was estimated to be 2 to 3 h. Taken together, the above results suggested that a substantial decrease of HSP90 might be associated with a decline in sperm motility during cooling of boar spermatozoa. PMID:10729022

  19. Sperm DNA methylation analysis in swine reveals conserved and species-specific methylation patterns and highlights an altered methylation at the GNAS locus in infertile boars.

    Science.gov (United States)

    Congras, Annabelle; Yerle-Bouissou, Martine; Pinton, Alain; Vignoles, Florence; Liaubet, Laurence; Ferchaud, Stéphane; Acloque, Hervé

    2014-12-01

    Male infertility is an increasing health issue in today's society for both human and livestock populations. In livestock, male infertility slows the improvement of animal selection programs and agricultural productivity. There is increasing evidence that epigenetic marks play an important role in the production of good-quality sperm. We therefore screened for specific or common epigenetic signatures of livestock infertility. To do so, we compared DNA methylation level in sperm DNA from fertile and infertile boars. We evaluated first the global level of sperm DNA methylation and found no difference between the two groups of boars. We then selected 42 loci of interest, most of them known to be imprinted in human or mice, and assessed the imprinting status of five of them not previously described in swine tissues: WT1, CNTN3, IMPACT, QPCT, and GRB10. DNA methylation level was then quantified in fertile and infertile boars at these 42 loci. Results from fertile boars indicated that the methylation level of the selected loci is highly conserved between pig, human, and mice, with a few exceptions, including the POU5F1 (OCT4) promoter and RTL1. Comparison between fertile and infertile boars revealed that one imprinted region, the GNAS locus, shows an increase in sperm DNA methylation in three out of eight infertile boars with low semen quality. This increase in DNA methylation is associated with an altered expression of the genes belonging to the GNAS locus, suggesting a new role for GNAS in the proper formation of functional gametes. PMID:25320151

  20. Studies on the effect of supplementing boar semen cryopreservation media with different avian egg yolk types on in vitro post-thaw sperm quality.

    Science.gov (United States)

    Bathgate, R; Maxwell, W M C; Evans, G

    2006-02-01

    Fertility after insemination of cryopreserved boar semen is currently below that of fresh semen. In an attempt to improve the post-thaw motility and acrosome integrity of boar sperm, semen was frozen using an adapted Westendorf method in which the chicken egg yolk was replaced by either duck or quail egg yolk. The different composition of the yolk types, particularly the amount of cholesterol, fatty acids and phospholipids, were thought to potentially afford a greater level of protection to sperm against damage during freezing and thawing. Sperm frozen in medium containing chicken egg yolk displayed higher motility immediately after thawing, but there was no difference in the motility of sperm frozen with different types of egg yolk 3 or 6 h after thawing and maintenance at 37 degrees C. Sperm frozen in media containing chicken or duck egg yolk had a higher proportion of intact acrosomes immediately after thawing than sperm frozen in medium containing quail egg yolk, but 6 h after thawing and maintenance at 37 degrees C the sperm that had been frozen in medium containing chicken egg yolk had a higher proportion of intact acrosomes than the sperm frozen in media containing duck or quail egg yolk. Analysis of the composition of the different yolk types showed that the basic components of the yolks were similar, but the ratios of fatty acids and phospholipid classes differed. Duck egg yolk had more monounsaturated fatty acids (MUFA) than chicken egg yolk, which had more MUFA than quail egg yolk. Duck egg yolk contained more phosphotidylinositol (PI) than chicken or quail egg yolks and quail egg yolk contained more phosphotidylserine than either chicken or duck egg yolks. The differences in post-thaw motility and acrosome integrity of boar sperm when frozen in media containing the different types of egg yolk may be due to the variation in composition. PMID:16420332

  1. Effect of cooling to different sub-zero temperatures on boar sperm cryosurvival

    Directory of Open Access Journals (Sweden)

    Angelica Garcia-Olivares

    2016-03-01

    Conclusions: Cooling of pig sperm to −7 °C (no freezing damaged sperm function and structure; in contrast, cooling to either −3 °C or −5 °C did not change pig sperm survival after freeze-thawing.

  2. Comparison of methods for assessing integrity of equine sperm membranes.

    Science.gov (United States)

    Foster, M L; Love, C C; Varner, D D; Brinsko, S P; Hinrichs, K; Teague, S; Lacaze, K; Blanchard, T L

    2011-07-15

    Sperm membrane integrity (SMI) is thought to be an important measure of stallion sperm quality. The objective was to compare three methods for evaluating SMI: flow cytometry using SYBR-14/propidium iodide (PI) stain; an automated cell counting device using PI stain; and eosin-nigrosin stain. Raw equine semen was subjected to various treatments containing 20 to 80% seminal plasma in extender, with differing sperm concentrations, to simulate spontaneous loss of SMI. The SMI was assessed immediately, and after 1 and 2 d of cooled storage. Agreement between methods was determined according to Bland-Altman methodology. Eosin-nigrosin staining yielded higher (2%) overall mean values for SMI than did flow cytometry. Flow cytometry yielded higher (6%) overall mean values for SMI than did the automated cell counter. As percentage of membrane-damaged sperm increased, agreement of SMI measurement between methods decreased. When semen contained 50-79% membrane-intact sperm, the 95% limits of agreement between SMI determined by flow cytometry and eosin-nigrosin staining were greater (range = -26.9 to 24.3%; i.e., a 51.2% span) than for SMI determined by flow cytometry and the automated cell counter (range = -3.1 to 17.0%; 20.1% span). When sperm populations contained range = -35.9 to 19.0%; 54.9% span) than for SMI determined by flow cytometry and the automated cell counter (range = -11.6 to 28.7%; 40.3% span). We concluded that eosin-nigrosin staining assessments of percent membrane-intact sperm agreed less with flow cytometry when <80% of sperm had intact membranes, whereas automated cell counter assessments of percent membrane-intact sperm agreed less with flow cytometry when <30% of sperm had intact membranes. PMID:21496902

  3. Field study analysis of the influences of deworming regimens and housing conditions on parasites and sperm output in 21 European boar studs.

    Science.gov (United States)

    Schulze, Martin; Ammon, Christian; Nürnberg, Gerd; Rüdiger, Karin; Jung, Markus; Demeler, Janina

    2016-03-01

    The current study reports the parasitological results of a quality control audit in 21 European boar studs. Field investigations were performed over a 2-year period (2012-2013) during the winter and spring. From each stud, an average of 30 (range, 25-33) individual faecal samples and ejaculates from 615 randomly selected Pietrain boars were analysed. Statistical analysis revealed a significant effect (P model indicated a significant effect (P = 0.0262) of DR × H × A on the presence of parasites. Sperm output was significantly affected (P < 0.0001) by the DR. Based on this study, recommendations for deworming AI boars are proposed. PMID:26831176

  4. Characterization of sperm surface protein patterns of ejaculated and capacitated boar sperm, with the detection of ZP binding candidates

    Czech Academy of Sciences Publication Activity Database

    Zigo, Michal; Jonáková, Věra; Šulc, Miroslav; Maňásková-Postlerová, Pavla

    2013-01-01

    Roč. 61, oct (2013), s. 322-328. ISSN 0141-8130 R&D Projects: GA ČR GAP503/12/1834 Institutional research plan: CEZ:AV0Z50520701 Institutional support: RVO:61388971 Keywords : Sperm surface protein * Zona pellucida-binding receptors * PKDREJ protein Subject RIV: CE - Biochemistry Impact factor: 3.096, year: 2013

  5. Characterization of proteins isolated from the sperm surface of boarsperm-zona pellucida binding receptors

    Czech Academy of Sciences Publication Activity Database

    Zigo, Michal; Jonáková, Věra; Šulc, Miroslav; Postlerová, Pavla

    Praha: Biotechnologický ústav, 2013 - (Pěknicová, J.). s. 19 [XIX. Symposium imunologie a biologie reprodukce s mezinárodní účastí. 23.05.2013-25.05.2013, Třešť] R&D Projects: GA ČR(CZ) GAP503/12/1834 Institutional research plan: CEZ:AV0Z50520701 Keywords : Zona pellucida * Sperm surface protein * 2D-electrophoresis * Glycoproteins Subject RIV: CE - Biochemistry

  6. Sperm sorting procedure induces a redistribution of Hsp70 but not Hsp60 and Hsp90 in boar spermatozoa.

    Science.gov (United States)

    Spinaci, Marcella; Volpe, Sara; Bernardini, Chiara; de Ambrogi, Marco; Tamanini, Carlo; Seren, Eraldo; Galeati, Giovanna

    2006-01-01

    Heat shock proteins, besides their protective function against stresses, have been recently indicated as key factors for sperm fertilizing ability. Since sexing sperm by high-speed flow-cytometry subjects them to different physical, mechanical, and chemical stresses, the present study was designed to verify, by immunofluorescence and Western blot, whether the sorting procedure induces any modification in the amount and cellular distribution of heat shock proteins 60, 70, and 90 (Hsp60, Hsp70, Hsp90). Immunolocalization and Western blot quantification of both Hsp60 and Hsp90 did not reveal differences between unsorted and sorted semen. On the contrary, a redistribution of Hsp70 immunoreactivity from the equatorial subsegment toward the equator of sperm cells was recorded after sorting; this relocation suggests capacitation-like changes of sperm membrane. This modification seems to be caused mainly by incubation with Hoechst 33342, while both passage of sperm through flow cytometer and laser beam represent only minor stimuli. A further Hsp70 redistribution seems to be due to the final steps of sperm sorting, charging, and deflection of drops, and to the dilution during collection. On the other hand, staining procedure and mechanical stress seem to be the factors most injurious to sperm viability. Moreover, Hsp70 relocation was deeply influenced by the storage method. In fact, storing sexed spermatozoa, after centrifugation, in a small volume in presence of seminal plasma induced a reversion of Hsp70 redistribution, while storage in the diluted catch fluid of collection tubes caused Hsp70 relocation in most sorted spermatozoa. PMID:16870948

  7. The effect of extender, method of thawing, and duration of storage on in vitro fertility measures of frozen-thawed boar sperm.

    Science.gov (United States)

    Knox, R V; Ringwelski, J M; McNamara, K A; Aardsma, M; Bojko, M

    2015-08-01

    Frozen-thawed boar sperm (FTS) has reduced in vitro and in vivo life span compared to liquid semen. Experiments tested whether extenders, thawing procedures, and storage temperatures could extend the fertile life span of FTS. Experiment 1 tested the effect of six extenders on postthaw motility (MOT) and viability (VIA). Straws from boars (n = 6) were thawed, diluted into each extender, and evaluated at 20, 60, and 120 minutes. There was a trend (P = 0.08) for an extender-by-time interaction for MOT and effect of extender and time for MOT (P extender (P = 0.10) and time (P boar ejaculates (n = 15) were thawed at 50 °C for 10, 20, or 30 seconds or at 70 °C for 5, 10, or 20 seconds and evaluated at 5, 30, and 60 minutes. There was an effect of thawing treatment on MOT, VIA, and ACR (viable sperm with intact acrosomes, P extenders, thawing, and storage. PMID:25913276

  8. Supplementation of different concentrations of Orvus Es Paste (OEP) to ostrich egg yolk lipoprotein extender improves post-thaw boar semen quality.

    Science.gov (United States)

    Fraser, L; Jasiewicz, E; Kordan, W

    2014-01-01

    This study aimed to compare post-thaw quality of boar semen following freezing in an ostrich egg yolk lipoprotein (LPFo) extender supplemented with 0%, 0.25% and 0.50% Orvus Es Paste (OEP). Sperm assessments included total motility (TMOT), mitochondrial function (MF), plasma membrane integrity (PMI) and acrosome integrity (normal apical ridge, NAR). Considerable variations among boars and OEP treatments had a significant effect (P egg yolk lipoproteins, could have varying effects on post-thaw sperm survival. PMID:24988847

  9. Egg Yolk Protective Effect in Boar Spermatozoa Cooled at 5ºC

    OpenAIRE

    Alexandru-Vasile Rusu; Vasile Miclea; Marius Zahan

    2011-01-01

    Nowadays, many boar reproduction researches are directed to improve extenders and to increase cold shock protection of semen. Little research is focused on the influence of egg yolk combined with alternative cold shock protective media. Egg yolk could interfere with other compounds present in the extender composition. The influence of egg yolk addition was assessed in boar sperm cells, cooled at 5ºC, to elucidate its effect on motility and membrane integrity. Flow Cytometry and Computer Assis...

  10. Lactose-egg yolk diluent supplemented with N-acetyl-D-glucosamine affect acrosome morphology and motility of frozen-thawed boar sperm.

    Science.gov (United States)

    Yi, Y J; Im, G S; Park, C S

    2002-12-16

    These experiments were carried out to investigate the effect of N-acetyl-D-glucosamine, and to obtain additional information about the effect of orvus es paste (OEP) and egg yolk concentration in the freezing of boar sperm in the maxi-straw. The highest post-thaw acrosomes of normal apical ridge (NAR) and motility were obtained with 0.025 or 0.05% N-acetyl-D-glucosamine concentration in the first diluent. However, there were no effects of N-acetyl-D-glucosamine among the diluents with or without N-acetyl-D-glucosamine at the second dilution. The N-acetyl-D-glucosamine in the first and second diluents was added at room temperatures (20-23 degrees C) and 5 degrees C, respectively. It is suggested that the temperature of N-acetyl-D-glucosamine addition is important for the effect of boar sperm protection during freezing and thawing. When the 0.05% N-acetyl-D-glucosamine was supplemented in the first diluent, the optimum final OEP content was 0.5%. The optimum content of egg yolk in the diluent with 0.05% N-acetyl-D-glucosamine concentration was 20% and egg yolk was one of the main cryoprotective agents. In conclusion, we found out that the diluent with 0.025 or 0.05% soluble N-acetyl-D-glucosamine in the first diluent, 0.5% final orvus es paste concentration and 20% egg yolk concentration significantly enhanced NAR acrosomes and motility of boar sperm after freezing and thawing. PMID:12417120

  11. Differences in the ability of spermatozoa from individual boar ejaculates to withstand different semen-processing techniques.

    Science.gov (United States)

    Parrilla, Inma; del Olmo, David; Sijses, Laurien; Martinez-Alborcia, María J; Cuello, Cristina; Vazquez, Juan M; Martinez, Emilio A; Roca, Jordi

    2012-05-01

    The present study aimed to evaluate the ability of spermatozoa from individual boar ejaculates to withstand different semen-processing techniques. Eighteen sperm-rich ejaculate samples from six boars (three per boar) were diluted in Beltsville Thawing Solution and split into three aliquots. The aliquots were (1) further diluted to 3×10(7) sperm/mL and stored as a liquid at 17°C for 72 h, (2) frozen-thawed (FT) at 1×10(9) sperm/mL using standard 0.5-mL straw protocols, or (3) sex-sorted with subsequent liquid storage (at 17°C for 6 h) or FT (2×10(7) sperm/mL using a standard 0.25-mL straw protocol). The sperm quality was evaluated based on total sperm motility (the CASA system), viability (plasma membrane integrity assessed using flow cytometry and the LIVE/DEAD Sperm Viability Kit), lipid peroxidation (assessed via indirect measurement of the generation of malondialdehyde (MDA) using the BIOXYTECH MDA-586 Assay Kit) and DNA fragmentation (sperm chromatin dispersion assessed using the Sperm-Sus-Halomax(®) test). Data were normalized to the values assessed for the fresh (for liquid-stored and FT samples) or the sorted semen samples (for liquid stored and the FT sorted spermatozoa). All of the four sperm-processing techniques affected sperm quality (Psemen donor, with reduced percentages of motile and viable sperm and increased MDA generation and percentages of sperm with fragmented DNA. Significant (Pboar (effect of boars within each semen-processing technique) and intra-boar (effect of semen-processing techniques within each boar) differences were evident for all of the sperm quality parameters assessed, indicating differences in the ability of spermatozoa from individual boars to withstand the semen-processing techniques. These results are the first evidence that ejaculate spermatozoa from individual boars can respond in a boar-dependent manner to different semen-processing techniques. PMID:22554791

  12. Effects of hypothermic liquid storage and cryopreservation on basal and induced plasma membrane phospholipid disorder and acrosome exocytosis in boar spermatozoa.

    Science.gov (United States)

    Guthrie, H D; Welch, G R

    2005-01-01

    Flow cytometry was utilised to determine whether short-term (Day 1) or long-term hypothermic liquid storage (Day 5), or cryopreservation of boar spermatozoa (1) caused changes in plasma membrane phospholipid disorder (MPLD) and acrosome exocytosis (AE), indicative of an advanced stage of capacitation or acrosome status, and (2) facilitated or inhibited the induction of capacitation and the acrosome reaction. Merocyanine with Yo-Pro-1 and peanut agglutinin-fluorescein isothiocyanate with propidium iodide were used to identify MPLD and AE, respectively, in viable spermatozoa. The incidence of basal sperm MPLD and AE in fresh semen was very low (1.1 and 2.2%, respectively) and was increased (P semen (3-8%). Compared to no bicarbonate, incubation with bicarbonate increased MPLD, but the response was greatest (P semen than for Day 1 (45%) and Day 5 (57%) semen. In summary, hypothermic liquid storage and cryopreservation of boar spermatozoa did not advance capacitation or acrosome status in viable spermatozoa, but did alter their responses to induction of capacitation and the acrosome reaction. PMID:15899159

  13. A functional assessment of the sperm membrane: a multi-species approach

    OpenAIRE

    Murphy, Craig

    2014-01-01

    peer-reviewed A functional sperm membrane is essential for many of the processes which lead to the fertilisation of an oocyte, but this membrane is susceptible to oxidative damage with increasing duration of storage in liquid semen or during the cryopreservation process. The objectives of this thesis were to examine the effects of storage temperature, catalase supplementation and sperm number on the membrane function of liquid stored bull semen, sperm membrane protein profil...

  14. Ophiobolin A from Bipolaris oryzae perturbs motility and membrane integrities of porcine sperm and induces cell death on mammalian somatic cell lines.

    Science.gov (United States)

    Bencsik, Ottó; Papp, Tamás; Berta, Máté; Zana, Annamária; Forgó, Péter; Dombi, György; Andersson, Maria A; Salkinoja-Salonen, Mirja; Vágvölgyi, Csaba; Szekeres, András

    2014-09-01

    Bipolaris oryzae is a phytopathogenic fungus causing a brown spot disease in rice, and produces substance that strongly perturbs motility and membrane integrities of boar spermatozoa. The substance was isolated from the liquid culture of the fungal strain using extraction and a multi-step semi-preparative HPLC procedures. Based on the results of mass spectrometric and 2D NMR techniques, the bioactive molecule was identified as ophiobolin A, a previously described sesterterpene-type compound. The purified ophiobolin A exhibited strong motility inhibition and viability reduction on boar spermatozoa. Furthermore, it damaged the sperm mitochondria significantly at sublethal concentration by the dissipation of transmembrane potential in the mitochondrial inner membrane, while the plasma membrane permeability barrier remained intact. The study demonstrated that the cytotoxicity of ophiobolin A toward somatic cell lines is higher by 1-2 orders of magnitude compared to other mitochondriotoxic mycotoxins, and towards sperm cells unique by replacing the progressive motility by shivering tail beating at low exposure concentration. PMID:25251540

  15. Ophiobolin A from Bipolaris oryzae Perturbs Motility and Membrane Integrities of Porcine Sperm and Induces Cell Death on Mammalian Somatic Cell Lines

    Directory of Open Access Journals (Sweden)

    Ottó Bencsik

    2014-09-01

    Full Text Available Bipolaris oryzae is a phytopathogenic fungus causing a brown spot disease in rice, and produces substance that strongly perturbs motility and membrane integrities of boar spermatozoa. The substance was isolated from the liquid culture of the fungal strain using extraction and a multi-step semi-preparative HPLC procedures. Based on the results of mass spectrometric and 2D NMR techniques, the bioactive molecule was identified as ophiobolin A, a previously described sesterterpene-type compound. The purified ophiobolin A exhibited strong motility inhibition and viability reduction on boar spermatozoa. Furthermore, it damaged the sperm mitochondria significantly at sublethal concentration by the dissipation of transmembrane potential in the mitochondrial inner membrane, while the plasma membrane permeability barrier remained intact. The study demonstrated that the cytotoxicity of ophiobolin A toward somatic cell lines is higher by 1–2 orders of magnitude compared to other mitochondriotoxic mycotoxins, and towards sperm cells unique by replacing the progressive motility by shivering tail beating at low exposure concentration.

  16. Comparison of semen variables, sperm DNA damage and sperm membrane proteins in two male layer breeder lines.

    Science.gov (United States)

    M, Shanmugam; T R, Kannaki; A, Vinoth

    2016-09-01

    Semen variables are affected by the breed and strain of chicken. The present study was undertaken to compare the semen quality in two lines of adult chickens with particular reference to sperm chromatin condensation, sperm DNA damage and sperm membrane proteins. Semen from a PD3 and White Leghorn control line was collected at 46 and 47 weeks and 55 weeks of age. The semen was evaluated for gross variables and sperm chromatin condensation by aniline blue staining. Sperm DNA damage was assessed by using the comet assay at 47 weeks of age and sperm membrane proteins were assessed at 55 weeks of age. The duration of fertility was studied by inseminating 100 million sperm once into the hens of the same line as well as another line. The eggs were collected after insemination for 15days and incubated. The eggs were candled on 18th day of incubation for observing embryonic development. The White Leghorn control line had a greater sperm concentration and lesser percentage of morphologically abnormal sperm at the different ages where assessments occurred. There was no difference in sperm chromatin condensation, DNA damage and membrane proteins between the lines. Only low molecular weight protein bands of less than 95kDa were observed in samples of both lines. The line from which semen was used had no effect on the duration over which fertility was sustained after insemination either when used in the same line or another line. Thus, from the results of the present study it may be concluded that there was a difference in gross semen variables between the lines that were studied, however, the sperm chromatin condensation, DNA damage, membrane proteins and duration over which fertility was sustained after insemination did not differ between the lines. PMID:27470200

  17. Assessment of boar sperm intracellular Ca²⁺ level and motility characters by flow cytometry and CASA. Optimizing the protocol for Fluo-4 assessment of sperm intracellular Ca²⁺level by flow cytometry and investigation of relationships between storage time, intracellular Ca²⁺ and sperm motility characters

    OpenAIRE

    Khezri, Abdolrahman

    2013-01-01

    Porcine production industry demand effective artificial insemination for future challenges. Variation in field fertility and litter size not only caused by sows or farm management but also affected by semen and boar related parameters. Assessment of semen using objective methods provides an effective tool for better and precise assessment of sperm quality during the collection until consumption and leads to prediction of field fertility and genetic selection. In order to eva...

  18. Inter- and intra-breed comparative study of sperm motility and viability in Iberian and Duroc boar semen during long-term storage in MR-A and XCell extenders.

    Science.gov (United States)

    Martín-Hidalgo, D; Barón, F J; Robina, A; Bragado, M J; Llera, A Hurtado de; García-Marín, L J; Gil, M C

    2013-06-01

    During boar semen liquid preservation, extender is one of the factors that influence storage tolerance of spermatozoa. However, there are few studies about intra-breed variation in the preservation of semen quality during storage in different extenders. Similarly, boar breed is generally not considered a possible factor influencing variation in the semen storage tolerance in a particular extender. The aim of this study was to compare boar semen storage potential, in terms of the ability to maintain sperm viability and motility, of two currently used long-term extenders, MR-A and XCell. Extended semen from two breeds, Iberian and Duroc that had been stored at 17°C for up to 7 days was used. Intra- and inter-breed effect was studied. On Days 1, 4 and 7 (Day 0=day of semen collection), motility parameters and the percentage of total motile sperm and progressively motile sperm using a CASA system was evaluated. Viability (SYBR-14/PI) was evaluated by flow cytometry. Within each breed and for each storage day, there were differences between extenders, although semen tolerance to preservation was more influenced by the extender in the Iberian than in the Duroc breed. Neither breed nor extender influenced the percentage of viable spermatozoa during the storage time. Moreover, differences in motility parameters were observed between breeds, although the differences were greater when the XCell extender was used. In conclusion, both extender and breed influence motility characteristics of liquid-stored boar semen, so both aspects have to be considered in the design of comparative studies about stored boar semen quality from different breeds or with different extenders. Further studies are needed to corroborate these findings. PMID:23660365

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1978-05-01

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

  20. Lipid domains in the ram sperm plasma membrane demonstrated by differential scanning calorimetry.

    OpenAIRE

    Wolf, D. E.; Maynard, V M; McKinnon, C A; Melchior, D L

    1990-01-01

    Mammalian sperm plasma membranes, in contrast to those of mammalian somatic cells, exhibit a significant fraction of lipid that does not diffuse laterally in the plane of the membrane. This nondiffusing fraction results from lipid-lipid interactions. Similar nondiffusing fractions are found in mixed-lipid model systems that contain coexistent gel and fluid domains. These results suggest that the sperm plasma membrane may also exhibit lateral phase segregations of lipids and may contain signif...

  1. Identification and purification of a sperm surface protein with a potential role in sperm-egg membrane fusion.

    Science.gov (United States)

    Primakoff, P; Hyatt, H; Tredick-Kline, J

    1987-01-01

    Sperm-egg plasma membrane fusion during fertilization was studied using guinea pig gametes and mAbs to sperm surface antigens. The mAb, PH-30, strongly inhibited sperm-egg fusion in a concentration-dependent fashion. When zona-free eggs were inseminated with acrosome-reacted sperm preincubated in saturating (140 micrograms/ml) PH-30 mAb, the percent of eggs showing fusion was reduced 75%. The average number of sperm fused per egg was also reduced by 75%. In contrast a control mAb, PH-1, preincubated with sperm at 400 micrograms/ml, caused no inhibition. The PH-30 and PH-1 mAbs apparently recognize the same antigen but bind to two different determinants. Both mAbs immunoprecipitated the same two 125I-labeled polypeptides with Mr 60,000 (60 kD) and Mr 44,000 (44 kD). Boiling a detergent extract of sperm severely reduced the binding of PH-30 but had essentially no effect on the binding of PH-1, indicating that the two mAbs recognize different epitopes. Immunoelectron microscopy revealed that PH-30 mAb binding was restricted to the sperm posterior head surface and was absent from the equatorial region. The PH-30 and PH-1 mAbs did not bind to sperm from the testis, the caput, or the corpus epididymis. PH-30 mAb binding was first detectable on sperm from the proximal cauda epididymis, i.e., sperm at the developmental stage where fertilization competence appears. After purification by mAb affinity chromatography, the PH-30 protein retained antigenic activity, binding both the PH-30 and PH-1 mAbs. The purified protein showed two polypeptide bands of 60 and 44 kD on reducing SDS PAGE. The two polypeptides migrated further (to approximately 49 kD and approximately 33 kD) on nonreducing SDS PAGE, showing that they do not contain interchain disulfide bonds, but probably have intrachain disulfides. 44 kD appears not to be a proteolytic fragment of 60 kD because V8 protease digestion patterns did not reveal related peptide patterns from the 44- and 60-kD bands. In the absence of

  2. Effect of semen extenders on frozen-thawed boar sperm characteristics and distribution in the female genital tract after deep intrauterine insemination in sows.

    Science.gov (United States)

    Noguchi, Michiko; Yoshioka, Koji; Hikono, Hirokazu; Suzuki, Chie; Kikuchi, Kazuhiro

    2015-12-01

    We compared the effects of extenders of frozen-thawed semen on post-thaw sperm characteristics and the distribution of frozen-thawed spermatozoa in the female genital tract after fixed-timed deep intrauterine insemination (DIUI) in sows. Frozen semen samples were thawed and diluted in either modified Modena solution (mMS) or porcine fertilization medium (PFM) containing theophylline, adenosine and cysteine. Sperm quality, assessed in vitro based on motility using a computer-assisted sperm analyzer and the integrity of the plasma and acrosomal membranes using flow cytometry, was evaluated at 0.5, 1.5, 3 and 6h after thawing. Progressive motility and the percentage of spermatozoa with damaged acrosomal membranes in PFM were significantly better than in mMS throughout the 6h. Sows with estrus synchronized using prostaglandin F2 alpha, equine chorionic gonadotropin and human chorionic gonadotropin (hCG) were inseminated once with mMS- or PFM-diluted 5 × 10(8) frozen-thawed spermatozoa by DIUI at 34 h after the hCG injection. At 4h after DIUI, reproductive tracts were recovered from 30 sows. There were significantly fewer polymorphonuclear leukocytes (PMNs) and more spermatozoa outside PMNs in the uterine horn after PFM treatment than with mMS. When 22 sows were administered DIUI with 10 × 10(8) frozen-thawed spermatozoa at 36 h after hCG, the pregnancy rates did not differ significantly between the mMS- (36%) and PFM- (64%) treated groups. Thus, PFM enhanced progressive sperm motility but increased sperm membrane damage compared with mMS; it also suppressed the migration of PMNs into the uterine lumen. PMID:26588890

  3. 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.

  4. Egg Yolk Protective Effect in Boar Spermatozoa Cooled at 5ºC

    Directory of Open Access Journals (Sweden)

    Alexandru-Vasile Rusu

    2011-05-01

    Full Text Available Nowadays, many boar reproduction researches are directed to improve extenders and to increase cold shock protection of semen. Little research is focused on the influence of egg yolk combined with alternative cold shock protective media. Egg yolk could interfere with other compounds present in the extender composition. The influence of egg yolk addition was assessed in boar sperm cells, cooled at 5ºC, to elucidate its effect on motility and membrane integrity. Flow Cytometry and Computer Assisted Semen Analysis (CASA were used to determine the rate of sperm with intact plasma and acrosomal membrane, respectively the sperm cells motility. Statistical analyses (T-Test were performed using GraphPad Prism version 5.00. Androhep Plus supplemented with 20% egg yolk (AhPlus+20%EY indicated a higher cold shock protection in progressive motility (93.9±2.64% and membrane integrity (79.78±4.14%, rather than the extender without egg yolk (p<0.01, respectively p<0.05. The results of the this study showed that egg yolk addition to AhPlus do not interfere with its compounds, the data being in a close range with those obtained by using the standard Lactose Egg Yolk extender (p>0.05. The combination egg yolk-AhPlus seems to be an alternative to standard extenders, conferring stability in boar sperm cells against cold shock.

  5. Investigation of pig sperm plasma membrane reorganization using progesterone-albumin-fluorescein probes

    Institute of Scientific and Technical Information of China (English)

    Alfredo Medrano; Paul F Watson; William V Holt

    2012-01-01

    Objective:To relate semen susceptibility in cooling protocols to sperm plasma membrane properties.Methods:A series of experiments was performed using the fluorescent markers, progesterone-BSA-FITC andBSA-FITC.Results:These experiments indicated that both progesterone-BSA-FITC andBSA-FITC bound to specific sperm plasma membrane domains, thus producing four different binding patterns, revealing probable changes in membrane organization during capacitation and during cooling.Those patterns seem to make a sequence progressing from non-capacitated status to capacitated status.The proportion of each pattern was different during incubation than during cooling, showing the latter had a higher proportion of dead sperm than the former.Conclusions:At this stage, the association of sperm plasma membrane alterations was revealed byBSA-FITC probes and cryosensitivity remains unclear.

  6. A molecular approach on sperm changes during epididymal maturation, ejaculation and in vitro capacitation of boar spermatozoa

    OpenAIRE

    Fàbrega Coll, Anna

    2012-01-01

    Mammalian spermatozoa acquire functionality during epididymal maturation and ability to penetrate and fertilize the oocyte during capacitation. Sperm quality results indicated that both epididymal maturation and ejaculation are key events for further capacitation, because only ejaculated spermatozoa are capable to undergo the set of changes leading to capacitation. Epididymal maturation is associated with a progressive loss of phosphotyrosine residues followed by a subtle increase after in vi...

  7. Electrophoretic and zymographic characterization of proteins isolated by various extraction methods from ejaculated and capacitated boar sperms

    Czech Academy of Sciences Publication Activity Database

    Zigo, Michal; Jonáková, Věra; Maňásková-Postlerová, Pavla

    2011-01-01

    Roč. 32, č. 11 (2011), s. 1309-1318. ISSN 0173-0835 R&D Projects: GA ČR(CZ) GA303/09/1285; GA MZd(CZ) NS10009; GA MŠk(CZ) 1M06011 Institutional research plan: CEZ:AV0Z50520701 Keywords : Extraction methods * Sperm proteins * Substrate zymography Subject RIV: CE - Biochemistry Impact factor: 3.303, year: 2011

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

    OpenAIRE

    L. S. L. S. Reis; A.A. Ramos; A.S. Camargos; E. Oba

    2016-01-01

    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 e...

  9. Effect of Conjugated Linoleic Acid on Boar Semen Quality After Long-term Refrigeration at 17°C.

    Science.gov (United States)

    Teixeira, Smp; Chaveiro, A; Moreira da Silva, F

    2015-08-01

    In this study, the effect of conjugated linoleic acid (10 trans, 12 cis) (CLA) on refrigerated boar sperm quality parameters up to 14 days at 17°C was assessed. Semen was extended in Androhep and divided into four treatments supplemented with CLA (25, 50, 100 and 200 μm) and control group, then kept for 2 h at 22°C. Afterwards an aliquot of each treatment was removed, and mitochondrial activity, viability, lipid membrane peroxidation (LPO) and stability of the sperm plasma membrane were assessed by flow cytometry. The remaining extended semen was maintained at 17°C until 336 h, repeating the same analysis every 48 h. Regarding percentage of live spermatozoa, no statistical differences were observed among treatments up to 96 h. After this time, viability decreased significantly (p boar A presented better results when compared with the other boars, especially at concentrations of 50 and 100 μm boar B showed significantly higher results (p boar spermatozoa. PMID:25976112

  10. Cryodamage to plasma membrane integrity in head and tail regions of human sperm

    Institute of Scientific and Technical Information of China (English)

    Wei-JieZHU; Xue-GaoLIU

    2000-01-01

    Aim: To investigate the effect of cryopreservation on the plasma membrane integrity in the head and tail regions of individual sperm, and the relationship between intact cryopreserved sperm and its motility and zona-free hamster oocyte penetration rate. Methods: The eosin Y exclusion and the hypoosmotic swelling tests were combined to form a single test (HOS-EY test) to identify the spermatozoa with four types of membrane integrity. Results: After cryopreservation, there was a marked decline in the percentage of spermatozoa with Type IV membrane integrity (head membrane intact/tail membrane intact), and a significant increase in those with Type Ⅰ (head membrane damaged/tail membrane damaged) and Type Ⅲ (head membrane damaged/tail membrane intact) membrane integrity (n = 50, P0.05). Conclusion: (1) The HOS-EY test has the advantage of showing four patterns of membrane integrity in individual spermatozoon; (2) Cryopreservation causes a significant membrane rupture in the head and tail regions of spermatozoa; Type IT[ is the main transitional state of membrane cryodamage; (3) Cryodamage to head and tail membrane may occur independently; the presence of an intact tail membrane does not necessarily indicate the intactness of head membrane. (4) Intact membranes am closely related to postthaw motility, but do not reflect the fertilizing potential.

  11. Sperm membrane proteome in wild Japanese macaque (Macaca fuscata) and Sika deer (Cervus nippon).

    Science.gov (United States)

    Kawase, Osamu; Cao, Shinuo; Xuan, Xuenan

    2015-01-01

    Whereas recent advances in proteome-related techniques have accumulated a lot of information about sperm proteins in model animals, the information in non-model wildlife species is absolutely deficient, although this knowledge would be valuable to regulate wildlife overabundance. To characterize the repertoires of sperm membrane proteins in Japanese overpopulated wildlife, our study focuses on the following two species: Macaca fuscata and Cervus nippon. We enriched sperm membrane proteins by the phase partitioning with Triton X-114, and then separated them by two-dimensional electrophoresis, and, finally, they were comprehensively identified by peptide mass fingerprinting. Sperm membrane proteins were successfully enriched. They included some proteins with unknown function and fertility-related proteins that work in sperm development, motility, capacitation, transport, protection, acrosome reaction, and fertilization. Additionally, beta-defensin 126 and epithelial chloride channel were strongly detected in M. fuscata but not in C. nippon, whereas lactadherin and NADH-cytochrome b5 reductase 1 were strongly detected in C. nippon alone. This study is an initiative case showing that the sperm of wildlife conserve major fertility-related proteins, but express some proteins in a species-specific manner. In the development of a practical method for fertility control, this aspect may be taken into consideration. PMID:25277530

  12. Effect of a pre-freezing treatment with cholesterol-loaded cyclodextrins on boar sperm longevity, capacitation dynamics, ability to adhere to porcine oviductal epithelial cells in vitro and DNA fragmentation dynamics.

    Science.gov (United States)

    Tomás, C; Blanch, E; Fazeli, A; Mocé, E

    2013-01-01

    The aim of this work was to examine how a pre-freezing treatment with cholesterol-loaded cyclodextrins (CLC) affects boar sperm longevity, capacitation dynamics, ability to bind to a porcine telomerase-immortalised oviductal epithelial cell line (TERT-OPEC) in vitro and DNA integrity dynamics after freeze-thawing. Although the samples treated with CLC exhibited lower sperm quality than the control samples (P0.05) after long-term incubation (26h at 37 or 16°C). Additionally, the CLC-treated spermatozoa underwent similar capacitation and DNA fragmentation dynamics as the control spermatozoa (P>0.05). However, CLC-treated spermatozoa were better able to bind to TERT-OPEC in vitro (POPEC in vitro, which could have an effect on the establishment of the sperm reservoir in the ampullary--isthmic junction in vivo. Additionally, frozen-thawed spermatozoa can be stored at 16°C for at least 6h without a significant observable decline in sperm quality, which could be beneficial for the transport of thawed diluted doses of spermatozoa from the laboratory to the farm. PMID:23036662

  13. Modification of trout sperm membranes associated with activation and cryopreservation. Implications for fertilizing potential.

    Science.gov (United States)

    Purdy, P H; Barbosa, E A; Praamsma, C J; Schisler, G J

    2016-08-01

    We investigated the effects of two trout sperm activation solutions on sperm physiology and membrane organization prior to and following cryopreservation using flow cytometry and investigated their impact on in vitro fertility. Overall, frozen-thawed samples had greater phospholipid disorder when compared with fresh samples (high plasma membrane fluidity; P < 0.0001) and sperm activated with water also had high plasma membrane fluidity when compared to sperm activated with Lahnsteiner solution (LAS; P < 0.0001). Following cryopreservation water activated samples had membranes with greater membrane protein disorganization compared with LAS but the membrane protein organization of LAS samples was similar to samples prior to freezing (P < 0.0001). Post-thaw water activation resulted in significant increases in intracellular calcium compared to LAS (P < 0.002). In vitro fertility trials with frozen-thawed milt and LAS activation resulted in greater fertility (45%) compared to water activated samples (10%; P < 0.0001). Higher fertility rates correlated with lower intracellular calcium with water (R(2) = -0.9; P = 0.01) and LAS (R(2) = -0.85; P = 0.03) activation. Greater plasma membrane phospholipid (R(2) = -0.89; P = 0.02) and protein (R(2) = -0.84; P = 0.04) disorder correlated with lower water activation fertility rates. These membrane organization characteristics only approached significance with LAS activation in vitro fertility (P = 0.09, P = 0.06, respectively). Potentially the understanding of sperm membrane reorganizations and the physiology associated with activation following cryopreservation may enable users in a repository or hatchery setting to estimate the fertilizing potential of a sample and determine its value. PMID:27234987

  14. Na+-permeable channels of human sperm membrane re- assembled into giant liposome

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Previous data showed that a Na+-transmembrane flux was accompanied with acrosome reaction of sperm. However, the electrophysiological recording and characterization of Na+ current in human sperm membrane have not been yet reported. In the present investigation, membrane proteins extracted from human sperms were reassembled into liposome bilayer, and then the liposomes were fused by dehydration-rehydration into giant liposomes with the diameter of more than 10 mm. By patch clamping the giant liposomes two kinds of single channel currents were recorded in a NaCl solution system. Both of them were Na+-carried, TTX-sensitive and strongly rectifying, but with different unit conductance and open probability. Moreover, bursting activity and channel-substates as well as two open time constants were observed in the larger channel.

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

    Directory of Open Access Journals (Sweden)

    Farzaneh Eskandari

    2016-01-01

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

  16. Controlled cooling during semen cryopreservation does not induce capacitation of spermatozoa from two portions of the boar ejaculate.

    Science.gov (United States)

    Saravia, F; Hernández, M; Wallgren, M; Johannisson, A; Rodríguez-Martínez, H

    2007-12-01

    Cryopreservation imposes dramatic changes in boar sperm survivability but it is as yet unclear which part of the process affects the spermatozoa the most. The present study monitored, along the entire process of cryopreservation, the stability (PMS) of the architecture of the lipid plasma membrane and its integrity (PMI), as well as the kinetics of the processed spermatozoa using two portions from the boar ejaculate (P1 = the first 10 mL of the sperm-rich fraction, SRF; P2 = the rest of the ejaculate), frozen in a recently developed package, the MiniFlatPack (MFPs, 0.5 x 10(9) sperm/dose). Evaluation was made at four specific stages, viz. S1 = after collection (suspended in Beltsville thawing solution, BTS); S2 = at 15 degrees C (suspended in lactose-egg yolk, LEY); S3 = at 5 degrees C (suspended in LEY plus glycerol); and S4 = post-thaw. Both sperm kinetics (using computer-assisted sperm analysis, CASA) and PMS [i.e. the degree of lipid disorder and of the exteriorization of phosphatidylserine (PS) in the plasma membrane, measured by flow cytometry using Merocyanine-540 (M-540), and Annexin-V (AV) respectively], as well as plasma membrane integrity [PMI, i.e. the degree of membrane damage, measured using Yo-Pro-1 or propidium iodide (PI)] were assessed after incubation in BTS at 38 degrees C. Moreover, spermatozoa were challenged by incubation in modified Brackett-Oliphant medium (mBO+) with 37 mm of bicarbonate at 38 degrees C for 30 min, and their PMS and PMI further explored. Total sperm motility was significantly higher in P1 than in P2 along the entire process (S1-S4; p boar spermatozoa suspended in BTS (S1), LEY (S2) or LEY plus glycerol (S3) were maintained during controlled cooling but were altered by thawing, showing more characteristics of cell injury than of sperm capacitation. The spermatozoa were able to capacitate but the bicarbonate challenge destabilized the plasma membrane during initial cooling and accelerated membrane changes post-thaw. We

  17. Cryopreservation of boar semen and its future importance to the industry.

    Science.gov (United States)

    Bailey, Janice L; Lessard, Christian; Jacques, Joannie; Brèque, Christelle; Dobrinski, Ina; Zeng, Wenxian; Galantino-Homer, Hannah L

    2008-11-01

    Whereas AI has arguably been the most important management tool leading to improved herd productivity, long-term storage of semen brings forth additional advantages to producers of agriculturally important animals and the AI industry. Semen cryopreservation greatly facilitates the distribution of agriculturally desirable genes, rapidly increasing herd productivity. Of particular importance to the pig industry, the use of frozen semen would help to control transmission of certain pathogens, thereby protecting the health status of the herd. Moreover, a reserve of cryopreserved semen would minimize the effects of a sudden outbreak of a contagious illness or a natural disaster. Successful cryopreservation of boar semen is necessary for international sales. Finally, effective gene banking depends on the availability of functional, cryopreserved germplasm. Despite these potential advantages of long-term semen storage, porcine sperm are notoriously sensitive to cold temperatures, and frozen-thawed semen is not routinely used by the industry. The objective of our laboratories is to develop protocols for efficient long-term storage of porcine semen using cryopreservation. We hypothesize that since the sperm plasma membrane is the primary site of cold-induced damage, reinforcing the membranes with molecules having particular properties, such as cholesterol, will improve the ability of boar sperm to withstand cold temperatures and cryopreservation protocols. Based on our data, such approaches should help alleviate the problems with sperm function after cooling, thereby resulting in better survival and motility characteristics, and reduced non-regulated capacitation and spontaneous acrosome reactions. PMID:18653225

  18. Supramolecular organization of the sperm plasma membrane during maturation and capacitation

    Institute of Scientific and Technical Information of China (English)

    Roy Jones; Peter S. James; Liz Howes; Andreas Bruckbauer; David Klenerman

    2007-01-01

    Aim: In the present study, a variety of high resolution microscopy techniques were used to visualize the organization and motion of lipids and proteins in the sperm's plasma membrane. We have addressed questions such as the presence of diffusion barriers, confinement of molecules to specific surface domains, polarized diffusion and the role of cholesterol in regulating lipid rafts and signal transduction during capacitation. Methods: Atomic force microscopy identified a novel region (EqSS) within the equatorial segment of bovine, porcine and ovine spermatozoa that was enriched in constitutively phosphorylated proteins. The EqSS was assembled during epididymal maturation. Fluorescence imaging techniques were then used to follow molecular diffusion on the sperm head. Results: Single lipid molecules were freely exchangeable throughout the plasma membrane and showed no evidence for confinement within domains. Large lipid aggregates, however, did not cross over the boundary between the post-acrosome and equatorial segment suggesting the presence of a molecular filter between these two domains. Conclusion: A small reduction in membrane cholesterol enlarges or increases lipid rafts concomitant with phosphorylation of intracellular proteins. Excessive removal of cholesterol, however, disorganizes rafts with a cessation of phosphorylation. These techniques are forcing a revision of long-held views on how lipids and proteins in sperm membranes are assembled into larger complexes that mediate recognition and fusion with the egg.

  19. COMPARISON OF CD46 EXPRESSION ON THE INNER ACROSOMAL MEMBRANE OF SPERMS FROM NORMOSPERMIC AND ASTHENOSPERMIC INDIVIDUALS

    Directory of Open Access Journals (Sweden)

    M NASR ESFAHANI

    2003-03-01

    Full Text Available Introduction: CD46 is a membrane cofactor protein (MCP of complement system wich is present on the membrane of all somatic cells except RBC. It is also present on the inner acrosmal membrane of human sperm. Thus, the aim of this study was to compare the expression of this prote, in on the inner acrosmal membrane of sperms from normospermic and asthenospermic individuals. Method: Semen from 6 normospermic and 17 asthenospermic individuals were examined for CD46 expression. After solublization of sperms in solublizing detergent, the solublized sperm membrane was separated from the rest of cell organelles by centrifugation. Solublized sperm membrane were divide to equal parts and SOS-PAGE gel was canied out in paired on the same gel for each sample. Western blot was carried out on half of the gel and then the nitrocellose papers were stained by a monocolonal Ab and HRP conjugate Ab. The other half were stained by silver stain for identification of MW. Results: After scoring the stained nitrocellose papen in each groups, no statistical significant difference was observed for C046 expression between the two groups. However, a significant Spearmen correlation was observed between CD46 expression and sperm motility (r=0.597, P=0.003. The MW of C046 was between 36 to 45 KD. with a mean of 42 KD. Discussion: This is the first report of a positive Spearmen correlation between sperm CD46 expression and sperm motility which suggest that there might be relation between CD46 expression and sperm motility.

  20. Membrane permeability parameters for freezing of stallion sperm as determined by Fourier transform infrared spectroscopy.

    Science.gov (United States)

    Oldenhof, Harriëtte; Friedel, Katharina; Sieme, Harald; Glasmacher, Birgit; Wolkers, Willem F

    2010-08-01

    Cellular membranes are one of the primary sites of injury during freezing and thawing for cryopreservation of cells. Fourier transform infrared spectroscopy (FTIR) was used to monitor membrane phase behavior and ice formation during freezing of stallion sperm. At high subzero ice nucleation temperatures which result in cellular dehydration, membranes undergo a profound transition to a highly ordered gel phase. By contrast, low subzero nucleation temperatures, that are likely to result in intracellular ice formation, leave membrane lipids in a relatively hydrated fluid state. The extent of freezing-induced membrane dehydration was found to be dependent on the ice nucleation temperature, and showed Arrhenius behavior. The presence of glycerol did not prevent the freezing-induced membrane phase transition, but membrane dehydration occurred more gradual and over a wider temperature range. We describe a method to determine membrane hydraulic permeability parameters (E(Lp), Lpg) at subzero temperatures from membrane phase behavior data. In order to do this, it was assumed that the measured freezing-induced shift in wavenumber position of the symmetric CH(2) stretching band arising from the lipid acyl chains is proportional to cellular dehydration. Membrane permeability parameters were also determined by analyzing the H(2)O-bending and -libration combination band, which yielded higher values for both E(Lp) and Lpg as compared to lipid band analysis. These differences likely reflect differences between transport of free and membrane-bound water. FTIR allows for direct assessment of membrane properties at subzero temperatures in intact cells. The derived biophysical membrane parameters are dependent on intrinsic cell properties as well as freezing extender composition. PMID:20553897

  1. Effects of cationic antimicrobial peptides on liquid-preserved boar spermatozoa.

    Directory of Open Access Journals (Sweden)

    Martin Schulze

    Full Text Available Antibiotics are mandatory additives in semen extenders to control bacterial contamination. The worldwide increase in resistance to conventional antibiotics requires the search for alternatives not only for animal artificial insemination industries, but also for veterinary and human medicine. Cationic antimicrobial peptides are of interest as a novel class of antimicrobial additives for boar semen preservation. The present study investigated effects of two synthetic cyclic hexapeptides (c-WFW, c-WWW and a synthetic helical magainin II amide derivative (MK5E on boar sperm during semen storage at 16 °C for 4 days. The standard extender, Beltsville Thawing Solution (BTS containing 250 µg/mL gentamicin (standard, was compared to combinations of BTS with each of the peptides in a split-sample procedure. Examination revealed peptide- and concentration-dependent effects on sperm integrity and motility. Negative effects were more pronounced for MK5E than in hexapeptide-supplemented samples. The cyclic hexapeptides were partly able to stimulate a linear progressive sperm movement. When using low concentrations of cyclic hexapeptides (4 µM c-WFW, 2 µM c-WWW sperm quality was comparable to the standard extender over the course of preservation. C-WFW-supplemented boar semen resulted in normal fertility rates after AI. In order to investigate the interaction of peptides with the membrane, electron spin resonance spectroscopic measurements were performed using spin-labeled lipids. C-WWW and c-WFW reversibly immobilized an analog of phosphatidylcholine (PC, whereas MK5E caused an irreversible increase of PC mobility. These results suggest testing the antimicrobial efficiency of non-toxic concentrations of selected cyclic hexapeptides as potential candidates to supplement/replace common antibiotics in semen preservation.

  2. Genome-wide association study for sperm membrane integrity in frozen-thawed semen of Holstein-Friesian bulls.

    Science.gov (United States)

    Kamiński, Stanisław; Hering, Dorota M; Oleński, Kamil; Lecewicz, Marek; Kordan, Władysław

    2016-07-01

    The aim of the study was to screen the entire bull genome to identify SNP markers and propose candidate genes potentially involved in the variation of sperm membrane integrity in Holstein-Friesian bulls. Two hundred eighty eight bulls kept in one AI center were included in the study. Each bull was genotyped for 54.001 Single Nucleotide Polymorpisms (SNP) by the Illumina BovineSNP50 BeadChip. Commercial straws of frozen-thawed semen were used for the evaluation of sperm plasma membrane integrity (SYBR-14/PI staining) and sperm mitochondrial function (JC1/PI staining). An additive model for Linear Regression Analysis was applied to estimate the effect of SNP marker for sperm membrane integrity (by the use of GoldenHelix SVS7 software). Five significant markers (encompassing 2,2 MB region located on chromosome 6) for SYBR-14/PI were found. Among them one marker-rs41570391 passed Bonferroni correction test. Within approximately 3 Mb genomic region including significant markers three candidate genes: SGMS2 (Sphingomyelin Synthase 2), TET2 (Methylcytosine dioxygenase 2) and GSTCD genes (Gluthatione S-transferase C terminal domain) were proposed as potentially involved in sperm membrane integrity in frozen-thawed semen of Holstein-Friesian bulls. PMID:27236378

  3. Bivalent response to long-term storage in liquid-preserved boar semen: a flow cytometric analysis.

    Science.gov (United States)

    Henning, Heiko; Petrunkina, Anna M; Harrison, Robin A P; Waberski, Dagmar

    2012-07-01

    The fertility of liquid-preserved boar semen declines during storage at 17°C, insemination trials even indicating early losses in fertilizing ability within the first 24-48 h of storage. Standard semen parameters barely reflect these changes in semen quality, and new approaches for assessment of functional changes in stored spermatozoa are needed. Capacitation, the essential prefertilization step for spermatozoa in the female genital tract, is specifically induced in vitro by bicarbonate. Therefore, we have investigated changes in responsiveness of boar spermatozoa to bicarbonate during storage. Ejaculates of 14 boars were diluted in Beltsville thawing solution, cooled to 17°C and stored for 12, 24, 72, 120, and 168 h before investigation. At each time, basic semen quality was characterized by sperm motility and viability. Subsequently, washed subsamples were incubated in variants of an in vitro fertilization (IVF) medium and assessed for kinetic changes of viability (plasma membrane integrity) and intracellular calcium concentration using flow cytometry in combination with propidium iodide and Fluo-3. By this means, it was possible to determine specific effects of bicarbonate and calcium on sperm subpopulations over incubation time. During storage, standard semen parameters remained on a high level. However, flow cytometric analysis of sperm responses to capacitating and control media revealed two opposing effects of storage. There was a loss of response to bicarbonate in part of the live sperm population but an increasing degree of instability in the rest. Assessment of response to capacitating media by flow cytometry appears a markedly more sensitive way of monitoring sperm functionality during storage than the standard semen parameters of motility and viability. PMID:22573481

  4. New Approaches to Boar Semen Evaluation, Processing and Improvement.

    Science.gov (United States)

    Sutovsky, P

    2015-07-01

    The improvement of boar reproductive performance may be the next frontier in reproductive management of swine herd in Unites States, facilitated by better understanding of boar sperm function and by the introduction of new advanced instrumentation in the andrology field. Objective single ejaculate evaluation and individual boar fertility prediction may be possible by introducing automated flow cytometric semen analysis with vital stains (e.g. acrosomal integrity and mito-potential), DNA fragmentation analysis and biomarkers (ubiquitin, PAWP, ALOX15, aggresome) associated with normal or defective sperm phenotypes. Measurement of sperm-produced reactive oxygen species (ROS) is a helpful indicator of normal semen sample. Semen ROS levels could be managed by the addition of ROS-scavenging antioxidants. Alternative energy regeneration substrates and sperm stimulants such as inorganic pyrophosphate and caffeine could increase sperm lifespan in extended semen and within the female reproductive system. Such technology could be combined with timed sperm release in the female reproductive system after artificial insemination. Sperm phenotype analysis by the image-based flow cytometry will go hand in hand with the advancement of swine genomics, linking aberrant sperm phenotype to the fertility influencing gene polymorphisms. Finally, poor-quality ejaculates could be rescued and acceptable ejaculates improved by semen purification methods such as the nanoparticle-based semen purification and magnetic-activated sperm sorting. Altogether, these scientific and technological advances could benefit swine industry, provided that the challenges of new technology adoption, dissemination and cost reduction are met. PMID:26174914

  5. The role of ubiquitin–proteasome pathway in the regulation of activity of two sperm surface proteins the aqn1 spermadhesin And the acrosin inhibitor in boar fertilization

    Czech Academy of Sciences Publication Activity Database

    Jonáková, Věra; Postlerová, Pavla; Yi, Y.J.; Sutovsky, P.; Pěknicová, Jana

    Praha: Biotechnologický ústav, 2013 - (Pěknicová, J.). s. 17-18 [XIX. Symposium imunologie a biologie reprodukce s mezinárodní účastí. 23.05.2013-25.05.2013, Třešť] R&D Projects: GA ČR GAP503/12/1834 Institutional research plan: CEZ:AV0Z50520701 Keywords : Ubiquitin-proteasome pathway * Sperm surface protein * Spermadhesin * Fertilization * Acrosin Subject RIV: CE - Biochemistry

  6. 31P MR Spectroscopy of the Testes and Immunohistochemical Analysis of Sperm 
of Transgenic Boars Carried N terminal Part of Human Mutated Huntingtin

    Czech Academy of Sciences Publication Activity Database

    Jozefovičová, M.; Herynek, V.; Jírů, F.; Dezortová, M.; Juhásová, Jana; Juhás, Štefan; Klíma, Jiří; Bohuslavová, Božena; Motlík, Jan; Hájek, M.

    2015-01-01

    Roč. 78, Suppl 2 (2015), s. 28-33. ISSN 1210-7859. [Conference on Animal Models for neurodegenerative Diseases /3./. Liblice, 08.11.2015-10.11.2015] R&D Projects: GA MŠk ED2.1.00/03.0124 Institutional support: RVO:67985904 Keywords : Huntington´s disease * testes * sperm Subject RIV: FH - Neurology Impact factor: 0.165, year: 2014

  7. Studies on the mechanism of capacitation: albumin-mediated changes in plasma membrane lipids during in vitro incubation of rat sperm cells.

    OpenAIRE

    Davis, B. K.; Byrne, R.; Bedigian, K

    1980-01-01

    Plasma membrane isolated from rat sperm cells after incubation in vitro had a significantly lower cholesterol/phospholipid mole ratio when the medium contained serum albumin. Transfer of albumin-bound phospholipids to the membrane can largely account for this effect. The result is broadly consistent with a previously proposed model for albumin-induced destabilization of sperm membrane (capacitation) and its reversal by seminal plasma membrane vesicles. Albumin also decreased sialic acid and, ...

  8. Effect of supplementation of butylated hydroxytoluene on post-thaw sperm viability, motility and membrane integrity of Hariana bulls

    Directory of Open Access Journals (Sweden)

    Akhil Patel

    2015-06-01

    Full Text Available Aim: This study was aimed to see the beneficial effect of butylated hydroxytoluene (BHT as a semen additive of Hariana bull semen. Materials and Methods: The study was carried out in Hariana bulls. Twenty-four ejaculates from two bulls were used for this study. Each ejaculate was extended with standard glycerolated egg yolk tris extender and supplemented with BHT at two concentrations as 0.5 mM (T1 and 1.0 mM (T2. After dilution, equilibration and 24 h of cryopreservation, the samples were analyzed for progressive motility, sperm viability and membrane integrity. Results: Progressive motility, sperm viability and sperm membrane integrity were significantly (p<0.05 increased in the samples fortified with BHT as compared to the control during the process of cryopreservation and thawing. The BHT concentration of 1 mM revealed better results as compared to 0.5 mM. Conclusion: Addition of 1.0 mM BHT was found better in cryopreservation of Hariana bull semen compared to 0.5 mM BHT and control samples. The addition of BHT has improved the sperm quality by acting as an antioxidant thereby reducing the lipid peroxidation of the sperms.

  9. Soluble products of Escherichia coli induce mitochondrial dysfunction-related sperm membrane lipid peroxidation which is prevented by lactobacilli.

    Directory of Open Access Journals (Sweden)

    Arcangelo Barbonetti

    Full Text Available Unidentified soluble factors secreted by E. coli, a frequently isolated microorganism in genitourinary infections, have been reported to inhibit mitochondrial membrane potential (ΔΨm, motility and vitality of human spermatozoa. Here we explore the mechanisms involved in the adverse impact of E. coli on sperm motility, focusing mainly on sperm mitochondrial function and possible membrane damage induced by mitochondrial-generated reactive oxygen species (ROS. Furthermore, as lactobacilli, which dominate the vaginal ecosystem of healthy women, have been shown to exert anti-oxidant protective effects on spermatozoa, we also evaluated whether soluble products from these microorganisms could protect spermatozoa against the effects of E. coli. We assessed motility (by computer-aided semen analysis, ΔΨm (with JC-1 dye by flow cytometry, mitochondrial ROS generation (with MitoSOX red dye by flow cytometry and membrane lipid-peroxidation (with the fluorophore BODIPY C11 by flow cytometry of sperm suspensions exposed to E. coli in the presence and in the absence of a combination of 3 selected strains of lactobacilli (L. brevis, L. salivarius, L. plantarum. A Transwell system was used to avoid direct contact between spermatozoa and microorganisms. Soluble products of E. coli induced ΔΨm loss, mitochondrial generation of ROS and membrane lipid-peroxidation, resulting in motility loss. Soluble factors of lactobacilli prevented membrane lipid-peroxidation of E. coli-exposed spermatozoa, thus preserving their motility. In conclusion, sperm motility loss by soluble products of E. coli reflects a mitochondrial dysfunction-related membrane lipid-peroxidation. Lactobacilli could protect spermatozoa in the presence of vaginal disorders, by preventing ROS-induced membrane damage.

  10. Influence of enterococci on human sperm membrane in vitro%肠球菌体外对人精子膜的影响

    Institute of Scientific and Technical Information of China (English)

    H.Qiang; M.S.Jiang; J.Y.Lin; W.M.He

    2007-01-01

    Aim:To study the influence of enterococci on human sperm membrane in vitro. Methods: Ejaculated human sperm were artificially infected with β-hemolytic or non-β-hemolytic enterococci at the bacteria: sperm ratio of 50:1 at 37℃.Sperm membrane integrity was examined after incubation for 1, 3 and 5 h by hypoosmotic swelling (HOS) test and electron microscopy. Results: Sperm infected with β-hemolytic enterococci had lower HOS scores compared with non-β-hemolytic strains or uninfected control (P < 0.01). The HOS test scores of sperm infected with β-hemolytic enterococci increased in the presence of phosphatidylcholine, an inhibitor of hemolysin. Non-β-hemolytic strains showed no significant difference in swelling rate, compared with the control group (P > 0.05). It was shown by electron microscopy that β-hemolytic enterococci caused significant rupture of human sperm membrane. Conclusion: β-hemolytic enterococci caused human sperm membrane injury, and might be mediated by the hemolysin of enterococci.

  11. Effects of L-Carnitine and Pentoxifylline on Carbohydrate Distribution of Mouse Testicular Sperm Membrane

    Directory of Open Access Journals (Sweden)

    Elham Aliabadi

    2013-06-01

    Full Text Available Background: The glycoconjugate content of sperms indicates their physiological and fertility properties. Lectin reactivity is indicative of intact, capacitated, and acrosome-reacted sperms. In the epididymis, sperms experience maturation, glycoconjugate modification, and simultaneously, higher L-carnitine (LC concentrations. The aim of this project was to evaluate the effects of LC and Pentoxifylline (PF on the integrity, capacitation, and acrosomal reaction of sperms by studying their lectin reactivity.Methods: Mouse testicular sperm samples were divided into three parts. Each sample was added Ham’s F10 (control or media containing 1.76 mM LC or PF. At 30 and 90 minutes after incubation, sperm motility was assessed. Peanut agglutinin (PNA, wheat germ agglutinin (WGA, and Concanavalin A (Con A were used to detect non-acrosome-reacted, non-capacitated, and acrosome-reacted sperms, respectively and the frequency was evaluated by flow cytometry. Statistical analysis was performed using the ANOVA. Results: Sperm motility increased after 30 and 90 minutes of incubation in the LC- and PF-treated cultures (P=0.001. LC administration created a significant increase in the percentage of the non-acrosome-reacted sperms compared to the control sperms after 30 and 90 minutes (P=0.02 and P=0.03, respectively. The frequency of the non-capacitated sperms in the LC-treated group increased compared to the control sperms after 30 minutes significantly (P=0.01. Conclusion: Although the administration of LC and PF enhanced sperm motility, LC also impacted glycoconjugates on the sperm surface. Glycoconjugates are involved in the interaction between the sperm and the zona pellucida and subsequently fertilization, thereby probably influencing the male fertility state.

  12. INFLUENCE OF CERTAIN BIOACTIVE PREPARATIONS ON THE DURATION OF BOAR SEMEN PRESERVATION

    Directory of Open Access Journals (Sweden)

    V. HAREA

    2013-07-01

    Full Text Available The experiences were held on the boar sperm. There were studied the bioactive substances with the role of antioxidizer made at the Institute of Genetic of Science Academy of Republic of Moldova. The bioactive substances (GL-2 were used as a structure dilution GHTS what is used for boars sperm dilution with the concentration of 0,1 – 1%. The experimental researches showed that the studied substances were not toxic for sperm used in the structure of GHTS dilution with the concentration of 0,1-1 whit gave the possibility to increase the period of boar sperm stoking till 168 hours, keeping the sperms mobility at the level of standard of artificial insemination.

  13. Nitric oxide induces caspase activity in boar spermatozoa.

    Science.gov (United States)

    Moran, J M; Madejón, L; Ortega Ferrusola, C; Peña, F J

    2008-07-01

    Nitric oxide (NO) is a highly reactive free radical that plays a key role in intra- and intercellular signaling. Production of radical oxygen species and an apoptotic-like phenomenon have recently been implicated in cryodamage during sperm cryopreservation. The objective of the present study was to evaluate the effect of sodium nitroprusside (SNP), an NO donor, on boar sperm viability. Semen samples were pooled from four boars that were routinely used for artificial insemination. Flow cytometry was used to compare semen incubated with SNP to control semen. Specifically, NO production was measured using the NO indicator dye diaminofluorescein diacetate, and caspase activity was determined using the permeable pan-caspase inhibitor Z-VAD linked to FITC. SNP induced a significant increase in the percentage of sperm cells showing caspase activity, from 9.3% in control samples to 76.2% in SNP-incubated samples (Pboar sperm damage. PMID:18433854

  14. The photon emission, ATP level and motility of boar spermatozoa during liquid storage.

    Science.gov (United States)

    Gogol, Piotr; Szcześniak-Fabiańczyk, Barbara; Wierzchoś-Hilczer, Agnieszka

    2009-03-01

    Changes were studied in induced photon emission (as an indicator of oxidative stress), ATP level and sperm motility during seven day-storage of boar semen at 15 degrees C extended with the use of BTS extender. Photon emission was measured using a luminometer equipped with a cooled photomultiplier with a spectral response range from 370 to 620 nm. The time of storage had a significant effect on luminescence parameters (integral and peak max), intracellular ATP level and percentage of motile spermatozoa. The increase in luminescence parameters was paralleled by a decrease in ATP level and sperm motility. A significant correlation was found between the percentage of motile spermatozoa and integral (r=-0.27) and peak max (r=-0.31). ATP level was correlated with integral (r=-0.25) but not with peak max. Our results suggest that reactive oxygen species and products of cell membrane lipid peroxidation have a negative effect on ATP level and sperm motility. Induced luminescence assessment in combination with sperm motility and ATP level can give valuable information about the status and function of spermatozoa which may be relevant for predicting the fertilizing potential of the semen. PMID:19352416

  15. Expression of Human Sperm Membrane Protein in the Recombinant Salmonella Typhimurium Vaccine

    Institute of Scientific and Technical Information of China (English)

    匡颖; 胡菁华; 翟玉梅; 缨时英; 王琳芳; 严缘昌

    1999-01-01

    A 550 bp cDNA fragment of HSD-Ⅰ coding for an extracellular domain of hu-man sperm membrane protein(hSMP-1)was ligated with an Adapter containing the universal stop codon,and the ligated fragment cDNA was then cloned into the MAS of pUC19.The desired plasmid with correct open reading frame was obtained, and was cut with EcoR Ⅰ.The insert was purified and then cloned into the two asd+ Salmonella expression vectors(the low copy number plasmid-pYA292 and the high copy number plasmid-pYA3137).The recombinant plasmid containing the insert with the correct orientation was selected by restriction enzyme digestion analysis. The recom-binant plasmids were transferred into the non-pathogenic Salmonella typhimurium X4550,which was deletion of the △cya, △crp and △asd genes.Western-blot analysis of the whole cell lysate of the two recombinants of S.typhimurium showed a predomi-nant protein band at 21 KD,which reacted with the anti-hSMP-1 antiserum. The re-sult indicated that two recombinants of S.typhimurium containing the 550 bp cDNA of HSD-Ⅰ were constructed and the characteristics of their growth in vitro were deter-mined. They may be used as new potential mucosalanti fertility.

  16. Methyl glycol, methanol and DMSO effects on post-thaw motility, velocities, membrane integrity and mitochondrial function of Brycon orbignyanus and Prochilodus lineatus (Characiformes) sperm.

    Science.gov (United States)

    Viveiros, Ana T M; Nascimento, Ariane F; Leal, Marcelo C; Gonçalves, Antônio C S; Orfão, Laura H; Cosson, Jacky

    2015-02-01

    The aim of this study was to use more accurate techniques to investigate the effects of cryoprotectants (CPAs) and extenders on post-thaw sperm quality of Brycon orbignyanus and Prochilodus lineatus. Six freezing media comprising the combination of three CPAs (DMSO, methanol and methyl glycol) and two extenders (BTS and glucose) were used. Sperm was diluted in each medium, loaded into 0.5-mL straws, frozen in a nitrogen vapor vessel (dry-shipper), and stored in liquid nitrogen at -196 °C. Post-thaw sperm motility rate and velocities (curvilinear = VCL; straight line = VSL; average path = VAP) were evaluated using a computer-assisted sperm analyzer. Membrane integrity and mitochondrial function were determined using fluorochromes. Post-thaw quality was considered high when samples presented the following minimum values: 60 % motile sperm, 140 µm/s of VCL, 50 % intact sperm membrane and 50 % mitochondrial function integrity. High post-thaw quality was observed in B. orbignyanus sperm frozen in BTS-methyl glycol and in P. lineatus sperm frozen in BTS-methyl glycol, glucose-methyl glycol and glucose-methanol. All samples frozen in DMSO yielded low quality. The presence of ions in the BTS extender affected post-thaw sperm quality positively in B. orbignyanus and negatively in P. lineatus. Methyl glycol was the most suitable CPA for both fish species, leading to a good protection of cell membrane, mitochondrial function and motility apparatus during the cryopreservation process. For an improved protection, B. orbignyanus sperm should be frozen in an ionic freezing medium. PMID:25433690

  17. Cryopreservation of boar semen

    OpenAIRE

    Eriksson, Bengt

    2000-01-01

    The world’s pig population is consistently being upgraded through the international trade of superior genetics. The two major systems that are used for this purpose are the transport of live animals and the export of frozen boar semen. The main limiting factors for a wider use of frozen-thawed (FT) boar semen are low fertility levels of FT in comparison with liquid semen, and between-boar variation in freezing success. Consequently, there is a need for improved boar semen freezing methods. Th...

  18. Intra- and interboar variability in flow cytometric sperm sex sorting.

    Science.gov (United States)

    Alkmin, Diego V; Parrilla, Inmaculada; Tarantini, Tatiana; Parlapan, Laura; Del Olmo, David; Vazquez, Juan M; Martinez, Emilio A; Roca, Jordi

    2014-08-01

    To improve the efficiency of porcine sperm sex sorting using flow cytometry, the aims of the present study were to determine the relevance of inter- and intraboar variability in sperm sortability and to evaluate the significance of ejaculate semen characteristics in such variability. In addition, the variability among boars in the ability of sex-sorted spermatozoa to survive liquid storage at 15 °C to 17 °C was also evaluated. In total, 132 ejaculates collected from 67 boars of different breeds that were housed at an artificial insemination center were used in three experiments. X- and Y-chromosome-bearing sperm were simultaneously separated according to the Beltsville sperm-sorting technology using a high-speed flow cytometer. In the first experiment, interboar variability in the ability of the ejaculated spermatozoa to undergo the flow-based sex-sorting procedure was observed; the ejaculates of nearly 15% of the boars (n = 67) did not exhibit well-defined X- and Y-chromosome-bearing spermatozoa peaks in the histogram, and the ejaculate sperm concentration demonstrated good predictive value for explaining this variation, as indicated by the area under the receiver operating characteristics curve (0.88, P boars that showed poor sperm sortability (measured according to the presence or not a well-defined split together with sperm sortability parameters) in the first ejaculate (n = 3). In contrast, boars classified as having good sperm sortability in the first ejaculate (n = 5) maintained this condition in five ejaculates collected over the subsequent 5 months. In the third experiment, sex-sorted spermatozoa from boars with good sperm sortability (n = 5) remained viable and motile (above 70% in all boars) after 48 hours of storage at 15 °C to 17 °C, which may facilitate the commercial application of sex-sorted spermatozoa in swine artificial insemination programs. PMID:24930604

  19. Improvement of bovine semen quality by removal of membrane-damaged sperm cells with DNA aptamers and magnetic nanoparticles.

    Science.gov (United States)

    Farini, Veronica L; Camaño, Carla V; Ybarra, Gabriel; Viale, Diego L; Vichera, Gabriel; Yakisich, Juan S; Radrizzani, Martín

    2016-07-10

    In cattle, cryopreservation of semen and sex-sorting kill up to 50% of spermatozoa and decrease the success of assisted insemination (AI). Therefore, significant efforts are being carried out to improve the quality of semen prior to AI. In this work we used the Cell-SELEX technique to select single strand DNA aptamers able to recognize with high affinity and specificity damaged sperm cells generated by heat-treatment. We first isolated aptamers with a conserved two motifs of 6 nucleotides of length that bind to the membrane of heat-treated spermatozoa. Then, we used synthetic biotin-labeled aptamers containing the conserved motif to recognize membrane-damaged cells and separate them from viable cells by the use of avidin-coated superparamagnetic iron oxide nanoparticles (SPION). This procedure improved the quality of semen by significantly increasing the percentage of healthy sperm cells without affecting the rate of blastocyst cleavage. This technique was successfully applied to both unsorted and sex-sorted sperm suspension. PMID:27164256

  20. Osmotic effects on volume and motility of bull sperm exposed to membrane permeable and nonpermeable agents.

    Science.gov (United States)

    Liu, Z; Foote, R H

    1998-11-01

    Factorially arranged experiments were designed to study prefreeze packed cell volume (PCV) changes and associated percentages of motile and unstained bull sperm in simple macromolecule-free Tyrode's solution and egg yolk-Tris (EYT), varying in osmolarity, and with addition of rapidly permeating cryoprotectants, glycerol and 1,2-propanediol, and nonpermeating substances, sucrose and NaCl. The percentage of motile and unstained sperm was assessed after resuspending sperm in 300 mOsm/L Tyrode's solution. At 25 degreesC PCV increased in Tyrode's solution as osmolarity was decreased from 250 to 150 mOsm/L and decreased as Tyrode's solution was increased to 400 mosmol/L. The relationship of PCV to the reciprocal of the osmolarity was essentially linear over the range of 150 to 400 mOsm/L, but PCV did not decrease further in solutions ranging from 500 to 1000 mOsm/L. The percentage of motile sperm declined to zero in Tyrode's solution at 700 mOsm/L, but 40% of the sperm were still unstained in 1000 mOsm/L solutions. The addition of glycerol or 1,2-propanediol had little effect on PCV. With glycerol or 1,2-propanediol added to 308 mOsm/L Tyrode's solution to give a total of 1267 mOsm/L, there were 49 and 56% motile sperm, respectively, compared to 1% with NaCl added to give 787 mOsm/L. The PCV and percentage of motile sperm suspended in EYT responded to osmotic changes similar to those reported for Tyrode's solution at both 25 and 5 degreesC. Some sperm remained motile after initial exposure to 800 mOsm/L solutions. These findings may have application in improving bull sperm cryopreservation. PMID:9787066

  1. Characterization of cDNA encoding a human sperm membrane protein related to A4 amyloid protein.

    OpenAIRE

    Yan, Y C; Bai, Y.(Institute of High Energy Physics, Chinese Academy of Sciences, Beijing, China); Wang, L.F.; Miao, S Y; Koide, S S

    1990-01-01

    A rat testis lambda gt11 cDNA library was screened with a monoclonal antibody raised against a human sperm membrane protein designated YWK-II. A clone was found with a cDNA insert composed of 1837 base pairs that contained an open reading frame coding for 191 amino acid residues. The deduced polypeptide contained a segment with high homology to the transmembrane-cytoplasmic domains of the A4 amyloid protein found in brain plaques of Alzheimer disease patients. A sequence of basic amino acid r...

  2. Beneficial effects of relaxin on motility characteristics of stored boar spermatozoa

    OpenAIRE

    Feugang, Jean M; Rodríguez-Muñoz, Juan C; Dillard, Darby S; Crenshaw, Mark A; Willard, Scott T.; Ryan, Peter L.

    2015-01-01

    Background Relaxin is detected in seminal plasma of many species and its association with sperm motility may be beneficial in some aspects of assisted reproduction. Here, we immunolocalized relaxin receptors and investigated the effects of exogenous relaxin on motility characteristics, viability, and cAMP content of boar spermatozoa after storage. Methods Commercial doses of boar semen were obtained on the collection day (Day 0) and kept in shipping containers at room temperature for up to 4 ...

  3. The effect of melatonin on the quality of extended boar semen after long-term storage at 17 °C.

    Science.gov (United States)

    Martín-Hidalgo, D; Barón, F J; Bragado, M J; Carmona, P; Robina, A; García-Marín, L J; Gil, M C

    2011-05-01

    Melatonin (MLT) is an efficient antioxidant that protects cells and tissues and initiates a host of receptor-mediated effects. In order to enhance the life span of refrigerated boar semen, our aim was to evaluate the effects of addition of 1 μM MLT to commercially produced pig semen (33 seminal doses from 14 boars) that had been preserved at 17 °C for 7 days. Samples without MLT served as controls. On Days 1, 4 and 7, we evaluated motility parameters and the percentage of total motile and progressively motile spermatozoa by a computer-aided sperm analysis system. Viability (SYBR-14/PI), acrosomal status (FITC-PNA/PI), membrane fluidity (M-540/YoPro-1) and mitochondrial membrane potential status (JC-1) were evaluated by flow cytometry. MLT treatment significantly enhanced the percentage of static spermatozoa after 7 days of storage and significantly reduced the percentage of progressively motile spermatozoa on Day 7. The velocity characteristics (VCL, VSL and VAP) were significantly higher for MLT-treated samples on Day 1 and were their lowest on Day 7. With regard to flow cytometry results, the percentage of viable spermatozoa with an intact acrosome was higher in MLT samples throughout the entire storage period. In addition, there was a significantly higher proportion of live spermatozoa on Day 7 in the samples that had not been treated with MLT. The proportion of spermatozoa showing a high mitochondrial membrane potential remained at similar levels (P > 0.05) throughout the trial. Although the findings of the present study revealed that 1 μM MLT increased the proportion of live sperm with an intact acrosome, this treatment did not enhance the spermatic quality of refrigerated boar semen. PMID:21320723

  4. Sperm membrane integrity in fresh and frozen-thawed canine semen samples: a comparison of vital stains with the NucleoCounter SP-100.

    Science.gov (United States)

    Daub, L; Geyer, A; Reese, S; Braun, J; Otzdorff, C

    2016-07-15

    The objective of this study was to assess sperm membrane integrity in canine semen samples using three different vital stains and the NucleoCounter SP-100 (NC). In addition, the occurrence of half-stained sperm heads, the influence of investigator, and storage-related artifacts using stained smears were examined. Forty fresh (30 dogs) and 40 frozen-thawed (28 dogs) canine semen samples were analyzed. The vital stains eosin (E), eosin-nigrosin (EN), and bromphenolblue-nigrosin (BN) were compared. Two smears per stain were prepared and a total of 200 sperm per slide were classified using bright field microscopy. Each slide was examined twice by three investigators. Spermatozoa with completely red (E, EN) or blue (BN) stained sperm heads were classified as "dead". Half-stained sperm heads were counted separately. Sperm concentration and viability were determined using the NC. The NC works with a built-in fluorescence microscope using propidium iodide as a fluorescence dye. Statistical analysis for comparison of results was made using mean values with standard deviation, Bland-Altman plot and coefficient of variation (CV). Staining with E led to a significant higher percentage of dead sperm compared with EN and BN (P < 0.05), which gave comparable results. Vital stains revealed higher CVs (range 8.8%-32.1%) than the NC (<6.5%). Interobserver viability ranged from 17.5% to 45.4% and was within the same range between stains. If only completely stained sperm heads were considered, best agreement was found between the E and the NC. In case of EN and BN, inclusion of half-stained sperm heads reduced the difference compared with NC. In general, the agreement between methods was better in samples with a low percentage of dead spermatozoa. In smears of fresh semen stored up to 3 months, no increase in the percentage of dead spermatozoa could be observed. In some smears of frozen-thawed samples stained with E (n = 12) or BN (n = 2), all previously unstained spermatozoa

  5. Rooster sperm plasma membrane protein and phospholipid organization and reorganization attributed to cooling and cryopreservation

    Science.gov (United States)

    Cholesterol to phospholipid ratio is used as a representation for membrane fluidity, and predictor of cryopreservation success but results are not consistent across species and ignore the impact of membrane proteins. Therefore, this research explored the modulation of membrane fluidity and protein ...

  6. Effect of natural betaine on estimates of semen quality in mature AI boars during summer heat stress.

    Science.gov (United States)

    Cabezón, F A; Stewart, K R; Schinckel, A P; Barnes, W; Boyd, R D; Wilcock, P; Woodliff, J

    2016-07-01

    This study evaluated the effect of supplemental dietary betaine at three concentrations (0.0%, 0.63% and 1.26%) on semen characteristics, quality and quality after storage on boars. The trial was conducted between 22 July and 1 October 2014 in a boar stud located in Oklahoma. Boars were blocked by age within genetic line and randomly allotted to receive 0% (CON, n (line T)=22, n (line L)=10), 0.63% (BET-0.63%, n (line T)=21, n (line L)=6) or 1.26% (BET-1.26%, n (line T)=23, n (line L)=7). The diets containing betaine were fed over 10 weeks, to ensure supplemental betaine product (96% betaine) daily intakes of 16.34 and 32.68g, for the BET-0.63% and BET-1.26% diets, respectively. Serum homocysteine concentrations were less for animals with betaine treatments (P=0.016). Rectal temperatures of the boars were unaffected by betaine diets. Betaine tended to increase total sperm in the ejaculates when collectively compared with data of the control animals (P=0.093). Sperm morphology analysis indicated there was a greater percent of sperm with distal midpiece reflex (P=0.009) and tail (P=0.035) abnormalities in boars fed the BET-1.26% than boars fed the BET-0.63% diet. Betaine concentration in the seminal plasma was greater in boars with betaine treatments, with animals being fed the 0.63% and 1.26% diets having 59.2% and 54.5% greater betaine concentrations in seminal plasma as compared with boars of the control group (P=0.046). In conclusion, betaine supplementation at 0.63% and 1.26% tended to increase sperm concentration in the ejaculates by 6% and 13%, respectively, with no negative impacts on semen quality when 0.63% of betaine was included in the diet. PMID:27095614

  7. EFFECT OF REDUCING THE NUMBER OF SPERM CELLS (PER INSEMINATION, INCREASING ENERGY AND CRYOPROTECTING CONCENTRATIONS ON MOTION CHARACTERISTICS AND MEMBRANE INTEGRITY IN FROZEN THAWED BUFFALO SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    A.Abbas, S. M. H. Andrabiand N. Ahmad

    2001-07-01

    Full Text Available The objective of this study was to find out the minimum number of sperm cells per dose of insemination that will not affect the conception rate, if levels of egg yolk and glycerol are increased. For this purpose, sperm motion characteristics and plasma membrane integrity in three concentrations of sperm cells per dose, each with two levels of egg yolk and glycerol were compared. Semen was collected from buffalo bulls using an artificial vagina. Split pool ejaculates possessing more than 60% visual sperm motility were diluted in Tris-citric acid (TCA extender at 37°C, either in (1 30-6-20 (Number of sperm cells in millions/0.5 ml insemi1:1ation dose-Glycerol%-Egg yolk %,(2 15-6-20, (3 7.5-6-20, (4 30-10- 20, (5 15-10-20, (6 7.5-10-20, (7 30-6-30, (8 15-6-30, (9 7.5-6-30, (10 30-10-30, (11 15-10-30 and (12 7.5-10-30. Semen was cooled to 4°C in 2 hours, equilibrated at 4aC for 4 hours, filled in 0.5 ml straws and frozen in a programmable cell freezer ( + 4 to -15°C @ -3°C/minute and then to -80°C@ -10°C/minute before plunging them into liquid nitrogen (-196°C. Thawing of frozen semen was performed after 24 hours at 37°C for 15 seconds. Sperm motion characteristics, including motilities (computer-assisted, linear and circular, velocities (straight-line, average path and curvilinear, and lateral head displacement (LHD were assessed using computer assisted semen analyzer (CASA. Plasma membrane integrity was determined by using Hypo-Osmotic Swelling assay (HOS. Analysis of variance revealed that visual motility, computer-assisted motility, linear motility, circular motility, and plasma membrane integrity did not vary significantly when cell number was reduced from 30x106/dose to 15x106/dose. However, visual motility, computer-assisted motility and plasma membrane integrity were reduced significantly (P<0.05 when cell number was decreased to 7.5x106/dose. The velocities (straight- line, average path, curvilinear and LHD did not vary

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

    Directory of Open Access Journals (Sweden)

    J. L. Yániz

    2013-03-01

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

  9. Heat shock protein A8 stabilizes the bull sperm plasma membrane during cryopreservation: Effects of breed, protein concentration, and mode of use.

    Science.gov (United States)

    Holt, W V; Del Valle, I; Fazeli, A

    2015-09-15

    Heat shock protein A8 (HSPA8) is a highly conserved member of the Hsp70 family, which is expressed in oviductal cells, translocated into oviductal fluid, and becomes attached to the sperm surface during sperm transport. Previous research has shown that HSPA8 supports mammalian sperm viability during in vitro incubation at both 5 °C and body temperature. The present series of experiments was designed to explore the possibility that bovine recombinant HSPA8 might therefore protect bull spermatozoa during cryopreservation through its beneficial effects on the sperm plasma membrane. Soy-based cryopreservation media were used in these experiments. The effects of HSPA8 addition before freezing were examined at concentrations ranging from 0.2 to 6.4 μg/mL, whereas the effects of postthaw HSPA8 addition were tested between 0.2 and 12.8 μg/mL. When bull spermatozoa (from beef and dairy breeds) were frozen in the presence of HSPA8, beneficial but complex effects on postthaw viability were observed. Low HSPA8 concentrations (0.2 and 0.4 μg/mL) resulted in significantly reduced postthaw sperm viability, but concentrations above 0.8 μg/mL improved plasma membrane integrity. If HSPA8 was added to spermatozoa after thawing, outcomes were also biphasic and beneficial effects on viability were only seen if the HSPA8 concentration exceeded 3.2 μg/mL. Beneficial effects were significantly more apparent with beef rather than dairy breeds. When HSPA8 was used in combination with cholesterol-loaded cyclodextrin, spermatozoa from the beef breeds showed significantly lower apoptotic effects. This was not observed with the dairy breeds. PMID:26047707

  10. THE ADDITION OF CAFFEINE IN EARLE’S BALANCED SALT SOLUTION MEDIA WITH WASHING UP METHOD INCREASE MEMBRANE INTEGRITY AND ACROSOMAL SPERM

    Directory of Open Access Journals (Sweden)

    B. K. Satriyasa

    2014-12-01

    Full Text Available Background: caffeine, a methylxanthine derivate, appears to inhibit phosphodiesterase, thereby inhibiting the break down of cAMP and increasing its concentration inside cell. This study aims to assess the effect of caffeine addition in Earles’s Balanced Salt Solution (EBSS on the increase in membrane integrity and acrosome reaction of spermatozoa using swim up method. Methods: This study was carried out at the Clinic of Sexology and Andrology, Sanglah Public Hospital at Denpasar Bali-Indonesia. This study was an experimental study using the design of pre and post test paired control group design. The samples were sperm specimens of eighteen infertile couple male or volunteers who were infertile with age ranged between 20-40 years old. The samples   were divided into two groups: treatment group (caffeine + EBSS and control group (EBSS. The data were analysed statistically by normality test (Kolmogorov - Smirnov Goodness of Fit Test, Homogeneity test, and Paired Student’s t test.  Results: The results showed that the caffeine addition in EBSS medium could increase significantly (p<0.05.  The integrity of the sperm membrane obtained were from 81.30 % to 86.60 % and acrosomal reaction from 82.60% to 89.60% evaluated by hypo-osmotic swelling test (HOS. The conclusion of this study is that addition of caffeine in EBSS medium increases significantly membrane integrity and acrosomal reaction of the human sperm.

  11. Arachidonate 15-lipoxygenase and ubiquitin as fertility markers in boars.

    Science.gov (United States)

    Lovercamp, K W; Safranski, T J; Fischer, K A; Manandhar, G; Sutovsky, M; Herring, W; Sutovsky, P

    2007-03-01

    Accurate semen analysis is an important issue in the swine industry. We evaluated two candidate fertility marker proteins associated with sperm cytoplasmic droplet (CD), including 15-lipoxygenase (15-LOX) and ubiquitin (UBI) in a controlled single-sire artificial insemination (AI) trial. Ejaculates (n=116) were collected from 18 fertile Large White boars monthly for 8 mo, and analyzed by semi-quantitative, densitometry-based Western blotting and flow cytometry with antibodies against 15-LOX and UBI. Data were correlated with farrowing rates (FR) and total numbers of piglets born (TNB) from 1754 AI services by 13 of 18 boars, and compared with a conventional microscopic semen analysis. In semi-quantitative Western blotting, both 15-LOX and UBI were correlated with seasonal changes in the percentage of normal (r=-0.38, Pflow cytometry, UBI and 15-LOX levels showed seasonal changes coinciding with seasonal changes of FR and TNB, representing 13 boars, 88 ejaculates and 1,232 AI services. There were correlations between flow cytometric values of UBI and FR (r=0.31; PFlow cytometric measurements of 15-LOX correlated negatively with TNB (r=-0.33; Pboar fertility estimation could be achieved within a group of fertile boars by the use of objectively measurable fertility markers. Flow cytometry appeared more informative and more practical than semi-quantitative Western blotting. This technology could be further optimized for the selection of the most fertile sires in an artificial insemination program. PMID:17116325

  12. Characterizing the glycocalyx of poultry spermatozoa; semen cryopreservation methods alter the carbohydrate component of rooster sperm membrane glycoconjugates

    Science.gov (United States)

    The carbohydrate-rich zone on the sperm surface is essential for inmunoprotection in the female tract and early gamete interactions. We recently have shown the glycocalyx of chicken sperm to be extensively sialylated and contain residues of mannose, glucose, galactose, fucose, N-acetyl-galactosamine...

  13. Nutritional plans for boars

    Directory of Open Access Journals (Sweden)

    Charles Kiefer

    2012-06-01

    Full Text Available The objective of the present study was to evaluate nutritional plans for boars. Four hundred animals of 67 to 135 days of age and initial weight of 27.75±1.61 kg were distributed in a randomized block design with seven nutritional plans for boars (9.0-8.0; 9.0-9.0; 10.0-9.0; 10.0-10.0; 11.0-10.0; 11.0-11.0 and 12.0-11.0 g/kg of digestible lysine from 67 to 107 days and from 108 to 135 days, respectively with four repetitions and a control plan for barrows (11.0-10.0 g/kg of digestible lysine with eight repetitions and ten animals each. Uncastrated male swine presented better feed conversion; however they showed a lower marbling degree in relation to barrows, regardless of the nutritional plan. The nutritional plan that corresponds to the sequence of 11.0-10.0 g/kg of digestible lysine from the 67 to the 107 days and from the 108 to the 135 days, respectively, meets the nutritional needs of boars.

  14. Development of an in vitro index to characterize fertilizing capacity of boar ejaculates.

    Science.gov (United States)

    Schulze, M; Ruediger, K; Mueller, K; Jung, M; Well, C; Reissmann, M

    2013-07-01

    The aim of this research was the selection of spermatozoa parameters related to boar fertility performance and their combination into an in vitro index. A first set (data set 1) of 36 Pietrain boars with 138 ejaculates from two seasons with 5083 single-sire inseminations from 34 farms was used to determine correlations between in vitro sperm quality parameters and fertility performance. 2970 ejaculates representing a second set (data set 2) served calculation of seasonal and age effects on semen quality. Morphological spermatozoa parameters were estimated manually with a phase contrast microscope on the day of semen collection, whereas mitochondrial activity and viability were analyzed by double-staining with rhodamine123/propidium iodide on day 2 of semen storage using flow cytometry. Sperm motility was tested on day 7 by thermoresistance (TRT) after 30min (TRT1) and 300min (TRT2) incubation at 38̊C using computer-assisted semen analysis (CASA). Correlations revealed four independent sperm quality parameters qualifying as relevant predictors of boar fertility: (i) percentage of spermatozoa with proximal cytoplasmic droplets, (ii) percentage of spermatozoa with active mitochondria, (iii) beat cross frequency of progressively motile spermatozoa in TRT1, and (iv) oscillation measure of the actual path of progressively motile spermatozoa in TRT2. There were no significant effects of sperm concentration, ejaculate volume, and total number of sperm cells per ejaculate on litter size (LS) and on pregnancy rate (PR). Our findings suggest the usefulness of sperm quality parameters based on adjusted range of methods and enable the construction of an in vitro index as a means to predicting boar fertility. PMID:23773327

  15. Application of Antioxidant During Boar Semen Cryopreservation%抗氧化剂在公猪精液冷冻保存中的应用

    Institute of Scientific and Technical Information of China (English)

    张伟; 易康乐; 燕海峰; 王春强; 周虚

    2011-01-01

    In recent years, the application of antioxidant during the cryopreservation of boar semen in order toimprove quality of post-thaw semen attracted considerable research efforts.The data revealed that the effect of supplemented glutathione,superoxide and catalase,alpha-tocopherol,L-cysteine and L-glutamine, Chinese herbal medicine in extender could effectively prevented damage,improved kinematics parameters, protected sperm acrosome integrity and plasma membrane integrity, and increasing the sperm fertilizability after freezed-thawed.This review will contribute to make a better understanding of these antioxidants on the mechanisms of resistance to sperm cryodamage, develop an objective evaluation of effectiveness and estimate the applied prospect during the cryopreservation of boar semen.%近几年来,在公猪精液冷冻稀释液中添加抗氧化剂以提高冷冻精液质量的研究受到广泛的关注.添加谷胱甘肽、超氧化物歧化酶和过氧化物酶、维生素E、L-半胱氨酸和L-谷氨酰胺、中草药成分等抗氧化剂可以有效地防止氧化损伤,提高解冻后精子活率、精子成活力、质膜与顶体完整性等,进而提高冷冻保存精液的受精能力.作者综述了抗氧化剂的抗精子冷冻损伤的作用机制及不同抗氧化剂的应用效果,并对其在公猪精液冷冻保存中的应用前景进行了展望.

  16. Mutation in the porcine SERPINA7 gene and its association with boar fertility

    Institute of Scientific and Technical Information of China (English)

    REN Dongren; REN Jun; XING Yuyun; MA Junwu; WU Yanbo; GUO Yuanmei; HUANG Lusheng

    2006-01-01

    The porcine SERPINA7 gene is considered as a positional candidate gene responsible for testis size for its location on X chromosome and its biologically critical role in the development of testis. A nonsynonymous polymorphism (His226Asn or C678A) in the ligand-binding domain of SERPINA7 has been identified, which alters SERPINA7' s affinity to thyroxine and is closely associated with testis size. In this study, a primer mutagenesis strategy was developed to genotype this polymorphism in Chinese indigenous pigs and some western commercial pigs. The C allele existed in all tested Chinese indigenous and wild pigs, while the A allele is specific for western commercial breeds, indicating the occurrence of the mutation is of western origin. The correlation of this polymorphism with different boar fertility traits was assessed using a White Duroc × Erhualian intercross which included 110 F2 mature boars. The results showed that the C678A polymorphism was closely associated with testis weight and epididymis weight( P<0.0001 and P = 0. 0016, respectively) with significant heavier testis weight and epididymis weight in boars carrying the A allele than boars with the C allele. A significant correlation was also observed between this polymorphism and total sperm in the ejaculate ( P<0.01 ) as well as semen volume ( P<0.05). No statistically significant association of the C678A polymorphism with sperm concentration and sperm motility was found.

  17. Age-related changes in semen quality characteristics and expectations of reproductive longevity in Duroc boars.

    Science.gov (United States)

    Huang, Yu Hung; Lo, Ling Ling; Liu, Shyh Hwa; Yang, Tien Shuh

    2010-08-01

    Quadratic fitting was used to regress semen characteristics of 1441 samples consisting of 12-month collection from 58 Duroc boars against animal age varied from 10 to 80 months. Data was divided into two groups of cool (14.0-22.7 degrees C, RH 81.5%) and hot season (22.9-29.9 degrees C, RH 86.6%), to test effects of age, season and their interactions. Results revealed that young boars of around 1 year old could endure the hot season. The endurance gradually diminished as animals grew. In the hot season animals exhibited peak performance at age around 33 month and it remained for 1 month, while cool-season kept boars could last for 48 months from 16 months old onward. The reproductive longevity should be 51 month in a subtropical environment and it may extend to 70 month if heat stress can be avoided. The estimated total sperm contribution of a Duroc boar would be 1.8 times more when kept below 22 degrees C than in a natural subtropical environment. It is concluded that to maintain Duroc boars as semen donor to at least 4 years of age is feasible in a subtropical environment and boar longevity could reach 6 years old if well kept in a temperate region. PMID:20662811

  18. Metabolic incorporation of unsaturated fatty acids into boar spermatozoa lipids and de novo formation of diacylglycerols

    DEFF Research Database (Denmark)

    Svetlichnyy, V.; Müller, P.; Günther-Pomorski, Thomas;

    2014-01-01

    Lipids play an important role in the maturation, viability and function of sperm cells. In this study, we examined the neutral and polar lipid composition of boar spermatozoa by thin-layer chromatography/mass spectrometry. Main representatives of the neutral lipid classes were diacylglycerols...

  19. Mitochondria functionality and sperm quality.

    Science.gov (United States)

    Amaral, Alexandra; Lourenço, Bárbara; Marques, Mónica; Ramalho-Santos, João

    2013-01-01

    Although mitochondria are best known for being the eukaryotic cell powerhouses, these organelles participate in various cellular functions besides ATP production, such as calcium homoeostasis, generation of reactive oxygen species (ROS), the intrinsic apoptotic pathway and steroid hormone biosynthesis. The aim of this review was to discuss the putative roles of mitochondria in mammalian sperm function and how they may relate to sperm quality and fertilisation ability, particularly in humans. Although paternal mitochondria are degraded inside the zygote, sperm mitochondrial functionality seems to be critical for fertilisation. Indeed, changes in mitochondrial integrity/functionality, namely defects in mitochondrial ultrastructure or in the mitochondrial genome, transcriptome or proteome, as well as low mitochondrial membrane potential or altered oxygen consumption, have been correlated with loss of sperm function (particularly with decreased motility). Results from genetically engineered mouse models also confirmed this trend. On the other hand, increasing evidence suggests that mitochondria derived ATP is not crucial for sperm motility and that glycolysis may be the main ATP supplier for this particular aspect of sperm function. However, there are contradictory data in the literature regarding sperm bioenergetics. The relevance of sperm mitochondria may thus be associated with their role in other physiological features, particularly with the production of ROS, which in controlled levels are needed for proper sperm function. Sperm mitochondria may also serve as intracellular Ca²⁺ stores, although their role in signalling is still unclear. PMID:23901129

  20. Cryopreservation of sea urchin (Evechinus chloroticus) sperm.

    Science.gov (United States)

    Adams, Serean L; Hessian, Paul A; Mladenov, Philip V

    2004-01-01

    A method was developed for cryopreserving sperm of the sea urchin, Evechinus chloroticus. Sperm fertilisation ability, mitochondrial function and membrane integrity were assessed before and after cryopreservation. Highest post-thaw fertilisation ability was achieved with lower concentrations (2.5%-7.5%) of dimethyl sulphoxide (DMSO). In contrast, post-thaw mitochondrial function and membrane integrity were higher for sperm frozen in intermediate and high DMSO concentrations (5%-15%). Surprisingly, some sperm frozen in seawater only, without DMSO, were able to survive post-thawing, although the fertilisation ability (10(6) sperm/ml; approximately 50% fertilisation), mitochondrial function and membrane integrity of these sperm were notably lower than of sperm frozen with DMSO (10(6) sperm cells/ml; 2.5%-7.5% DMSO; >85% fertilisation) at the concentrations tested. Amongst sperm from individual males, fertilisation ability varied before and after cryopreservation for both males frozen with and without cryoprotectant. Specific differences among males also varied. Sperm mitochondrial function and membrane integrity was similar among males before cryopreservation but differed considerably after cryopreservation. Cryopreserved sperm were able to fertilise eggs and develop to pluteus stage larvae. This study has practical applications and will provide benefits such as reduced broodstock conditioning costs, control of parental input and opportunities for hybridisation studies. PMID:15375439

  1. Profiling of relaxin and its receptor proteins in boar reproductive tissues and spermatozoa

    OpenAIRE

    Feugang, Jean M; Greene, Jonathan M; Sanchez-Rodríguez, Hector L; Stokes, John V; Crenshaw, Mark A; Willard, Scott T.; Ryan, Peter L.

    2015-01-01

    Background Relaxin levels in seminal plasma have been associated with positive effects on sperm motility and quality, and thus having potential roles in male fertility. However, the origin of seminal relaxin, within the male reproductive tract, and the moment of its release in the vicinity of spermatozoa remain unclear. Here, we assessed the longitudinal distribution of relaxin and its receptors RXFP1 and RXFP2 in the reproductive tract, sex accessory glands, and spermatozoa of adult boars. M...

  2. Effect of dietary administration of oil extract from rosemary on reproductive efficiency in boars

    Directory of Open Access Journals (Sweden)

    A. Bonomi

    2010-01-01

    Full Text Available A decrease in reproductive performance in boars during and immediately after hot summer weather has been previously reported (Park and Yi, 2002. High temperature causes germ-cell destruction and results in a temporary decrease in sperm production and fertility. The increase of metabolic activity following thermic stress matches with a higher production of free radicals that impairs cells, such as spermatozoa, particularly rich in polyunsatured fatty acids and poor in antioxidants systems.

  3. Effect of dietary administration of oil extract from rosemary on reproductive efficiency in boars

    OpenAIRE

    A. Bonomi; Beretti, V.; Talarico, L.; P. Superchi

    2010-01-01

    A decrease in reproductive performance in boars during and immediately after hot summer weather has been previously reported (Park and Yi, 2002). High temperature causes germ-cell destruction and results in a temporary decrease in sperm production and fertility. The increase of metabolic activity following thermic stress matches with a higher production of free radicals that impairs cells, such as spermatozoa, particularly rich in polyunsatured fatty acids and poor in antioxidants systems.

  4. Seminal plasma and sperm surface proteins in reproduction

    Czech Academy of Sciences Publication Activity Database

    Jonáková, Věra; Postlerová, Pavla; Davidová, Nina; Tichá, M.; Šutovský, P.; Pěknicová, Jana

    Praha: BTO-N, 2009. s. 31-32. [XV. Symposium českých reprodukčních imunologů s mezinárodní účastí. 29.05.2009-31.05.2009, Žďár nad Sázavou] R&D Projects: GA MŠk(CZ) 1M06011; GA ČR GA303/09/1285; GA ČR GD523/08/H064 Institutional research plan: CEZ:AV0Z50520701 Keywords : sperm surface protein * ubiqutin C-terminal hydrolase * boar spermadhesin * AQN 1 * boar seminal plasma Subject RIV: EC - Immunology

  5. The Correlation of Sperm Chromatin Decondensation Following In Vitro Exposure to Heparin and Sperm Penetration Rates

    OpenAIRE

    Carrell, Douglas T.; Emery, Benjamin R.; Peterson, C. Matthew

    1998-01-01

    Purpose:The aim of this study was to evaluate the possible correlation of low-dose heparin-induced decondensation of sperm chromatin with sperm concentration, motility, morphology, membrane hypoosmotic response, ejaculate volume, and the ability of sperm to penetrate zona-free hamster oocytes.

  6. Detecting subtle changes in sperm membranes in veterinary andrology%兽医男科学中精子膜细微变化的检测

    Institute of Scientific and Technical Information of China (English)

    F. J. Pe(n)a

    2007-01-01

    在过去十年中由于流式细胞仪在兽医精子学中得到越来越多的应用,许多新的细胞膜完整性检测方法被开发出来.这些检测方法比精子的运动性监测等其它的检测更有助于预测生殖能力,因而显得十分重要.精子质量评估的主要部分演变为能在较早时期检测到精子损伤的检测.利用磷脂移位测试法,能通过荧光探针结合流式细胞仪评估大量精子的细胞膜渗透性和流动性早期的变化.%Thanks to the increasing use of flow cytometry in research in veterinary spermatology, many new membrane integrity assays have been developed over the past decade. These assays are important because of their superior ability to forecast fertility when compared with other tests, such as sperm motility. This major component of the sperm quality assessment has generated new investigations with the aim of developing tests that can detect membrane damage in a very early state. Using phospholipid transposition tests, early changes in membrane permeability and fluidity can be assessed in a large number of spermatozoa using fluorescent probes in combination with flow cytometry.

  7. Cationic synthetic peptides: assessment of their antimicrobial potency in liquid preserved boar semen.

    Directory of Open Access Journals (Sweden)

    Stephanie Speck

    Full Text Available Various semen extender formulas are in use to maintain sperm longevity and quality whilst acting against bacterial contamination in liquid sperm preservation. Aminoglycosides are commonly supplemented to aid in the control of bacteria. As bacterial resistance is increasing worldwide, antimicrobial peptides (AMPs received lively interest as alternatives to overcome multi-drug resistant bacteria. We investigated, whether synthetic cationic AMPs might be a suitable alternative for conventional antibiotics in liquid boar sperm preservation. The antibacterial activity of two cyclic AMPs (c-WWW, c-WFW and a helical magainin II amide analog (MK5E was studied in vitro against two Gram-positive and eleven Gram-negative bacteria. Isolates included ATCC reference strains, multi-resistant E. coli and bacteria cultured from boar semen. Using broth microdilution, minimum inhibitory concentrations were determined for all AMPs. All AMPs revealed activity towards the majority of bacteria but not against Proteus spp. (all AMPs and Staphylococcus aureus ATCC 29213 (MK5E. We could also demonstrate that c-WWW and c-WFW were effective against bacterial growth in liquid preserved boar semen in situ, especially when combined with a small amount of gentamicin. Our results suggest that albeit not offering a complete alternative to traditional antibiotics, the use of AMPs offers a promising solution to decrease the use of conventional antibiotics and thereby limit the selection of multi-resistant strains.

  8. Classification of ostrich sperm characteristics.

    Science.gov (United States)

    Smith, A M J; Bonato, M; Dzama, K; Malecki, I A; Cloete, S W P

    2016-05-01

    The success of assisted reproduction techniques is dependent on a sound foundation of understanding sperm characteristics to evaluate so as to improve semen processing. This study offers a descriptive basis for ostrich semen quality in terms of sperm function characteristics (SFC) that include motility, measured by computer assisted sperm analysis CASA (SCA(®)), viability (SYBR14/PI) and membrane integrity (hypo-osmotic swelling test). Relationships among these SFC's were explored and described by correlations and regressions. Certain fixed effects including the dilution of semen, season, year and male associated with semen collection were interpreted for future applications. The seasonal effect on sperm samples collected throughout the year suggested that it is prudent to restrict collections to spring and summer when SFC's and sperm concentration are maximized, compared to winter when these aspects of sperm quality are suppressed. Dilution of ejaculates helped to maintain important SFC's associated with fertilization success. The SFC's and sperm concentration varied among males, with specific males, having greater values for the percentage of motile (MOT) and progressively motile (PMOT) sperm, as well as sperm velocity (VCL, VSL, VAP) and linearity (LIN) variables. Males may thus be screened on these variables for inclusion in an artificial insemination (AI) programme to optimize fertility success rates. PMID:27039985

  9. Season of ejaculate collection influences the freezability of boar spermatozoa.

    Science.gov (United States)

    Barranco, Isabel; Ortega, Maria D; Martinez-Alborcia, Maria J; Vazquez, Juan M; Martinez, Emilio A; Roca, Jordi

    2013-12-01

    The aim of this retrospective study was to evaluate whether the season of ejaculate collection influences the freezability of porcine sperm. A total of 434 ejaculates were collected from boars of six different breeds over three years (2008-2011) and throughout the four seasons of the year identified in the northern hemisphere (winter, spring, summer and autumn). The ejaculates were cryopreserved using a standard 0.5 mL straw freezing protocol. Sperm quality was assessed before (fresh semen samples kept 24h at 17°C) and after freezing and thawing (at 30 and 150 min post-thawing in semen samples kept in a water bath at 37 °C), according to the percentages of total motility, as assessed by the CASA system, and viability, as assessed by flow cytometry after staining with SYBR-14, PI and PE-PNA. The data, in percentages, on sperm motility and viability after freezing and thawing were obtained at each evaluation time (recovered) and were normalized to the values before freezing (normalized). The season of ejaculate collection influenced (Pboar. Sperm quality was lower in summer, both in terms of motility and viability, and in autumn, in terms of motility, than in winter and spring. Seasonality in the normalized data indicates that the season of ejaculate collection influences sperm freezability, regardless of the season's influence on sperm quality before freezing. Consequently, the spermatozoa from ejaculates collected during summer and, to a lesser extent, also in autumn, are more sensitive to cryopreservation than those from ejaculates collected during winter and spring. PMID:24045067

  10. Compartmentalization of membrane trafficking, glucose transport, glycolysis, actin, tubulin and the proteasome in the cytoplasmic droplet/Hermes body of epididymal sperm.

    Science.gov (United States)

    Au, Catherine E; Hermo, Louis; Byrne, Elliot; Smirle, Jeffrey; Fazel, Ali; Kearney, Robert E; Smith, Charles E; Vali, Hojatollah; Fernandez-Rodriguez, Julia; Simon, Paul H G; Mandato, Craig; Nilsson, Tommy; Bergeron, John J M

    2015-08-01

    Discovered in 1909 by Retzius and described mainly by morphology, the cytoplasmic droplet of sperm (renamed here the Hermes body) is conserved among all mammalian species but largely undefined at the molecular level. Tandem mass spectrometry of the isolated Hermes body from rat epididymal sperm characterized 1511 proteins, 43 of which were localized to the structure in situ by light microscopy and two by quantitative electron microscopy localization. Glucose transporter 3 (GLUT-3) glycolytic enzymes, selected membrane traffic and cytoskeletal proteins were highly abundant and concentrated in the Hermes body. By electron microscope gold antibody labelling, the Golgi trafficking protein TMED7/p27 localized to unstacked flattened cisternae of the Hermes body, as did GLUT-3, the most abundant protein. Its biogenesis was deduced through the mapping of protein expression for all 43 proteins during male germ cell differentiation in the testis. It is at the terminal step 19 of spermiogenesis that the 43 characteristic proteins accumulated in the nascent Hermes body. PMID:26311421

  11. 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 (Psemen. A higher (Pboar 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. PMID:21893053

  12. DNA fragmentation and membrane damage of bocachico Prochilodus magdalenae (Ostariophysi: Prochilodontidae sperm following cryopreservation with dimethylsulfoxide and glucose

    Directory of Open Access Journals (Sweden)

    José Gregorio Martínez

    2012-09-01

    Full Text Available The endangered bocachico Prochilodus magdalenae is a native freshwater fish of Colombia, the most captured species locally and one of the most important species for ex-situ conservation (germplasm banks. The aim of this study was to examine the effect of three concentrations of Dimethylsulfoxide (DMSO (5%, 10%, 15% and three of glucose (305, 333, 361 mM in the extender on spermatic DNA fragmentation (F-DNA (by Halomax®, Chromatin dispersion and membrane damage (D-Me (by eosin-nigrosin staining. After assessment of sperm quality by computer analysis of motility, one part of semen from males was diluted separately with three parts of extender and filled into 0.5 ml straws. Freezing was carried out in liquid nitrogen vapor dry shipper for 30 minutes and thawed at 60ºC for 8 seconds in a water bath and evaluated for the percentage of cells found with F-DNA and D-Me. The results demonstrated that cryopreservation causes greater F-DNA (13.62 ± 1.6% to 28.91 ± 3.25 and D-Me (24.27 ± 1.1% to 58.33 ± 2.81% when compared with pre-freezing semen (PFS (6.71 ± 1.54% and 2.34 ± 0.5%, respectively for F-DNA and D-Me. A significant interaction was found between DMSO and glucose concentration in this experiment. Use of extender: 10% DMSO + 305 mM glucose + 12% chicken egg yolk and, 10% DMSO + 333 mM glucose + 12% chicken egg yolk, allow for lower F-DNA and D-Me during cryopreservation of bocachico semen. A high correlation between F-DNA and D-Me was found (r = 0.771.O curimba Prochilodus magdalenae, é uma espécie nativa de água doce da Colômbia ameaçada de extinção, sendo a mais capturada localmente e uma das mais importantes para a conservação ex-situ (bancos de germoplasma. O objetivo deste estudo foi avaliar o efeito de três concentrações de dimetilsulfóxido (DMSO (5%, 10%, 15% e três de glicose (305, 333, 361 mM no diluente sobre a fragmentação do ADN espermático (F-DNA (através de Halomax®, dispersão da cromatina e danos em

  13. Antioxidant effects of cultured wild ginseng root extracts on the male reproductive function of boars and guinea pigs.

    Science.gov (United States)

    Yun, Suk Jun; Bae, Gui-Seck; Park, Jae Hawn; Song, Tae Ho; Choi, Ahreum; Ryu, Buom-Yong; Pang, Myung-Geol; Kim, Eun Joong; Yoon, Minjung; Chang, Moon Baek

    2016-07-01

    The main objective of this study was to investigate the effects of cultured wild ginseng root extracts (cWGRE) on the sperm of boars and the reproductive system of guinea pigs. Firstly, semen collected from boars (n=10) were incubated in 38°C for 1h with xanthine and xanthine oxidase to generate ROS. The cWGRE was added to the sperm culture system to test its antioxidant effect on the boar sperm. The amount of Reactive Oxygen Species (ROS) was measured by a chemiluminescence assay using luminol. The results indicated that the addition of cWGRE to boar sperm culture inhibited xanthine and xanthine oxidase-induced ROS concentrations. Treatment with cWGRE also had a positive effect on maintaining sperm motility. Effects of cWGRE administration on vitamin C-deficient guinea pigs were further investigated. Hartley guinea pigs (n=25) at 8 weeks of age were randomly divided into five groups. With the exception of the positive control group, each group was fed vitamin C-deficient feed for 21days (d). Respective groups were also orally administered cWGRE, ginseng extract, or mixed ginsenosides for 21 days. In comparison to the control group, oral administration of cWGRE reduced (P<0.05) amount of lipid peroxidation and increased (P<0.05) both glutathione peroxidase concentrations and the trolox equivalent antioxidant capacity. In addition, administration of cWGRE induced increases (P<0.05) in body weight, testosterone concentrations, and spermatid populations. The results of the present study support our hypothesis that cWGRE has positive effects on male reproductive functions via suppression of ROS production. PMID:27068520

  14. TRIXcell+, a new long-term boar semen extender containing whey protein with higher preservation capacity and litter size

    Directory of Open Access Journals (Sweden)

    B.M. van den Berg

    2014-02-01

    Full Text Available It was the aim of the present study to test whey as protective protein for the sperm cell in the long-term boar semen preservation medium TRIXcell. Analyses of sperm cell motility using computer-assisted semen analysis (CASA indicated that the whey protein Porex has a similar protective effect as bovine serum albumin (BSA in maintaining viability of stored boar sperm. Boar sperm diluted in TRIXcell+ maintains commercially acceptable motility (>60% for 10 days, while swine sperm diluted in the semen preservation medium Beltsville Thawing Solution (BTS maintains commercially acceptable motility (>60% for 3-5 days for most boars. To test the on-farm fertility performance of TRIXcell+ compared to BTS, inseminations were started on 35 commercial pig production farms in the summer of 2006. During the period of July 2006 until July 2012 for each farm and each calendar year the mean farrowing rate and litter size for semen diluted in TRIXcell+ and stored for 3-5 days was found higher than that of semen stored for 1-2 days in BTS. Based on data gained from a total of 583.749 sows inseminated through the years 2006-2012, the mean farrowing rate for semen diluted in TRIXcell+ and BTS was 90.4 ± 4.0 and 87.9 ± 3.6, respectively, which is not significantly different. Based on the same data, the mean total number of piglets born alive for semen diluted in TRIXcell+ and BTS was 14.2 ± 0.7 and 13.6 ± 0.6, respectively, which is significantly different. We conclude that whey protein can effectively be used in the long-term preservation medium TRIXcell resulting in a higher litter size.

  15. The adult boar testicular and epididymal transcriptomes

    Directory of Open Access Journals (Sweden)

    Guyonnet Benoît

    2009-08-01

    Full Text Available Abstract Background Mammalians gamete production takes place in the testis but when they exit this organ, although spermatozoa have acquired a specialized and distinct morphology, they are immotile and infertile. It is only after their travel in the epididymis that sperm gain their motility and fertility. Epididymis is a crescent shaped organ adjacent to the testis that can be divided in three gross morphological regions, head (caput, body (corpus and tail (cauda. It contains a long and unique convoluted tubule connected to the testis via the efferent ducts and finished by joining the vas deferens in its caudal part. Results In this study, the testis, the efferent ducts (vas efferens, VE, nine distinct successive epididymal segments and the deferent duct (vas deferens, VD of four adult boars of known fertility were isolated and their mRNA extracted. The gene expression of each of these samples was analyzed using a pig generic 9 K nylon microarray (AGENAE program; GEO accession number: GPL3729 spotted with 8931 clones derived from normalized cDNA banks from different pig tissues including testis and epididymis. Differentially expressed transcripts were obtained with moderated t-tests and F-tests and two data clustering algorithms based either on partitioning around medoid (top down PAM or hierarchical clustering (bottom up HCL were combined for class discovery and gene expression analysis. Tissue clustering defined seven transcriptomic units: testis, vas efferens and five epididymal transcriptomic units. Meanwhile transcripts formed only four clusters related to the tissues. We have then used a specific statistical method to sort out genes specifically over-expressed (markers in testis, VE or in each of the five transcriptomic units of the epididymis (including VD. The specific regional expression of some of these genes was further validated by PCR and Q-PCR. We also searched for specific pathways and functions using available gene ontology

  16. Presence and function of dopamine transporter (DAT in stallion sperm: dopamine modulates sperm motility and acrosomal integrity.

    Directory of Open Access Journals (Sweden)

    Javier A Urra

    Full Text Available Dopamine is a catecholamine with multiple physiological functions, playing a key role in nervous system; however its participation in reproductive processes and sperm physiology is controversial. High dopamine concentrations have been reported in different portions of the feminine and masculine reproductive tract, although the role fulfilled by this catecholamine in reproductive physiology is as yet unknown. We have previously shown that dopamine type 2 receptor is functional in boar sperm, suggesting that dopamine acts as a physiological modulator of sperm viability, capacitation and motility. In the present study, using immunodetection methods, we revealed the presence of several proteins important for the dopamine uptake and signalling in mammalian sperm, specifically monoamine transporters as dopamine (DAT, serotonin (SERT and norepinephrine (NET transporters in equine sperm. We also demonstrated for the first time in equine sperm a functional dopamine transporter using 4-[4-(Dimethylaminostyryl]-N-methylpyridinium iodide (ASP(+, as substrate. In addition, we also showed that dopamine (1 mM treatment in vitro, does not affect sperm viability but decreases total and progressive sperm motility. This effect is reversed by blocking the dopamine transporter with the selective inhibitor vanoxerine (GBR12909 and non-selective inhibitors of dopamine reuptake such as nomifensine and bupropion. The effect of dopamine in sperm physiology was evaluated and we demonstrated that acrosome integrity and thyrosine phosphorylation in equine sperm is significantly reduced at high concentrations of this catecholamine. In summary, our results revealed the presence of monoamine transporter DAT, NET and SERT in equine sperm, and that the dopamine uptake by DAT can regulate sperm function, specifically acrosomal integrity and sperm motility.

  17. Effect of estrogens on boar sperm capacitation in vitro

    Czech Academy of Sciences Publication Activity Database

    Děd, Lukáš; Dostálová, Pavla; Dorosh, Andriy; Dvořáková-Hortová, K.; Pěknicová, Jana

    2010-01-01

    Roč. 8, - (2010), ---. ISSN 1477-7827 R&D Projects: GA MŠk(CZ) 1M06011; GA ČR(CZ) GD523/08/H064; GA ČR(CZ) GA523/09/1793 Institutional research plan: CEZ:AV0Z50520701 Keywords : capacitation * acrosome reaction * monoclonal antibody * estrogen * flow cytometry Subject RIV: EI - Biotechnology ; Bionics Impact factor: 1.695, year: 2010

  18. Computer-assisted sperm analysis (CASA): capabilities and potential developments.

    Science.gov (United States)

    Amann, Rupert P; Waberski, Dagmar

    2014-01-01

    Computer-assisted sperm analysis (CASA) systems have evolved over approximately 40 years, through advances in devices to capture the image from a microscope, huge increases in computational power concurrent with amazing reduction in size of computers, new computer languages, and updated/expanded software algorithms. Remarkably, basic concepts for identifying sperm and their motion patterns are little changed. Older and slower systems remain in use. Most major spermatology laboratories and semen processing facilities have a CASA system, but the extent of reliance thereon ranges widely. This review describes capabilities and limitations of present CASA technology used with boar, bull, and stallion sperm, followed by possible future developments. Each marketed system is different. Modern CASA systems can automatically view multiple fields in a shallow specimen chamber to capture strobe-like images of 500 to >2000 sperm, at 50 or 60 frames per second, in clear or complex extenders, and in process data apparently are available. PMID:24274405

  19. A NEW MODEL OF BOAR SEMEN EVALUATION AND THE IMPACT OF CRYOGENIC FACTOR ON SPERMATIC CELLS

    Directory of Open Access Journals (Sweden)

    M. ZĂHAN

    2009-05-01

    Full Text Available Nowadays, sperm evaluation is mostly used to predict fertility and freezability. Theaim of this study is to evaluate the possibility of investigating the effects of thecryogenic agent on boar spermatozoa, by identifying a set of laboratory tests for arapid and efficient evaluation of semen quality. Usual sperm analysis such as spermconcentration, motility and spermatozoa morphology are not able to show subtleabnormalities, which are having a basic role in the fertilizing ability. Moreover, itseems that other sperm characteristics, involved in the fertilizing ability, can interferewith the freezing-thawing processes, being not evaluated or maybe not known.Morphological (microscopic analysis of stained spermatozoa, functional (motilityanalysis and hypo-osmotic swelling test and chromatin integrity (Acridine OrangeTest and Comet Assay analysis were performed aiming to show the differences inspermatozoon integrity and functionality, caused by the cryogenic factor

  20. Proteomics of ionomycin-induced ascidian sperm reaction: Released and exposed sperm proteins in the ascidian Ciona intestinalis.

    Science.gov (United States)

    Nakazawa, Shiori; Shirae-Kurabayashi, Maki; Otsuka, Kei; Sawada, Hitoshi

    2015-12-01

    Sperm proteins mediating sperm-egg interaction should be exhibited on the sperm surface, or exposed or released when sperm approach an egg. In ascidians (protochordates), sperm undergo a sperm reaction, characterized by enhanced sperm motility and mitochondrial swelling and shedding on contact with the vitelline coat (VC) or by treatment with Ca(2+) ionophore. Here, proteomic analysis was conducted on sperm exudates and sperm surface proteins using ionomycin-induced sperm reaction and cell-impermeable labeling in Ciona intestinalis type A (C. robusta). In the exudate from sperm treated with ionomycin, membrane proteins including a possible VC receptor CiUrabin were abundant, indicating the release of membranous compartments during sperm reaction. Among the surface proteins XP_009859314.1 (uncharacterized protein exhibiting homology to HrTTSP-1) was most abundant before the sperm reaction, but XP_004227079.1 (unknown Ig superfamily protein) appears to be most abundantly exposed by the sperm reaction. Moreover, proteins containing a notable set of domains, astacin-like metalloprotease domain and thrombospondin type 1 repeat(s), were found in this fraction. Possible roles in fertilization as well as localizations and behaviors of these proteins are discussed. PMID:26223815

  1. Sertoli Cell Differentiation in Pubertal Boars

    Science.gov (United States)

    Meishan boars experience puberty at a younger age than crossbred (BX) boars in association with earlier cessation of Sertoli cell proliferation and smaller post pubertal testicular size. The current study defined changes in expression, assessed by immunohistochemistry, of anti-Mullerian hormone (AMH...

  2. High resolution DNA content measurements of mammalian sperm

    Energy Technology Data Exchange (ETDEWEB)

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

    1982-01-01

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

  3. Reduction of boar taint - the practical way

    OpenAIRE

    Jensen, Bent Borg; Maribo, Hanne; Thomsen, Rikke

    2013-01-01

    The aim of organic pig production is to ensure high animal welfare and natural products. Banning castration is thus a logical step forward, but the risk of boar taint in the meat is a major barrier for marketing meat from entire male pigs. Is it possible to use genetic tools and breeding strategies to prevent boar taint? What is the effect of feeding, management, housing and hygiene? Is it possible to process the meat to minimize the risk of boar taint? These issues will be discussed based on...

  4. Sperm function test.

    Science.gov (United States)

    Talwar, Pankaj; Hayatnagarkar, Suryakant

    2015-01-01

    With absolute normal semen analysis parameters it may not be necessary to shift to specialized tests early but in cases with borderline parameters or with history of fertilization failure in past it becomes necessary to do a battery of tests to evaluate different parameters of spermatozoa. Various sperm function tests are proposed and endorsed by different researchers in addition to the routine evaluation of fertility. These tests detect function of a certain part of spermatozoon and give insight on the events in fertilization of the oocyte. The sperms need to get nutrition from the seminal plasma in the form of fructose and citrate (this can be assessed by fructose qualitative and quantitative estimation, citrate estimation). They should be protected from the bad effects of pus cells and reactive oxygen species (ROS) (leukocyte detection test, ROS estimation). Their number should be in sufficient in terms of (count), structure normal to be able to fertilize eggs (semen morphology). Sperms should have intact and functioning membrane to survive harsh environment of vagina and uterine fluids (vitality and hypo-osmotic swelling test), should have good mitochondrial function to be able to provide energy (mitochondrial activity index test). They should also have satisfactory acrosome function to be able to burrow a hole in zona pellucida (acrosome intactness test, zona penetration test). Finally, they should have properly packed DNA in the nucleus to be able to transfer the male genes (nuclear chromatic decondensation test) to the oocyte during fertilization. PMID:26157295

  5. Sperm function test

    Directory of Open Access Journals (Sweden)

    Pankaj Talwar

    2015-01-01

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

  6. Sperm Competition, Sperm Numbers and Sperm Quality in Muroid Rodents

    OpenAIRE

    Laura Gómez Montoto; Concepción Magaña; Maximiliano Tourmente; Juan Martín-Coello; Cristina Crespo; Juan José Luque-Larena; Montserrat Gomendio; Roldan, Eduardo R. S.

    2011-01-01

    Sperm competition favors increases in relative testes mass and production efficiency, and changes in sperm phenotype that result in faster swimming speeds. However, little is known about its effects on traits that contribute to determine the quality of a whole ejaculate (i.e., proportion of motile, viable, morphologically normal and acrosome intact sperm) and that are key determinants of fertilization success. Two competing hypotheses lead to alternative predictions: (a) sperm quantity and qu...

  7. Measurements of Boar Spermatozoa Motility Using PFG NMR Method

    International Nuclear Information System (INIS)

    The evaluation of spermatozoa motility, viability and morphology is an essential parameter in the examination of sperm quality and in the establishment of correlations between sperm quality and fertility. Until now, assessment of sperm quality has been based on subjective evaluation of parameters, such as motility and viability, and on objective parameters, such as semen concentration and morphology abnormalities. When subjective optical microscopic evaluation was used in humans and animals, variations of 30 to 60% have been reported in the estimation of the motility parameters of the same ejaculates. To overcome this variability, different systems have been proposed such as turbidimetry, laser-Doppler spectroscopy, and photometric methods. Other accurate techniques, such as flow cytometry, which allows the evaluation of concentration, and cellulose-acetate/nitrate filter measure only a single semen parameter. The more recent track semen analysis system, based on individual spermatozoon evaluation, offers an accurate calculation of different semen parameters. Although some interesting results have already been obtained, many questions remain, which have to be answered to allow for further development in veterinary medicine, clinical fertility settings, physiological, and toxicology research activities. Pulsed field gradient nuclear magnetic resonance (PFG NMR) techniques have been presented demonstrating the potential to study flow and transport processes in complex systems. By PFG NMR, the molecular displacement can be measured that occurs during a time interval D, between two consecutive magnetic field gradient pulses. In this poster we present the results of PFG-NMR obtained for a number of samples of boar spermatozoa with varying motility and discuss whether this method can be useful for fast and reliable spermatozoa motility evaluation. (author)

  8. Evaluation of CD52 positive sperms in subfertile human semen samples: Is there any relationship with main semen parameters?

    OpenAIRE

    Roshanak Aboutorabi; Fatemeh Mazani; Laleh Rafiee

    2014-01-01

    Background: Sperm maturation and sperm membrane integration are the most important elements in male fertility. CD52 is one of the antigens. CD52 is a GPI (glycosylphosphatidylinositol) anchored that express on lymphocytes and epididymal cells. This antigen bind to sperm membrane during transition sperm from epididymal duct as well as its relationship with semenogelins in human seminal plasma. The aim of this study was to obtain any association between the percentage of CD52 positive sperms wi...

  9. Study on the Vesiculation during Mouse Sperm Acrosome Reaction

    Institute of Scientific and Technical Information of China (English)

    林家豪; 周作民; 胡志刚; 王黎熔; 林敏; 张适

    1994-01-01

    The location of the mono-membrane and the bi-membrane vesicles of mouse sperm was identified using Con A in conjugation with the colloidal gold. The observation showed that both mono-membrane vesicfes and outer layer of the hi-membrane vesicles come from the outer acrosome membrane. The inner membrane layer of the bi-member vesicles and residual membrane distributed among the vesicles are really the ptasmatemma. It is suggested that the outer acrosome membrane did not fuse with the pfasmafemma during mouse sperm acrosome reaction and that both the mono-membrane and the bi-membrane vesicles of mouse sperm were formed due to winding of the outer acrosome membrane.

  10. Aquaporin3 is a sperm water channel essential for postcopulatory sperm osmoadaptation and migration

    Institute of Scientific and Technical Information of China (English)

    Qi Chen; Hongying Peng; Li Lei; Ying Zhang; Haibin Kuang; Yujing Cao; Qi-xian Shi; Tonghui Ma; Enkui Duan

    2011-01-01

    In the journey from the male to female reproductive tract,mammalian sperm experience a natural osmotic decrease (e.g.,in mouse,from ~415 mOsm in the cauda epididymis to ~310 mOsm in the uterine cavity). Sperm have evolved to utilize this hypotonic exposure for motility activation,meanwhile efficiently silence the negative impact of hypotonic cell swelling. Previous physiological and pharmacological studies have shown that ion channel-controlled water influx/efflux is actively involved in the process of sperm volume regulation; however,no specific sperm proteins have been found responsible for this rapid osmoadaptation. Here,we report that aquaporin3 (AQP3) is a sperm water channel in mice and humans. Aqp3-deficient sperm show normal motility activation in response to hypotonicity but display increased vulnerability to hypotonic cell swelling,characterized by increased tail bending after entering uterus. The sperm defect is a result of impaired sperm volume regulation and progressive cell swelling in response to physiological hypotonic stress during male-female reproductive tract transition. Time-lapse imaging revealed that the cell volume expansion begins at cytoplasmic droplet,forcing the tail to angulate and form a hairpin-like structure due to mechanical membrane stretch. The tail deformation hampered sperm migration into oviduct,resulting in impaired fertilization and reduced male fertility. These data suggest AQP3 as an essential membrane pathway for sperm regulatory volume decrease (RVD) that balances the "trade-off" between sperm motility and cell swelling upon physiological hypotonicity,thereby optimizing postcopulatory sperm behavior.

  11. 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 (P<0.05) total motility. The spermatozoa plasma membrane integrity and mitochondrial activity were improved at four different concentrations: 0.4, 0.6, 0.8, 1.0mg/mL. The addition of alginate 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 (P<0.05). The freezing extenders with the presence of alginate led to higher SOD and GSH-Px activities and lower MDA levels, in comparison to the control (P<0.05). In summary, alginate 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. PMID:24814905

  12. Palliative therapy of osteochondrosis dessicans in a Duroc boar

    OpenAIRE

    Oomah, Kevin

    2008-01-01

    A 2-year-old, 210-kg, Duroc boar manifested with a grade II–III left front lameness. The boar was treated systemically with isolfupredone acetate and a 5-week course of ketoprofen. The lameness resolved and the ketoprofen was discontinued; however, the lameness returned and the boar was euthanized humanely. Postmortem examination was consistent with osteochondrosis dessicans.

  13. Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm.

    Science.gov (United States)

    Gürler, H; Malama, E; Heppelmann, M; Calisici, O; Leiding, C; Kastelic, J P; Bollwein, H

    2016-07-15

    The objective was to examine if there are relationships between alterations in sperm viability, reactive oxygen species (ROS) synthesis, and DNA integrity induced by cryopreservation of bovine sperm. Four ejaculates were collected from each of six bulls. Each ejaculate was diluted and divided into two aliquots; one was incubated for 24 hours at 37 °C, and the other frozen, thawed, and incubated for 24 hours at 37 °C. Analyses of quality of sperm were performed after 0, 3, 6, 12, and 24 hours of incubation. Progressive motile sperm was determined with computer assisted sperm analysis. Percentages of plasma membrane- and acrosome-intact sperm, sperm with a high mitochondrial membrane potential, sperm showing a high degree of DNA fragmentation (%DFI), and their reactive oxygen species content were assessed with dichlorofluorescein-diacetate, dihydrorhodamine, diaminofluorescein diacetate, and mitochondrial superoxide indicator using flow cytometry. Although all other sperm parameters showed alterations (P  0.05, 0.91 ± 0.23) in nonfrozen sperm. Cryopreservation induced changes of all sperm parameters (P < 0.05). In contrast to all other sperm parameters, dichlorofluorescein-diacetate-fluoroescence indicating the synthesis of H2O2 showed a similar exponential rise (P < 0.05) like the %DFI values in frozen sperm. In conclusion, changes of DNA integrity in frozen sperm seem to be related to synthesis of H2O2 but not to sperm viability and synthesis of other reactive oxygen species. PMID:27039074

  14. Porcine embryos produced after intracytoplasmic sperm injection using xenogeneic pig sperm from neonatal testis tissue grafted in mice.

    Science.gov (United States)

    Honaramooz, Ali; Cui, Xiang-Shun; Kim, Nam-Hyung; Dobrinski, Ina

    2008-01-01

    Embryo development after homologous intracytoplasmic sperm injection (ICSI) with sperm from testis tissue xenografts from pigs or any other farm animal species has not been evaluated critically. Here, we report development of porcine embryos in vitro following ICSI with sperm retrieved from xenografted neonatal pig testis. Small pieces of testis tissue from newborn piglets were grafted under the back skin of castrated immunodeficient mice (n = 4) and the xenografts were collected 8 months after grafting. Spermatozoa were recovered by mincing of the grafted tissue. For comparison, testicular, epididymal and ejaculated spermatozoa were also collected from mature boars. Oocytes injected with xenogeneic spermatozoa were either fixed to determine fertilisation processes (n = 89 in five replicates) or allowed to develop in vitro (n = 143 in four replicates). Xenogeneic porcine spermatozoa were fertilisation competent (24% v. 58%, 68%, 62% or 0% for xenogeneic v. control testicular, epididymal and ejaculated spermatozoa or no spermatozoa, respectively) and embryos developed to the blastocyst stage (8% v. 22%, 27%, 25% or 0%, respectively). These results demonstrate that porcine spermatozoa derived from immature testis tissue xenografted into mice are fertilisation competent, albeit at a lower rate than testicular, epididymal or ejaculated spermatozoa from control boars, and support embryo development after ICSI. PMID:18842182

  15. Sperm cells as vectors in the production of transgenic animals

    Energy Technology Data Exchange (ETDEWEB)

    Prince, R.M.

    1993-04-28

    Transgenic animals are used in industry and in biomedical research in order to provide in vivo experimental model systems. Sperm cells have been reported used as vectors in the production of transgenic animals before, however no approach has of yet proven to be successful. Fertilizing eggs with genetically modified sperm would be advantageous in that sperm are readily accessible and stable, and eggs can be fertilized by modified sperm cells in vivo. Recent elucidations regarding the unique manner of DNA packaging in sperm chromatin by protamines has provided us with the insight for developing a method of introducing foreign DNA into sperm which is likely to succeed where others have failed. We have developed a method for mimicking the in vivo system of sperm chromatin toroid subunits in vitro, concentrating these toroids, and fluorescent visualization. Our present work concerns development of a method to successfully deliver DNA across the cell membranes and into the nucleus.

  16. Characterizing the Glycocalyx of Poultry Spermatozoa: II. Low Temperature Storage of Turkey Semen and Sperm Mobility Phenotype Impact the Carbohydrate Component of Membrane Glycoconjugates

    Science.gov (United States)

    The turkey sperm glycocalyx is known to contain residues of sialic acid, alpha-mannose/alpha-glucose, alpha- and beta-galactose, alpha-fucose, alpha- and beta-N-acetyl-galactosamine, monomers and dimers of N-acetyl-glucosamine and N-acetyl-lactosamine. Potential changes in these carbohydrates during...

  17. Predictive capacity of sperm quality parameters and sperm subpopulations on field fertility after artificial insemination in sheep.

    Science.gov (United States)

    Santolaria, P; Vicente-Fiel, S; Palacín, I; Fantova, E; Blasco, M E; Silvestre, M A; Yániz, J L

    2015-12-01

    This study was designed to evaluate the relevance of several sperm quality parameters and sperm population structure on the reproductive performance after cervical artificial insemination (AI) in sheep. One hundred and thirty-nine ejaculates from 56 adult rams were collected using an artificial vagina, processed for sperm quality assessment and used to perform 1319 AI. Analyses of sperm motility by computer-assisted sperm analysis (CASA), sperm nuclear morphometry by computer-assisted sperm morphometry analysis (CASMA), membrane integrity by acridine orange-propidium iodide combination and sperm DNA fragmentation using the sperm chromatin dispersion test (SCD) were performed. Clustering procedures using the sperm kinematic and morphometric data resulted in the classification of spermatozoa into three kinematic and three morphometric sperm subpopulations. Logistic regression procedures were used, including fertility at AI as the dependent variable (measured by lambing, 0 or 1) and farm, year, month of AI, female parity, female lambing-treatment interval, ram, AI technician and sperm quality parameters (including sperm subpopulations) as independent factors. Sperm quality variables remaining in the logistic regression model were viability and VCL. Fertility increased for each one-unit increase in viability (by a factor of 1.01) and in VCL (by a factor of 1.02). Multiple linear regression analyses were also performed to analyze the factors possibly influencing ejaculate fertility (N=139). The analysis yielded a significant (Psemen variables to predict field fertility was analyzed using receiver operating characteristic (ROC) curve analysis. Sperm viability and VCL showed significant, albeit limited, predictive capacity on field fertility (0.57 and 0.54 Area Under Curve, respectively). The distribution of spermatozoa in the different subpopulations was not related to fertility. PMID:26507945

  18. Effects of mechanical stresses on sperm function and fertilization rate in mice.

    Science.gov (United States)

    Shi, Xiao; Wang, Ting; Qiu, Zhuo Lin; Li, Ke; Li, Liu; Chan, Carol Pui Shan; Chan, Si Mei; Li, Tian-Chiu; Quan, Song

    2016-01-01

    In this study, we investigated whether any of the observed changes in mouse sperm function tests secondary to mechanical stresses (centrifugation and pipetting) correlate with sperm fertilization ability. Chinese Kunming mice were used as sperm and oocyte donors. Sperm samples were allocated evenly into centrifugation, pipette, and control groups. Sperm plasma membrane integrity (PMI), mitochondrial membrane permeability (MMP), baseline and stimulated intracellular ROS, and sperm fertilization ability were measured by hypo-osmotic swelling, flow cytometry, and fertilization tests. Parallel studies were conducted and all tests were repeated six times. Our results showed that after centrifugation, the progressive motility, average path velocity, and overall sperm motility and PMI decreased significantly (p < 0.05). In addition, the MMP level decreased significantly in viable sperm when the centrifugation condition reached 1,400 g × 15 minutes (p < 0.05). When pipetting was performed two or more times, progressive motility, average path velocity, and overall sperm motility decreased significantly (p < 0.05); when it was performed four or more times, sperm membrane integrity and intracellular basal ROS level of viable sperm was also significantly decreased (p < 0.05). In conclusion, various mechanical stresses seem to affect sperm function, however this does not appear to alter fertilization rate. Laboratory handling steps should be minimized to avoid unnecessary mechanical stresses being applied to sperm samples. PMID:26889695

  19. Regulating wild boar populations is "somebody else's problem"! - Human dimension in wild boar management.

    Science.gov (United States)

    Keuling, Oliver; Strauß, Egbert; Siebert, Ursula

    2016-06-01

    As a part of the ongoing game survey of the German federal state of Lower Saxony (WTE), we conducted inquiries into wild boar management and distribution, as well as hunters' attitudes, in order to determine the reasons for the increase of wild boar populations and to inform our game management strategy. According to hunters' reports within the WTE, increases in distribution and population continue and a reduction of the wild boar population has been deemed necessary on a large scale. In the home region, however, it seems to be "somebody else's problem" (SEP), according to hunters' opinions. The majority of hunters are not able to regulate the population and this could be a reason that wild boar numbers continue to increase. Cooperation and comprehensive hunting with efficient hunting methods seems to be the most promising solution, as non-hunting methods are unpopular amongst hunters. The hunters seem to be aware of the problems, solutions and contributing factors; however, most hunters do not feel responsible and see the management of wild boar, again, as a SEP. Regional conditions, as well as hunters' willingness and capacity to manage wild boar will have to be incorporated into management concepts. PMID:26956178

  20. Intracytoplasmic Sperm Injection (ICSI)

    Science.gov (United States)

    ... male partner produces too few sperm to do artificial insemination (intrauterine insemination [IUI]) or IVF. • The sperm may ... birth defects may actually be due to the infertility and not the treatments used to overcome the ...

  1. Snail sperm production characteristics vary with sperm competition risk

    OpenAIRE

    Oppliger, A.; Hosken, D J; Ribi, G

    1998-01-01

    Sperm competition is widespread and influences both male investment in spermatogenic tissue and ejaculate characteristics. Sperm competition models assume trade-offs between sperm size and number, although such trade-offs may be difficult to detect. This study examines the effects of sperm competition risk on the sperm production characteristics of the freshwater snail Viviparus ater. In this prosobranch, females mate frequently and store sperm, generating sperm competition. Males produce two...

  2. Larger sperm outcompete smaller sperm in the nematode Caenorhabditis elegans.

    OpenAIRE

    LaMunyon, C W; Ward, S.

    1998-01-01

    Sperm competition is generally thought to drive the evolution of sperm miniaturization. Males gain advantage by transferring more sperm, which they produce by dividing limited resources into ever smaller cells. Here, we describe the opposite effect of size on the competitiveness of amoeboid sperm in the hermaphroditic nematode Caenorhabditis elegans. Larger sperm crawled faster and displaced smaller sperm, taking precedence at fertilization. Larger sperm took longer to produce, however, and s...

  3. Sperm midpiece length predicts sperm swimming velocity in house mice

    OpenAIRE

    Firman, Renée C.; Simmons, Leigh W.

    2010-01-01

    Evolutionary biologists have argued that there should be a positive relationship between sperm size and sperm velocity, and that these traits influence a male's sperm competitiveness. However, comparative analyses investigating the evolutionary associations between sperm competition risk and sperm morphology have reported inconsistent patterns of association, and in vitro sperm competition experiments have further confused the issue; in some species, males with longer sperm achieve more compe...

  4. 鸵鸟卵黄低密度脂蛋白对猪精子冻后品质的影响%Effects of Ostrich Egg Yolk LDL on Boar Spermatozoa Quality Following Freezing-Thawing

    Institute of Scientific and Technical Information of China (English)

    侯丽鹏; 石芳萍; 卜书海; 李青旺; 胡建宏

    2012-01-01

    在家畜精液冷冻中,卵黄被广泛应用,且其中的低密度脂蛋白(LDL)对精子起主要保护作用.本研究利用含6%、7%、8%和9%鸵鸟卵黄LDL配制的稀释液制作猪细管冷冻精液,分析鸵鸟卵黄LDL对冷冻-解冻后猪精子质量参数的影响.结果表明:在含不同浓度鸵鸟卵黄LDL的稀释液中,8%LDL的稀释液冷冻效果最好,冻后精子活率平均可达52.13%,显著高于其他组(P<0.05);精子顶体完整率平均为58.33%,质膜完整率为72.38%,与其他处理组相比差异显著(P<0.05).但与鸡蛋卵黄LDL和鸽子蛋卵黄LDL处理组相比,鸵鸟卵黄LDL处理组冷冻-解冻后猪精子质量参数相对较低.本研究表明,虽然鸵鸟卵黄LDL在冷冻过程中对猪精子具有一定的保护作用,但相对于鸽子蛋和鸡蛋卵黄LDL效果并不理想.%Egg yolk has been widely used in boar frozen semen dilution and the protective action of yolk is largely attributed to the low-density lipoprotein (LDL). In this study, LDL of ostrich egg yolk was added at concentrations of 6-9% to the extenders used to freeze boar semen and its effects on the quality of frozen-thawed sperm were assessed. The results indicated that supplementation of LDL at 8% LDL resulted in significantly higher spermatozoa motility (52.13%) than that of other groups(P< 0.05), and there was higher acrosome integrity (58.33%) and membrane integrity(72.38%) (P < 0.05) compared with other treatment groups. But the quality parameters of boar sperm after freezing-thawing following the LDL of ostrich egg yolk is lower compared with the extender containing LDL of egg yolk and pigeon yo!k. According to all measured parameters, the extender containing 8% LDL of ostrich egg yolk showed beneficial cryoprotective effects on frozen-thawed boar spermatozoa. But its effect is not best compared with 9% egg yolk LDL and 8% pigeon egg yolk LDL.

  5. The role of complement component C3b and its receptors in sperm-oocyte interaction.

    OpenAIRE

    Anderson, D. J.; Abbott, A F; Jack, R M

    1993-01-01

    Previous studies have shown that human sperm that have undergone the acrosome reaction express a unique tissue-specific variant of the complement component 3 (C3)-binding molecule membrane cofactor protein (MCP, CD46) and that damaged or dead sperm activate the alternative pathway of complement and bind C3 catabolites. In this study we provide evidence that MCP on sperm that have undergone the acrosome reaction specifically binds dimeric C3b and that human sperm acrosomal proteases released d...

  6. Sperm navigation along helical paths in 3D chemoattractant landscapes

    Science.gov (United States)

    Jikeli, Jan F.; Alvarez, Luis; Friedrich, Benjamin M.; Wilson, Laurence G.; Pascal, René; Colin, Remy; Pichlo, Magdalena; Rennhack, Andreas; Brenker, Christoph; Kaupp, U. Benjamin

    2015-08-01

    Sperm require a sense of direction to locate the egg for fertilization. They follow gradients of chemical and physical cues provided by the egg or the oviduct. However, the principles underlying three-dimensional (3D) navigation in chemical landscapes are unknown. Here using holographic microscopy and optochemical techniques, we track sea urchin sperm navigating in 3D chemoattractant gradients. Sperm sense gradients on two timescales, which produces two different steering responses. A periodic component, resulting from the helical swimming, gradually aligns the helix towards the gradient. When incremental path corrections fail and sperm get off course, a sharp turning manoeuvre puts sperm back on track. Turning results from an `off' Ca2+ response signifying a chemoattractant stimulation decrease and, thereby, a drop in cyclic GMP concentration and membrane voltage. These findings highlight the computational sophistication by which sperm sample gradients for deterministic klinotaxis. We provide a conceptual and technical framework for studying microswimmers in 3D chemical landscapes.

  7. USE OF ALTERNATIVE EXTENDERS AND TEMPERATURES IN LONG TERM STORAGE OF BOAR SEMEN

    Directory of Open Access Journals (Sweden)

    Lina Raquel Santos Araújo

    2016-01-01

    Full Text Available The use of appropriate extenders is important for the success of an artificial insemination program. The objective of this study was to evaluate the efficiency of alternative extenders for swine semen at different temperatures (17 to 10 °C. The following extenders were used: Beltsville Thawing Solution (BTS, powdered coconut water (ACP-103®, and skimmed milk powder (LPD. The 50 ejaculates were analyzed daily, in natura and after dilution, during the 5-day period of semen preservation  (D0 to D4, regarding spermatic vigor and motility. Acrosome integrity and sperm viability were evaluated on D0 and D4. Data were analyzed using Mann-Whitney, Students, Tukey and chi-square tests (p<0.05. The LPD extender at 10 °C presented higher motility and sperm vigor compared to BTS and ACP until D2, and to treatments stored at 17 °C. Acrosome vitality and integrity remained higher (p<0.001 with LPD at 10 °C on D0 and D4. LPD showed to be a good extender for the swine semen at lower temperature (10 °C. Furthermore, it provided better protection to sperm cells, by allowing greater integrity and vitality of the acrosome. Keywords: coconut water; conservation; skimmed milk; semen boar.

  8. Sperm Selection in ART

    Directory of Open Access Journals (Sweden)

    Montag M

    2012-01-01

    Full Text Available Selection of sperm is a crucial part in assisted reproductive treament (ART. Sperm preparation methods do mainly differentiate according to sperm motility and are indispensable for therapies like intrauterine insemination, in-vitro fertilization (IVF and intracytoplasmic sperm injection (ICSI. Although in the beginning of the era of ICSI andrology was thought to play a minor role, ICSI has offered new options by correlating the treatment outcome to parameters of the individual applied spermatozoon. Hence the possibility for selecting spermatozoa has shifted from parameters which characterize the entire sperm cohort to a single-sperm specific assessment technology. Consequently, sperm selection is a topic which is intensively discussed nowadays. This article gives a comprehensive overview of the technologies which can be applied today and give a prospective on future techniques.

  9. Trichinellosis in farmed wild boar: meat inspection findings and seroprevalence

    OpenAIRE

    Sukura A; Näreaho A.; Veijalainen P.; Oivanen I.

    2001-01-01

    A reflection of highly prevalent endemic wildlife trichinellosis is seen in wild boar farming in Finland. During the last five years, 0.7 % (15/2265) of wild boars undergoing official meat inspection have been determined to be Trichinella-positive. These findings originate from six different farms. In Finland, T. spiralis and T. pseudospiralis have been discovered in meat inspection of wild boars. ELISA showed 11 out of 9 9 serum samples (11 %) as having specific antibodies for T. spiralis cr...

  10. Evolution of sperm size in nematodes: sperm competition favours larger sperm.

    OpenAIRE

    LaMunyon, C W; Ward, S.

    1999-01-01

    In the free-living rhabditid nematode Caenorhabditis elegans, sperm size is a determinant of sperm competitiveness. Larger sperm crawl faster and physically displace smaller sperm to take fertilization priority, but not without a cost: larger sperm are produced at a slower rate. Here, we investigate the evolution of sperm size in the family Rhabditidae by comparing sperm among 19 species, seven of which are hermaphroditic (self-fertile hermaphrodites and males), the rest being gonochoristic (...

  11. Boar semen bacterial contamination in Italy and antibiotic efficacy in a modified extender

    Directory of Open Access Journals (Sweden)

    Carla Bresciani

    2014-02-01

    Full Text Available The aims of the study were to identify microbial flora in boar semen under field conditions in northern Italy, to investigate antibiotic resistance and sensitivity of isolated bacteria, and to evaluate elimination of bacteria after storage in two types of extenders added with different antibiotics (amikacin vs gentamicin. A total of 60 boars were collected in 13 pig farms. Bacteriological and mycological investigations were performed immediately on raw semen samples, then at 48 and 120 h of storage on semen diluted randomly in a new short-term modified extender (ME-S or in a commercial one (CRONOSTM. Bacterial contamination was found in 63% of raw semen samples and different bacterial species were isolated: E.coli, Serratia marcescens, Staphylococcus epidermidis and aureus, Proteus spp., Streptococcus spp. and Pseudomonas aeruginosa. E. coli was the most isolated contaminant (53%; Pseudomonas aeruginosa was found only in one semen sample. The analysis of variance of factors affecting contamination levels was significant for the farm of origin (P<0.05 and not significant for the breed. Antibiotic resistance of these bacteria was assessed using different antibiotics. Significant differences (P<0.05 between observed and expected frequencies of bacterial isolates resistant or not to the antibiotics contained in the extenders were found. At 48 h of storage a reduction of aerobic contamination was found after ME-S dilution by 85.3% and after CRONOSTM by 63.8%. This paper proved the presence of pathogenic bacteria in semen. We thus believe it is highly advisable to perform periodic microbiological screening of boar semen in the swine industry to avoid the use of low sperm quality.

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

    Science.gov (United States)

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

    2015-10-01

    The possibility of including amino acids for cryopreservation of ram semen to improve the quality of frozen semen was explored in this study in sheep model. 24 samples were collected in triplicate from 8 rams of 2-3 year old Bannur cross bred rams maintained at the Institute Experimental Livestock Unit. Semen was diluted in tris-egg yolk glycerol diluent and made into 7 aliquots as follows: aliquot 1 served as control, "l-alanine" was added at 100 and 135mM in the aliquots 2 and 3, "l-glutamine" was added at 20 and 25mM in the aliquots 4 and 5 and "l-proline" was added at 25 and 50mM in the aliquots 6 and 7, respectively. Diluted semen was filled in 0.25ml French straws and frozen in LN2. Inclusion of "l-proline" and "l-glutamine" in the diluent increased the percent live sperm (Psemen additive to freeze ram semen as they prevented cryoinjuries to sperm and improved the pre-freeze and post-thaw semen characteristics. PMID:26362050

  13. Techniques for sperm evaluation using fluorescent probes

    Directory of Open Access Journals (Sweden)

    Andrielle Thainar Mendes Cunha

    2015-12-01

    Full Text Available  A variety of laboratory tests were developed to obtain more reliable results of sperm evaluation and increase the accuracy of sperm fertility predictions. These tests detected damage of sperm specific compartments or organelles, which cannot be detected in routine sperm analysis. The use of fluorescent probes and detection using fluorescent microscopy or flow cytometry is an important tool but a more precise and accurate laboratory test is needed. Propidium iodide and 6-carboxyfluorescein diacetate are used for evaluations of plasmatic membrane integrity. Fluorescein isothiocyanate, associated with conjugated lecithin Psium sativum or Arachis hypogaea, are used for evaluations of acrosome integrity. Two probes, MitoTracker or Rhodamine123, are generally used to measure the absence or presence of mitochondrial potential. However, a better option is 5,5’; 6,6’ - tetrachloro - 1,1’; 3,3’ -tetraetilbenzimidazolil-carbocyanine (JC-1 dye, which assesses not only the presence of mitochondrial potential and distinguished spermatozoa with poorly and highly functional mitochondria. Two techniques, TUNEL or COMETA, and the Acridine Orange Test (AOT dye are used to evaluate chromatin integrity. A fluorescence technique based on chlortetracycline (CTC or Merocyanine 540 is used to estimate whether sperm pass by or through the capacitation process. This review focuses on the fluorescent probes that are most widely used to evaluate plasma membrane integrity, capacitation, acrosome integrity, chromatin integrity and mitochondrial potential.

  14. ENVIRONMENTAL CONTAMINANTS IN WILD BOARS FROM CALABRIA

    Directory of Open Access Journals (Sweden)

    F. Naccari

    2011-01-01

    Full Text Available The aim of this study was to determine heavy metals (Cd, Cu, Pb and Zn organochlorine pesticides (POCs and polychlorinated biphenyl (PCBs in some samples (heart, kidney, liver, lung, muscle tissue and spleen of wild boars (utilized as “bioindicator” from various areas from Calabria. Quantitative determination of POCs and PCBs were carried out using GC-ECD and confirmed with GC-MS. The concentrations of heavy metals were determined by a Varian Atomic Absorption Spectroscopy instrument. Our data have shown low residual levels of OCs, heavy metals and the absence of PCBs in all samples analyzed and therefore the boar meat products are not dangerous for the consumer. Moreover, results obtained deserve particular attention not only for their significance but especially because they were recorded in Calabria, a region a low risk of environmental pollution due to the shortage of industries and the traditional agricultural activity.

  15. Porcine hokovirus in wild boar in Portugal.

    Science.gov (United States)

    Miranda, Carla; Coelho, Catarina; Vieira-Pinto, Madalena; Thompson, Gertrude

    2016-04-01

    Porcine hokovirus (PHoV), also referred to as porcine parvovirus 4 (P-PARV4), a recently discovered parvovirus of swine that is closely related to human parvovirus 4/5 (H-PARV4/5), was first described in Hong Kong. To evaluate the occurrence of P-PARV4 in Portuguese wild boars in the hunting season of 2011/2012, liver and serum samples were tested. P-PARV4 was detected in 24 % of the wild boars analyzed. Phylogenetic analysis showed a close relationship between the P-PARV4 isolates and other P-PARV4 reference strains. This virus appears to be emerging, with yet unknown implications for public health. PMID:26711454

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

    OpenAIRE

    Gomendio Montserrat; Tourmente Maximiliano; Roldan Eduardo RS

    2011-01-01

    Abstract Background The influence of sperm competition upon sperm size has been a controversial issue during the last 20 years which remains unresolved for mammals. The hypothesis that, when ejaculates compete with rival males, an increase in sperm size would make sperm more competitive because it would increase sperm swimming speed, has generated contradictory results from both theoretical and empirical studies. In addition, the debate has extended to which sperm components should increase i...

  17. ENVIRONMENTAL CONTAMINANTS IN WILD BOARS FROM CALABRIA

    OpenAIRE

    Naccari, F.; E. Palma; C. Giofrè; P. Licata; F. Giofrè; Rotiroti, D

    2011-01-01

    The aim of this study was to determine heavy metals (Cd, Cu, Pb and Zn) organochlorine pesticides (POCs) and polychlorinated biphenyl (PCBs) in some samples (heart, kidney, liver, lung, muscle tissue and spleen) of wild boars (utilized as “bioindicator”) from various areas from Calabria. Quantitative determination of POCs and PCBs were carried out using GC-ECD and confirmed with GC-MS. The concentrations of heavy metals were determined by a Varian Atomic Absorption Spectroscopy in...

  18. Acrosin inhibitor detecting along the boar epididymis

    Czech Academy of Sciences Publication Activity Database

    Maňásková-Postlerová, Pavla; Cozlová, Nina; Dorosh, Andriy; Šulc, Miroslav; Guyonet, B.; Jonáková, Věra

    2016-01-01

    Roč. 82, Jan 2016 (2016), s. 733-739. ISSN 0141-8130 R&D Projects: GA ČR(CZ) GAP503/12/1834; GA MŠk(CZ) ED1.1.00/02.0109; GA ČR GA14-05547S Institutional support: RVO:86652036 ; RVO:61388971 Keywords : Acrosin inhibitor * Boar epididymis * Spermatozoa Subject RIV: CE - Biochemistry Impact factor: 2.858, year: 2014

  19. Use of Silver Nitrate for the Assessment of Sperm Measurements in Selected Farm and Free-Living Animal Species

    Directory of Open Access Journals (Sweden)

    Andraszek Katarzyna

    2014-10-01

    Full Text Available The study was conducted on spermatozoa of selected farm and free-living animal species, isolated post mortem from the tail of the epididymis, and stained with silver nitrate - AgNO3. The material was collected from pigs, goats, wild boar, and European roe deer. Twenty morphologically normal spermatozoa randomly selected from each animal and well visible under the microscope, were analysed. The following measurements were considered: head length, width, perimeter and area, acrosome area, mid-piece length, tail length, and overall sperm length. AgNO3 staining differentiated the acrosomal (light hue and distal (dark hue part of the sperm head, and a light-hued mid-piece was visible within the sperm tail. Silver nitrate staining revealed species and variety-related differences, particularly in reference to the sperm head. Clear-cut differentiation within the head and tail area made it possible to perform detailed morphometric measurements of the spermatozoa.

  20. Sperm preparation for fertilization

    OpenAIRE

    Gadella, B.M.

    2014-01-01

    Description This book contains 19 chapters that discuss theoretical and applied andrology for domestic, zoo and wild animals. Topics include semen and its constituents; sperm production and harvest; determinants of sperm morphology; sperm preparation for fertilization; practical aspects of semen cryopreservation; evaluation of semen in the andrology laboratory; genetic aspects of male reproduction; emerging techniques and future development of semen evaluation and handling and applied androlo...

  1. A role for the WH-30 protein in sperm-sperm adhesion during rouleaux formation in the guinea pig.

    Science.gov (United States)

    Flaherty, S P; Swann, N J; Primakoff, P; Myles, D G

    1993-03-01

    Mammalian spermatozoa participate in specific cell adhesion phenomena during their development and functional lifespan; this includes interaction with Sertoli cells, the zona pellucida, and the oolemma. In some species such as the guinea pig, an additional sperm-sperm adhesion occurs during epididymal maturation which results in the formation of rouleaux in which the sperm heads are stacked one upon the other and the periacrosomal plasma membranes of adjacent sperm are linked by periodic cross-bridges. In this study, we have used a monoclonal antibody to investigate the role of the WH-30 protein on the sperm surface in the formation of the junctional zones between adjacent guinea pig sperm in rouleaux. WH-30 monoclonal antibodies caused a dose- and time-dependent dissociation of rouleaux and an increase in the percentage of single, acrosome-intact sperm; there were no effects on sperm motility (maintained at 80-90%) or ultrastructure during the 120-min incubations. The maximal effect of about 80% single sperm was obtained with a 1:4 dilution of the WH-30 hybridoma supernatant or 5-50 micrograms/ml of purified WH-30 IgG. In contrast, incubation of sperm in AH-20 IgG, myeloma cell supernatants, or purified, nonspecific mouse IgG1 had no effect on rouleaux. Treatment of sperm with a WH-30 Fab fragment resulted in almost complete dissociation of rouleaux without any observed effect on sperm motility or acrosomal status. Surface labeling of sperm followed by immunoprecipitation and SDS-PAGE revealed that the WH-30 antibody recognizes a single polypeptide of 43-45 kDa. Using immunofluorescence, the WH-30 protein was localized over the entire surface of the sperm head (whole-head pattern), and immunogold labeling showed that WH-30 is localized in the glycocalyx on both the dorsal and ventral surfaces of the periacrosomal and postacrosomal plasma membranes. These results indicate that the WH-30 protein on the sperm surface is a cell adhesion protein which is involved in

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

    Science.gov (United States)

    ... of the American Society for Reproductive Medicine Sperm morphology (shape): Does it affect fertility? How is a ... motility of the sperm (percentage of moving sperm), morphology of the sperm (percentage of normally shaped sperm), ...

  3. 二甲基亚砜对猪精液冷冻保存效果研究%Effects of Dimethyl Sulfoxide on Boar Semen Cryopreservation

    Institute of Scientific and Technical Information of China (English)

    马丽; 李青旺; 吴民耀

    2014-01-01

    concentration of 30 mL/L,respec-tively marked for group A,B ,C,D and E,and then the boar semen cryopreservation were compared and the optimum proportion were selected.The results showed that by adding 60 mL/L DMSO,after being frozen-thawed,the sperm abnormal rates,the plasma membrane integrity and sperm acrosomes integrity were remarkably better than that in the control group and other groups (P 0.05),it had no significant difference.The sperm acro-somes integrity and the plasma membrane integrity were 53% and 52%,which were remarkably higher than groups added with 40,50,70,80 mL/L DMSO and other compatible groups (P 0.05).The sperm viability that was 37% was remarkably lower than that in the control group (P < 0.05),but remarkably higher than that in other DMSO added groups and the compatible groups (P < 0.05).So when adding DMSO sepa-rately,the optimum concentration was 60 mL/L,and the best proportion for the mixed cryoprotectants a-gent,of which the final concentration was 30 mL/L,was 3∶1 (glycerin to DMSO).

  4. Sperm Proteases that May Be Involved in the Initiation of Sperm Motility in the Newt, Cynops pyrrhogaster

    Directory of Open Access Journals (Sweden)

    Misato Yokoe

    2014-08-01

    Full Text Available A protease of sperm in the newt Cynops pyrrhogaster that is released after the acrosome reaction (AR is proposed to lyse the sheet structure on the outer surface of egg jelly and release sperm motility-initiating substance (SMIS. Here, we found that protease activity in the sperm head was potent to widely digest substrates beneath the sperm. The protease activity measured by fluorescein thiocarbamoyl-casein digestion was detected in the supernatant of the sperm after the AR and the activity was inhibited by 4-(2-aminoethyl benzenesulfonyl fluoride (AEBSF, an inhibitor for serine or cysteine protease, suggesting the release of serine and/or cysteine proteases by AR. In an in silico analysis of the testes, acrosins and 20S proteasome were identified as possible candidates of the acrosomal proteases. We also detected another AEBSF-sensitive protease activity on the sperm surface. Fluorescence staining with AlexaFluor 488-labeled AEBSF revealed a cysteine protease in the principal piece; it is localized in the joint region between the axial rod and undulating membrane, which includes an axoneme and produces powerful undulation of the membrane for forward sperm motility. These results indicate that AEBSF-sensitive proteases in the acrosome and principal piece may participate in the initiation of sperm motility on the surface of egg jelly.

  5. Effect of Ostrich Egg Yolk on Boar Spermatozoa Quality after Freezing-thawing%鸵鸟卵黄对猪精子冷冻后质量的影响

    Institute of Scientific and Technical Information of China (English)

    马玲琴; 贾永宏; 马国际; 袁建民

    2012-01-01

    为了提高猪冷冻精液品质和精子抵抗低温打击的能力,本研究以5%、10%、15%、20%和25%等不同浓度的鸵鸟卵黄作为冷冻保护剂,以20%的鸡蛋卵黄和20%的鸽蛋卵黄为对照,将冷冻-解冻后的精子活率、质膜完整率和顶体完整率作为评价指标,分析鸵鸟卵黄对猪精子的抗冷冻保护作用。结果表明:稀释液中添加20%鸽蛋卵黄时,精子活率、顶体完整率和质膜完整性分别为52.11%、55.62%和54.94%,显著高于其他组(P〈0.05)。虽然稀释液中添加15%鸵鸟卵黄时,冷冻-解冻后精子活率、顶体完整率和质膜完整率显著高于5%、10%、20%和25%鸵鸟卵黄组,但仍然显著低于稀释液中添加20%鸽蛋卵黄处理组。本研究表明,鸵鸟卵黄在冷冻过程中对猪精子具有一定的保护作用,但相对于鸽子蛋和鸡蛋卵黄效果并不理想。%In order to improve the frozen semen quality and sperm resistance to cold shock,the different concentrations of 5%,10%,15%,20% and 25% ostrich egg yolk were added in extender as cryoprotectants,and 20% egg yolk and 20% pigeon egg yolk were control.The cryoprotective effects of ostrich egg yolk on the boar spermatozoa were analyzed and spermatozoa motility,acrosome integrity and membrane integrity after freezing-thawing were evaluated.The results indicated that the sperm motility,acrosome integrity,membrane integrity were 52.11%,55.62% and 54.94% respectively while the extender was supplied with 20% pigeon egg yolk,which was higher than that of other groups(P0.05).Although sperm motility,acrosome integrity and membrane integrity in the extender supplemented with 15% ostrich egg yolk were significantly higher than that of 5%,10%,20% and 25% ostrich egg yolk groups,they were significantly lower than that of 20% pigeon egg yolk group.According to our study,the ostrich egg yolk could protect the boar spermatozoa during the frozen-thawed,but is not as effective as pigeon egg yolk and egg

  6. The biopsy of the boar testes using ultrasonographic examination

    Directory of Open Access Journals (Sweden)

    Laima Liepa

    2014-03-01

    Full Text Available The biopsy of live animal testes is an important clinical manipulation to control spermatogenesis and reproductive system pathologies. The aim was to develop a method of boar testes biopsy using a biopsy gun with ultrasound guidance and to investigate the influence of this procedure on the boar testes parenchyma and quality of ejaculate. The biopsy was carried out in six 8-month-old boars. Fourteen days prior to and 21 days after biopsy, the quality of ejaculate was examined (weight of ejaculate; concentration and motility of spermatozoa with a seven-day intervals. Ultrasound images of the testes parenchyma were recorded three times: directly before and 15 minutes after the biopsy, then 21 days after the procedure. The testes biopsies of generally anesthetized boars were performed with the biopsy gun for needle biopsy with a 12cm long, disposable 16-gauge needle 1.8mm in diameter (Vitesse through 1cm skin incision in the depth of 1.2-1.6cm of parenchyma. Fifteen minutes after the biopsy, macroscopic injures of the parenchyma of all the boar testes were not detected in the ultrasound image. Twenty one days after biopsy, the hyperechogenic line 0.1-0.2cm in diameter was seen in the testes parenchyma of six boars in the depth of 1.2-1.6cm. The biopsy of boar testes did not influence the quality of boars ejaculate. The ultrasonographic examination of boar testicles before the biopsy reduced possibilities to traumatize large blood vessels of the testes. A perfect boar testicular biopsy was easy to perform using ultrasonographic examination in the pigsty conditions.

  7. Membraner

    DEFF Research Database (Denmark)

    Bach, Finn

    2009-01-01

    Notatet giver en kort introduktion til den statiske virkemåde af membraner og membrankonstruktioner......Notatet giver en kort introduktion til den statiske virkemåde af membraner og membrankonstruktioner...

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

    DEFF Research Database (Denmark)

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

    2003-01-01

    Multiple insemination induces sperm competition and may select for longer, faster moving sperm in species where sperm is short-lived and egg fertilization takes place almost immediately after ejaculation. Here we report the first detailed analysis of sperm length in social insects with long...

  9. Interactions between zona pellucida glycoproteins and sperm proacrosin/acrosin during fertilization.

    Science.gov (United States)

    Howes, Liz; Jones, Roy

    2002-01-01

    Fertilization is one of the most specific and carefully regulated cell-cell interactions in the animal body and is determined to a large extent by compatibility between ligand and receptor molecules on the surface of each gamete. On the zona pellucida (ZP), sperm receptor activity is associated with glycoproteins ZP3 (primary receptor for acrosome-intact sperm) and ZP2 (secondary receptor for acrosome-reacted sperm) but their complementary binding proteins on sperm are less well defined. In this communication we review the evidence for proacrosin as a secondary ZP binding protein. Proacrosin/acrosin binds non-enzymically to ZP glycoproteins. Binding is a strong ionic interaction between polysulphate groups on ZP glycoproteins (probably on their carbohydrate moieties) and basic residues on the surface of proacrosin. The stereochemistry of the reactants is crucial and determines to a large extent the affinity of binding. Site-directed mutagenesis and a 3D-structural analysis of boar and ram acrosin have identified 2 clusters of basic residues potentially involved in binding. A polysulphonated anticancer drug, suramin, has been shown to bind strongly to proacrosin/acrosin and to inhibit sperm-egg binding in vitro. In the mouse model, 125I-ZP2 and 3H-suramin bind approximately 65% less effectively to acrosin 'null' sperm than to wild-type sperm. Neither ZP2 nor suramin bind to acrosome intact sperm and can, therefore, only exert their effects after exposure of the acrosomal contents. Overall, this combination of biochemical, genetic and functional data supports the hypothesis that proacrosin is a multifunctional protein with a significant role in retaining acrosome-reacted sperm on the ZP surface long enough to enable ZP penetration to begin. PMID:11730915

  10. Wild Boar Research – A Never Ending Story?

    Directory of Open Access Journals (Sweden)

    O. Keuling

    2013-12-01

    Full Text Available Wild boar science is changing a lot. The species wild boar (Sus scrofa, once threatened, is one of the latest domesticated species. Wild boar is so successful that currently it causes strong economic and ecological damages all over the world. The interest in Sus scrofa continues to grow rapidly, not only within its native range, but also in all other continents where wild boar and feral pigs have been introduced. Environmentally sensitive and adaptative management plus conservation of wild boar, feral pigs and other suids is of increasing concern to conservation biologists, wildlife managers, veterinarians, policy makers and the general public. Important advances in research may help managing wild boar as a pest and other suids as threatened species. Also a good exchange with stakeholders is of huge importance within wildlife management. In this special issue of Wildlife Biology in Practice some results from the 9th International Symposium on Wild Boar and other Suids as well as additional publications on wild boar are centralised. All together 110 participants from 24 countries took part at the 9th ISWB in Hannover, Germany. The main part of the 59 presentations focused on wild boar management and monitoring (29 contributions. These numbers points out the importance of wild boar in all parts of its current distribution area. Everywhere populations are increasing (with some very few exceptions. In many of these regions economic problems, mainly by agricultural damages, road accidents and animal diseases are the main drivers for scientific interests. Recently many researchers try to establish, or even to create, reliable and practical census methods. Only with reliable data on numbers, reproduction, im- and emigration as well as mortality rates, managers will be able to know the efficiency of management methods. Even if a lot of effort is done, it looks like we are still far away from successful control of wild boar or feral pigs’ populations

  11. Differences in the fatty-acid composition of rodent spermatozoa are associated to levels of sperm competition

    Directory of Open Access Journals (Sweden)

    Javier delBarco-Trillo

    2015-03-01

    Full Text Available Sperm competition is a prevalent phenomenon that drives the evolution of sperm function. High levels of sperm competition lead to increased metabolism to fuel higher sperm velocities. This enhanced metabolism can result in oxidative damage (including lipid peroxidation and damage to the membrane. We hypothesized that in those species experiencing high levels of sperm competition there are changes in the fatty-acid composition of the sperm membrane that makes the membrane more resistant to oxidative damage. Given that polyunsaturated fatty acids (PUFAs are the most prone to lipid peroxidation, we predicted that higher sperm competition leads to a reduction in the proportion of sperm PUFAs. In contrast, we predicted that levels of sperm competition should not affect the proportion of PUFAs in somatic cells. To test these predictions, we quantified the fatty-acid composition of sperm, testis and liver cells in four mouse species (genus Mus that differ in their levels of sperm competition. Fatty-acid composition in testis and liver cells was not associated to sperm competition levels. However, in sperm cells, as predicted, an increase in sperm competition levels was associated with an increase in the proportion of saturated fatty-acids (the most resistant to lipid peroxidation and by a concomitant decrease in the proportion of PUFAs. Two particular fatty acids were most responsible for this pattern (arachidonic acid and palmitic acid. Our findings thus indicate that sperm competition has a pervasive influence in the composition of sperm cells that ultimately may have important effects in sperm function.

  12. Oocyte specific oolemmal SAS1B involved in sperm binding through intra-acrosomal SLLP1 during fertilization

    OpenAIRE

    Sachdev, Monika; Mandal, Arabinda; Mulders, Sabine; Digilio, Laura C.; Panneerdoss, Subbarayalu; Suryavathi, Viswanadhapalli; Pires, Eusebio; Klotz, Kenneth L.; Hermens, Laura; Herrero, Maria Belen; Flickinger, Charles J.; van Duin, Marcel; Herr, John C.

    2011-01-01

    Molecular mechanisms by which fertilization competent acrosome-reacted sperm bind to the oolemma remain uncharacterized. To identify oolemmal binding partner(s) for sperm acrosomal ligands, affinity panning was performed with mouse oocyte lysates using sperm acrosomal protein, SLLP1 as a target. An oocyte specific membrane metalloproteinase, SAS1B (Sperm Acrosomal SLLP1 Binding), was identified as a SLLP1 binding partner. cDNA cloning revealed six SAS1B splice variants, each containing a zinc...

  13. Effect of astaxanthin on human sperm capacitation.

    Science.gov (United States)

    Donà, Gabriella; Kožuh, Ivana; Brunati, Anna Maria; Andrisani, Alessandra; Ambrosini, Guido; Bonanni, Guglielmo; Ragazzi, Eugenio; Armanini, Decio; Clari, Giulio; Bordin, Luciana

    2013-06-01

    In order to be able to fertilize oocytes, human sperm must undergo a series of morphological and structural alterations, known as capacitation. It has been shown that the production of endogenous sperm reactive oxygen species (ROS) plays a key role in causing cells to undergo a massive acrosome reaction (AR). Astaxanthin (Asta), a photo-protective red pigment belonging to the carotenoid family, is recognized as having anti-oxidant, anti-cancer, anti-diabetic and anti-inflammatory properties and is present in many dietary supplements. This study evaluates the effect of Asta in a capacitating buffer which induces low ROS production and low percentages of acrosome-reacted cells (ARC). Sperm cells were incubated in the presence or absence of increasing concentrations of Asta or diamide (Diam) and analyzed for their ROS production, Tyr-phosphorylation (Tyr-P) pattern and percentages of ARC and non-viable cells (NVC). Results show that Asta ameliorated both sperm head Tyr-P and ARC values without affecting the ROS generation curve, whereas Diam succeeded in enhancing the Tyr-P level but only of the flagellum without increasing ARC values. It is suggested that Asta can be inserted in the membrane and therefore create capacitation-like membrane alteration which allow Tyr-P of the head. Once this has occurred, AR can take place and involves a higher numbers of cells. PMID:23736766

  14. Effect of Astaxanthin on Human Sperm Capacitation

    Directory of Open Access Journals (Sweden)

    Luciana Bordin

    2013-06-01

    Full Text Available In order to be able to fertilize oocytes, human sperm must undergo a series of morphological and structural alterations, known as capacitation. It has been shown that the production of endogenous sperm reactive oxygen species (ROS plays a key role in causing cells to undergo a massive acrosome reaction (AR. Astaxanthin (Asta, a photo-protective red pigment belonging to the carotenoid family, is recognized as having anti-oxidant, anti-cancer, anti-diabetic and anti-inflammatory properties and is present in many dietary supplements. This study evaluates the effect of Asta in a capacitating buffer which induces low ROS production and low percentages of acrosome-reacted cells (ARC. Sperm cells were incubated in the presence or absence of increasing concentrations of Asta or diamide (Diam and analyzed for their ROS production, Tyr-phosphorylation (Tyr-P pattern and percentages of ARC and non-viable cells (NVC. Results show that Asta ameliorated both sperm head Tyr-P and ARC values without affecting the ROS generation curve, whereas Diam succeeded in enhancing the Tyr-P level but only of the flagellum without increasing ARC values. It is suggested that Asta can be inserted in the membrane and therefore create capacitation-like membrane alteration which allow Tyr-P of the head. Once this has occurred, AR can take place and involves a higher numbers of cells.

  15. Effect of various commercial buffers on sperm viability and capacitation.

    Science.gov (United States)

    Andrisani, Alessandra; Donà, Gabriella; Ambrosini, Guido; Bonanni, Guglielmo; Bragadin, Marcantonio; Cosmi, Erich; Clari, Giulio; Armanini, Decio; Bordin, Luciana

    2014-08-01

    A wide variety of sperm preparation protocols are currently available for assisted conception. They include density gradient separation and washing methods. Both aim at isolating and capacitating as much motile sperm as possible for subsequent oocyte fertilization. The aim of this study was to examine the effects of four commercial sperm washing buffers on sperm viability and capacitation. Semen samples from 48 healthy donors (normal values of sperm count, motility, morphology, and volume) were analyzed. After separation (density gradient 40/80%), sperm were incubated in various buffers then analysed for reactive oxygen species (ROS) production, viability, tyrosine phosphorylation (Tyr-P), cholera toxin B subunit (CTB) labeling, and the acrosome reaction (AR). The buffers affected ROS generation in various ways resulting either in rapid cell degeneration (when the amount of ROS was too high for cell survival) or the inability of the cells to maintain correct functioning (when ROS were too few). Only when the correct ROS generation curve was maintained, suitable membrane reorganization, evidenced by CTB labeling was achieved, leading to the highest percentages of both Tyr-P- and acrosome-reacted-cells. Distinguishing each particular pathological state of the sperm sample would be helpful to select the preferred buffer treatment since both ROS production and membrane reorganization can be significantly altered by commercial buffers. PMID:24673547

  16. Effect of sperm concentration on characteristics and fertilization capacity of rooster sperm frozen in the presence of the antioxidants catalase and vitamin E.

    Science.gov (United States)

    Moghbeli, Morteza; Kohram, Hamid; Zare-Shahaneh, Ahmad; Zhandi, Mahdi; Sharideh, Hossein; Sharafi, Mohsen

    2016-10-01

    The objective of this study conducted was to determine the influence of different levels of sperm concentration, including catalase (CAT) and vitamin E (VitE) in rooster semen extender on postthawed quality and fertility of rooster semen. Semen was collected twice a week from six roosters (Arian) and diluted according to experimental treatments consisting of sperm suspensions containing different sperm concentrations (200, 400, and 600 × 106 sperm/mL) without antioxidant supplementation as control (Con) groups (Con200, Con400, and Con600, respectively), sperm suspensions containing different sperm concentrations (200, 400, and 600 × 106 sperm/mL) supplemented with 5-μg/mL VitE (VitE200, VitE400, and VitE600, respectively) and different sperm concentrations (200, 400, and 600 × 106 sperm/mL) supplementation with 100 IU/mL CAT (CAT200, CAT400, and CAT600, respectively). After thawing; sperm motility, membrane integrity, and mitochondrial function were assessed. Fertility and hatchability rates were determined by using 100 artificially inseminated hens. The percentage of total motility (TM) and activity of mitochondria decreased (P  0.05) on fertility and hatchability rates. In conclusion, although adding VitE and CAT in extender with different levels of sperm concentration improved postthawed quality of rooster semen, but adding VitE and CAT in the extender have no effect on fertility rate. PMID:27444422

  17. Conception Rate and Litter Size in Multiparous Sows after Intrauterine Insemination Using Frozen-Thawed Boar Semen in a Commercial Swine Herd in Thailand

    OpenAIRE

    Chanapiwat, Panida; OLANRATMANEE, Em-on; Kaeoket, Kampon; Tummaruk, Padet

    2014-01-01

    ABSTRACT The aim of the present study was to determine the conception rate and litter size in sows after fixed time intra-uterine insemination using frozen-thawed boar semen in a commercial swine herd in Thailand. Sixty-nine Landrace multiparous sows were randomly allocated into two groups, including control (n=36) and treatment (n=33). The control sows were inseminated with extended fresh semen (3 × 109 motile sperm/dose, 100 ml) at 24, 36 and 48 hr after the onset of estrus. The treatment s...

  18. Fertilization Rate and Number of Embryos on Day 2 after Intrauterine and Deep Intrauterine Insemination Using Frozen-Thawed Boar Semen in Multiparous Sows

    OpenAIRE

    Kakanang Buranaamnuay; Yodchai Panyaboriban; Padet Tummaruk; Mongkol Techakumphu

    2011-01-01

    The present study determines fertilization rate and number of embryos on Day 2 after intrauterine insemination (IUI) and deep intrauterine insemination (DIUI) using frozen-thawed (FT) boar semen in multiparous sows. Twelve crossbred Landrace × Yorkshire multiparous sows were included. The sows were inseminated at 24 h after oestrus detection and reinseminated every 12 h until ovulation took place. The inseminations were conducted using IUI with 2 × 109 FT sperm per dose (n = 6) and DIUI with ...

  19. Standardisation of a novel sperm banking kit - NextGen(®) - to preserve sperm parameters during shipment.

    Science.gov (United States)

    Agarwal, A; Sharma, R; Singh, A; Gupta, S; Sharma, R

    2016-08-01

    Many male patients diagnosed with cancer are within their reproductive years. These men are advised to freeze their spermatozoa prior to the start of cancer treatment. Very often, sperm banking facilities may not be readily available and patients may be required to travel to distant sperm bank centres. Our objective was to design and standardise a remote home shipping sperm kit that allows patients to collect a semen sample at home and ship it overnight to a sperm bank. A total of 21 semen samples and two transport media (refrigeration media and human tubal fluid) and five different combinations of ice packs were tested for maintaining desired shipping temperature. Ten semen samples were assessed for pre- and post-shipment changes in sperm motility, membrane integrity, total motile spermatozoa and recovery of motile spermatozoa. Even though motility, membrane integrity and total motile spermatozoa declined both in samples examined under simulated shipped conditions and in overnight-shipped samples, the observed motility and total motile spermatozoa were adequate for use with assisted reproductive techniques. Using refrigeration media, cooling sleeve and ice packs, adequate sperm motility can be maintained utilising NextGen(®) kit and these spermatozoa can be used for procreation utilising ART techniques such as intracytoplasmic sperm injection. PMID:26564753

  20. Prevalence of infection in hunted wild boars () in Germany

    OpenAIRE

    Reiner, Gerald; Fresen, Christina; Bronnert, Sebastian; Haack, Ingo; Willems, Hermann

    2010-01-01

    International audience is the etiological agent of Glässer's disease, often involved in pneumonia, and also an early colonizer of the upper respiratory tract of healthy domestic pigs. Little information is available on in wild boars. The aim of the present study was to evaluate infection in wild boars in Germany. Tissue samples from the lungs and tonsils of 531 wild boars from 52 hunts during the hunting seasons 2004/2005 to 2006/2007 were examined independently for by PCR because is a fas...

  1. Daily sperm production

    DEFF Research Database (Denmark)

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

    2013-01-01

    We investigated the influence of maternal airway exposure to nanoparticulate titanium dioxide (TiO2, UV-Titan) and carbon black (CB, Printex90), on male reproductive function in the two following generations. Time-mated C57BL/6J mice were exposed by inhalation to UV-Titan, or by intratracheal...... exposure did not affect DSP statistically significantly in the F1 generation, although TiO2 tended to reduce sperm counts. Overall, time-to-first F2 litter increased with decreasing sperm production. There was no effect on sperm production in the F2 generation originating after TiO2 exposure. F2 offspring...

  2. A comparative study of sperm morphometric subpopulations in cattle, goat, sheep and pigs using a computer-assisted fluorescence method (CASMA-F).

    Science.gov (United States)

    Vicente-Fiel, S; Palacín, I; Santolaria, P; Yániz, J L

    2013-06-01

    This study was designed to compare the sperm nuclear morphometric subpopulations of four species of domestic artiodactyls (cattle, sheep, goat and pigs). Samples from 20 males of each species were collected. After semen collection, sperm concentration and motility were measured and samples prepared for morphometric determinations. Smears were fixed with 2% glutaraldehyde, stained with Hoechst 33342 and photographed. At least 200 spermatozoa per sample were processed using the Image J analysis open software. Clustering procedures were performed to identify sperm subpopulations using the morphometric data obtained from each species. Results of the present study show that, applying the computer-assisted sperm morphometry analyisis-fluorescence (CASMA-F) technology and multivariate cluster analyses, it was possible to determine the subpopulations of spermatozoa with different morphometric characteristics in the four species studied. Bulls and boars had two clearly differentiated size categories: large and small. However, the final sperm subpopulations were four in the bull (large-round, large-elongated, small-round, and small-elongated) and only three in the boar (large, small-elongated and small-round). In small ruminant species, three sperm nuclei size categories were established: large, average sized and small. Two of these subpopulations were also elongated in goat bucks, with three subpopulations (large-round, small-elongated and average size-elongated). In the ram three morphometric subpopulations were also obtained (large, small and average size-round), but none was elongated. When comparing among species, sperm subpopulations were smaller in the buck and less elliptical and elongated in the ram than those in the other species studied. Male variability was identified in the distribution of sperm subpopulations described in the four species studied. It was concluded that the combination of CASMA-F technology with multivariate cluster analyses allow the study of

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

    OpenAIRE

    J. L. Yániz; I. Palacín; S. Vicente-Fiel; J. Gosalvez; C. López-Fernández; Santolaria, P.

    2013-01-01

    This study was designed to develop a semiautomatic computer assisted methodology to evaluate the membrane integrity of ram spermatozoa using a commercial kit based on acridine orange/propidium iodide (AO/PI) labelling and ImageJ software. The study was divided into two experiments. In the first trial, the new computer-assisted method was validated by mixing fresh semen samples with different volumes of freeze killed spermatozoa to determine proportions of damaged spermatozoa in the final samp...

  4. THE ADDITION OF CAFFEINE IN EARLE’S BALANCED SALT SOLUTION MEDIA WITH WASHING UP METHOD INCREASE MEMBRANE INTEGRITY AND ACROSOMAL SPERM

    OpenAIRE

    B. K. Satriyasa; and A. N. Mahendra

    2014-01-01

    Background: caffeine, a methylxanthine derivate, appears to inhibit phosphodiesterase, thereby inhibiting the break down of cAMP and increasing its concentration inside cell. This study aims to assess the effect of caffeine addition in Earles’s Balanced Salt Solution (EBSS) on the increase in membrane integrity and acrosome reaction of spermatozoa using swim up method. Methods: This study was carried out at the Clinic of Sexology and Andrology, Sanglah Public Hospital at Denpasar Bali-Indones...

  5. Influence of ω-3 Polyunsaturated Fatty Acids on Boar Semen Quality and Their Mechanisms%ω-3多不饱和脂肪酸对公猪精液质量的影响及其机制

    Institute of Scientific and Technical Information of China (English)

    曾凡熙; 牟永斌; 靳露; 董国忠

    2012-01-01

    在规模化养猪生产中,公猪的精液品质直接影响母猪的繁殖性能,这对猪场的生产成绩非常重要.多不饱和脂肪酸(PUFA)对精子质膜的流动性和脂质过氧化反应的敏感性有重要作用,同时也影响精子的授精能力.研究表明,在公猪饲粮中添加ω-3 PUFA能提高公猪精液品质,但效果重复性还不稳定;公猪饲粮中添加ω-3 PUFA能增加猪精子中ω-3 PUFA的含量和延长精液的保存时间.本文主要综述公猪饲粮中添加ω-3 PUFA对公猪精液品质的影响及其主要的作用机制,为其在现代规模化养猪生产中有效提高精液品质、猪场生产成绩和经济效益提供参考.%In modern large-scale pig production, semen quality has a direct impact on reproductive performance of sows and plays a critical role in pig farm' s production efficiency. Polyunsaturated fatty acids (PUFA) play a key role in the sperm membrane fluidity and susceptibility to lipid peroxidation. Moreover, PUFA influence the sperm fertilization. Many studies have shown that dietary ω-3 PUFA supplementation improves semen quality , but the reproducibility is not stable. Dietary ω-3 PUFA supplementation can extend the storage time of semen and exert positive effects on sperm fatty acid profile. This paper reviews the effects of dietary ω-3 PUFA supplementation on boar semen quality, and analyzes the respective mechanisms, which can help to improve semen quality, increase productivity and enhance economic return in modern large-scale pig production.

  6. Calcium Clearance Mechanisms of Mouse Sperm

    OpenAIRE

    Wennemuth, Gunther; Babcock, Donner F; Hille, Bertil

    2003-01-01

    The spermatozoon is specialized for a single vital role in fertilization. Past studies show that Ca2+ signals produced by the opening of plasma membrane entry channels initiate several events required for the sperm to reach and enter the egg but reveal little about how resting [Ca2+]i is maintained or restored after elevation. We examined these homeostatic mechanisms by monitoring the kinetics of recovery from depolarizing stimuli under conditions intended to inhibit candidate mechanisms for ...

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

    Directory of Open Access Journals (Sweden)

    Sara Keshtgar

    2016-05-01

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

  8. Dual roles for ubiquitination in the processing of sperm organelles after fertilization

    OpenAIRE

    Hajjar, Connie; Sampuda, Katherine M; Boyd, Lynn

    2014-01-01

    Background The process of fertilization involves a cell fusion event between the sperm and oocyte. Although sperm contain mitochondria when they fuse with the oocyte, paternal mitochondrial genomes do not persist in offspring and, thus, mitochondrial inheritance is maternal in most animals. Recent evidence suggests that paternal mitochondria may be eliminated via autophagy after fertilization. In C. elegans, sperm-specific organelles called membraneous organelles (MO) cluster together with pa...

  9. Physiological and biochemical factors responsible for boar taint

    OpenAIRE

    Chen, Gang

    2007-01-01

    We used entire male pigs to: 1) compare different methods to analyse boar taint compounds; 2) evaluate the effects of raw potato starch (RPS), high amylose barley cultivar (Karmosé), sire selection and live weight on the levels of boar taint compounds; 3) investigate the relationship between indolic compounds and testicular steroids by using an hCG injection model; 4) evaluate the effect of incubation with steroids and indolic compounds on CYP2A6 protein expression in hepatocytes; 5) investig...

  10. The control of classical swine fever in wild boar

    OpenAIRE

    Moennig, Volker

    2015-01-01

    Classical swine fever (CSF) is a viral disease with severe economic consequences for domestic pigs. Natural hosts for the CSF virus (CSFV) are members of the family Suidae, i.e., Eurasian wild boar (sus scrofa) are also susceptible. CSF in wild boar poses a serious threat to domestic pigs. CSFV is an enveloped RNA virus belonging to the pestivirus genus of the Flaviviridae family. Transmission of the infection is usually by direct contact or by feeding of contaminated meat products. In recent...

  11. Extensive infanticide in enclosed European wild boars (Sus scrofa)

    OpenAIRE

    Andersson, Annelie; Valros, Anna; Rombin, Johan; Jensen, Per

    2011-01-01

    Infanticidal behaviour is wide-spread among animals of various taxonomic groups, but has not previously been reported in European wild boars, which are commonly kept in enclosures in Sweden and Finland for meat and recreation purposes. We studied the behaviour of wild boars in one enclosure during three reproductive seasons. Non-maternal infanticide was documented in 14 out of 22 litters, causing the deaths of all piglets in all but one affected litters. Infanticide was typically performed du...

  12. The control of classical swine fever in wild boar

    Directory of Open Access Journals (Sweden)

    Volker eMoennig

    2015-11-01

    Full Text Available Classical swine fever (CSF is a viral disease with severe economic consequences for domestic pigs. Natural hosts for the CSF virus (CSFV are members of the family Suidae, i.e. Eurasian wild boar (sus scrofa are also susceptible. CSF in wild boar poses a serious threat to domestic pigs. CSFV is an enveloped RNA virus belonging to the pestivirus genus of the Flaviviridae family. Transmission of the infection is usually by direct contact or by feeding of contaminated meat products. In recent decades CSF has been successfully eradicated from Australia, North America, and the European Union. In areas with dense wild boar populations CSF tends to become endemic whereas it is often self-limiting in small, less dense populations. In recent decades eradication strategies of CSF in wild boar have been improved considerably. The reduction of the number of susceptible animals to a threshold level where the basic reproductive number is R0<1 is the major goal of all control efforts. Depending on the epidemiological situation, hunting measures combined with strict hygiene may be effective in areas with a relatively low density of wild boar. Oral immunization was shown to be highly effective in endemic situations in areas with a high density of wild boar.

  13. Evolution and function of mammalian binder of sperm proteins.

    Science.gov (United States)

    Plante, Geneviève; Prud'homme, Bruno; Fan, Jinjiang; Lafleur, Michel; Manjunath, Puttaswamy

    2016-01-01

    Binder of sperm (BSP) proteins are ubiquitous among mammals and have been extensively investigated over the last three decades. They were first characterized in bull seminal plasma and have now been identified in more than 15 different mammalian species where they represent a superfamily. In addition to sharing a common structure, BSP proteins share many characteristics. They are expressed by seminal vesicles and epididymides, interact with similar ligands and bind to the outer leaflet of sperm membranes via an interaction with choline phospholipids. In addition to playing a major role in sperm capacitation, they are implicated as molecular chaperones in sperm motility and viability, in the formation of the oviductal sperm reservoir, in the regulation of cell volume and possibly in the interaction between sperm and oocytes, making them crucial multifunctional proteins. Furthermore, BSP proteins can bind to egg yolk low-density lipoproteins and milk components, an interaction important for the protection of sperm during semen preservation in liquid or frozen state. Our current knowledge of BSP proteins strongly emphasizes their fundamental importance in male fertility and in the optimization of semen preservation techniques. Much work is still ahead in order to fully understand all the mysteries of BSP proteins. PMID:26386584

  14. Loss of heat shock protein 70 from apical region of buffalo (Bubalus bubalis) sperm head after freezing and thawing.

    Science.gov (United States)

    Varghese, Tincy; Divyashree, Bannur C; Roy, Sudhir C; Roy, Kajal S

    2016-03-15

    The post-thaw fertility of frozen-thawed mammalian spermatozoa is substantially low as compared with that of fresh sperm. Furthermore, the post-thaw fertility of the cryopreserved buffalo sperm has been reported to be poor as compared with that of cattle sperm. Recently, heat shock protein 70 (HSP70) has been found to play a critical role in mammalian fertilization and early embryonic development in boar and cattle. However, the presence of such fertility-related HSP70 in buffalo sperm and its status after cryopreservation has not been reported so far. Thus, a study was conducted to determine the effect of cryopreservation on the level and distribution pattern of HSP70 molecule in buffalo sperm after cryopreservation. Buffalo semen samples, after dilution in semen extender, were aliquoted in straws and divided into two groups. One group was not cryopreserved, and the other group was cryopreserved for 60 days. Sperm proteins were extracted from both non-cryopreserved (NC) and cryopreserved (C) sperm and subjected to Western blot analysis for detection of HSP70 using a monoclonal anti-HSP70 antibody. The distribution pattern of these proteins in buffalo sperm was also monitored before and after cryopreservation using indirect immunofluorescence technique. A prominent 70-kDa protein band of HSP70 protein was detected in protein extracts of both NC and C buffalo sperm. Densitometry analysis revealed that the intensity of 70-kDa HSP70 protein band of cryopreserved sperm decreased significantly (P sperm. However, the level of HSP70 in cryopreserved extended seminal plasma (ESP) did not change as compared with that of NC samples indicating a possible degradation of HSP70 in the spermatozoa itself rather than leakage of the protein into the ESP. Furthermore, Western blot also confirmed that several HSP70 immunoreactive protein bands detected in the ESP were contributed by the egg yolk that was added to the extender. Immunocytochemistry revealed that HSP70 proteins were

  15. Proteomics of boar seminal plasma - current studies and possibility of their application in biotechnology of animal reproduction.

    Science.gov (United States)

    Strzeżek, Jerzy; Wysocki, Paweł; Kordan, Władysław; Kuklińska, Magdalena; Mogielnicka, Marzena; Soliwoda, Daniel; Fraser, Leyland

    2005-11-01

    Proteomics is critical to identify the properties and functions of proteins involved in the mechanism regulating the male reproductive tract function. This approach is important in male fertility assessment and clinical diagnosis of the physiological state of individual reproductive organs. Proteomics also provides a tool to understand the interactions of seminal plasma proteins with spermatozoa, which could provide a useful model for studying ligand-cell interaction occurring at the sperm cell surface. This review covers a selection of advances in the realm of functional proteomics of boar seminal plasma proteins and is focused on some fundamental proteomic technologies. Also, this review explores key themes in proteomics and their application in animal reproductive techniques. PMID:16372045

  16. Subversive practices of sperm donation - globalizing Danish sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

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

  17. Assessment of sperm damages during different stages of cryopreservation in water buffalo by fluorescent probes.

    Science.gov (United States)

    Kumar, Dharmendra; Kumar, Pradeep; Singh, Pawan; Yadav, S P; Yadav, P S

    2016-05-01

    The present study was designed to investigate the sperm damages occurring in acrosome, plasma membrane, mitochondrial activity, and DNA of fresh, equilibrated and frozen-thawed buffalo semen by fluorescent probes. The stability of sperm acrosome and plasma membrane stability, mitochondrial activity and DNA status were assessed by fluorescein conjugated lectin Pisum sativum agglutinin, Annexin-V/propidium iodide, JC-1 and TUNEL assay, respectively, under the fluorescent microscope. The damages percentage of acrosome integrity was significantly increased during equilibration and freezing-thawing process. The stability of sperm plasma membrane is dependent on stability of phosphatidylserine (PS) on the inner leaflet of plasma membrane. The frozen-thawed sperm showed externalization of PS leading to significant increase in apoptotic, early necrotic and necrotic changes and lowered high mitochondrial membrane potential as compared with the fresh sperm but all these parameters were not affected during equilibration. However, the DNA integrity was not affected during equilibration and freezing-thawing procedure. In conclusion, the present study revealed that plasma membrane and mitochondria of buffalo sperm are more susceptible to damage during cryopreservation. Furthermore, the use of fluorescent probes to evaluate integrity of plasma and acrosome membranes, as well as mitochondrial membrane potential and DNA status increased the accuracy of semen analyses. PMID:25373338

  18. In vitro capacitation and acrosome reaction in sperm of the phyllostomid bat Artibeus jamaicensis.

    Science.gov (United States)

    Álvarez-Guerrero, Alma; González-Díaz, Francisco; Medrano, Alfredo; Moreno-Mendoza, Norma

    2016-04-01

    Sperm capacitation occurs during the passage of sperm through the female reproductive tract. Once the sperm binds to the pellucid zone, the acrosome reaction to enable penetration of the oocyte is completed. In this study, sperm of Artibeus jamaicensis bat was used to evaluate both capacitation status and the acrosome reaction under in vitro conditions, incubating sperm at 32 and 37°C with and without progesterone. Sperm was incubated at different times to assess sperm cells' functionality in terms of capacitation and acrosome reaction, using the chlortetracycline staining, lectin fluoresceinisocyanate conjugate-Pisum sativum agglutinin (FITC-PSA), and transmission electron microscopy. Sperm cells that presented uniform fluorescence throughout the head and mid-piece were classified as non-capacitated. Subsequently, sperm cells, which were observed with fluorescence only in the anterior portion of the head and mid-piece, were classified as capacitated. Sperm cells with no fluorescence in the head, but fluorescence in the mid-piece, were categorized as sperm cells that have carried out the acrosome reaction. During the acrosome reaction, sperm cells showed changes in their morphology, so it was not possible to distinguish the plasma and acrosomal membranes. Around the entire head, it was not possible to distinguish the fusion points between these membranes that made it possible for the acrosomal reaction to take place and thus to release the enzymes necessary to penetrate the pellucid zone. In conclusion, under appropriate in vitro conditions and by supplementing the culture medium with progesterone, A. jamaicensis bat sperm cells are able to be capacitated in a period from 6 to 8 h and to carry out the acrosome reaction. PMID:26744028

  19. Fertilization Rate and Number of Embryos on Day 2 after Intrauterine and Deep Intrauterine Insemination Using Frozen-Thawed Boar Semen in Multiparous Sows

    Directory of Open Access Journals (Sweden)

    Kakanang Buranaamnuay

    2011-01-01

    Full Text Available The present study determines fertilization rate and number of embryos on Day 2 after intrauterine insemination (IUI and deep intrauterine insemination (DIUI using frozen-thawed (FT boar semen in multiparous sows. Twelve crossbred Landrace × Yorkshire multiparous sows were included. The sows were inseminated at 24 h after oestrus detection and reinseminated every 12 h until ovulation took place. The inseminations were conducted using IUI with 2×109 FT sperm per dose (n=6 and DIUI with 1×109 FT sperm per dose (n=6. The sows were slaughtered at 45.1±7.2 h after ovulation. Embryos and unfertilized oocytes were flushed from the oviducts. IUI yielded a better fertilization rate than DIUI (66.0% versus 31.0%, P<.001. The number of embryos was 13.5±2.7 and 6.6±3.2 embryos/sow in IUI and DIUI groups, respectively (P=.08. The proportion of sows having unilateral fertilization was higher in the DIUI (3/5 than the IUI group (1/6. In conclusion, IUI with at least 2×109 total number of FT boar spermatozoa is recommended.

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

    Science.gov (United States)

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

    2006-11-01

    Abnormal chromatin condensation is not detected using classical techniques for sperm analysis. SCSA has demonstrated its usefulness in sperm chromatin analysis in several species (human, bull, stallion and boar). In this work, we studied sperm samples from red deer, ram and dog to analyze the differentiation of chromatin structure applying SCSA in epididymal and ejaculated spermatozoa. Epididymal samples were obtained from the caput, corpus and cauda by means of cuts, and ejaculated ones were obtained by electroejaculation (deer), artificial vagina (ram) and digital manipulation (dog). SCSA results suggested different critical points in sperm maturation (spermatozoa with loose chromatin to more condensed chromatin) among species: from corpus to cauda in ram and from caput to corpus in deer and dog. Moreover, we also detected differences in ruminants and dog, reflected in the appearance of SCSA plots. Indeed, ram and deer samples rendered two peaks within the sperm main population (sperm with condensed chromatin), whereas only one was detected in dog. Although some differences were observed between cauda and ejaculated samples, SCSA parameters indicated good chromatin condensation, making these samples suitable for germplasm banking. Some species-dependent modifications in the analysis of the results may be necessary to take full advantage of its analytical power. PMID:16790270

  1. Morphometric changes in goat sperm heads induced by cryopreservation.

    Science.gov (United States)

    Marco-Jiménez, F; Viudes-de-Castro, M P; Balasch, S; Mocé, E; Silvestre, M A; Gomez, E A; Vicente, J S

    2006-04-01

    Two experiments were designed to evaluate the effect of cryopreservation on morphometric characteristics of the goat sperm head. To address this question, we evaluated the size of the sperm head in fresh control cells, post-cooling cells after equilibration with the glycerol preservation solution, and post-thawing cells. Assessment was by automated morphometric sperm head analysis (ASMA) using phase-contrast microscopy without staining. In the first experiment, ASMA was performed on heterospermic pooled samples (fresh, post-cooling after equilibration with the glycerol preservation solution and post-thawing): length, width, area and perimeter were measured. In the second experiment, sperm viability was assessed by Hoechst staining and head morphometry was carried out as before, simultaneously during the cryopreservation process, and the head size was identified for both live and dead spermatozoa. The data were analysed by principal component analysis (PCA). The purpose of PCA is to derive a small number of linear combinations (principal components) from a set of variables (length, width, area and perimeter), that retain as much of the information in the original variables as possible. The main findings that have emerged from this study are that (i) a simple procedure has been developed for measuring spermatozoa heads without staining, which minimises the possibility that sperm head dimensions were influenced by procedural artefacts; (ii) the dimensions of goat sperm heads after cryopreservation in skimmed milk-glucose medium were smaller than in fresh sperm, but this was due to the equilibration phase with the cryoprotectant and not to the cryopreservation process itself; and (iii) dead spermatozoa showed smaller heads than live sperm, consequent upon the loss of membrane function. No differences were observed between post-cooling cells after equilibration with the glycerol preservation solution and post-thawing spermatozoa and only minor osmotic differences

  2. Subversive practices of sperm donation - globalizing Danish sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    inquires into how the bending of boundaries by “inappropriate parents”, fertility travellers, private sperm banks and fertility clinics have been part in negotiating the changes of the legislation in practice, and thus been part of developing a Danish industry of sperm banking. The presentation is based on......During the past two decades, Denmark has developed in to an important destination for fertility travellers in need of donor sperm. Furthermore, two of the largest sperm banks in Europe have been established in Denmark, exporting sperm globally. This development has taken place at the same time as...... a multi-sited ethnography drawing on ethnographic research including observations and interviews from fertility clinics and sperm banks in Denmark during 2002/2003 and 2011- 2013, legislative documents and websites of fertility clinics and sperm banks. The presentation is methodologically inspired...

  3. Optical tweezers and non-ratiometric fluorescent-dye-based studies of respiration in sperm mitochondria

    International Nuclear Information System (INIS)

    The purpose of this study is to investigate how the mitochondrial membrane potential affects sperm motility using laser tweezers and a non-ratiometric fluorescent probe, DiOC6(3). A 1064 nm Nd:YVO4 continuous wave laser was used to trap motile sperm at a power of 450 mW in the trap spot. Using customized tracking software, the curvilinear velocity (VCL) and the escape force from the laser tweezers were measured. Human (Homo sapiens), dog (Canis lupis familiaris) and drill (Mandrillus leucophaeus) sperm were treated with DiOC6(3) to measure the membrane potential in the mitochondria-rich sperm midpieces. Sperm from all three species exhibited an increase in fluorescence when treated with the DiOC6(3). When a cyanide inhibitor (CCCP) of aerobic respiration was applied, sperm of all three species exhibited a reduction in fluorescence to pre-dye levels. With respect to VCL and escape force, the CCCP had no effect on dog or human sperm, suggesting a major reliance upon anaerobic respiration (glycolysis) for ATP in these two species. Based on the preliminary study on drill sperm, CCCP caused a drop in the VCL, suggesting potential reliance on both glycolysis and aerobic respiration for motility. The results demonstrate that optical trapping in combination with DiOC6(3) is an effective way to study sperm motility and energetics

  4. Efficiency of different selection strategies against boar taint in pigs.

    Science.gov (United States)

    Haberland, A M; Luther, H; Hofer, A; Tholen, E; Simianer, H; Lind, B; Baes, C

    2014-01-01

    The breeding scheme of a Swiss sire line was modeled to compare different target traits and information sources for selection against boar taint. The impact of selection against boar taint on production traits was assessed for different economic weights of boar taint compounds. Genetic gain and breeding costs were evaluated using ZPlan+, a software based on selection index theory, gene flow method and economic modeling. Scenario I reflected the currently practiced breeding strategy as a reference scenario without selection against boar taint. Scenario II incorporated selection against the chemical compounds of boar taint, androstenone (AND), skatole (SKA) and indole (IND) with economic weights of -2.74, -1.69 and -0.99 Euro per unit of the log transformed trait, respectively. As information sources, biopsy-based performance testing of live boars (BPT) was compared with genomic selection (GS) and a combination of both. Scenario III included selection against the subjectively assessed human nose score (HNS) of boar taint. Information sources were either station testing of full and half sibs of the selection candidate or GS against HNS of boar taint compounds. In scenario I, annual genetic gain of log-transformed AND (SKA; IND) was 0.06 (0.09; 0.02) Euro, which was because of favorable genetic correlations with lean meat percentage and meat surface. In scenario II, genetic gain increased to 0.28 (0.20; 0.09) Euro per year when conducting BPT. Compared with BPT, genetic gain was smaller with GS. A combination of BPT and GS only marginally increased annual genetic gain, whereas variable costs per selection candidate augmented from 230 Euro (BPT) to 330 Euro (GS) or 380 Euro (both). The potential of GS was found to be higher when selecting against HNS, which has a low heritability. Annual genetic gain from GS was higher than from station testing of 4 full sibs and 76 half sibs with one or two measurements. The most effective strategy to reduce HNS was selecting against

  5. Characterisation of Streptococcus suis isolates from wild boars (Sus scrofa).

    Science.gov (United States)

    Sánchez del Rey, Verónica; Fernández-Garayzábal, José F; Mentaberre, Gregorio; Briones, Víctor; Lavín, Santiago; Domínguez, Lucas; Gottschalk, Marcelo; Vela, Ana Isabel

    2014-06-01

    Wild boar are widely distributed throughout the Iberian Peninsula and can carry potentially virulent strains of Streptococcus suis. The objective of this study was to determine the prevalence of S. suis in wild boars from two large geographical regions of Spain. Serotypes 1, 2, 7 and 9 identified were further genetically characterised by virulence-associated genotyping, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) to determine the population structure of S. suis carried by these animals. Streptococcus suis was isolated from 39.1% of the wild boars examined: serotype 9 was the most frequently isolated (12.5%), followed by serotype 1 (2.5%). Serotype 2 was rarely isolated (0.3%). Eighteen additional serotypes were identified indicating wide diversity of this pathogen within the wild boar population. This heterogeneity was confirmed by PFGE and MLST analyses and the majority of isolates exhibited the virulence-associated genotype mrp-/epf-/sly-. The results of this study highlight that the carriage of S. suis by wild boars is commonplace. However, MLST data indicate that these isolates are not related to prevalent clonal complexes ST1, ST16, ST61 and ST87 typically associated with infection of pigs or humans in Europe. PMID:24726078

  6. Feasibility of boar taint classification using a portable Raman device.

    Science.gov (United States)

    Liu, Xiaoye; Schmidt, Heinar; Mörlein, Daniel

    2016-06-01

    The feasibility of Raman spectroscopy for boar taint detection and classification was investigated using tainted and untainted backfat samples of 46 boars. For this exploratory study, backfat samples were selected according to their levels of androstenone and skatole as determined by gas chromatography and their sensory score by a trained panel. Raman spectra were collected with a portable device at freshly cut surfaces of frozen-thawed samples. Both inner and outer layers of subcutaneous fat were studied. Their varying level of unsaturation was reflected in the Raman spectra. Partial least squares regression discriminant analysis (PLS-DA) was applied to the spectra together with various pre-processing methods. A model using only spectra obtained at the inner layer resulted in the highest classification accuracy for boar taint (81% of samples correctly classified). The discrimination is shown to reflect differences in the degree of fatty acid saturation between tainted and untainted boars. In conclusion, the findings suggest that with further development Raman spectroscopy may be used to classify boar taint. PMID:26882212

  7. A role for carbohydrate recognition in mammalian sperm-egg binding

    Energy Technology Data Exchange (ETDEWEB)

    Clark, Gary F., E-mail: clarkgf@health.missouri.edu

    2014-08-01

    Highlights: • Mammalian sperm-egg binding as a carbohydrate dependent species recognition event. • The role of carbohydrate recognition in human, mouse and pig sperm-egg binding. • Historical perspective and future directions for research focused on gamete binding. - Abstract: Mammalian fertilization usually requires three sequential cell–cell interactions: (i) initial binding of sperm to the specialized extracellular matrix coating the egg known as the zona pellucida (ZP); (ii) binding of sperm to the ZP via the inner acrosomal membrane that is exposed following the induction of acrosomal exocytosis; and (iii) adhesion of acrosome-reacted sperm to the plasma membrane of the egg cell, enabling subsequent fusion of these gametes. The focus of this review is on the initial binding of intact sperm to the mammalian ZP. Evidence collected over the past fifty years has confirmed that this interaction relies primarily on the recognition of carbohydrate sequences presented on the ZP by lectin-like egg binding proteins located on the plasma membrane of sperm. There is also evidence that the same carbohydrate sequences that mediate binding also function as ligands for lectins on lymphocytes that can inactivate immune responses, likely protecting the egg and the developing embryo up to the stage of blastocyst hatching. The literature related to initial sperm-ZP binding in the three major mammalian models (human, mouse and pig) is discussed. Historical perspectives and future directions for research related to this aspect of gamete adhesion are also presented.

  8. MECHANISMS UNDERLYING THE MICRON-SCALE SEGREGATION OF STEROLS AND GM1 IN LIVE MAMMALIAN SPERM

    OpenAIRE

    Selvaraj, Vimal; Asano, Atsushi; Buttke, Danielle E.; Sengupta, Prabuddha; Weiss, Robert S.; Travis, Alexander J.

    2009-01-01

    We demonstrate for the first time that a stable, micron-scale segregation of focal enrichments of sterols exists at physiological temperature in the plasma membrane of live murine and human sperm. These enrichments of sterols represent microheterogeneities within this membrane domain overlying the acrosome. Previously, we showed that cholera toxin subunit B (CTB), which binds the glycosphingolipid, GM1, localizes to this same domain in live sperm. Interestingly, the GM1 undergoes an unexplain...

  9. Genetics Research and Advance on Development and Utilization of Wild Boars

    Institute of Scientific and Technical Information of China (English)

    LIU Chunlong; LIU Di; LI Zhongqiu

    2011-01-01

    Wild boar is one of the most important beast resources. It plays an important role in the maintenance of biological diversity. The genetic resources of wild boar can not only protect the genetic resources, but also improve the formation of new breeds in pigs. This paper summarized the advance on the main biological characteristics of wild boars, evolutionary origin between wild boars and domesticated pigs, and development and utilization of wild boars aimed to provide further insight into wild boar's genetic research and its resource protection.

  10. Sperm surface protein PH-20 is bifunctional: one activity is a hyaluronidase and a second, distinct activity is required in secondary sperm-zona binding.

    Science.gov (United States)

    Hunnicutt, G R; Primakoff, P; Myles, D G

    1996-07-01

    In previous studies, we have found that the sperm membrane protein PH-20 acts during two different stages of fertilization. On acrosome-intact sperm, PH-20 has a hyaluronidase activity that is required for sperm penetration through the cumulus cell layer that surrounds the oocyte. On acrosome-reacted sperm, PH-20 has a required function in sperm-zona binding (secondary binding). Because hyaluronic acid (HA) has been detected in the zona pellucida, secondary sperm-zona adhesion could depend on repetitive binding and hydrolysis of HA by PH-20 acting as a hyaluronidase. Alternatively, PH-20 may be bifunctional and have a second, different activity required for secondary binding. To distinguish between these two possibilities, in this study we used reagents that inhibit either PH-20's function in sperm-zona binding or its hyaluronidase activity. We found that an anti-PH-20 monoclonal antibody that inhibited sperm-zona binding (approximately 90%) had no effect on hyaluronidase activity. Conversely, apigenin, a hyaluronidase inhibitor, blocked PH-20 hyaluronidase activity 93% without inhibiting sperm-zona binding. Similarly, another anti-PH-20 monoclonal antibody that inhibited hyaluronidase activity 95% only partially inhibited sperm-zona binding (approximately 45%). We also extensively pretreated oocytes with hyaluronidase to remove all accessible HA on or in the zona pellucida and found little or no effect on secondary sperm-zona binding. Our results suggest that PH-20 is bifunctional and has two activities: a hyaluronidase activity and a second, separate activity required for secondary sperm-zona binding. PMID:8793062

  11. Meat quality characteristics of DurocxYorkshire, DurocxYorkshirexwild boar and wild boar

    Directory of Open Access Journals (Sweden)

    Ivanović Snežana D.

    2013-01-01

    Full Text Available Chemical composition, pH value, fatty acids profile, cholesterol content, color and sensory analysis of pork meat from Duroc x Yorkshire (D x Y, Duroc x Yorkshire x wild boar (D x Y x WB crossbreeds and wild boars (WB was investigated. Samples for all tests were taken from m. longissimus dorsi. Chemical composition and pH value were tested by ISO methods. Fatty acid and cholesterol determination was performed by gas chromatography technique with external standard method. Color was determined instrumentally using the thristimulus colourimeter. The overall sensoric quality (appearance, texture and smell of samples of raw meat was evaluated. In evaluation of results the scoring system was used. In chemical composition (moisture, fat, protein, ash and pH values statistically significant difference was noted (p<0,05 between each of the examined groups. Also, among all the examined groups statistically significant difference (p<0,05 was found for fatty acids and cholesterol content. Measurment of the color of meat from all three groups showed that the L*, a * b *, Chroma and Hue angle were also statistically significantly different (p<0,01.

  12. Organochlorine pesticides and PCBs in wild boars from Calabria (Italy).

    Science.gov (United States)

    Naccari, F; Giofrè, F; Licata, P; Martino, D; Calò, M; Parisi, N

    2004-01-01

    At present, there are no specific studies on the evaluation of environmental toxicological risks in Calabria (south of Italy) and on the presence of contaminants in the fauna of this region. The aim of the present research was to investigate the levels of contamination by OC pesticides and PCBs in some organs and tissues of wild boars (utilized as 'biological indicator') from various areas of Calabria. Quantitative determinations of organochlorines were carried out using GC-ECD and confirmed with GC-MS in 154 samples from wild boars (heart, liver, lung, kidney, muscle tissue and spleen) during the hunting season from 2000 to 2002. The results indicate the low residual levels of DDE in 8 samples and DDT in 4 samples and PCBs residues (Aroclor 1232) below the detection limits were found. Therefore, these results show that the Calabria region is not at contamination risk from organochlorines and moreover is free from health problems for the consumer of boar meat. PMID:15327158

  13. Shedding patterns of endemic Eurasian wild boar (Sus scrofa) pathogens.

    Science.gov (United States)

    González-Barrio, David; Martín-Hernando, María Paz; Ruiz-Fons, Francisco

    2015-10-01

    The Eurasianwild boar has experienced aworldwide demographic explosion that increases awareness on shared pathogens. However, shedding routes of relevant wild boar pathogens are unknown. Previous observations on sex- and age-related differences in Aujeszky's disease virus (ADV) exposure led us to hypothesize that shedding patterns of endemicwild boar pathogens may be influenced by individual traits.We investigated shedding routes of ADV, porcine parvovirus (PPV), porcine circovirus type 2 (PCV2) and Coxiella burnetii and analysed the effect of host sex and age on pathogen shedding patterns. The presence of pathogen antibodies in serumand of pathogen DNA in oral, nasal, genital and rectal swabswas analysed by ELISA and PCR, respectively. The influence of sex and age in pathogen shedding prevalencewas tested statistically.Main routes of ADV, PPV, PCV2 and C. burnetii shedding were identified but the hypothesis of sex- and/or age-related shedding patterns couldn't be confirmed. PMID:26412545

  14. Radiocesium burden in wild boar: causes and trends until today

    International Nuclear Information System (INIS)

    The examination of nearly 500 samples showed, that in certain circumstances the radiocesium contamination of wild boars is depending on the supply with seeds of oaks and beeches. If this regionally very important nourishment is missing, the boars are forced to gain feed by rooting the upper soil-layers. The radiocesium-contamination of soil in forests will decrease very slowly, because cesium remains nearly completely in the upper layers. So diminution is given mainly by the half-life of 137Cs (30 years). In principle the result of this study is transferable to all (noxious) components which are existing in those soil-layers. The dependance between acorn-mast and the contamination of wild boars requires appropriate care, especially if trend-studies are carried out. (orig.)

  15. First TBEV serological screening in Flemish wild boar

    Science.gov (United States)

    Roelandt, Sophie; Suin, Vanessa; Van der Stede, Yves; Lamoral, Sophie; Marche, Sylvie; Tignon, Marylène; Saiz, Juan Carlos; Escribano-Romero, Estela; Casaer, Jim; Brochier, Bernard; Van Gucht, Steven; Roels, Stefan; Vervaeke, Muriel

    2016-01-01

    In the frame of a Flemish wildlife surveillance in 2013, a serological screening was performed on sera from wild boar (Sus scrofa; n=238) in order to detect tick-borne encephalitis virus (TBEV)-specific antibodies. Neutralising antibodies were titrated with a seroneutralisation test (SNT), using two cut-off titres (1/10–1/15). Seven wild boars were found TBEV-seropositive and showed moderate (>1/15) to high (>1/125) SNT-titres; three individuals had borderline results (1/10–1/15). This study demonstrated the presence of TBEV-specific antibodies in wild boar and highlighted potential TBEV-foci in Flanders. Additional surveillance including direct virus testing is now recommended. PMID:27087689

  16. An Antioxidant Davallialactone from Phellinus baumii Enhances Sperm Penetration on In Vitro Fertilization of Pigs

    Science.gov (United States)

    Lee, In-Kyoung; Lee, Sang-Myeong

    2016-01-01

    Davallialactone (DAVA) is a hispidin analogue derived from the medicinal fungus Phellinus baumii. We examined the effect of DAVA on in vitro fertilization (IVF) of pigs. Boar spermatozoa were incubated in fertilization medium with varying concentrations of DAVA, then sperm motility and reactive oxygen species (ROS) level were evaluated. Higher sperm motility was found following the addition of 0.5 or 1 µM DAVA after incubation than addition of other concentrations or controls. ROS level decreased significantly with the addition of DAVA. The rate of normal fertilization was higher in the presence of 1 µM DAVA (65.1%) than were those of other concentrations or controls (45.4~59.4%), and the highest total fertilization rate (mono- and polyspermic oocytes) was observed at 1 µM DAVA (83%). In conclusion, addition of DAVA to fertilization medium improved sperm motility, and reduced ROS level so as to potentially improve sperm-oocyte binding in IVF, suggesting the potential of a compound isolated from mushrooms in assisted reproductive technology for humans and animals. PMID:27103855

  17. Phenotypic engineering of sperm-production rate confirms evolutionary predictions of sperm competition theory

    OpenAIRE

    Sekii, Kiyono; Vizoso, Dita B.; Kuales, Georg; De Mulder, Katrien; Ladurner, Peter; Schärer, Lukas

    2013-01-01

    Sperm production is a key male reproductive trait and an important parameter in sperm competition theory. Under sperm competition, paternity success is predicted to depend strongly on male allocation to sperm production. Furthermore, because sperm production is inherently costly, individuals should economize in sperm expenditure, and conditional adjustment of the copulation frequency according to their sperm availability may be expected. However, experimental studies showing effects of sperm ...

  18. Rogue sperm indicate sexually antagonistic coevolution in nematodes.

    Directory of Open Access Journals (Sweden)

    Ronald E Ellis

    2014-07-01

    Full Text Available Intense reproductive competition often continues long after animals finish mating. In many species, sperm from one male compete with those from others to find and fertilize oocytes. Since this competition occurs inside the female reproductive tract, she often influences the outcome through physical or chemical factors, leading to cryptic female choice. Finally, traits that help males compete with each other are sometimes harmful to females, and female countermeasures may thwart the interests of males, which can lead to an arms race between the sexes known as sexually antagonistic coevolution. New studies from Caenorhabditis nematodes suggest that males compete with each other by producing sperm that migrate aggressively and that these sperm may be more likely to win access to oocytes. However, one byproduct of this competition appears to be an increased probability that these sperm will go astray, invading the ovary, prematurely activating oocytes, and sometimes crossing basement membranes and leaving the gonad altogether. These harmful effects are sometimes observed in crosses between animals of the same species but are most easily detected in interspecies crosses, leading to dramatically lowered fitness, presumably because the competitiveness of the sperm and the associated female countermeasures are not precisely matched. This mismatch is most obvious in crosses involving individuals from androdioecious species (which have both hermaphrodites and males, as predicted by the lower levels of sperm competition these species experience. These results suggest a striking example of sexually antagonistic coevolution and dramatically expand the value of nematodes as a laboratory system for studying postcopulatory interactions.

  19. Quality decline and oxidative damage in sperm of freshwater crab Sinopotamon henanense exposed to lead.

    Science.gov (United States)

    Li, Na; Hou, Yu-Hua; Jing, Wei-Xin; Dahms, Hans-Uwe; Wang, Lan

    2016-08-01

    Lead (Pb) induces male infertility in vertebrates. Whether lead is related to reproductive abnormalities in aquatic invertebrates remains uncertain. In this work, effects of Pb on the sperm quality and oxidative stress of the freshwater crab Sinopotamon henanense were investigated after 3, 5 and 7d exposure to different Pb concentrations (0, 3.675, 7.35, 14.7, 29.4 and 58.8mg/L). Sperm quality indices including sperm plasma-membrane integrity and acrosomal-membrane integrity were measured by flow cytometry. DNA integrity was measured by fluorescence microscopy. The results showed that Pb levels in sperm increased significantly upon Pb exposure in most treated groups, sperm plasma-membrane integrity, acrosomal-membrane integrity, and DNA integrity were reduced at higher concentrations after 5 d and 7d. Oxidative stress of sperm induced by Pb was reflected in significant up-regulation of reactive oxygen species (ROS) levels after 3, 5 and 7d. A significant reduction of the total antioxidant capacity levels occurred after exposure to 14.7mg/L Pb and above at 7d compared to the control. The results of oxidative damage to lipids, proteins and DNA of sperm showed that malondialdehyde, protein carbonylation and DNA-protein crosslinks were increased in a concentration- and time-dependent manner. Our findings document that Pb can induce harmful effects on several reproductive endpoints in a freshwater crab. PMID:27123971

  20. Unraveling Sarcocystis miescheriana and Sarcocystis suihominis infections in wild boar.

    Science.gov (United States)

    Coelho, Catarina; Gomes, Jacinto; Inácio, João; Amaro, Ana; Mesquita, João Rodrigo; Pires, Isabel; Lopes, Ana Patrícia; Vieira-Pinto, Madalena

    2015-09-15

    Sarcocystis species are worldwide spread cyst-forming protozoa that can infect wild boar but little is known about the prevalence of these parasites. In this study we assessed the prevalence of Sarcocystis spp. infections in wild boars from northeastern Portugal, for which novel PCR testing assays targeting Sarcocystis genus, S. miescheriana and S. suihominis were implemented, and risk factors potentially associated with these infections were evaluated. Samples from muscle tissue, namely diaphragm (n=102), oesophagus (n=96) and heart (n=101), were collected from a total of 103 wild boar hunted between October 2011 and February 2012. Diaphragm muscle was used for the PCR detection of Sarcocystis nucleic acids since a higher proportion of samples showed the presence of cysts during histological examination. PCR assay targeting Sarcocystis genus yielded a 73.8% infection rate, which indicate a high level of exposure to these protozoan parasites among wild boars. These samples showed to be positive with the S. miescheriana-specific PCR assay and no sample was positive with the S. suihominis-specific assay, suggesting that a single species infecting wild boar is circulating in Portugal. These results were confirmed by the partial sequencing of the 18S rRNA gene amplified from selected samples from different geographic regions. Adults, young adults and female wild boars were found to be more likely infected. Hunters have an important role in the life cycle of S. miescheriana since potentially infected viscera and carcasses can be left behind promoting the protozoan dissemination to the scavenging final hosts. If hunting dogs bite and ingest infected meat they can perpetuate the life cycle of Sarcocystis spp. spreading oocysts or sporocysts in the environment. PMID:26319199

  1. Biochemical studies on proacrosin and acrosin from epididymal boar spermatozoa: in vitro translation of boar testicular proacrosin mRNA

    International Nuclear Information System (INIS)

    An inactive form of acrosin was extracted from epididymal boar spermatozoa at an acid pH. When subjected to activation in alkaline environment, this form turns into an enzymatically active species, which exhibits closely related electrophoretic characteristics. Both the precursor and the activated species, when incubated in the presence of thermolysin, give rise to two fast moving acrosin molecular forms. In order to establish the nature of the true acrosin zymogen, poly(A+)-RNA was isolated from boar testicles, performed its translation in vitro in the presence of [35S]-methionine and reticulocyte lysate, immunoprecipitated the translation products with anti-boar acrosin antibody, and analyzed them by SDS-polyacrylamide gel electrophoresis and autoradiography. A single translation product of molecular weight 55,000 was detected

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

    Institute of Scientific and Technical Information of China (English)

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

    2012-01-01

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

  3. Single layer centrifugation (SLC) improves sperm quality of cryopreserved Blanca-Celtibérica buck semen.

    Science.gov (United States)

    Jiménez-Rabadán, P; Morrell, J M; Johannisson, A; Ramón, M; García-Álvarez, O; Maroto-Morales, A; Alvaro-García, P J; Pérez-Guzmán, M D; Fernández-Santos, M R; Garde, J J; Soler, A J

    2012-12-01

    The aim of the present study was to evaluate the effect of sperm selection by means of single layer centrifugation (SLC) on sperm quality after cryopreservation, either when SLC is used before freezing or after thawing, using Blanca-Celtibérica buck semen collected by electroejaculation (EE). Ejaculates from six bucks were collected by EE and divided into two aliquots. One of them (unselected) was diluted with Biladyl(®) by the two-step method and frozen over nitrogen vapor. The other aliquot was selected by the SLC technique and subsequently frozen in the same way as the unselected samples (SLC before freezing). In a further treatment, two unselected straws were thawed and SLC was carried out (SLC after thawing). At thawing, sperm motility of all samples ((i) unselected; (ii) selected before freezing and (iii) selected after thawing) was evaluated by CASA. In addition, integrity of the plasma membrane, mitochondrial membrane potential, ROS production and DNA fragmentation index were assessed by flow cytometry. Most of the sperm parameters were improved (P≤0.001) in samples selected by SLC after thawing in relation to unselected or selected by SLC before freezing. The percentage of progressive motile spermatozoa was greater (86%) for sperm samples selected after thawing compared with unselected (58%) or selected before freezing (54%). Moreover, percentages of spermatozoa with intact plasma membrane and spermatozoa with high mitochondrial membrane potential (hMMP) were also greater for sperm samples selected after thawing compared to sperm samples unselected or selected before freezing (spermatozoa with intact plasma membrane: 80% vs. 32% vs. 12%; spermatozoa with hMMP: 54% vs. 1% vs. 15%; respectively). Therefore, sperm quality after cryopreservation is improved in Blanca-Celtibérica buck ejaculates collected by EE when a sperm selection technique such as SLC is carried out after thawing. PMID:23084569

  4. Relative testis size and sperm morphometry across mammals: no evidence for an association between sperm competition and sperm length.

    OpenAIRE

    Gage, Matthew J.g.; Robert P Freckleton

    2003-01-01

    Understanding why there is extensive variation in sperm form and function across taxa has been a challenge because sperm are specialized cells operating at a microscopic level in a complex environment. This comparative study collates published data to determine whether the evolution of sperm morphometry (sperm total length and separate component dimensions) is associated with sperm competition (when different males' sperm mix and compete for a female's ova) across 83 mammalian species. We use...

  5. Sperm function test

    OpenAIRE

    Pankaj Talwar; Suryakant Hayatnagarkar

    2015-01-01

    With absolute normal semen analysis parameters it may not be necessary to shift to specialized tests early but in cases with borderline parameters or with history of fertilization failure in past it becomes necessary to do a battery of tests to evaluate different parameters of spermatozoa. Various sperm function tests are proposed and endorsed by different researchers in addition to the routine evaluation of fertility. These tests detect function of a certain part of spermatozoon and give ins...

  6. Cytometry of mammalian sperm

    Energy Technology Data Exchange (ETDEWEB)

    Gledhill, B.L.

    1983-10-11

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

  7. Cytometry of mammalian sperm

    International Nuclear Information System (INIS)

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

  8. Role of Ion Channels in the Sperm Acrosome Reaction.

    Science.gov (United States)

    Beltrán, Carmen; Treviño, Claudia L; Mata-Martínez, Esperanza; Chávez, Julio C; Sánchez-Cárdenas, Claudia; Baker, Mark; Darszon, Alberto

    2016-01-01

    The acrosome reaction (AR) is a unique exocytotic process where the acrosome, a single membrane-delimited specialized organelle, overlying the nucleus in the sperm head of many species, fuses with the overlying plasma membrane. This reaction, triggered by physiological inducers from the female gamete, its vicinity, or other stimuli, discharges the acrosomal content modifying the plasma membrane, incorporating the inner acrosomal membrane, and exposing it to the extracellular medium. The AR is essential for sperm-egg coat penetration, fusion with the eggs' plasma membrane, and fertilization. As in most exocytotic processes Ca(2+) is crucial for the AR, as well as intracellular pH and membrane potential changes. Thus, among the required processes needed for this reaction, ion permeability changes involving channels are pivotal. In spite of the key role ion channels play in the AR, their identity and regulation is not fully understood. Though molecular and pharmacological evidence indicates that various ionic channels participate during the AR, such as store-operated Ca(2+) channels and voltage-dependent Ca(2+) channels, whole cell patch clamp recordings have failed to detect some of them until now. Since sperm display a very high resistance and a minute cytoplasmic volume, very few channels are needed to achieve large membrane potential and concentration changes. Functional detection of few channels in the morphologically complex and tiny sperm poses technical problems, especially when their conductance is very small, as in the case of SOCs. Single channel recordings and novel fluorescence microscopy strategies will help to define the participation of ionic channels in the intertwined signaling network that orchestrates the AR. PMID:27194349

  9. Surgical sperm retrieval: Techniques and their indications

    OpenAIRE

    Shah, Rupin

    2011-01-01

    Men with azoospermia can father a child through intra-cytoplasmic sperm injection if sperm can be retrieved from their epididymis or testis. Several percutaneous and open surgical procedures have been described to retrieve sperm. The various techniques and their merits are discussed in this review. In men with obstructive azoospermia, epididymal sperm can usually be retrieved by percutaneous epididymal sperm aspiration (PESA). If PESA fails then testicular sperm are obtained by needle aspirat...

  10. Wild boar hunters profile in Shimane prefecture, western Japan

    Directory of Open Access Journals (Sweden)

    Ueda, G.

    2005-12-01

    Full Text Available Wild boars have been expanding their range and seriously damage agricultural crops all over Japan. Such situation is obvious in Shimane Prefecture, western end of Honshu Island, where most of its territory is mountainous. Populaton control is strongly expected by farmers and administration. However, the number of hunters has been drastically decreasing since the 1970’s. To maintain and increase hunters, we must investigate their activities and attitudes to clarify the problems. Questionnaires were conducted in 2001 on 310 hunters who renewed their hunting license at local office. The response rate was 80.0%. Wild boar hunters accounted for 61.6%, and the others were mostly bird hunters (32.5%. The objective of wild boar hunting was predominantly nuisance control, and very few hunted for money despite of its high commercial value. Most of them were farmers (35.8% and/or farm village dwellers (53.6%, and used the leg snare (61.4%. Despite the stable number of hunters, the number of hunters using guns is decreasing. Hunters do not to appear to be interested in maintaining the local hunting society. Leisure is the most pursued objective rather nuisance control. Therefore, actions should be taken to stimulate hunting as a leisure activity thus maintaining an important tool for wild boar management.

  11. Parasitic infections in wild ruminants and wild boar

    Directory of Open Access Journals (Sweden)

    Ilić Tamara

    2011-01-01

    Full Text Available Wild ruminants and wild boar belong to the order Artiodactyla, the suborders Ruminantia and Nonruminantia and are classified as wild animals for big game hunting, whose breeding presents a very important branch of the hunting economy. Diseases caused by protozoa are rarely found in wild ruminants in nature. Causes of coccidiosis, cryptosporidiosis, toxoplasmosis, sarcocystiosis, giardiasis, babesiosis, and theileriosis have been diagnosed in deer. The most significant helminthoses in wild ruminants are fasciosis, dicrocoeliasis, paramphistomosis, fascioloidosis, cysticercosis, anoplocephalidosis, coenurosis, echinococcosis, pulmonary strongyloidiasis, parasitic gastroenteritis, strongyloidiasis and trichuriasis, with certain differences in the extent of prevalence of infection with certain species. The most frequent ectoparasitoses in wild deer and doe are diseases caused by ticks, mites, scabies mites, and hypoderma. The most represented endoparasitoses in wild boar throughout the world are coccidiosis, balantidiasis, metastrongyloidiasis, verminous gastritis, ascariasis, macracanthorhynchosis, trichinelosis, trichuriasis, cystecercosis, echinococcosis, and less frequently, there are also fasciolosis and dicrocoeliasis. The predominant ectoparasitoses in wild boar are ticks and scabies mites. Knowledge of the etiology and epizootiology of parasitic infections in wild ruminants and wild boar is of extreme importance for the process of promoting the health protection system for animals and humans, in particular when taking into account the biological and ecological hazard posed by zoonotic infections.

  12. Effects of the Czech Propolis on Sperm Mitochondrial Function

    Science.gov (United States)

    Cedikova, Miroslava; Miklikova, Michaela; Stachova, Lenka; Grundmanova, Martina; Tuma, Zdenek; Vetvicka, Vaclav; Zech, Nicolas; Kralickova, Milena; Kuncova, Jitka

    2014-01-01

    Propolis is a natural product that honeybees collect from various plants. It is known for its beneficial pharmacological effects. The aim of our study was to evaluate the impact of propolis on human sperm motility, mitochondrial respiratory activity, and membrane potential. Semen samples from 10 normozoospermic donors were processed according to the World Health Organization criteria. Propolis effects on the sperm motility and mitochondrial activity parameters were tested in the fresh ejaculate and purified spermatozoa. Propolis preserved progressive motility of spermatozoa in the native semen samples. Oxygen consumption determined in purified permeabilized spermatozoa by high-resolution respirometry in the presence of adenosine diphosphate and substrates of complex I and complex II (state OXPHOSI+II) was significantly increased in the propolis-treated samples. Propolis also increased uncoupled respiration in the presence of rotenone (state ETSII) and complex IV activity, but it did not influence state LEAK induced by oligomycin. Mitochondrial membrane potential was not affected by propolis. This study demonstrates that propolis maintains sperm motility in the native ejaculates and increases activities of mitochondrial respiratory complexes II and IV without affecting mitochondrial membrane potential. The data suggest that propolis improves the total mitochondrial respiratory efficiency in the human spermatozoa in vitro thereby having potential to improve sperm motility. PMID:25104965

  13. Mouse egg integrin alpha 6 beta 1 functions as a sperm receptor.

    Science.gov (United States)

    Almeida, E A; Huovila, A P; Sutherland, A E; Stephens, L E; Calarco, P G; Shaw, L M; Mercurio, A M; Sonnenberg, A; Primakoff, P; Myles, D G; White, J M

    1995-06-30

    Binding between sperm and egg plasma membranes is an essential step in fertilization. Whereas fertilin, a mammalian sperm surface protein, is involved in this crucial interaction, sperm receptors on the egg plasma membrane have not been identified. Because fertilin contains a predicted integrin ligand domain, we investigated the expression and function of integrin subunits in unfertilized mouse eggs. Polymerase chain reactions detected mRNAs for alpha 5, alpha 6, alpha v, beta 1, beta 3, and beta 5. Immunofluorescence revealed alpha 6 beta 1 and alpha v beta 3 on the plasma membrane. GoH3, a function-blocking anti-alpha 6 monoclonal antibody, abolished sperm binding, but a nonfunction-blocking anti-alpha 6 monoclonal antibody, a function-blocking anti-alpha v beta 3 polyclonal antibody, and an RGD peptide had no effect. Somatic cells bound sperm avidly, but only if they expressed alpha 6 beta 1. A peptide analog of the fertilin integrin ligand domain inhibited sperm binding to eggs and alpha 6 beta 1+ cells and diminished GoH3 staining of eggs. Our results indicate a novel role for the integrin alpha 6 beta 1 as a cell-cell adhesion receptor that mediates sperm-egg binding. PMID:7600577

  14. Comparison between different markers for sperm quality in the cat: Diff-Quik as a simple optical technique to assess changes in the DNA of feline epididymal sperm

    OpenAIRE

    Mota, Paula C.; Ramalho-Santos, João

    2006-01-01

    The majority of wild felids, as well as some domestic cats, have low sperm concentration in their ejaculates, and a high proportion of abnormal spermatozoa. We have employed several possible semen quality markers to further characterize cat epididymal sperm. Methods included possible apoptotic reporters, such as the annexin V assay to monitor exposure of phosphatidylserine (PS) on the outer leaflet of the plasma membrane, as well as cell integrity; and the TUNEL assay to quantify DNA breaks. ...

  15. Endocannabinoids and Human Sperm Cells

    Directory of Open Access Journals (Sweden)

    Giovanna Zolese

    2010-10-01

    Full Text Available N-acylethanolamides (NAEs are naturally occurring signaling lipids consisting of amides and esters of long-chain polyunsaturated fatty acids. Usually they are present in a very small amounts in many mammalian tissues and cells, including human reproductive tracts and fluids. Recently, the presence of N-arachidonoylethanolamide (anandamide, AEA, the most characterised member of endocannabinoids, and its congeners palmitoylethanolamide (PEA and oleylethanolamide (OEA in seminal plasma, oviductal fluid, and follicular fluids was demonstrated. AEA has been shown to bind not only type-1 (CB1 and type-2 (CB2 cannabinoid receptors, but also type-1 vanilloid receptor (TRPV1, while PEA and OEA are inactive with respect to classical cannabinoid CB1 and CB2 but activate TRPV1 or peroxisome proliferator activate receptors (PPARs. This review concerns the most recent experimental data on PEA and OEA, endocannabinoid-like molecules which appear to exert their action exclusively on sperm cells with altered features, such as membrane characteristics and kinematic parameters. Their beneficial effects on these cells could suggest a possible pharmacological use of PEA and OEA on patients affected by some forms of idiopathic infertility.

  16. [CO-CULTURE OF BOAR SPERMATOGONIAL CELLS WITH SERTOLI CELLS].

    Science.gov (United States)

    Savchenkova, I P; Vasil'eva, S A

    2016-01-01

    In the present study, we developed in vitro culture conditions using co-culture of boar spermatogonial cells with Sertoli cells. Testes from 60-day-old crossbred boar were used. A spermatogonia-enriched culture was achieved by enzymatic digestion method and purification by density gradient centrifugation using a discontinuous Percoll gradient and differentiated adherence technique. Lipid drops were detected in isolated Sertoli cells by Oil Red O staining. We have found that the cultivation of boar spermatogonia in the presence of Sertoli cells (up to 35 days) leads to their differentiation as well as in vivo in testis. Association of cells in groups, formation of chains and suspension clusters of the spermatogenic cells were observed on the 10th day. Spermatogonial cellular colonies were noted at the same time. These cellular colonies were analyzed for the expression of genes: Nanog and Plzf in RT PCR. The expression of the Nanog gene in the experimental cellular clones obtained by short-term culture of spermatogonial cells in the presence of Sertoli cells was 200 times higher than the expression of this gene in the freshly isolated spermatogonial cells expression was found in freshly isolated germ cells and in cellular clones derived in vitro. We have found that, in the case of longer cultivation of these cells on Sertoli cells, in vitro process of differentiation of germ cells and formation of single mobile boar spermatozoa occurs at 30-33 days. Cellular population is heterogeneous at this stage. Spermatogenic differentiation in vitro without Sertoli cells stays on the 7th day of cultivation. The results show that co-culture of boar spermatogonia-enriched cells with Sertoli cells can induce their differentiation into spermatozoa in vitro and facilitate obtaining of porcine germ cell culture. PMID:27228660

  17. Sperm competition experiments between lines of crickets producing different sperm lengths.

    OpenAIRE

    Morrow, E. H.; Gage, M J

    2001-01-01

    Sperm numbers can be important determinants of fertilization success in sperm competition. However, the importance of variation in sperm size is less well understood. Sperm size varies significantly both between and within species and comparative studies have suggested that some of this variance can be explained by sperm competition. In this study we examine whether variation in sperm length has consequences for fertilization precedence using controlled sperm competition experiments in the fi...

  18. Viable offspring obtained from Prm1-deficient sperm in mice

    Science.gov (United States)

    Takeda, Naoki; Yoshinaga, Kazuya; Furushima, Kenryo; Takamune, Kazufumi; Li, Zhenghua; Abe, Shin-ichi; Aizawa, Shin-ichi; Yamamura, Ken-ichi

    2016-01-01

    Protamines are expressed in the spermatid nucleus and allow denser packaging of DNA compared with histones. Disruption of the coding sequence of one allele of either protamine 1 (Prm1) or Prm2 results in failure to produce offspring, although sperm with disrupted Prm1 or Prm2 alleles are produced. Here, we produced Prm1-deficient female chimeric mice carrying Prm1-deficient oocytes. These mice successfully produced Prm1+/− male mice. Healthy Prm1+/− offspring were then produced by transferring blastocysts obtained via in vitro fertilization using zona-free oocytes and sperm from Prm1+/− mice. This result suggests that sperm lacking Prm1 can generate offspring despite being abnormally shaped and having destabilised DNA, decondensed chromatin and a reduction in mitochondrial membrane potential. Nevertheless, these mice showed little derangement of expression profiles. PMID:27250771

  19. RNA in human sperm

    Institute of Scientific and Technical Information of China (English)

    Rui Pires Martins; Stephen A. Krawetz

    2005-01-01

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

  20. Sperm preparation for ART

    Directory of Open Access Journals (Sweden)

    Schill Wolf-Bernhard

    2003-11-01

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

  1. Influence of chicory roots (Cichorium intybus L) on boar taint in entire male and female pigs

    OpenAIRE

    Hansen, L L; Mejer, H.; Thamsborg, S.M.; Burne, D.V.; Roepstorff, A.; Karlsson, A H; Hansen-Møller, J.; Jensen, M.T.; Tuomola, M.

    2005-01-01

    Boar taint is an off-flavour of pork caused primarily by a microbial breakdown product, skatole and a testicular steroid, androstenone. As skatole is produced in the large intestine from tryptophan, it is possible that some "bioactive" ingredients could modify protein fermentation and, in the process, diminish boar taint. The aim of this study was to examine the effect of inulin-rich chicory roots (Cichorium intybus L.) on boar taint. In the first of three trials individually penned, entire m...

  2. SEROLOGIC AND VIROLOGICAL EVIDENCE OF PARVOVIRUS CIRCULATION IN WILD BOARS IN PORTUGAL

    OpenAIRE

    Duarte, Margarida; Fevereiro, Miguel

    2011-01-01

    Wild boars (Sus scrofa) are indigenous species in many countries and can act as reservoirs for important infectious diseases in domestic pigs. The increase in wild boar population observed in several European Countries such as Portugal, Spain, Italy and Germany higher the risk of transmission of diseases between these species particularly in free-ranging production regions. Our objective was to infer about the actual epidemiology of Porcine Parvovirus infection in the wild boar population...

  3. First isolation of Trichinella britovi from a wild boar (Sus scrofa) in Belgium.

    OpenAIRE

    Schynts, F; van der Giessen, Joke; Tixhon, S; Pozio, E.; Dorny, P; Borchgrave, J de

    2006-01-01

    Since 1992, when the European Union Council Directive requires that wild boars (Sus scrofa) hunted in EU for commercial purpose should be examined for Trichinella, the infection has not been detected in wild boars from Belgium, despite serological evidence of the presence of anti-Trichinella antibodies in wildlife and previous reports of Trichinella larvae in this host species. In November 2004, Trichinella larvae were detected in a wild boar hunted near Mettet, Namur province (Southern Belgi...

  4. A survey of porcine reproductive and respiratory syndrome among wild boar populations in Korea

    OpenAIRE

    Choi, Eun-Jin; Lee, Chang-Hee; Hyun, Bang-Hun; Kim, Jae-Jo; Lim, Seong-In; Song, Jae-Young; Shin, Yeun-Kyung

    2012-01-01

    No information is currently available on porcine reproductive and respiratory syndrome virus (PRRSV) infection in wild boars (Sus scrofa) in Korea. In this study, the status of PRRS in wild boars was investigated. Blood samples were collected from 267 wild boars from eight provinces in Korea. Four of the samples tested (1.5%) were positive for PRRSV antibodies and eight (3.0%) were positive for antigens. Of the virus-positive samples, three and five samples were typed as containing European (...

  5. Sperm variables as predictors of fertility in Black Castellana roosters; use in the selection of sperm donors for genome resource banking purposes

    Energy Technology Data Exchange (ETDEWEB)

    Santiago Moreno, J.; Lopez Sebastian, A.; Castano, C.; Coloma, M. A.; Gomez Brunet, A.; Toledano Diaz, A.; Prieto, M. T.; Campo, J. L.

    2009-07-01

    Semen was collected from 10 Black Castellana roosters and the classic sperm variables (ejaculate volume, sperm concentration and sperm motility) examined. In addition, the hypo-osmotic swelling test was used to investigate sperm cell membrane integrity, and acidic aniline blue staining used to screen for morphological abnormalities (including acrosome integrity) and to examine the condensation status of the chromatin. The latter was also examined by Gram staining. Large and small semen volumes were associated high and low sperm concentrations respectively (R2=0.04, P<0.05). The percentage of motile spermatozoa correlated strongly with the percentage of sperm cells showing an intact acrosome (R2=0.13, P<0.001) and with the percentage of morphologically normal spermatozoa (R2=0.04, P<0.05). The percentage of Gram positive spermatozoa was positively correlated with semen appearance (R2=0.12, P<0.05), sperm cell concentration (R2=0.13, P<0.05), and with the sperm motility variables studied (R2=0.14, P<0.05 for percentage mobility, and R2=0.12, P<0.05 for quality of movement). Only three of the 10 roosters, all with fertilisation potentials of 80-90%, were considered potential sperm donors for genome resource banking purposes. The remaining birds were all of low fertility (. 50%); in fact, some produced semen volumes too small to perform fertility tests. Semen volume and membrane integrity were found to be the best variables for predicting the fertilisation potential of rooster ejaculates. (Author) 37 refs.

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

    Science.gov (United States)

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

    2016-04-01

    Action of reactive oxygen species, protamination failures and apoptosis are considered the most important etiologies of sperm DNA fragmentation. This study evaluated the effects of induced lipid peroxidation susceptibility on native semen profile and identified the mechanisms involved in sperm DNA fragmentation and testicular antioxidant defense on Santa Ines ram sperm samples. Semen was collected from 12 adult rams (Ovis aries) performed weekly over a 9-week period. Sperm analysis (motility, mass motility, abnormalities, membrane and acrosome status, mitochondrial potential, DNA fragmentation, lipid peroxidation and intracellular free radicals production); protamine deficiency; PRM1, TNP1 and TNP2 gene expression; and determination of glutathione peroxidase (GPx), glutathione reductase, catalase (CAT) and superoxide dismutase activity and immunodetection in seminal plasma were performed. Samples were distributed into four groups according to the sperm susceptibility to lipid peroxidation after induction with ascorbate and ferrous sulfate (low, medium, high and very high). The results were analyzed by GLM test and post hoc least significant difference. We observed an increase in native GPx activity and CAT immunodetection in groups with high susceptibility to induced lipid peroxidation. We also found an increase in total sperm defects, acrosome and membrane damages in the group with the highest susceptibility to induced lipid peroxidation. Additionally, the low mitochondrial membrane potential, susceptible to chromatin fragmentation and the PRM1 mRNA were increased in the group showing higher susceptibility to lipid peroxidation. Ram sperm susceptibility to lipid peroxidation may compromise sperm quality and interfere with the oxidative homeostasis by oxidative stress, which may be the main cause of chromatin damage in ram sperm. PMID:26811546

  7. Cryopreservation of aoudad (Ammotragus lervia sahariensis) sperm obtained by transrectal ultrasound-guided massage of the accessory sex glands and electroejaculation.

    Science.gov (United States)

    Santiago-Moreno, J; Castaño, C; Toledano-Díaz, A; Esteso, M C; López-Sebastián, A; Guerra, R; Ruiz, M J; Mendoza, N; Luna, C; Cebrián-Pérez, J A; Hildebrandt, T B

    2013-01-15

    This study examines (1) the effectiveness of transrectal, ultrasound-guided massage of the accessory sex glands (TUMASG) combined with electroejaculation for obtaining aoudad (Ammotragus lervia sahariensis) sperm samples for cryopreservation, and (2) the effectiveness of a Tris-citric acid-glucose-based medium (TCG; usually used for freezing ibex sperm) and a TES-Tris-glucose-based medium (TTG; typically used in the cryopreservation of mouflon sperm) as sperm extenders. After TUMASG, just one to three electrical pulses were required for ejaculation to occur in five of the six animals studied; one ejaculated after TUMASG alone. Transrectal, ultrasound-guided massage of the accessory sex glands would therefore appear to be useful in obtaining sperm samples from this species, requiring few subsequent electrical electroejaculation stimuli and sometimes none at all. After thawing, the membrane integrity (assessed by nigrosin-eosin staining) of sperm extended with TTG was greater than that of sperm extended with TCG (P < 0.05). The total percentage of sperm showing an intact acrosome, as assessed by fluorescein isothiocyanate-conjugated peanut (Arachis hypogea) agglutinin, was also higher in the TTG-extended sperm (P < 0.05), and the percentage of dead sperm with a damaged acrosome was lower (P < 0.05). No differences were seen between TCG and TTG in terms of apoptotic manifestations (DNA damage, caspase activity, mitochondrial membrane potential, and plasmalemma stability). Therefore, TTG appears to be a better extender than TCG for cryopreserving aoudad sperm. PMID:23158213

  8. Post-testicular sperm maturation and identification of an epididymal protein in the Japanese quail (Coturnix coturnix japonica).

    Science.gov (United States)

    Nixon, Brett; Ewen, Katherine A; Krivanek, Klara M; Clulow, John; Kidd, Grahame; Ecroyd, Heath; Jones, Russell C

    2014-03-01

    The role of the avian epididymis in post-testicular development and capacitation was examined to assess whether avian spermatozoa undergo any processes similar to those characteristic of mammalian sperm development. We found no evidence of a need for quail sperm to undergo capacitation and 90% of testicular sperm could bind to a perivitelline membrane and acrosome react. However, computer-assisted sperm analysis showed that 20% of testicular sperm from the quail were capable of movement and only about 12% of the motile sperm would have a curvilinear velocity greater than the mean for sperm from the distal epididymis. Nevertheless, epididymal transit was associated with increases in mean sperm velocity and the proportion of motile sperm. Together, these findings explain why earlier workers have achieved some fertilizations following inseminations of testicular spermatozoa and also demonstrate the need for some epididymal maturation of avian spermatozoa. Analysis of the electrophoretic profile of quail epididymal luminal proteins revealed that only one major protein (∼16 kDa) is secreted by the epididymis and it was virtually the only protein secreted by the ipsilateral epididymis following unilateral orchidectomy. Mass spectrometry showed that this protein is hemoglobin; this finding was confirmed using anti-hemoglobin antibodies. It is suggested that hemoglobin may support sperm metabolism in the quail epididymis, aid in motility, and/or serve as an antioxidant. PMID:24298048

  9. Influence of anaesthetic drugs on the epididymal sperm quality in domestic cats.

    Science.gov (United States)

    Jiménez, E; Pérez-Marín, C C; Millán, Y; Agüera, E

    2011-02-01

    The present study investigated the effect of different anaesthetic agents commonly used in cats on the fresh and frozen-thawed epididymal sperm. Seventeen male domestic cats were castrated using pentobarbital, ketamine HCl or isoflurane. Sperm samples were recovered from epididymides and evaluated before and after freezing, determining the vigor, motility, morphology, acrosome status, sperm viability and functional membrane integrity. Fresh epididymal sperm was influenced by the drugs used, noting that motility features, i.e. vigor (p≤0.05) and progressive motility (p≤0.05), were higher for the inhalation anaesthetic while the others did not showed statistical differences. In frozen-thawed sperm samples, cats treated with barbiturics showed lower values for acrosome status (p≤0.05) and integrity and functionality of membrane (p≤0.05 and p≤0.01, respectively) than in the others groups. Results suggested that drugs used for castration in cats could affect the sperm quality and this should be considered when implementing sperm cryopreservation in the feline. PMID:21288668

  10. Effect of dietary vitamin E on the sperm quality of turbot ( Scophthalmus maximus)

    Science.gov (United States)

    Xu, Houguo; Huang, Lina; Liang, Mengqing; Zheng, Keke; Wang, Xinxing

    2015-08-01

    A 3-month feeding experiment was conducted in an in-door seawater system to investigate the effect of dietary vitamin E (Ve) on the sperm quality of turbot ( Scophthalmus maximus). D-α-tocopherol acetate was supplemented to the basal (control) diet (65.14 mg kg-1 Ve) to obtain low and high levels of dietary Ve (244.60 mg kg-1, LVe; 721.60 mg kg-1, HVe). Compared with the control, sperm concentration was significantly increased in Ve-supplemented groups (LVe and HVe); while relative sperm volume and testis-somatic index were significantly increased in group HVe only. Sperm motility duration was significantly longer in group HVe than in the control, but no significant difference was observed in percent motility among groups. Sperm size, the uniformity of mitochondrial size, and the integrity of mitochondria cristae and plasma membrane were improved by dietary Ve, especially in HVe. The content of Ve in testis and liver as well as polyunsaturated fatty acids in sperm increased with dietary Ve. These results suggested that dietary Ve, especially at the high level (721.60 mg kg-1), significantly improved sperm concentration and motility duration and maintained normal sperm morphology of turbot.

  11. Diffusion and regionalization in membranes of maturing ram spermatozoa

    OpenAIRE

    1984-01-01

    An essential feature of the "fluid mosaic model" (Singer, S. J., and G. L. Nicolson , 1972, Science (Wash. DC)., 175:720-731) of the cell plasma membrane is the ability of membrane lipids and proteins to diffuse laterally in the plane of the membrane. Mammalian sperm are capable of overcoming free random diffusion and restricting specific membrane components, both lipid and protein, to defined regions of the sperm's surface. The patterns of these regionalizations evolve with the processes of ...

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

    Institute of Scientific and Technical Information of China (English)

    KausikiChakrabarti; SulagnaPal; AsokK.Bhattacharyya

    2003-01-01

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

  13. Lipid Peroxidation and Nitric Oxide Levels in Male Smokers' Spermatozoa and their Relation with Sperm Motility

    OpenAIRE

    Ghaffari, Mohammad Ali; Rostami, Morad

    2012-01-01

    Background Nitric oxide (NO) is synthesized from L-arginine by a family of enzymes known as nitric oxide synthases. Low concentrations of NO is essential in biology and physiology of spermatozoa, but high amounts of NO is toxic and has negative effects on sperm functions. Moreover, sperm membrane contains high concentrations of polyunsaturated fatty acids that are highly susceptible to oxidative damage that interferes with fertilization ability. Therefore, we investigated the correlation betw...

  14. Microdissection testicular sperm extraction: an update

    OpenAIRE

    Dabaja, Ali A; Schlegel, Peter N.

    2012-01-01

    Patients with non-obstructive azoospermia (NOA) were once considered to be infertile with few treatment options due to the absence of sperm in the ejaculate. In the last two decades, the advent of intracytoplasmic sperm injection (ICSI), and the application of various testicular sperm retrieval techniques, including fine needle aspiration (FNA), conventional testicular sperm extraction (TESE) and microdissection testicular sperm extraction (micro-TESE) have revolutionized treatment in this gr...

  15. Human sperm rheotaxis: a passive physical process

    OpenAIRE

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

    2016-01-01

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

  16. Fluctuating food resources influence developmental plasticity in wild boar.

    Science.gov (United States)

    Gamelon, Marlène; Douhard, Mathieu; Baubet, Eric; Gimenez, Olivier; Brandt, Serge; Gaillard, Jean-Michel

    2013-10-23

    To maximize long-term average reproductive success, individuals can diversify the phenotypes of offspring produced within a reproductive event by displaying the 'coin-flipping' tactic. Wild boar (Sus scrofa scrofa) females have been reported to adopt this tactic. However, whether the magnitude of developmental plasticity within a litter depends on stochasticity in food resources has not been yet investigated. From long-term monitoring, we found that juvenile females produced similar-sized fetuses within a litter independent of food availability. By contrast, adult females adjusted their relative allocation to littermates to the amount of food resources, by providing a similar allocation to all littermates in years of poor food resources but producing highly diversified offspring phenotypes within a litter in years of abundant food resources. By minimizing sibling rivalry, such a plastic reproductive tactic allows adult wild boar females to maximize the number of littermates for a given breeding event. PMID:23904566

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

    OpenAIRE

    Oliver Vöcking; Gabriele Uhl; Peter Michalik

    2013-01-01

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

  18. Parasitic infections in wild ruminants and wild boar

    OpenAIRE

    Ilić Tamara; Stojanov Igor; Dimitrijević Sanda

    2011-01-01

    Wild ruminants and wild boar belong to the order Artiodactyla, the suborders Ruminantia and Nonruminantia and are classified as wild animals for big game hunting, whose breeding presents a very important branch of the hunting economy. Diseases caused by protozoa are rarely found in wild ruminants in nature. Causes of coccidiosis, cryptosporidiosis, toxoplasmosis, sarcocystiosis, giardiasis, babesiosis, and theileriosis have been diagnosed in deer. The most ...

  19. Antibiotic resistances of intestinal lactobacilli isolated from wild boars.

    Science.gov (United States)

    Klose, Viviana; Bayer, Katharina; Kern, Corinna; Goelß, Florian; Fibi, Silvia; Wegl, Gertrude

    2014-01-10

    Acquired antibiotic resistances have been reported in lactobacilli of various animal and food sources, but there are no data from wild boar. The objective was a preliminary examination of the antibiotic resistance prevalence of intrinsically vancomycin-resistant lactobacilli isolated from wild boar intestines and analysis of the genetic determinants implicated. Out of three wild boars, 121 lactobacilli were recovered and grouped according to their whole cell protein patterns. Initial phenotypic screening revealed that all were susceptible to erythromycin (2 μg/ml), but 30 were resistant to tetracycline (32 μg/ml). Based on Randomly Amplified Polymorphic DNA-PCR clustering, 64 strains were selected as representative genotypes for identification and minimum inhibitory concentration (MIC) determination. Partial 16S rRNA gene sequencing identified four species: (i) L. mucosae (n=57), (ii) L. reuteri (n=47), (iii) L. fermentum (n=12), and (iv) L. murinus (n=5). Most heterofermentative strains displayed low MICs for ampicillin (AMP), chloramphenicol (CHL), streptomycin (STR), kanamycin (KAN), gentamicin (GEN), erythromycin (ERY), quinupristin/dalfopristin (Q/D), and clindamycin (CLI). Atypical MICs were found mainly in L. mucosae and L. reuteri for TET, KAN, STR, AMP and CHL, but except the TET MICs of L. mucosae mostly at low level. L. murinus strains revealed atypical MICs for aminoglycosides, and/or CHL, AMP, CLI. PCR screening detected tet(W) in 12 and tet(M) in one of heterofermentative strains, as well as the aph(3')-III kanamycin gene in L. murinus. This is the first report showing acquired antibiotic resistance determinants in intestinal lactobacilli of wild boar origin. PMID:24326231

  20. Expression and localization of acrosin inhibitor in boar reproductive tract

    Czech Academy of Sciences Publication Activity Database

    Davidová, Nina; Jonáková, Věra; Maňásková-Postlerová, Pavla

    2009-01-01

    Roč. 338, č. 2 (2009), s. 303-311. ISSN 0302-766X R&D Projects: GA ČR GA303/09/1285; GA MŠk 1M06011; GA ČR GD523/08/H064 Institutional research plan: CEZ:AV0Z50520701 Keywords : Acrosin inhibitor * Reproductive tract * Spermatozoa * Boar Subject RIV: CE - Biochemistry Impact factor: 2.308, year: 2009

  1. Fluctuating food resources influence developmental plasticity in wild boar

    OpenAIRE

    Gamelon, Marlène; Douhard, Mathieu; Baubet, Eric; Gimenez, Olivier; Brandt, Serge; Gaillard, Jean-Michel

    2013-01-01

    To maximize long-term average reproductive success, individuals can diversify the phenotypes of offspring produced within a reproductive event by displaying the ‘coin-flipping’ tactic. Wild boar (Sus scrofa scrofa) females have been reported to adopt this tactic. However, whether the magnitude of developmental plasticity within a litter depends on stochasticity in food resources has not been yet investigated. From long-term monitoring, we found that juvenile females produced similar-sized fet...

  2. Characterization of peptide proteinase inhibitors isolated from boar seminal plasma

    Czech Academy of Sciences Publication Activity Database

    Jelínková, Petra; Tichá, M.; Jonáková, Věra

    Praha : UOCHB AV ČR, 2003 - (Slaninová, J.; Collinsová, M.; Klasová, L.), s. 1-57 [Biologicky aktivní peptidy /8./. Praha (CZ), 23.04.2003-25.04.2003] R&D Projects: GA ČR GA303/99/0357; GA ČR GP303/02/P069; GA MZd NJ7463 Institutional research plan: CEZ:MSM 113100001 Keywords : boar seminal plasma proteins * proteinase inhibitors Subject RIV: CE - Biochemistry

  3. High prevalence of porcine Hokovirus in German wild boar populations

    Directory of Open Access Journals (Sweden)

    Ellerbrok Heinz

    2010-07-01

    Full Text Available Abstract Porcine Hokovirus (PHoV was recently discovered in Hong Kong. This new Parvovirus of pigs is closely related to the human Parvoviruses 4 and 5 (PARV4/5 and bovine Hokovirus (BHoV. So far, nothing is known about the presence and prevalence of PHoV in regions of the world other than Hong Kong. A study was initiated to investigate PHoV in German wild boars from five different geographical regions, using a newly established quantitative real-time PCR assay. Analysis of collected liver and serum samples revealed high overall prevalence (32.7%; 51/156 of PHoV in wild boars. The prevalence differed between the regions and increased with age. Two near full-length genomes and a large fragment for three additional isolates from different regions were sequenced and used for phylogenetic analysis. The German PHoV sequences from wild boars showed a close relationship with sequences of isolates from Hong Kong.

  4. Trichinellosis in farmed wild boar: meat inspection findings and seroprevalence

    Directory of Open Access Journals (Sweden)

    Sukura A.

    2001-06-01

    Full Text Available A reflection of highly prevalent endemic wildlife trichinellosis is seen in wild boar farming in Finland. During the last five years, 0.7 % (15/2265 of wild boars undergoing official meat inspection have been determined to be Trichinella-positive. These findings originate from six different farms. In Finland, T. spiralis and T. pseudospiralis have been discovered in meat inspection of wild boars. ELISA showed 11 out of 9 9 serum samples (11 % as having specific antibodies for T. spiralis crude antigen. Positive samples were from three out of the thirteen farms from which the sera were available. Most of the positive serum samples (8/11 originated from a farm where trichinellosis was also revealed in meat inspection, the other two seropositive farms were without previous Trichinella records. Over the last few decades, no reports have been made of human trichinellosis acquired in Finland. This indicates both efficient meat inspection as well as public awareness of high-risk foodstuff.

  5. Selection of Sperm Based on Hypo-Osmotic Swelling May Improve ICSI Outcome: A Preliminary Prospective Clinical Trial

    Directory of Open Access Journals (Sweden)

    Nasim Charehjooy

    2014-03-01

    Full Text Available Background: The intra-cytoplasmic sperm injection (ICSI technique selects sperm according to morphology and motility. However, these parameters cannot predict the chromatin integrity of sperm. Considering the detrimental effects of DNA-damaged sperm on reproductive outcomes, novel sperm selection procedures have been proposed to circumvent the possibility of inseminating DNA-damaged sperm. It has been shown that different potential hypo-osmotic swelling test (HOST patterns possess the potential to differentiate between sperm that have intact or damaged chromatin. Therefore, for the first time, this preliminary study evaluates the role of HOST as a sperm selection procedure in a clinical setting. Materials and Methods: In this preliminary prospective clinical trial study, we divided infertile couples diagnosed with male infertility into two groups. In the treatment group (n=39, half of the oocytes were inseminated by sperm selected following density gradient centrifugation (DGC group. The remaining oocytes from the treatment group were inseminated by sperm chosen according to HOST pattern (c, d or e following DGC processing (HOST group. In the control group (n=63, all oocytes were inseminated by sperm chosen after DGC. Results: There was a significantly higher percentage of embryos that had good quality, implantation, and chemical pregnancy rates in the HOST group compared to the DGC group (p≤0.05. Conclusion: This study has shown that selecting sperm according to membrane functionality (HOST pattern rather morphology and viability may open a new window in our approach for determining the appropriate sperm for ICSI, particularly in individuals with severe male infertility (Registration Number: IRCT201307087223N2.

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

    Directory of Open Access Journals (Sweden)

    Jaime Gosálvez

    2014-06-01

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

  7. Sperm competition games: optimal sperm allocation in response to the size of competing ejaculates

    OpenAIRE

    Engqvist, Leif; Reinhold, Klaus

    2007-01-01

    Sperm competition theory predicts that when males are certain of sperm competition, they should decrease sperm investment in matings with an increasing number of competing ejaculates. How males should allocate sperm when competing with differently sized ejaculates, however, has not yet been examined. Here, we report the outcomes of two models assuming variation in males' sperm reserves and males being faced with different amounts of competing sperm. In the first ‘spawning model’, two males co...

  8. Comparison of permeating and nonpermeating cryoprotectants for mouse sperm cryopreservation.

    Science.gov (United States)

    Sztein, J M; Noble, K; Farley, J S; Mobraaten, L E

    2001-02-01

    Mouse sperm has proven to be more difficult to cryopreserve than sperm of other mammalian species. Published reports show that only three cryoprotectant agents (CPAs), alone or combined, have been studied: glycerol and dimethyl sulfoxide (DMSO), as permeating agents, and raffinose, as a nonpermeating agent. To date, the most consistent results for mouse sperm cryopreservation have been achieved by use of raffinose/skim milk as cryoprotectant with rapid cooling at 20 degrees C per minute. In this study, we compared the cryoprotection provided by permeating (glycerol, formamide, propanediol, DMSO, adonitol) or nonpermeating (lactose, raffinose, sucrose, trehalose, d-mannitol) compounds for freezing mouse sperm. Different solutions were made using 3% skim milk solution as the buffer or extender in which all different cryoprotectant agents were dissolved at a concentration of 0.3 M, with a final osmolality of approx. 400 mOsm. Sperm samples from CB6F1 (hybrid) and C57BL/6J (inbred) mice collected directly into each CPA were frozen/thawed under identical conditions. After thawing and CPA elimination (centrifugation) raffinose (59%), trehalose (61%), and sucrose (61%) sustained the best motility (P = Membrane integrity was analyzed and showed that the simple exposure (prefreeze) to sugars was less harmful than the exposure to glycols. Coincidentally, sperm frozen in trehalose (41%), raffinose (40.5%), and sucrose (37.5%) were the samples less injured among all different postthawed CPA tested. The in vitro fertilization results demonstrated that hybrid mouse spermatozoa frozen with sugars (lactose 80%, raffinose 80%, trehalose 79% of two-cell embryos production) were more fertile than those frozen with glycols (glycerol 11%). PMID:11336487

  9. Study of sperm cell phosphorylating systems using nucleotide photoaffinity probes

    International Nuclear Information System (INIS)

    The major thrust of the research presented in this thesis was to identify specific nucleotide binding proteins and phosphoproteins of rat caput and cauda sperm. Also, the differences in these proteins between caput and cauda sperm were investigated as well as determination of the membrane sidedness of the proteins and their location in either the head or tail/mid-piece region. In addition, the effects of small molecular weight modifers such as cGMP, cAMP and Ca2+ on the detection of binding proteins and phosphorylated proteins was studied. The technique used to identify and locate nucleotide binding proteins was photoaffinity labeling using the proven 8-azidopurine nucleotide analogs of cAMP, ATP and GTP in radioactive form. The first study presented involved the use of [32P]8-N 3cAMP which showed that both caput and cauda sperm contained both type I and type II regulatory subunits (R/sub I/ and R/sub II/, respectively) of the cAMP dependent kinases and that the great majority of the regulatory subunits were located in the tail/mid-piece section and not in the sperm head. The second phase of this study involved the use of [γ32P]8-azidoadensosine triphosphate ([γ32P]8-N3ATP) and (γ32P)8-azidoguanosine triphosphate ([γ32P]8-N3GTP) to photolable specific ATP and GTP binding proteins and to phosphorylate specific phosphoproteins. Again, this was done on caput versus cauda sperm and the location of the majority of the photolabeled or phosphorylated proteins was shown to be in the tail/mid-piece fraction. In addition, considerable differences were found in both the phosphorylated and photolabeled proteins of caput versus cauda sperm

  10. Mitochondria functionality and sperm quality

    OpenAIRE

    Amaral, Alexandra; Lourenço, Bárbara; Marques, Mónica; Ramalho-Santos, J.

    2013-01-01

    Although mitochondria are best known for being the eukaryotic cell powerhouses, these organelles participate in various cellular functions besides ATP production, such as calcium homoeostasis, generation of reactive oxygen species (ROS), the intrinsic apoptotic pathway and steroid hormone biosynthesis. The aim of this review was to discuss the putative roles of mitochondria in mammalian sperm function and how they may relate to sperm quality and fertilisation ability, particularly in...

  11. The association between disease and profitability in individual finishing boars at a test station

    DEFF Research Database (Denmark)

    Jensen, Tina Birk; Baadsgaard, Niels Peter; Houe, Hans;

    2008-01-01

    Endemic diseases in finisher herds are considered to be costly for the pig producer. We investigated the effect of diseases on the profit margin using data from a Danish boar test station (n = 5777) collected from July 2002 to December 2004. Boars reaching a target slaughter weight of at least 80...

  12. Disease risks associated with free-ranging wild boar in Saskatchewan.

    Science.gov (United States)

    McGregor, Glenna F; Gottschalk, Marcelo; Godson, Dale L; Wilkins, Wendy; Bollinger, Trent K

    2015-08-01

    This study investigated the disease status of Saskatchewan's feral wild boar population. Whole carcasses, tissue samples, and/or serum from 81 hunter-killed boars from Saskatchewan were submitted to the Canadian Wildlife Health Cooperative (CWHC) between 2009 and 2014. Serological tests were negative for PRRS, H1N1, and H3N2 swine influenza, PCV-2, and TGE/PRCV in 22/22 boars and for Toxoplasma gondii and Mycoplasma hyopneumoniae in 20/20 boars. Of 20 boars whose sera were tested 20 were positive for Actinobacillus pleuropneumoniae, with 7 positive for, among other strains, serotype 14; 16 were positive for Lawsonia intracellularis, 1 was positive and 6 were suspicious for Salmonella spp. Polymerase chain reaction tests were negative for PRRS and PCV2 in 58/58 boars and positive for Torque teno virus in 1/8 boars. Digestion assays were negative for Trichinella spp. in 22/22 boars. The high seroprevalence of A. pleuropneumoniae serotype 14 is noteworthy as this serotype has not been previously reported in North America. PMID:26246630

  13. Effect of refractoriness to long photoperiod on sperm production and quality in tomcats.

    Science.gov (United States)

    Nuñez Favre, R; Bonaura, M C; Tittarelli, C M; Stornelli, M C; de la Sota, R L; Stornelli, M A

    2012-12-01

    The aim of this study was to assess whether refractoriness to long photoperiod (LP) could be reversed by subjecting tomcats to a period of short days. Our hypothesis was that photoperiod changes can avoid refractoriness and restore sperm quality and production to that before refractoriness. Tomcats (n = 6) were housed in a conditioned room with LP (12L: 12D) for 45 days of acclimation and then maintained under LP for 18 month. Then, tomcats were changed to a period of decreasing light at a rate of 8 min/day for 1 month. Tomcats stayed for 1 month with short photoperiod (SP; 8L: 16D) and then were switched back to a period of increasing light at a rate of 8 min/day for 1 month. The experiment was completed after tomcats remained in LP for 2 months. Toms were anaesthetized and semen samples were collected by electroejaculation every 2 weeks. Sperm parameters were evaluated in all ejaculates, and data were analysed by anova. Motility, velocity, volume, sperm concentration, total sperm count, viability, acrosome integrity, plasma membrane integrity and sperm morphology were higher during LP compared with a refractory LP (p 0.20).Whereas motility, velocity, viability, acrosome integrity and plasma membrane integrity were similar in a refractory LP compared with SP (p > 0.05), volume, sperm concentration, total sperm count and sperm morphology were lower in a refractory LP compared with SP (p < 0.05). In conclusion, refractoriness and reduced sperm production and quality induced by a prolonged LP of 18 month can be restored after placing tomcats to a SP. PMID:23279508

  14. Ion channels, phosphorylation and mammalian sperm capacitation

    Institute of Scientific and Technical Information of China (English)

    Pablo E Visconti; Dario Krapf; José Luis de la Vega-Beltrán; Juan José Acevedo; Alberto Darszon

    2011-01-01

    Sexually reproducing animals require an orchestrated communication between spermatozoa and the egg to generate a new individual. Capacitation, a maturational complex phenomenon that occurs in the female reproductive tract, renders spermatozoa capable of binding and fusing with the oocyte, and it is a requirement for mammalian fertilization. Capacitation encompasses plasma membrane reorganization, ion permeability regulation, cholesterol loss and changes in the phosphorylation state of many proteins. Novel tools to study sperm ion channels, image intracellular ionic changes and proteins with better spatial and temporal resolution, are unraveling how modifications in sperm ion transport and phosphorylation states lead to capacitation. Recent evidence indicates that two parallel pathways regulate phosphorylation events leading to capacitation, one of them requiring activation of protein kinase A and the second one involving inactivation of ser/thr phosphatases. This review examines the involvement of ion transporters and phosphorylation signaling processes needed for spermatozoa to achieve capacitation. Understanding the molecular mechanisms leading to fertilization is central for societies to deal with rising male infertility rates, to develop safe male gamete-based contraceptives and to preserve biodiversity through better assisted fertilization strategies.

  15. Anandamide induces sperm release from oviductal epithelia through nitric oxide pathway in bovines.

    Directory of Open Access Journals (Sweden)

    Claudia Osycka-Salut

    Full Text Available Mammalian spermatozoa are not able to fertilize an egg immediately upon ejaculation. They acquire this ability during their transit through the female genital tract in a process known as capacitation. The mammalian oviduct acts as a functional sperm reservoir providing a suitable environment that allows the maintenance of sperm fertilization competence until ovulation occurs. After ovulation, spermatozoa are gradually released from the oviductal reservoir in the caudal isthmus and ascend to the site of fertilization. Capacitating-related changes in sperm plasma membrane seem to be responsible for sperm release from oviductal epithelium. Anandamide is a lipid mediator that participates in the regulation of several female and male reproductive functions. Previously we have demonstrated that anandamide was capable to release spermatozoa from oviductal epithelia by induction of sperm capacitation in bovines. In the present work we studied whether anandamide might exert its effect by activating the nitric oxide (NO pathway since this molecule has been described as a capacitating agent in spermatozoa from different species. First, we demonstrated that 1 µM NOC-18, a NO donor, and 10 mM L-Arginine, NO synthase substrate, induced the release of spermatozoa from the oviductal epithelia. Then, we observed that the anandamide effect on sperm oviduct interaction was reversed by the addition of 1 µM L-NAME, a NO synthase inhibitor, or 30 µg/ml Hemoglobin, a NO scavenger. We also demonstrated that the induction of bull sperm capacitation by nanomolar concentrations of R(+-methanandamide or anandamide was inhibited by adding L-NAME or Hemoglobin. To study whether anandamide is able to produce NO, we measured this compound in both sperm and oviductal cells. We observed that anandamide increased the levels of NO in spermatozoa, but not in oviductal cells. These findings suggest that anandamide regulates the sperm release from oviductal epithelia probably by

  16. Surveillance of classical swine fever in wild boar in South Korea from 2010-2014.

    Science.gov (United States)

    Kim, Yong Kwan; Lim, Seong-In; Kim, Jae-Jo; Cho, Yoon-Young; Song, Jae-Young; Cho, In-Soo; Hyun, Bang-Hun; Choi, Sung-Hyun; Kim, Seung-Hoe; Park, Eun-Hye; An, Dong-Jun

    2016-01-01

    Classical swine fever (CSF) is a highly contagious systemic hemorrhagic viral disease of pigs. Wild boar plays a crucial role in the epidemiology of CSF. Between 2010 and 2014, samples were collected nationwide from 6,654 wild boars hunted in South Korea. Anti-CSF antibodies were identified in 0.59% (39 of 6,654) of the wild boar samples using a virus neutralization test and were primarily detected in wild boars living close to the demilitarized zone and the area of the Taebaek Mountains surroundings. The CSF virus (subgroup 2.1b) was isolated from two wild boars captured in a nearby border area. The criteria used to define high-risk areas for targeted CSF surveillance in South Korea should be further expanded to include other regions nationwide. PMID:26178821

  17. First detection of sarcoptic mange in free-ranging wild boar (Sus scrofa) in Switzerland.

    Science.gov (United States)

    Haas, C; Origgi, F C; Akdesir, E; Batista Linhares, M; Giovannini, S; Mavrot, F; Casaubon, J; Ryser-Degiorgis, M-P

    2015-05-01

    In Switzerland sarcoptic mange is frequent in free-ranging wild carnivores but until recent years no cases had been recorded in wild ungulates. Since 2010, cases have been observed in wild boar in the cantons of Solothurn, Tessin and Thurgau. Here, we report the detection of mange-like skin lesions in wild boars by photo-trapping and the post-mortem findings in 6 culled animals presenting different stages of the disease. Potential sources of infection include mangy red foxes, outdoor domestic pigs and wild boars from surrounding countries. Disease spread in the wild boar population may become relevant not only for wildlife but also for domestic pig health in the future if piggeries' biosecurity is insufficient to prevent interactions with wild boar. PMID:26753342

  18. Nicotiana ovule extracts in duce nuclear reconstitution of demembranated Xenopus sperm in cell-free system

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    s Nicotiana tabaccum ovule extracts induced nuclear reconstitution of demembranated Xenopus leavis sperm in a ceil-free system. Demembranated Xenopus sperm began to swell after 15 rmin of incubation with Nicotiana ovulde extracts. Accompanying the process of incubation,Xenopus sperm decondensed and their shapes changed gradually from long and ellipse to round. The completely decondensed chromatin was surrounded with membrane structure, which was a mixture envelope of a double membrane and a single membrane. Nucleosome assembly was verified by means of micrococcal nuclease digestion to reconstituted nuclei and DNA electrophoresis. Nicotiana ovule extracts supplied one more experimental model and system.The new system could promote powerfully the research on mechanisms of cell division and cell cycle regulation.

  19. A specific flagellum beating mode for inducing fusion in mammalian fertilization and kinetics of sperm internalization

    Science.gov (United States)

    Ravaux, Benjamin; Garroum, Nabil; Perez, Eric; Willaime, Hervé; Gourier, Christine

    2016-01-01

    The salient phases of fertilization are gamete adhesion, membrane fusion, and internalization of the spermatozoon into the oocyte but the precise timeline and the molecular, membrane and cell mechanisms underlying these highly dynamical events are far from being established. The high motility of the spermatozoa and the unpredictable location of sperm/egg fusion dramatically hinder the use of real time imaging optical techniques that should directly provide the dynamics of cell events. Using an approach based on microfluidics technology, the sperm/egg interaction zone was imaged with the best front view, and the timeline of the fertilization events was established with an unparalleled temporal accuracy from the onset of gamete contact to full sperm DNA decondensation. It reveals that a key element of the adhesion phase to initiate fusion is the oscillatory motion of the sperm head on the oocyte plasma membrane generated by a specific flagellum-beating mode. It also shows that the incorporation of the spermatozoon head is a two steps process that includes simultaneous diving, tilt, and plasma membrane degradation of the sperm head into the oocyte and subsequent DNA decondensation. PMID:27539564

  20. Nutrition and Reproductive Health: Sperm versus Erythrocyte Lipidomic Profile and ω-3 Intake

    Directory of Open Access Journals (Sweden)

    Gabriela Ruth Mendeluk

    2015-01-01

    Full Text Available Fatty acid analyses of sperm and erythrocyte cell membrane phospholipids in idiopathic infertile patients evidenced that erythrocyte contents of EPA, DHA, omega-6–omega-3 ratio and arachidonic acid provide a mathematical correspondence for the prediction of EPA level in sperm cells. The erythrocyte lipidomic profile of patients was significantly altered, with signatures of typical Western pattern dietary habits and no fish intake. A supplementation with nutritional levels of EPA and DHA and antioxidants was then performed for 3 months, with the follow-up of both erythrocyte and sperm cell membranes composition as well as conventional sperm parameters. Some significant changes were found in the lipidomic membrane profile of erythrocyte but not in sperm cells, which correspondently did not show significant parameter ameliorations. This is the first report indicating that membrane lipids of different tissues do not equally metabolize the fatty acid elements upon supplementation. Molecular diagnostic tools are necessary to understand the cell metabolic turnover and monitor the success of nutraceuticals for personalized treatments.

  1. Bovine binder-of-sperm protein BSP1 promotes protrusion and nanotube formation from liposomes

    Energy Technology Data Exchange (ETDEWEB)

    Lafleur, Michel, E-mail: michel.lafleur@umontreal.ca [Department of Chemistry, Center for Self-Assembled Chemical Systems, Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada); Courtemanche, Lesley [Department of Chemistry, Center for Self-Assembled Chemical Systems, Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada); Karlsson, Goeran; Edwards, Katarina [Department of Physical and Analytical Chemistry, Uppsala University, Box 579, S-751 23 Uppsala (Sweden); Schwartz, Jean-Louis [Department of Physiology, Groupe d' etude des Proteines Membranaires, Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada); Manjunath, Puttaswamy [Maisonneuve-Rosemont Hospital Research Center and Faculty of Medecine, Universite de Montreal, 5415 L' Assomption Blvd, Montreal, Quebec, Canada H1T 2M4 (Canada)

    2010-08-27

    Research highlights: {yields} Binder-of-sperm protein 1 (BSP1) modifies the morphology of lipidic vesicles inducing bead necklace-like and thread-like structures. {yields} In the presence of multilamellar liposomes, BSP1 leads to the formation of long nanotubes. {yields} The insertion of BSP1 in the external lipid leaflet of membranes induces local changes in bilayer curvature. -- Abstract: Binder-of-sperm (BSP) proteins interact with sperm membranes and are proposed to extract selectively phosphatidylcholine and cholesterol from these. This change in lipid composition is a key step in sperm capacitation. The present work demonstrates that the interactions between the protein BSP1 and model membranes composed with phosphatidylcholine lead to drastic changes in the morphology of the lipidic self-assemblies. Using cryo-electron microscopy and fluorescence microscopy, we show that, in the presence of the protein, the lipid vesicles elongate, and form bead necklace-like structures that evolve toward small vesicles or thread-like structures. In the presence of multilamellar vesicles, where a large reservoir of lipid is available, the presence of BSP proteins lead to the formation of long nanotubes. Long spiral-like threads, associated with lipid/protein complexes, are also observed. The local curvature of lipid membranes induced by the BSP proteins may be involved in lipid domain formation and the extraction of some lipids during the sperm maturation process.

  2. Bovine binder-of-sperm protein BSP1 promotes protrusion and nanotube formation from liposomes

    International Nuclear Information System (INIS)

    Research highlights: → Binder-of-sperm protein 1 (BSP1) modifies the morphology of lipidic vesicles inducing bead necklace-like and thread-like structures. → In the presence of multilamellar liposomes, BSP1 leads to the formation of long nanotubes. → The insertion of BSP1 in the external lipid leaflet of membranes induces local changes in bilayer curvature. -- Abstract: Binder-of-sperm (BSP) proteins interact with sperm membranes and are proposed to extract selectively phosphatidylcholine and cholesterol from these. This change in lipid composition is a key step in sperm capacitation. The present work demonstrates that the interactions between the protein BSP1 and model membranes composed with phosphatidylcholine lead to drastic changes in the morphology of the lipidic self-assemblies. Using cryo-electron microscopy and fluorescence microscopy, we show that, in the presence of the protein, the lipid vesicles elongate, and form bead necklace-like structures that evolve toward small vesicles or thread-like structures. In the presence of multilamellar vesicles, where a large reservoir of lipid is available, the presence of BSP proteins lead to the formation of long nanotubes. Long spiral-like threads, associated with lipid/protein complexes, are also observed. The local curvature of lipid membranes induced by the BSP proteins may be involved in lipid domain formation and the extraction of some lipids during the sperm maturation process.

  3. Is the hook of muroid rodent's sperm related to sperm train formation?

    Science.gov (United States)

    Tourmente, M; Zarka-Trigo, D; Roldan, E R S

    2016-06-01

    Competition between spermatozoa of rival males to gain fertilizations has led to a wide array of modifications in sperm structure and function. Sperm cells of most muroid rodents have hook-shaped extensions in the apical-ventral tip of the head, but the function of this structure is largely unknown. These 'hooks' may facilitate aggregation of spermatozoa in so-called 'trains', as an adaptation to sperm competition, because sperm in trains may swim faster than free-swimming cells. However, there is controversy regarding the role of the hook in train formation, and in relation to whether it is selected by sperm competition. We examined spermatozoa from muroid rodents with varying levels of sperm competition to assess whether (i) sperm aggregates are common in these taxa, (ii) presence of a hook relates to the formation of sperm aggregations, and (iii) formation of sperm aggregations is explained by sperm competition. Our analyses in 25 muroid species revealed that > 92% of spermatozoa swim individually in all species, with the exception of the wood mouse, Apodemus sylvaticus, which has ~50% spermatozoa swimming freely. Species with hooked spermatozoa had higher sperm competition levels and longer sperm than species whose sperm lack a hook. Neither the presence of hook nor sperm competition levels were related to the percentage of sperm in aggregations. Thus, (i) sperm aggregates in muroid rodents are an exceptional trait found only in a few species, (ii) evolution of the sperm hook is associated to sperm competition levels, but (iii) the hook is unlikely to be related to the formation of sperm aggregates. The evolutionary significance of the sperm head hook thus remains elusive, and future studies should examine potential roles of this pervasive structure in sperm's hydrodynamic efficiency and sperm-female tract interactions. PMID:26969911

  4. Potential Changes in Rat Spermatogenesis and Sperm Parameters after Inhalation of Boswellia papyrifera and Boswellia carterii Incense

    Directory of Open Access Journals (Sweden)

    Tajamul Hussain

    2013-02-01

    Full Text Available In this study the effect of Boswellia papyrifera (B. papyrifera and Boswellia carterii (B. carterii smoke exposure on spermatogenesis and sperm parameters in male albino rats was investigated. Rats (n = 11 were exposed daily in smoking chambers to smoke emanated by burning 4 g each of either B. papyrifera or B. carterii for 48 days. At the end of exposure duration rats were killed, and the testes were excised and analysed for histopathological and ultrastructural changes. Sperm analysis including total sperm count, motility, velocity and relative percentage of abnormal sperms were recorded. Rats exposed to B. papyrifera and B. carterii showed significant disturbances in spermatogenetic patterns and changes in sperm kinetics compared to unexposed rats. Atrophied seminiferous tubules with dynamic changes were also noticed. The boundaries of intercellular and intracellular vacuoles were seen in the Sertoli cells. Furthermore, in spermatids acrosomal vesicles were not fully formed. Degenerating spermatids were devoid of their nuclear membrane with electron dense matrix and vacuolization. Structural changes in Leydig cells were observed. Sperm analysis in exposed rats exhibited significant decrease in the sperm count, motility, speed and an increase in sperm anomalies when compare to controls. These findings demonstrate that the B. papyrifera and B. carterii smoke affects the process of spermatogenesis and sperm parameters and indicate the detrimental effects of these incense materials on human reproductive system.

  5. Molecular epidemiology of Mycoplasma hyopneumoniae from outbreaks of enzootic pneumonia in domestic pig and the role of wild boar.

    Science.gov (United States)

    Kuhnert, Peter; Overesch, Gudrun

    2014-11-01

    Mycoplasma hyopneumoniae is the major cause of enzootic pneumonia (EP) in domestic pigs, a disease with low mortality but high morbidity, having a great economic impact for producers. In Switzerland EP has been successfully eradicated, however, sporadic outbreaks are observed with no obvious source. Besides the possibility of recurrent outbreaks due to persisting M. hyopneumoniae strains within the pig population, there is suspicion that wild boars might introduce M. hyopneumoniae into swine herds. To elucidate possible links between domestic pig and wild boar, epidemiological investigations of recent EP outbreaks were initiated and lung samples of pig and wild boar were tested for the presence of specific genotypes by multilocus sequence typing (MLST). Despite generally different genotypes in wild boar, outbreak strains could be found in geographically linked wild boar lungs after, but so far not before the outbreak. Recurrent outbreaks in a farm were due to the same strain, indicating unsuccessful sanitation rather than reintroduction by wild boar. In another case outbreaks in six different farms were caused by the same strain never found in wild boar, confirming spread between farms due to hypothesized animal transport. Results indicate the presence of identical lineages of wild boar and domestic pig strains, and possible transmission of M. hyopneumoniae between wild boar and pig. However, the role of wild boar might be rather one as a recipient than a transmitter. More important than contact to wild boar for sporadic outbreaks in Switzerland is apparently persistence of M. hyopneumoniae within a farm as well as transmission between farms. PMID:25236987

  6. Altered miRNAs expression profiling in sperm of mice induced by fluoride.

    Science.gov (United States)

    Sun, Zilong; Zhang, Wen; Li, Sujuan; Xue, Xingchen; Niu, Ruiyan; Shi, Lei; Li, Baojun; Wang, Xiaowen; Wang, Jundong

    2016-07-01

    The reproductive toxicity of fluoride has become a major concern in the world. Fluoride can decrease the abilities of sperm capacitation, hyperactivation, chemotaxis, acrosome reaction and fertilization, but the studies on the responses of sperm small noncoding RNAs (sncRNAs), especially miRNAs, to fluoride exposure are lacking. miRNAs are demonstrated to influence sperm quality and male fertility by regulating gene expression at post-transcriptional levels or translational repression. The objective of this study is to analyze miRNA profiling in sperm of mice administrated with 25 and 100 mg L(-1) sodium fluoride (NaF) for 60 d using high-throughput sequencing technology. Along with reduced sperm concentration, survival, motility, and mitochondrial membrane potential, 31 differentially expressed known miRNAs were identified in fluoride groups, compared with the control group. 671 predicted target genes against the 16 altered miRNAs were mainly involved in protease inhibitor activity, apoptosis, ubiquitin mediated proteolysis, and signaling pathways of calcium, JAK-STAT, MAPK, p53, Wnt, which were proved to be directly related to sperm quality. These findings suggested that the altered sperm miRNAs could be potential biomarkers for fluoride reproductive toxicity. PMID:27108368

  7. Variability in sperm form and function in the context of sperm competition risk in two Tupinambis lizards

    OpenAIRE

    Blengini, Cecilia S.; Sergio, Naretto; Gabriela, Cardozo; Giojalas, Laura C.; Margarita, Chiaraviglio

    2014-01-01

    In polyandrous species, sperm morphometry and sperm velocity are under strong sexual selection. Although several hypotheses have been proposed to explain the role of sperm competition in sperm trait variation, this aspect is still poorly understood. It has been suggested that an increase in sperm competition pressure could reduce sperm size variation or produce a diversity of sperm to maximize male fertilization success. We aim at elucidating the variability of sperm morphometric traits and v...

  8. Acute effects of TCDD administration:special emphasis on testicular and sperm mitochondrial function

    Institute of Scientific and Technical Information of China (English)

    Paula C Mota; Renata S Tavares; Marlia Cordeiro; Susana P Pereira; Stephen J Publicover; Paulo J Oliveira; Joo Ramalho-Santos

    2012-01-01

    Objective: The goal of this study was to verify if 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) could have any effect on male germ cells mitochondria and in this way add new insights in how male reproductive alterations observed in other studies occur. Methods:In vivo and in vitro approaches using rat testis and human sperm as models were employed to evaluate TCDD effects on testicular and sperm mitochondria after 24 h of exposure. Results:Testicular mitochondria from TCDD-treated rats presented no differences in the bioenergetic parameters monitored except for a significantly higher electric membrane potential in the presence of ADP, corroborated when TCDD was directly added to testicular mitochondria from untreated rats. Nevertheless, sperm mitochondrial membrane potential, motility, viability, capacitation and acrosomal integrity did not change after TCDD treatment. Moreover, only few sperm cells exposed to TCDD increased their intracellular Ca2+concentration. Conlusions:TCDD can interact directly with rat testicular mitochondria inducing small changes. This effect, however, does not seem to occur in human sperm or it may be insufficient to induce significant alterations as observed by the maintenance of sperm function.

  9. Variation in sperm morphology among Afrotropical sunbirds

    Czech Academy of Sciences Publication Activity Database

    Omotoriogun, T. C.; Laskemoen, T.; Rowe, M.; Albrecht, Tomáš; Bowie, R. C. K.; Sedláček, O.; Hořák, D.; Ottosson, U.; Lifjeld, J. T.

    2016-01-01

    Roč. 158, č. 1 (2016), s. 155-166. ISSN 0019-1019 Institutional support: RVO:68081766 Keywords : comparative analysis * Nectariniidae * phylogenetic signal * sperm competition * sperm size Subject RIV: EG - Zoology Impact factor: 1.921, year: 2014

  10. Psychosocial counselling of identifiable sperm donors

    NARCIS (Netherlands)

    M. Visser; M.H. Mochtar; A.A. de Melker; F. van der Veen; S. Repping; T. Gerrits

    2016-01-01

    STUDY QUESTION: What do identifiable sperm donors feel about psychosocial counselling? SUMMARY ANSWER: Identifiable sperm donors found it important that psychosocial counselling focused on emotional consequences and on rules and regulations and they expected to have access to psychosocial counsellin

  11. A low conductivity culture medium suitable for the evaluation of sperm motility

    Science.gov (United States)

    Ma, Rui; Han, Chao; Sun, Zilong; Huang, Guoliang; Yu, Zhongyao; Zhou, Yuxiang; Wang, Jundong; Qiao, Jie; Cheng, Jing

    2008-12-01

    A novel culture medium of low conductivity suitable for dielectrophoretic selection of sperm was developed. Conventional IVF methods lack the capability of selecting the expected sperms and the embryonic development may be adversely affected to certain extent. Dielectrophoresis (DEP), a technique commonly applied in cell manipulation [1], may provide an alternative. However, the conventional IVF medium has a high conductivity, which may result in the unexpected heating effect during DEP causing damage to the gametes. The newly developed medium consists of sucrose, HEPES, BSA and low concentrations of ions. The conductivity of this medium is significantly lower than the conventional IVF medium. Motility and membrane integrality of the mouse sperm were tested in the low-conductivity medium, demonstrating an acceptable percent rate of motile sperm compared to the control groups.

  12. The effect of cryopreservation on sperm head morphometry in Florida male goat related to sperm freezability.

    Science.gov (United States)

    Hidalgo, M; Rodríguez, I; Dorado, J M

    2007-07-01

    The Sperm Class Analyzer was used to investigate the effect of freeze-thawing procedure on Florida buck sperm head morphometry, and to relate possible changes in sperm head dimensions to cryopreservation success. Semen samples (n=76) were frozen with tris and milk-based extenders and thawed. Sperm quality samples (motility, morphology, acrosome), and sperm head morphometric values (length, width, area, perimeter, ellipticity) were compared between fresh and frozen-thawed samples. Sperm freezability was judged according to the sperm quality parameters assessed. Fertility data was obtained after artificial insemination with cryopreserved semen. Cryopreservation success was different between freezing methods. Sperm head dimensions were significantly (pmilk spermatozoa respectively than in those of the fresh samples. The sperm head morphometric parameters that had changed after cryopreservation were lower in suitable semen samples after thawing and with successful pregnancies after artificial insemination. These data suggest that changes in sperm head morphometry might reflect spermatozoa injury occurred during cryopreservation. PMID:16904275

  13. Assembly of Lampbrush Chromosomes from Sperm Chromatin

    OpenAIRE

    Gall, Joseph G.; Murphy, Christine

    1998-01-01

    We have examined the behavior of demembranated sperm heads when injected into the germinal vesicle (GV) of amphibian oocytes. Xenopus sperm heads injected into Xenopus GVs swelled immediately and within hours began to stain with an antibody against RNA polymerase II (Pol II). Over time each sperm head became a loose mass of chromosome-like threads, which by 24–48 h resolved into individually recognizable lampbrush chromosomes (LBCs). Although LBCs derived from sperm are unreplicated single ch...

  14. High prevalence of porcine Hokovirus in German wild boar populations

    OpenAIRE

    Ellerbrok Heinz; Kaiser Marco; Adlhoch Cornelia; Pauli Georg

    2010-01-01

    Abstract Porcine Hokovirus (PHoV) was recently discovered in Hong Kong. This new Parvovirus of pigs is closely related to the human Parvoviruses 4 and 5 (PARV4/5) and bovine Hokovirus (BHoV). So far, nothing is known about the presence and prevalence of PHoV in regions of the world other than Hong Kong. A study was initiated to investigate PHoV in German wild boars from five different geographical regions, using a newly established quantitative real-time PCR assay. Analysis of collected liver...

  15. AB229. Effect of berberine on human sperm parameters in vitro

    Science.gov (United States)

    Chen, Liping; Wang, Tao; Liu, Jihong

    2016-01-01

    Objective Berberine is a natural compound isolated from plants with multiple pharmacological activities, like antioxidant and anti-inflammatory activities. The present study was to assess the effect of berberine with different concentration and at different time on sperm motility, viability and sperm membrane integrity of humans. Methods Semen samples were obtained from 30 patients (22–30 years of age) enrolled in this study, 15 of which were normospermic and 15 were asthenozoospermic. Semen were aseptically obtained by masturbation and prepared by swim-up method. After that they were incubated in Ham’s F-10 medium for 30 min at 37 °C to mimic the temperature inside the female reproductive tract. Semen samples were divided into aliquots. The negative control condition was supplemented with no drugs while 0.4 mmol/L L-carnitine was used as positive control condition. The treatment condition was supplemented with berberine at a concentration of 10-4, 10-5, 10-6, 10-7 mol/L. Measurements of sperm viability and motility were carried out at 10, 30, 60 and 120 min after incubation in all semen samples by computer-assisted sperm analysis (CASA). Results It showed that in both normal and asthenozoospermic samples, total and progressive sperm motility were maintained by in vitro treatment with berberine, whereas a significant decrease of these parameters occurred in parallel samples incubated in medium alone. Berberine also prevented the decrease of sperm kinetics but such an effect was highly significant only in asthenozoospermic samples. Moreover, berberine had similar effort as L-carnitine even at low concentration. Conclusions In vitro, berberine with suitable concentration exerted a direct protective effect on human sperm viability, motility rate and could modify plasma membrane functional integrity, especially in the asthenozoospermic patients. This could be a novel therapeutic drug for improving the function of sperm in vitro and treatment for male infertility.

  16. Human sperm rheotaxis: a passive physical process

    Science.gov (United States)

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

    2016-03-01

    A long-standing question in natural reproduction is how mammalian sperm navigate inside female reproductive tract and finally reach the egg cell, or oocyte. Recently, fluid flow was proposed as a long–range guidance cue for sperm navigation. Coitus induces fluid flow from oviduct to uterus, and sperm align themselves against the flow direction and swim upstream, a phenomenon termed rheotaxis. Whether sperm rheotaxis is a passive process dominated by fluid mechanics, or sperm actively sense and adapt to fluid flow remains controversial. Here we report the first quantitative study of sperm flagellar motion during human sperm rheotaxis and provide direct evidence indicating that sperm rheotaxis is a passive process. Experimental results show that there is no significant difference in flagellar beating amplitude and asymmetry between rheotaxis-turning sperm and those sperm swimming freely in the absence of fluid flow. Additionally, fluorescence image tracking shows no Ca2+ influx during sperm rheotaxis turning, further suggesting there is no active signal transduction during human sperm rheotaxis.

  17. Effects of indium chloride exposure on sperm morphology and DNA integrity in rats

    Directory of Open Access Journals (Sweden)

    Kuo-Hsin Lee

    2015-03-01

    Full Text Available Indium, a Group IIIA element of the periodic chart and a rare earth metal characterized by high plasticity, corrosion resistance, and a low melting point, is widely used in the electronics industry where released streams can contaminate the environment. Consequently, indium can reach humans mainly by natural ways, which could result in a health hazard. Although reproductive toxicities have been surveyed in some studies in animal models, the infertility effects of sperm function induced by indium compounds have not been greatly investigated. We designed a study to investigate the toxicities of subacute exposure to indium compounds on male sperm function and the process of spermatogenesis in a rodent model. Fourteen Sprague-Dawley rats on postnatal Day (PND 84 were randomly divided into exposure and control groups, and weekly received intraperitoneal injections of indium chloride (1.5 mg/kg body weight and normal saline, respectively, for 8 weeks. Cauda epididymal sperm count, motility, morphology, chromatin DNA integrity, mitochondrial membrane potential (MMP, reactive oxygen species (ROS generation, and testis DNA content were investigated. The indium chloride exposed group showed significant toxicity to sperm function, as well as an increased percentage of sperm morphological abnormality and chromatin DNA damage. Furthermore, positive correlations between abnormal sperm morphology, chromatin DNA damage, and superoxide anion generation were also noted. The results of this study demonstrated the toxic effect of subacute low dose indium exposure during sexual maturation on sperm function, resulting in sperm chromatin DNA damage through an increase in sperm ROS generation in a rodent model.

  18. Comparison of two different extenders for cryopreservation of epididymal dog sperm.

    Science.gov (United States)

    Martins, Mim; Justino, R C; Sant'anna, M C; Trautwein, L G C; Souza, F F

    2012-12-01

    The collection of epididymal sperm is an option for preservation of germplasm of genetically superior animals that need to be orchiectomized or have died. The extender type used to freeze sperm is important to avoid spermatozoal membrane damage and to preserve semen quality after cryopreservation. The objective of this study was to verify the effects of a commercial bovine extender (Bovimix(®); Nutricell, Campinas) and a traditional TRIS-citric acid-glucose-egg yolk-7% glycerol extender on cryopreservation of canine epididymal sperm. The testes of 13 adult dogs were kept at 5 °C for 24 h in saline solution, and epididymal sperm was recovered in Ringers solution without lactate and were evaluated for motility. Samples with ≥ 80% motility were pooled and then divided before dilution and packaging in 0.5 ml plastic straws, equilibration at 4 °C for 1 h, freezing in nitrogen vapour for 20 min and storing at -196 °C. The straws were thawed at 56 °C for 10 s and were evaluated for motility by computer assisted analysis (CASA). The semen parameters, sperm movement index, linearity, total motility and rapid progressive motility were statistically higher in Bovimix(®) than TRIS. In contrast, amplitude of lateral head displacement, slow sperm and static sperm were lower in Bovimix(®). Despite the high percentage of sperm defects in epididymal cells, regardless of the extender, we concluded that Bovimix(®) is a viable alternative for the freezing of canine epididymal sperm. PMID:23279522

  19. Concentração de lactato de cálcio e tempo de incubação sobre a capacidade de adesão e penetração de espermatozoides suínos na membrana perivitelina do ovo da galinha Calcium lactate concentration and incubation period on the binding and penetration capacity of swine sperm in the perivitelline membrane of chicken egg

    Directory of Open Access Journals (Sweden)

    Carine Dahl Corcini

    2012-01-01

    Full Text Available Este estudo testou o efeito de diferentes concentrações de lactato de cálcio e tempos de incubação sobre a capacidade de adesão e a penetração dos espermatozoides suínos nas membranas perivitelinas (MP de ovos de galinhas, na expectativa de desenvolver um teste para estimar o potencial de fertilidade de machos doadores de sêmen. No primeiro experimento, amostras de sêmen (n=12 foram incubadas em meio TCM com lactato de cálcio a 1,1 e 2,2µg mL-1. Posteriormente, após definida a concentração de 1,1µg mL-1 de lactato de cálcio, um segundo experimento comparou três períodos de incubação a 39°C: 10, 15 e 20min. As respostas testadas foram: a taxa de adesão (TA e o número de espermatozoides aderidos (NA na MP externa, e a taxa de penetração (TP e o número de orifícios (NO na MP interna. A TA na MP externa foi de 100%, independentemente da concentração e dos períodos testados. Com a concentração de lactato de cálcio de 1,1µg mL-1, o NA na membrana perivitelina externa e TP e NO na membrana perivitelina interna foram superiores (PThis study tested the effect of different calcium lactate concentrations and incubation periods on the binding and penetration capacity of swine sperm to the perivitelline layers (PL of chicken eggs, with the expectation of developing a test to estimate the potential fertility of boars used as sperm donors. In the first experiment, semen samples (n=12 were incubated with TCM medium ant two calcium lactate levels: 1.1 and 2.2µg mL-1. After the set up of fixed calcium lactate level (1.1µg mL-1, the second experiment compared three incubation periods: 10, 15 and 20min. The observed variables were binding rate (BR and number of bound sperm (NB in the outer PL, and penetration rate (PR and number of holes (NH in the inner PL. The BR to the outer MP was 100%, regardless of the tested concentrations and incubation periods. The NB to the outer perivitelline layer and the PR and the NH in the inner

  20. Prevalence of Haemophilus parasuis infection in hunted wild boars (Sus scrofa) in Germany

    OpenAIRE

    2010-01-01

    Abstract Haemophilus parasuis is the etiological agent of Glasser?s disease, often involved in pneumonia, and also an early colonizer of the upper respiratory tract of healthy domestic pigs. Little information is available on H. parasuis in wild boars. The aim of the present study was to evaluate H. parasuis infection in wild boars in Germany. Tissue samples from the lungs and tonsils of 531 wild boars from 52 hunts during the hunting seasons 2004/2005 to 2006/2007 were examined in...

  1. Trichinella species circulating in wild boar (Sus scrofa) populations in Poland ☆

    OpenAIRE

    Bilska-Zając, Ewa; Różycki, Mirosław; Chmurzyńska, Ewa; Marucci, Gianluca; Cencek, Tomasz; Karamon, Jacek; Bocian, Łukasz

    2013-01-01

    Hunting in Poland has a long tradition and became more popular after 1990. Each year over 60,000 wild boar are hunted. Some of them may act as Trichinella carriers thus all carcasses of wild boar are systematically sampled in game-handling establishments as part of the post-mortem examination. The aim of the study was to determine the species of Trichinella and to evaluate the year to year differences in the occurrence of those species in the populations of wild boar in Poland. Samples for th...

  2. Freeze-tolerance of Trichinella muscle larvae in experimentally infected wild boars

    DEFF Research Database (Denmark)

    Lacour, Sandrine A.; Heckmann, Aurelie; Mace, Pauline;

    2013-01-01

    Freeze-tolerance of encapsulated Trichinella muscle larvae (ML) is mainly determined by Trichinella species, but is also influenced by host species, the age of the infection and the storage time and temperature of the infected meat. Moreover, the freeze-tolerance of the encapsulated species appears...... Trichinella spiralis and Trichinella britovi ML in wild boar meat 24 weeks post inoculation (wpi). Three groups of four wild boars were infected with 200, 2000 or 20,000 ML of T. britovi (ISS 1575), respectively. Additionally, three wild boars were inoculated with 20,000 ML of T. spiralis (ISS 004) and two...

  3. Prevalence of Streptococcus suis Genotypes in Wild Boars of Northwestern Germany▿

    OpenAIRE

    Baums, Christoph G.; Verkühlen, Gerd Josef; Rehm, Thomas; Silva, Luciana M. G.; Beyerbach, Martin; Pohlmeyer, Klaus; Valentin-Weigand, Peter

    2006-01-01

    Invasive serotype 2 (cps2+) strains of Streptococcus suis cause meningitis in pigs and humans. Four case reports of S. suis meningitis in hunters suggest transmission of S. suis through the butchering of wild boars. Therefore, the objective of this study was to investigate the prevalence of potentially human-pathogenic S. suis strains in wild boars. S. suis was isolated from 92% of all tested tonsils (n = 200) from wild boars. A total of 244 S. suis isolates were genotyped using PCR assays fo...

  4. Thermodynamics of the interaction between bovine binder of sperm BSP1 and low-density lipoprotein from hen's egg yolk

    International Nuclear Information System (INIS)

    Research highlights: → Isothermal titration calorimetry revealed Binder-of-SPerm BSP1 protein has high affinity for egg yolk low density lipoproteins. → It is estimated that 104 BSP1 molecules could bind one LDL particle. → BSP1 has 2 phosphocholine binding sites and the BSP1/LDL ratio corresponds to 1.6 phosphatidylcholine per bound BSP1. → The strong interaction between LDL particles and BSP1 is proposed to be important for sperm protection by egg yolk extender. - Abstract: Egg yolk is used in extender to protect sperm from cold shock and freezing during preservation. It is the low-density lipoprotein (LDL) fraction of egg yolk that protects sperm. Even though essential for sperm capacitation, the major proteins from bull seminal plasma, the Binder of SPerm (BSP) proteins, are detrimental for sperm preservation because they induce a continual phospholipids and cholesterol efflux from sperm membranes. The BSP proteins were proposed to bind to egg yolk LDL, preventing the sperm membrane damage. We characterized the binding between the BSP proteins and the LDL by isothermal titration calorimetry, providing the thermodynamics and quantitative description of this putative association. The association between BSP1 (major BSP proteins) and LDL is characterized by an affinity constant (Ka) of 3.4 ± 0.4 μM-1. A protein/LDL ratio of 104 ± 5 was determined indicating that 104 molecules of BSP1 would bind to one LDL particle. This stoichiometry leads to proposing that the association involves 1.6 ± 0.1 phosphatidylcholines (PC) per BSP protein. This finding is satisfactorily consistent with the fact that each BSP1 protein has 2 binding sites for choline group. In conclusion, the formation of a high affinity complex between BSP1 and LDL is proposed to be important for the protection of sperm by egg yolk extender.

  5. Effects of the crude extract of Polygala tenuifolia Willd on human sperm in vitro

    Institute of Scientific and Technical Information of China (English)

    Yi QIU; Lei-guang WANG; Yi-fang JIA; Dan-tong YANG; Mei-hua ZHANG; Yan-ping ZHANG; Li-hong ZHANG; Ling GAI

    2011-01-01

    The aim of the present study is to analyze sperm membrane changes and the spermicidal effect in treatment with the crude extract from Polygala tenuifolia Willd (PTW) in vitro. The root of PTW was extracted in distilled water. Normal human spermatozoa were used to assess the spermicidal activity (Sander-Cramer assay) of the extract from the PTW root. The hypo-osmotic swelling (HOS) test and the eosin Y (EY) staining were used to detect the integrity of sperm membrane and vitality. The sperm chromatin dispersion (SCD) test was performed to determine sperm DNA integrity. N-9 was used as a reference standard and semen added to physiological saline was used as the control. Semen samples were donated by 42 healthy fertile men. The crude extract from the root of PTW could immobilize and kill 100% spermatozoa within 20 s in vitro at the concentrations of 20.0 and 10.0 mg/ml; at the concentration of 5.0 mg/ml, spermatozoa were immobilized in (39.5卤3.2) s. In the groups of the crude extract from the root of PTW and N-9 solution, the rate of the normal HOS (tails swollen) and the white head (unstained) was 0%, and the rate of the abnormal HOS (tails unswollen) and red head (stained) was 100%. Sperm DNA fragmentation showed no change in exposure to the crude extract from the root of PTW and N-9 solution. The sperm revival test did not show any spermatozoa that recovered their motilities. The rapid spermicidal activity of the crude extract from the root of PTW in vitro may occur by the disruption of the sperm membrane integrity.

  6. Sperm Ubiquitination Correlation with Human Semen Quality

    Directory of Open Access Journals (Sweden)

    MR Sadeghi

    2008-07-01

    Full Text Available Background: Ubiquitin, an 8.5 kDa peptide that marks other proteins for proteasomal degradation, tags defective sperm during epididymal passage. Thus, sperm ubiquitination is a universal marker for sperm defects and can be used as a sperm function test. The objective of the present study was to examine the relationships between sperm ubiquitination and clinical semen parameters, using simplified immunofluorescence assays in order to establish ubiquitin as a biomarker of male infertility. Methods: Semen samples from 100 couples attending Avicenna Infertility Clinic, Tehran, Iran, were collected and analyzed according to WHO criteria. Each sample was washed and adjusted at 106 sperm/ml concentration. Sperm were coated on slides, using cytospin centrifugation and were fixed in buffered formaldehyde. Subsequently ubiquitinated spermatozoa were evaluated by direct immunofluorescence microscopy using FITC-labeled anti-ubiquitin antibodies. After counting at least 200 sperm per sample, while employing light microscopy, the percentage of ubiquitinated spermatozoa was recorded on the same fields through epifluorescence microscopy. Results: Negative correlations were obtained between sperm ubiquitination and sperm count (r=-0.278, P< 0.001, sperm concentration (r=-0.37, P< 0.001, viability (r=-0.407, P< 0.001, sperm morphology (r=-0.509, P< 0.001, rapid progressive motility (a (r=-0.246, P< 0.001 and slow progressive motility (b (r=-0.474, P< 0.001. There was a positive correlation between ubiquitinated sperm and the percentage of immotile spermatozoa (r=0.486, P< 0.000. Conclusion: Increased sperm ubiquitination is inversely associated with good semen quality parameters, supporting the use of ubiquitin as a biomarker for evaluation of human sperm quality.

  7. Evaluation of possible mitigation measures to prevent introduction and spread of African swine fever virus through wild boar

    Directory of Open Access Journals (Sweden)

    European Food Safety Authority

    2014-03-01

    Full Text Available This scientific report has been prepared in response to a request for urgent scientific and technical assistance under Art 31 of Regulation (EC No 178/2002, in relation to possible mitigation measures to prevent introduction and spread of African swine fever virus (ASFV. It was requested to assess the feasibility to drastically reduce the wild boar population by hunting or by the use of traps, and to assess if prevention of movement of wild boars by feeding or by artificial physical barriers reduces the risk of spread of ASFV. No evidence was found in scientific literature proving that wild boar populations can be drastically reduced by hunting or trapping in Europe. The main reasons are the adaptive behaviour of wild boar, compensatory growth of the population and the possible influx of wild boar from adjacent areas. Thus, drastic hunting is not a tool to reduce the risk for introduction and spread of ASFV in wild boar populations. Furthermore, wild boar density thresholds for introduction, spread and persistence of ASFV in the wild boar populations are currently impossible to establish, due to the uncertainty regarding the extent of the spread and maintenance of ASFV, the biases in population datasets, the complex population structures and dynamics. Furthermore, attempts to drastically reduce wild boar populations may even increase transmission and facilitate progressive geographical spread of ASFV, since intensive hunting pressure on wild boar populations leads to dispersion of groups and individuals. Artificial feeding of wild boar might increase the risk of ASFV spread. Fencing can restrict wild boar movements, however further knowledge of the ASF epidemiology and spatial distribution of wild boar is required to identify the areas where fencing could be used as one possible element of a control programme and to assess the feasibility of its implementation.

  8. Effect of seminal plasma on post-thaw quality and functionality of corriedale ram sperm obtained by electroejaculation and artificial vagina.

    Science.gov (United States)

    Ledesma, A; Manes, J; Ríos, G; Aller, J; Cesari, A; Alberio, R; Hozbor, F

    2015-06-01

    We have already shown that seminal collection method affects seminal plasma composition and sperm quality in Corriedale rams. In this study, we evaluated the effect of seminal plasma collected by electroejaculation or artificial vagina on sperm resistance to cryodamage. Seminal plasma of five rams of the Corriedale breed collected by artificial vagina or electroejaculation was added before freezing to sperm cells collected by the two methods, and post-thaw quality parameters were evaluated. We found that seminal plasma has no effect on sperm resistance to cryodamage. However, we observed significantly higher percentages of sperm with intact and functional plasma membrane, intact acrosome and greater fertilizing potential after thawing in samples obtained by electroejaculation. This study demonstrates that sperm collected by electroejaculation are more resistant to damage caused by cryopreservation than those collected by artificial vagina. PMID:25684063

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

    Science.gov (United States)

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

    2016-05-01

    Head-to-head agglutination of ram spermatozoa is induced by dilution in the Tyrode's capacitation medium with albumin, lactate and pyruvate (TALP) and ameliorated by the addition of the thiol d-penicillamine (PEN). To better understand the association and disassociation of ram spermatozoa, we investigated the mechanism of action of PEN in perturbing sperm agglutination. PEN acts as a chelator of heavy metals, an antioxidant and a reducing agent. Chelation is not the main mechanism of action, as the broad-spectrum chelator ethylenediaminetetraacetic acid and the copper-specific chelator bathocuproinedisulfonic acid were inferior anti-agglutination agents compared with PEN. Oxidative stress is also an unlikely mechanism of sperm association, as PEN was significantly more effective in ameliorating agglutination than the antioxidants superoxide dismutase, ascorbic acid, α-tocopherol and catalase. Only the reducing agents cysteine and dl-dithiothreitol displayed similar levels of non-agglutinated spermatozoa at 0 h compared with PEN but were less effective after 3 h of incubation (37 °C). The addition of 10 µM Cu(2+) to 250 µM PEN + TALP caused a rapid reversion of the motile sperm population from a non-agglutinated state to an agglutinated state. Other heavy metals (cobalt, iron, manganese and zinc) did not provoke such a strong response. Together, these results indicate that PEN prevents sperm association by the reduction of disulphide bonds on a sperm membrane protein that binds copper. ADAM proteins are possible candidates, as targeted inhibition of the metalloproteinase domain significantly increased the percentage of motile, non-agglutinated spermatozoa (52.0% ± 7.8) compared with TALP alone (10.6% ± 6.1).Reproduction (2016) 151 1-10. PMID:26860122

  10. Characterisation of Caenorhabditis elegans sperm transcriptome and proteome

    OpenAIRE

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

    2014-01-01

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

  11. Detection of serum anti-sperm antibody in infertile couples with dot-immunogold filtration assay (DIGFA)

    International Nuclear Information System (INIS)

    Objective: To develop a new method for rapid detection of serum anti-sperm antibody in infertile couples. Methods: Human sperm antigen was prepared from pooled semen specimens of fertile males. Nitro-cellulose membrane was used as solid-phase carrier of the antigen. Colloidal gold pellet combined goat anti-human IgG was taken as labelled antibody. A dot-immunogold filtration assay system was established for test of serum anti-human sperm antibody. Serum specimens from 137 infertile couples were tested and the result compared with flat from ELISA. Results: The human sperm antigen would react with the anti-sperm antibody in the tested serum over the cellulose membrane through filtration and the result could be read with naked eye within 6 minutes. In this study of 137 infertile coupled, the anti-sperm antibody was positive in 21.9% of the female serum specimens and 13.19% of the males. Compared with the result from ELISA, the consistency rate was 96.1%. The sensitivity of the assay was 90.2% and specificity was 95.4%. The p reparation was stable after 6 months refrigerator storage. Conclusion: This newly developed DIGFA is very adequate for rap id detection of anti-sperm antibody and deserves popularization. (authors)

  12. Flow cytometric and near-infrared Raman spectroscopic investigation of quality in stained, sorted, and frozen-thawed buffalo sperm.

    Science.gov (United States)

    Li, Xiao-Xia; Wang, Meng; Chen, Huan-Hua; Li, Qing-Yang; Yang, Huan; Xu, Hui-Yan; Lu, Yang-Qing; Zhang, Ming; Yang, Xiao-Gan; Lu, Sheng-Sheng; Lu, Ke-Huan

    2016-07-01

    Flow cytometry and Laser Tweezers Raman spectroscopy have been used to investigate Nili-Ravi buffalo (Bubalus bubalis) sperm from different samples (fresh, stained, sorted and frozen-thawed) of the flow-sorting process to optimize sperm sex sorting procedures. During the sorting and freezing-thawing processes, the two detection methods both indicated there were differences in mitochondrial activity and membrane integrity. Moreover, a dispersive-type NIR (Near Infrared Reflection) use of the Raman system resulted in the ability to detect a variety of sperm components, including relative DNA, lipid, carbohydrates and protein contents. The use of the Raman system allowed for PCA (principal components analysis) and DFA (discriminant function analysis) of fresh, stained, sorted and frozen-thawed sperm. The methodology, therefore, allows for distinguishing sperm from different samples (fresh, stained, sorted and frozen-thawed), and demonstrated the great discriminative power of ANN (artificial neural network) classification models for the differentiating sperm from different phases of the flow-sorting process. In conclusion, the damage induced by sperm sorting and freezing-thawing procedures can be quantified, and in the present research it is demonstrated that Raman spectroscopy is a valuable technology for assessing sperm quality. PMID:27095613

  13. Sperm of the giant grouper: cryopreservation, physiological and morphological analysis and application in hybridizations with red-spotted grouper.

    Science.gov (United States)

    Tian, Yongsheng; Jiang, Jing; Wang, Na; Qi, Wenshan; Zhai, Jieming; Li, Bo; Liang, You; Chen, Youming; Yang, Chuanjun; Chen, Songlin

    2015-01-01

    In order to develop excellent germplasm resources for giant grouper (Epinephelus lanceolatus), cryopreservation of giant grouper sperm was examined in the present study. Firstly, 13 kinds of sperm dilution (ELS1-3, EM1-2, TS-2, MPRS, ELRS0-6) were prepared with physiological salt, sucrose, glucose and fetal bovine serum. The physiological parameters of ELRS3 (ratio of fast motion, ratio of slow motion, time of fast motion, time of slow motion, lifespan and motility) and ELS3 (sperm ratio of slow motion, time of slow motion and motility) were significantly higher than those of the other dilutions (P cryopreserved. Finally, the sperm of giant grouper cryopreserved with cryoprotectants (ELRS3 + 15% DMSO + 10% FBS) was used for electron-microscopic observation and crossbreeding with red-spotted groupers (Epinephelus akaara). The electron-microscopic observation revealed that part of the frozen-thawed sperm was cryodamaged, e.g., flagellum fracturing and mitochondria falling out, while the ultrastructure of sperm membrane, mitochondria and flagellum remained intact. Also, the fertilization and hatchability rates of giant grouper frozen sperm and red-spotted grouper eggs were as high as 94.56% and 75.56%, respectively. Thus, a technique for cryopreservation of giant grouper sperm was successfully developed and applied to crossbreeding with red-spotted grouper eggs. PMID:25985804

  14. Sperm abnormalities in exposed humans

    Czech Academy of Sciences Publication Activity Database

    Šrám, Radim; Rubeš, J.

    Cambridge : Issue in Toxicology, Royal Society of Chemistry Publ.,, 2007, s. 247-258. ISBN 978-0-85404-847-2 R&D Projects: GA MŽP SL/740/5/03 Institutional research plan: CEZ:AV0Z50390512 Keywords : air pollution exposure * sperm abnormalities * male reproductive health Subject RIV: DN - Health Impact of the Environment Quality

  15. Effect of cryopreservation on the efficiencies of goat sperm in picking up exogenous DNA and resulted transgenic embryos

    OpenAIRE

    Zhao, Yong-Ju; Wang, Yong; Jian WANG; Li, Fu-Bing; Wei, Hong

    2008-01-01

    The efficiency and the ability of fresh and frozen-thawed sperm in picking up exogenous DNA were investigated in this study. The fresh and frozen-thawed sperm was incubated with linearized end-labeled pEGFP-N1 plasmid DNA, and detected by in situ hybridization method. The in vitro producted embryos were screened by PCR assay. The ultrastructure of activated spermatozoa were observed by transmission electron microscope (TEM), and the integrity of sperm plasma membrane was evaluated with a comb...

  16. Patterns of sperm damage in Chernobyl passerine birds suggest a trade-off between sperm length and integrity

    OpenAIRE

    Hermosell, Ignacio G.; Laskemoen, Terje; Rowe, Melissah; Møller, Anders P.; Mousseau, Timothy A; Albrecht, Tomáš; Lifjeld, Jan T

    2013-01-01

    Interspecific variation in sperm size is enigmatic, but generally assumed to reflect species-specific trade-offs in selection pressures. Among passerine birds, sperm length varies sevenfold, and sperm competition risk seems to drive the evolution of longer sperm. However, little is known about factors favouring short sperm or constraining the evolution of longer sperm. Here, we report a comparative analysis of sperm head abnormalities among 11 species of passerine bird in Chernobyl, presumabl...

  17. Boar taint compound levels in back fat versus meat products: Do they correlate?

    Science.gov (United States)

    Wauters, Jella; Vercruysse, Vicky; Aluwé, Marijke; Verplanken, Kaat; Vanhaecke, Lynn

    2016-09-01

    Surgical castration of male pigs will soon be abandoned, turning a major advantage of this practice, the elimination of boar taint, into the biggest challenge for pig industry when raising intact male pigs becomes common practice. To map the (economical) consequences in relation to boar-taint consumer acceptance, as well as offer a processing strategy for tainted carcasses to stockholders, the current study investigated not only back fat boar taint levels, but additionally generated information on the levels of boar taint compounds recovered after the production of commercially relevant meat products using UHPLC-HRMS laboratory analysis. Our results demonstrate that levels of androstenone, skatole and indole in back fat and meat products tend to correlate strongly, particularly in fatty meat products (generally r>0.80). Concentration values in the edible (lean) meat fraction were significantly lower compared to back fat and fat sampled from fresh or processed meat (p<0.05). PMID:27041294

  18. First reports of Trichinella pseudospiralis in wild boars (Sus scrofa) of Italy.

    Science.gov (United States)

    Merialdi, Giuseppe; Bardasi, Lia; Fontana, Maria Cristina; Spaggiari, Brunella; Maioli, Giulia; Conedera, Gabriella; Vio, Denis; Londero, Mauro; Marucci, Gianluca; Ludovisi, Alessandra; Pozio, Edoardo; Capelli, Gioia

    2011-06-10

    Trichinella pseudospiralis is a non-encapsulated species infecting both mammals and birds. In Italy, this parasite was reported only in two night-birds of prey of Central Italy. In January 2010, Trichinella larvae were detected in three wild boars (Sus scrofa) of two regions of Northern Italy by enzymatic digestion. The parasites were identified as T. pseudospiralis by multiplex-PCR. The first infected wild boar was hunted in the Emilia Romagna region and the other two infected wild boars were bred outdoors in a small family farm of the Friuli Venezia Giulia region. These new epidemiological data reinforce the role of the wild boar as the main reservoir of T. pseudospiralis in Europe. PMID:21296503

  19. Comparative Bacteriological Study of Two Wild Boar Populations in Sierra Morena (Ja�n, Spain

    Directory of Open Access Journals (Sweden)

    Antonio NOTARIO

    2010-12-01

    Full Text Available The aim of this study was to investigate the presence of various bacterial species in the wild boar populations of Lugar Nuevo and Selladores-Contadero woodlands from Sierra Morena (Spain. Bacteriological analyses were carried out on a total of 229 wild boar individuals hunted in the period 2000-2003 in eleven experimental plots which are representative for the different biotopes of the area. The following species were detected: Brucella ovis, Clostridium sp., Corynebacterium sp., Streptococcus pneumoniae, Escherichia coli, Mycobacterium bovis, Mycobacterium avium subsp. paratuberculosis, Mannheimia haemolytica, Pasteurella multocida, Pseudomonas aeruginosa, Chlamydophila psittaci, Staphylococcus aureus and Staphylococcus sp. and the bacterial prevalence was estimated for each of them. The results provide useful indications of the health status of wild boar in both locations and highlight the potential of the wild boar populations to act as biological reservoirs of certain microorganisms that can be passed onto other vertebrate wild animals and humans.

  20. Seminal characteristics and cryopreservation of sperm from the squirrel monkey, Saimiri collinsi.

    Science.gov (United States)

    Oliveira, K G; Leão, D L; Almeida, D V C; Santos, R R; Domingues, S F S

    2015-09-15

    The Neotropical nonhuman primate squirrel monkey (Saimiri sp.) is one of the most commonly used species in research in several areas of knowledge. However, little progress has been reported in respect to techniques for preservation of their gametes. Thus, the main objectives of this study were (1) to describe testicular and seminal aspects of a new species, Saimiri collinsi, (2) to preserve semen of this species by cooling or freezing using ACP-118 (powdered coconut water), and (3) to test two glycerol (GLY) concentrations (1.5% or 3%) for semen freezing in the presence of ACP-118. The experimental group started with 14 captive males, but only 11 were suitable to collect ejaculates containing sperm. After anesthesia, both testes were evaluated: length, width, height, and testicular circumference. Semen was collected by electroejaculation and evaluated, followed by dilution, cooling, and freezing. Seminal parameters and sperm motility, vigor, plasma membrane integrity, and normal morphology were evaluated after each step; functionality was also checked in fresh and frozen-thawed sperm. Sperm motility, plasma membrane integrity, and normal sperm in cooled semen (n = 11) were 44.1 ± 34.0, 63.1 ± 15.6, and 73.8 ± 19.8, respectively, with vigor ranging of 2 to 3. Sperm motility, plasma membrane integrity, normal and functional sperm in frozen semen (n = 5) were 0.6 ± 1.3 (1.5% and 3% GLY); 4.4 ± 4.9 (1.5% GLY) and 6.6 ± 7.2 (3% GLY); 86.8 ± 3.0 (1.5% GLY) and 88.8 ± 5.1 (3% GLY); 13.3 ± 11.9 (1.5% GLY) and 14.3 ± 13.5 (3% GLY), respectively, and vigor 0 for both 1.5% and 3% GLY. No significant difference between GLY concentrations was observed. We concluded that electroejaculation was efficient for semen collection of S collinsi and tested the cooling protocol that allowed to recover a satisfactory percentage (63%) of membrane intact sperm. However, the freezing protocol was not appropriate to sperm preservation. PMID:26047706

  1. Factors influencing wild boar presence in agricultural landscape: a habitat suitability modelling approach

    OpenAIRE

    Morelle, Kevin; Lejeune, Philippe

    2012-01-01

    The wild boar (Sus scrofa L.) population has increased in most parts of Europe during the last three decades. As a consequence, population distribution ranges have shifted to more agricultural areas. These newly colonized areas are characterized by highly fragmented forest cover and a preponderance of crop fields and grasslands, thus increasing the risk of problematic interactions with farming activities and small game species. Although plasticity of wild boar to various environmental conditi...

  2. Controlling of CSFV in European wild boar using oral vaccination: a review

    OpenAIRE

    Sophie eRossi; Christoph eStaubach; Sandra eBlome; Vittorio eGuberti; HH eThulke; Ad eVos; Frank eKoenen; marie-frederique ele potier

    2015-01-01

    Classical swine fever (CSF) is among the most detrimental diseases for the swine industry worldwide. Infected wild boar populations can play a crucial role in CSF epidemiology and controlling wild reservoirs is of utmost importance for preventing domestic outbreaks. Oral mass vaccination (OMV) has been implemented to control CSF in wild boars and limit the spill over to domestic pigs. This retrospective overview of vaccination experiences illustrates the potential for that option. The C-strai...

  3. Influence of boar breeds or hybrid genetic composition on semen quality and seminal plasma biochemical variables.

    Science.gov (United States)

    Žaja, Ivona Žura; Samardžija, Marko; Vince, Silvijo; Majić-Balić, Ivanka; Vilić, Marinko; Đuričić, Dražen; Milinković-Tur, Suzana

    2016-01-01

    The enzyme concentrations of seminal plasma are important for spermatozoa metabolism and function in boars. The need has arisen for introducing a biochemical evaluation of semen, along with the usual standard semen analyses. There are no data on the influence of boar breeds on the seminal plasma biochemical variables investigated in this study. Therefore, the objective was to determine the influence of breed and hybrid genetic composition of boars on semen quality and seminal plasma biochemical variables. Semen samples of 27 boars (Swedish Landrace, German Landrace, Large White, Pietrain and Pig Improvement Company hybrid-PIC-hybrid), aged between 1.5 and 3 years, were collected. After evaluation of semen quality, the seminal plasma was separated from the spermatozoa by centrifugation of semen. The seminal plasma was subjected to spectrophotometric analysis to determine alkaline phosphatase (ALP), acid phosphatase (ACP), γ-glutamyltransferase (GGT), creatine kinase (CK) and lactate dehydrogenase (LDH) and to atomic absorption spectrophotometric analysis to measure the concentration of calcium and magnesium. Conventional semen quality variables differed depending on breed and PIC-hybrid genetic composition, though these differences were typically insignificant. In the seminal plasma, significant differences were determined in enzyme activity (ALP, GGT, CK and LDH) and in calcium concentration among boars of different breeds. There are, therefore, differences in semen quality and significant differences in the seminal plasma biochemical variables among boars of different breeds and PIC-hybrid genetic composition. The data and differences in semen variables detected in the present study provide knowledge for enhancing evaluation and monitoring of boar reproductive potential, semen quality and explain the potential causes of boar infertility. PMID:26692346

  4. Indirect versus direct detection methods of Trichinella spp. infection in wild boar (Sus scrofa)

    OpenAIRE

    Gómez-Morales, Maria Angeles; Ludovisi, Alessandra; Amati, Marco; Bandino, Ennio; Capelli, Gioia; Corrias, Franco; Gelmini, Luca; Nardi, Alberigo; Sacchi, Cristina; Cherchi, Simona; Lalle, Marco; Pozio, Edoardo

    2014-01-01

    Background Trichinella spp. infections in wild boar (Sus scrofa), one of the main sources of human trichinellosis, continue to represent a public health problem. The detection of Trichinella spp. larvae in muscles of wild boar by digestion can prevent the occurrence of clinical trichinellosis in humans. However, the analytical sensitivity of digestion in the detection process is dependent on the quantity of tested muscle. Consequently, large quantities of muscle have to be digested to warrant...

  5. Signaling Pathways Used by Ergot Alkaloids to Inhibit Bovine Sperm Motility

    Science.gov (United States)

    Ergot alkaloids exert their toxic or pharmaceutical effects through membrane receptor-mediated activities. This study investigated the signaling pathways involved in the in vitro inhibitory effects of both ergotamine (ET) and dihydroergotamine (DEHT) on bovine sperm motility using specific inhibitor...

  6. Human sperm immobilization effect of Carica papaya seed extracts: an in vitro study

    Institute of Scientific and Technical Information of China (English)

    NirmalKLohiya; LalitKKothari; BManivannan; PradyumnaKMishra; NeelamPathak

    2000-01-01

    Aim: To examine if the seed extracts of Carica papaya, which showed antispermatogenic/sperm immobilization properties in animal models, could cause human sperm immobilization in vitro. Methods: Chloroform extract, benzene chromatographic fraction of the chloroform extract, its methanol and ethyl acetate sub-fractions and the isolated compounds from the sub-fractions i.e., ECP 1 & 2 and MCP 1 & 2, of the seeds of Cadca papaya were used at concentrations of 0.1%, 0.5%, 1% and 2%. Sperm motility was assessed immediately after addition of extracts and every 5 minutes thereafter for 30 minutes. Results: There were dose-dependent spermicidal effects showing an instant fall in the sperm motility to less than 20 % at 2 % concentration. Isolated compounds ECP 1 & 2 were more effective inducing a motility of less than 10%. Many of the spermatozoa became vibratory on the spot. Total inhibition of motility was observed within 20 - 25 min at all concentrations of all products. Scanning and transmission electron microscopy revealed deleterious changes in the plasma membrane of the head and mid-piece of spermatozoa. Sperm viability test and the number of abnormal spermatozoa after completion of incubation suggested that the spermatozoa were infertile. The effects were spermicidal but not spermiostatic as revealed by the sperm revival test. Conclusion: The results reveal spermicidal activity in vitro of the seed extracts of Carica papaya.

  7. Detection of sperm-motivating factor in cervical mucus of ovulatory women

    Directory of Open Access Journals (Sweden)

    Anjana Ray Chaudhuri

    2013-07-01

    Full Text Available Objective: The objective of the present study was to detect the presence of any sperm selecting and / or motivating factor in the cervical mucus (CM of ovulatory women, which, in turn, might influence pregnancy rate. Materials & Methods: The study was conducted with 190 infertile couples, having indications for moderate seminopathy, enlisted for intra-uterine insemination (IUI, following routine ultrasound and laparoscopic evaluation at our centre, between April 2006 to August 2010. The female partners were in the age group 25-30 years and were randomly divided into 3 groups to receive sperm processed in 3 different methods, with or without CM. Pre and peri-ovulatory CM was subjected to paper electrophoresis on cellulose acetate membrane (CAM strip, to study the protein band pattern. The different procedures of sperm processing were compared primarily on the basis of sperm motility and quality in the swim-up layer and also the pregnancy outcome following IUI, with due emphasis on improvisation of current laboratory methodologies used in IUI. Results: The use of CM layer alongwith medium for swim-up proved to be the best regarding selection of sperm with maximum motility and normal morphology, as well as the pregnancy outcome. The CM of ovulatory women around ovulation showed the presence of a specific band in electrophoresis. Conclusion: The use of CM in sperm preparation technique proves to be beneficial to patients undergoing IUI.

  8. Genetic structure and subspecies divergence of wild boars Sus scrofa in mainland China based on the microsatellite variation analysis

    OpenAIRE

    ZHANG Bao-Wei; ZHANG Chen-Ling; CHEN Jian-Qin; Ding, Dong; LI Chong-Qi; Zhou, Kai-Ya; Chang, Qing

    2008-01-01

    In order to discuss the subspecies classification of wild boar Sus scrofa in China, We studied their genetic structure using 11 microsatellites, with emphasis on South China. High levels of genetic variationwere detected in South China (SC), North China (NC) and Northeast China (NE) wild boar populations. This was consistent with the huge population size of the wild boar and its status as a widely-distributed species. Weak genetic divergence was observed in all subspecies involved in this s...

  9. Epidemiological survey of swine influenza A virus in selected wild boar populations in Germany.

    Science.gov (United States)

    Kaden, Volker; Lange, Elke; Starick, Elke; Bruer, Wilhelm; Krakowski, Wolfgang; Klopries, Marlis

    2008-09-18

    The aim of this study was to evaluate the epidemiological situation of swine influenza virus (SIV) infections in different wild boar populations in Germany based on a serological surveillance in some Bundeslaender (federal states) in connection with virological investigations in wild boar shot in Northern Germany (Mecklenburg-Western Pomerania, district of Nordvorpommern). Altogether, 1245 sera from wild boar were tested using the hemagglutination inhibition test. The established seroprevalence rate was low (on average 5.2%). Antibodies were only detected against the subtypes H1N1 and H3N2 showing differences between wild boar populations and age classes. The virological investigation of samples derived from lungs of wild boar shot in Mecklenburg-Western Pomerania, district of Nordvorpommern (n=242), revealed that the virus prevalence (two virologically positive animals, 0.8%) was very low. Based on serological typing, the isolated SIV was identified as subtype H3N2. Molecular biological investigations of the hemagglutinin (HA) and neuraminidase (NA) genes confirmed this result. This study suggests that SIV infections in wild boar seem to be no serious threat for domestic pigs. PMID:18440732

  10. Wild boars meat as a potential source of human trichinellosis in Poland: current data.

    Science.gov (United States)

    Moskwa, Bożena; Cybulska, Aleksandra; Kornacka, Aleksandra; Cabaj, Władysław; Bień, Justyna

    2015-09-01

    Trichinellosis is an epidemiological problem with a global distribution. In Poland a substantial increase of the wild boar population has been observed since 2010, together with an increased incidence of trichinellosis after ingestion of raw or undercooked wild boar products containing Trichinella spp. larvae. However, the actual number of human cases remains particularly difficult to determine. The aim of the present study was to determine the current prevalence and spread of these parasites within wild boars. The diaphragm pillars and tongue from 833 wild boars were collected from 2010 to 2014, as well as one wild boar meat sausage known to be a source of infection. The samples were tested for Trichinella spp. using pepsin digestion. Recovered larvae were identified at species level by multiplex polymerase chain reaction (multiplex PCR). The overall prevalence in all examined samples was found to be 2.0% (17/833). Recovered larvae were identified as T. spiralis and T. britovi (9/18 and 5/18, respectively). T. spiralis larvae were isolated from the sausage. Mixed infection was confirmed only once. Three isolates were not identified. The results of our study confirm that the wild boar plays a key role in the maintenance of Trichinella nematodes through the sylvatic cycle. PMID:26204193

  11. Tenacity of Exogenous Human Papillomavirus DNA in Sperm Washing

    OpenAIRE

    Brossfield, Jeralyn E.; Chan, Philip J.; Patton, William C.; King, Alan

    1999-01-01

    Purpose:Sperm cells have been shown to take up exogenous DNA readily. The hypothesis was that sperm washing would remove exogenous viral DNA infecting sperm cells. The objective was to compare three types of sperm washing procedures for their capacity to remove exogenous human papillomavirus (HPV) DNA from infected sperm.

  12. On the origin of sperm epigenetic heterogeneity.

    Science.gov (United States)

    Laurentino, Sandra; Borgmann, Jennifer; Gromoll, Jörg

    2016-05-01

    The influence of epigenetic modifications on reproduction and on the function of male germ cells has been thoroughly demonstrated. In particular, aberrant DNA methylation levels in sperm have been associated with abnormal sperm parameters, lower fertilization rates and impaired embryo development. Recent reports have indicated that human sperm might be epigenetically heterogeneous and that abnormal DNA methylation levels found in the sperm of infertile men could be due to the presence of sperm populations with different epigenetic quality. However, the origin and the contribution of different germ cell types to this suspected heterogeneity remain unclear. In this review, we focus on sperm epigenetics at the DNA methylation level and its importance in reproduction. We take into account the latest developments and hypotheses concerning the functional significance of epigenetic heterogeneity coming from the field of stem cell and cancer biology and discuss the potential importance and consequences of sperm epigenetic heterogeneity for reproduction, male (in)fertility and assisted reproductive technologies (ART). Based on the current information, we propose a model in which spermatogonial stem cell variability, either intrinsic or due to external factors (such as endocrine action and environmental stimuli), can lead to epigenetic sperm heterogeneity, sperm epimutations and male infertility. The elucidation of the precise causes for epimutations, the conception of adequate therapeutic options and the development of sperm selection technologies based on epigenetic quality should be regarded as crucial to the improvement of ART outcome in the near future. PMID:26884419

  13. Sperm Morphology Assessment in Captive Neotropical Primates.

    Science.gov (United States)

    Swanson, W F; Valle, R R; Carvalho, F M; Arakaki, P R; Rodas-Martínez, A Z; Muniz, Japc; García-Herreros, M

    2016-08-01

    The main objective of this study was to evaluate sperm morphology in four neotropical primate species to compare the sperm morphological traits and the sperm morphometric parameters as a basis for establishing normative sperm standards for each species. Data from 80 ejaculates collected from four primate species, Callithrix jacchus, Callimico goeldii, Alouatta caraya and Ateles geoffroyi, were analysed for detection of sperm morphological alterations using subjective World Health Organization (WHO-2010) standards and Sperm Deformity Index (SDI) criteria, objective computer-assisted sperm morphometry analysis (CASMA) and subpopulation sperm determination (SSD) methods. There were multiple differences (p Area (A), Perimeter (P), Length (L), Width (W), Ellipticity, Elongation and Rugosity) (p ≤ 0.046). In conclusion, our findings using different evaluation methods indicate that pronounced sperm morphological variation exists among these four neotropical primate species. Because of the strong relationship observed among morphological and morphometric parameters, these results suggest that application of objective analysis methods could substantially improve the reliability of comparative studies and help to establish valid normative sperm values for neotropical primates. PMID:27260333

  14. Etiology and Evaluation of Sperm Chromatin Anomalies

    Directory of Open Access Journals (Sweden)

    Marziyeh Tavalaee

    2008-01-01

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

  15. CD81 and CD9 work independently as extracellular components upon fusion of sperm and oocyte

    Directory of Open Access Journals (Sweden)

    Naoko Ohnami

    2012-05-01

    When a sperm and oocyte unite into one cell upon fertilization, membranous fusion between the sperm and oocyte occurs. In mice, Izumo1 and a tetraspanin molecule CD9 are required for sperm-oocyte fusion as one of the oocyte factors, and another tetraspanin molecule CD81 is also thought to involve in this process. Since these two tetraspanins often form a complex upon cell-cell interaction, it is probable that such a complex is also formed in sperm-oocyte interaction; however, this possibility is still under debate among researchers. Here we assessed this problem using mouse oocytes. Immunocytochemical analysis demonstrated that both CD9 and CD81 were widely distributed outside the oocyte cell membrane, but these molecules were separate, forming bilayers, confirmed by immunobiochemical analysis. Electron-microscopic analysis revealed the presence of CD9- or CD81-incorporated extracellular structures in those bilayers. Finally, microinjection of in vitro-synthesized RNA showed that CD9 reversed a fusion defect in CD81-deficient oocytes in addition to CD9-deficient oocytes, but CD81 failed in both oocytes. These results suggest that both CD9 and CD81 independently work upon sperm-oocyte fusion as extracellular components.

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

    Directory of Open Access Journals (Sweden)

    Raycon Roberto Freitas Garcia

    2015-09-01

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

  17. Mechanism of infertility in male guinea pigs immunized with sperm PH-20.

    Science.gov (United States)

    Tung, K S; Primakoff, P; Woolman-Gamer, L; Myles, D G

    1997-05-01

    PH-20, a testis-specific protein first expressed in haploid germ cells, is present on the posterior head plasma membrane and inner acrosomal membrane of mature guinea pig sperm. PH-20 is bifunctional, having a hyaluronidase activity that allows sperm to penetrate the cumulus layer and a separate activity required for binding of acrosome-reacted sperm to the zona pellucida. The immunization of male guinea pigs with PH-20 reproducibly results in infertility with a duration of 6-12 mo or longer. In this study, we analyzed the immunopathology in the reproductive tract of PH-20-immunized males to probe the mechanism(s) responsible for the induced infertility and found two separate effects. Remarkably, in almost all infertile, PH-20-immunized males, the caudae epididymides were empty (contained no sperm) or contained only abnormal sperm. The complete loss of normal sperm in the epididymis apparently results in infertility. A second effect was the induction of experimental autoimmune orchitis (EAO), representing the first report of EAO induced by a purified testis/sperm molecule of known functions. PH-20-induced EAO differed from EAO induced by crude testis antigens in two respects: 1) an absence of epididymitis with abscess and granuloma and 2) the presence of antibody on germ cells within seminiferous tubules and inside the cauda epididymidis. The former suggests that crude testis antigens other than PH-20 are responsible for epididymitis, and the latter suggests a possible role of antibody in EAO pathogenesis and infertility induction. Return to fertility, after 6-12 mo, was accompanied by regression of EAO and reappearance of spermatozoa in the caudae epididymides. PMID:9160711

  18. Intracytoplasmic sperm injection outcomes with cryopreserved testicular sperm aspiration samples.

    Science.gov (United States)

    Roque, M; Valle, M; Marques, F; Sampaio, M; Geber, S

    2016-04-01

    Intracytoplasmic sperm injection (ICSI) may be performed with testicular frozen-thawed spermatozoa in patients with nonobstructive azoospermia (NOA). Sperm retrieval can be performed in advance of oocyte aspiration, as it may avoid the possibility of no recovery of spermatozoa on the day of oocyte pickup. There are few studies available in the literature concerning the use of frozen-thawed spermatozoa obtained from testicular sperm aspiration (TESA). To evaluate the effects and the outcomes of ICSI with frozen-thawed spermatozoa obtained by TESA, we performed a retrospective analysis of 43 ICSI cycles using frozen-thawed TESA. We obtained acceptable results with a fertilisation rate of 67.9%, an implantation rate (IR) of 17.1%, and clinical and ongoing pregnancy rates of 41.9% and 37.2% respectively. The results of this study suggest that performing ICSI using cryopreserved frozen-thawed testicular spermatozoa with TESA as a first option is a viable, safe, economic and effective method for patients with NOA. PMID:25998234

  19. Elucidation of the involvement of p14, a sperm protein during maturation, capacitation and acrosome reaction of caprine spermatozoa.

    Directory of Open Access Journals (Sweden)

    Pinki Nandi

    Full Text Available Mammalian sperm capacitation is an essential prerequisite to fertilization. Although progress is being made in understanding the physiology and biochemistry of capacitation, little has been yet explored about the potential role(s of individual sperm cell protein during this process. Therefore elucidation of the role of different sperm proteins in the process of capacitation might be of great importance to understand the process of fertilization. The present work describes the partial characterization of a 14-kDa protein (p14 detected in goat spermatozoa using an antibody directed against the purified protein. Confocal microscopic analysis reveals that the protein is present in both the intracellular and extracellular regions of the acrosomal and postacrosomal portion of caudal sperm head. Though subcellular localization shows that p14 is mainly cytosolic, however it is also seen to be present in peripheral plasma membrane and soluble part of acrosome. Immuno-localization experiment shows change in the distribution pattern of this protein upon induction of capacitation in sperm cells. Increased immunolabeling in the anterior head region of live spermatozoa is also observed when these cells are incubated under capacitating conditions, whereas most sperm cells challenged with the calcium ionophore A23187 to acrosome react, lose their labeling almost completely. Intracellular distribution of p14 also changes significantly during acrosome reaction. Interestingly, on the other hand the antibody raised against this 14-kDa sperm protein enhances the forward motility of caprine sperm cells. Rose-Bengal staining method shows that this anti-p14 antibody also decreases the number of acrosome reacted cells if incubated with capacitated sperm cells before induction of acrosome reaction. All these results taken together clearly indicate that p14 is intimately involved and plays a critical role in the acrosomal membrane fusion event.

  20. Sperm fucosyltransferase-5 mediates the sperm-oviductal epithelial cell interaction to protect human sperm from oxidative damage

    OpenAIRE

    Huang, Wenxin; 黃聞馨

    2013-01-01

    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. In mammals, including humans, oviduct functions as a sperm reservoir which is created by the binding of spermatozoa to the epithelial lining in the oviduct. Interaction between sperm and oviductal epithelial cells improves the fertilizing ability of and reduces chromatin damage in spermatozoa. However, the mechanism(s) ...

  1. No evidence of trade-offs in the evolution of sperm numbers and sperm size in mammals.

    Science.gov (United States)

    Tourmente, M; Delbarco Trillo, J; Roldan, E R S

    2015-10-01

    Post-copulatory sexual selection, in the form sperm competition, has influenced the evolution of several male reproductive traits. However, theory predicts that sperm competition would lead to trade-offs between numbers and size of spermatozoa because increased costs per cell would result in a reduction of sperm number if both traits share the same energetic budget. Theoretical models have proposed that, in large animals, increased sperm size would have minimal fitness advantage compared with increased sperm numbers. Thus, sperm numbers would evolve more rapidly than sperm size under sperm competition pressure. We tested in mammals whether sperm competition maximizes sperm numbers and size, and whether there is a trade-off between these traits. Our results showed that sperm competition maximizes sperm numbers in eutherian and metatherian mammals. There was no evidence of a trade-off between sperm numbers and sperm size in any of the two mammalian clades as we did not observe any significant relationship between sperm numbers and sperm size once the effect of sperm competition was taken into account. Maximization of both numbers and size in mammals may occur because each trait is crucial at different stages in sperm's life; for example size-determined sperm velocity is a key determinant of fertilization success. In addition, numbers and size may also be influenced by diverse energetic budgets required at different stages of sperm formation. PMID:26190170

  2. Gene expression during testis development in Duroc boars

    DEFF Research Database (Denmark)

    Lervik, Siri; Kristoffersen, Anja Bråthen; Conley, Lene;

    2015-01-01

    Androstenone is a steroid pheromone occurring in the pubertal Leydig cells. Breeding against androstenone can decrease pheromone odour in swine meat but appears to cause unwanted side effects such as delayed onset of puberty. To study causality, global gene expression in developing boar testes at...... 12, 16, 20 and 27 weeks was investigated using a porcine cDNA microarray. The morphological status and androgenic levels of the same individuals have been described in a previous publication. In the present paper, expression of genes and pathways has been analysed with reference to these findings....... Nine clusters of genes with significant differential expression over time and 49 functional charts were found in the analysed testis samples. Prominent pathways in the prepubertal testis were associated with tissue renewal, cell respiration and increased endocytocis. E-cadherines may be associated with...

  3. Sperm-egg fusion: a molecular enigma of mammalian reproduction.

    Science.gov (United States)

    Klinovska, Karolina; Sebkova, Natasa; Dvorakova-Hortova, Katerina

    2014-01-01

    The mechanism of gamete fusion remains largely unknown on a molecular level despite its indisputable significance. Only a few of the molecules required for membrane interaction are known, among them IZUMO1, which is present on sperm, tetraspanin CD9, which is present on the egg, and the newly found oolema protein named Juno. A concept of a large multiprotein complex on both membranes forming fusion machinery has recently emerged. The Juno and IZUMO1, up to present, is the only known extracellular receptor pair in the process of fertilization, thus, facilitating the essential binding of gametes. However, neither IZUMO1 nor Juno appears to be the fusogenic protein. At the same time, the tetraspanin is expected to play a role in organizing the egg membrane order and to interact laterally with other factors. This review summarizes, to present, the known molecules involved in the process of sperm-egg fusion. The complexity and expected redundancy of the involved factors makes the process an intricate and still poorly understood mechanism, which is difficult to comprehend in its full distinction. PMID:24933635

  4. Pregnancy rates after intracytoplasmic sperm injection in relation to sperm recovery techniques

    OpenAIRE

    Egbase, P. E.; Al-Sharhan, M.; Ing, R.; Grudzinskas, J G

    1997-01-01

    Purpose/Methods: Clinical outcome after intracytoplasmic sperm injection (ICSI) was evaluated in relation to three techniques of sperm recovery, mini-Percoll, simple concentration, and centrifugation and washing.

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

    Science.gov (United States)

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

    2016-01-01

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

  6. The effect of theophylline and dibutyryl cyclic AMP on the uptake of radioactive calcium and phosphate ions by boar and human spermatozoa

    International Nuclear Information System (INIS)

    Radioactive calcium uptake by suspensions of washed boar and human spermatozoa was inhibited by the mitochondrial uncoupling agent carbonylcyanide-p-trifluoromethoxy-phenylhydrazone (FCCP). Theophylline + dibutyryl cyclic AMP also inhibited calcium uptake in the presence or absence of FCCP. Uptake of low concentrations of calcium (0.1 mM) was inhibited by the calcium ionophore A23187, but at high calcium concentrations the ionophore stimulated calcium uptake. These observations are explained in terms of a mechanism for the regulation of calcium uptake in spermatozoa based on competing mitochondrial and plasma membrane pumps. Uptake of 32P was also inhibited. These effects provide evidence that cyclic AMP plays a role in the transport of ions across the plasma membrane of spermatozoa. (author)

  7. Nuclear microscopy of sperm cell elemental structure

    International Nuclear Information System (INIS)

    Theories suggest there is a link between protamine concentrations in individual sperm and male fertility. Previously, biochemical analyses have used pooled samples containing millions of sperm to determine protamine concentrations. These methods have not been able to determine what percentage of morphologically normal sperm are biochemically defective and potentially infertile. Nuclear microscopy has been utilized to measure elemental profiles at the single sperm level. By measuring the amount of phosphorus and sulfur, the total DNA and protamine content in individual sperm from fertile bull and mouse semen have been determined. These values agree with results obtained from other biochemical analyses. Nuclear microscopy shows promise for measuring elemental profiles in the chromatin of individual sperm. The technique may be able to resolve theories regarding the importance of protamines to male fertility and identify biochemical defects responsible for certain types of male infertility. (orig.)

  8. Astaxanthin Improves Human Sperm Capacitation by Inducing Lyn Displacement and Activation.

    Science.gov (United States)

    Andrisani, Alessandra; Donà, Gabriella; Tibaldi, Elena; Brunati, Anna Maria; Sabbadin, Chiara; Armanini, Decio; Alvisi, Gualtiero; Gizzo, Salvatore; Ambrosini, Guido; Ragazzi, Eugenio; Bordin, Luciana

    2015-09-01

    Astaxanthin (Asta), a photo-protective red pigment of the carotenoid family, is known for its multiple beneficial properties. In this study, the effects of Asta on isolated human sperm were evaluated. Capacitation involves a series of transformations to let sperm acquire the correct features for potential oocyte fertilization, including the generation of a controlled amount of reactive oxygen species (ROS), cholesterol depletion of the sperm outer membrane, and protein tyrosine phosphorylation (Tyr-P) process in the head region. Volunteers, with normal spermiogram values, were divided in two separate groups on the basis of their ability to generate the correct content of endogenous ROS. Both patient group (PG) and control group (CG) were analysed for Tyr-phosphorylation (Tyr-P) pattern and percentages of acrosome-reacted cells (ARC) and non-viable cells (NVC), in the presence or absence of Asta. In addition, the involvement of ROS on membrane reorganization and the presence of Lyn, a Src family kinase associated with lipid rafts, were investigated. Results show that Lyn is present in the membranes of human sperm, mainly confined in midpiece in resting conditions. Following capacitation, Lyn translocated to the head concomitantly with raft relocation, thus allowing the Tyr-P of head proteins. Asta succeeded to trigger Lyn translocation in PG sperm thus bypassing the impaired ROS-related mechanism for rafts and Lyn translocation. In this study, we showed an interdependence between ROS generation and lipid rafts and Lyn relocation leading the cells to undergo the successive acrosome reaction (AR). Asta, by ameliorating PG sperm functioning, may be utilised to decrease male idiopathic infertility. PMID:26308013

  9. Astaxanthin Improves Human Sperm Capacitation by Inducing Lyn Displacement and Activation

    Directory of Open Access Journals (Sweden)

    Alessandra Andrisani

    2015-08-01

    Full Text Available Astaxanthin (Asta, a photo-protective red pigment of the carotenoid family, is known for its multiple beneficial properties. In this study, the effects of Asta on isolated human sperm were evaluated. Capacitation involves a series of transformations to let sperm acquire the correct features for potential oocyte fertilization, including the generation of a controlled amount of reactive oxygen species (ROS, cholesterol depletion of the sperm outer membrane, and protein tyrosine phosphorylation (Tyr-P process in the head region. Volunteers, with normal spermiogram values, were divided in two separate groups on the basis of their ability to generate the correct content of endogenous ROS. Both patient group (PG and control group (CG were analysed for Tyr-phosphorylation (Tyr-P pattern and percentages of acrosome-reacted cells (ARC and non-viable cells (NVC, in the presence or absence of Asta. In addition, the involvement of ROS on membrane reorganization and the presence of Lyn, a Src family kinase associated with lipid rafts, were investigated. Results show that Lyn is present in the membranes of human sperm, mainly confined in midpiece in resting conditions. Following capacitation, Lyn translocated to the head concomitantly with raft relocation, thus allowing the Tyr-P of head proteins. Asta succeeded to trigger Lyn translocation in PG sperm thus bypassing the impaired ROS-related mechanism for rafts and Lyn translocation. In this study, we showed an interdependence between ROS generation and lipid rafts and Lyn relocation leading the cells to undergo the successive acrosome reaction (AR. Asta, by ameliorating PG sperm functioning, may be utilised to decrease male idiopathic infertility.

  10. A role for mouse sperm surface galactosyltransferase in sperm binding to the egg zona pellucida

    OpenAIRE

    1982-01-01

    Past studies have suggested that mouse sperm surface galactosyltransferase may participate during fertilization by binding N- acetylglucosamine (GlcNAc) residues in the zona pellucida. In this paper, we examined further the role of sperm surface galactosyltransferase in mouse fertilization. Two reagents that specifically perturb sperm surface galactosyltransferase activity both inhibit sperm-zona binding. The presence of the milk protein alpha- lactalbumin specifically modifies the substrate ...

  11. Effects of Sperm Conjugation and Dissociation on Sperm Viability In Vitro

    OpenAIRE

    Higginson, Dawn M.; Henn, Kali R. H.

    2012-01-01

    Sperm conjugation is an unusual variation in sperm behavior where two or more spermatozoa physically unite for motility or transport through the female reproductive tract. Conjugation has frequently been interpreted as sperm cooperation, including reproductive altruism, with some sperm advancing their siblings toward the site of fertilization while ostensibly forfeiting their own ability to fertilize through damage incurred during conjugate break-up. Conversely, conjugation has been proposed ...

  12. Sperm competition and sperm midpiece size: no consistent pattern in passerine birds

    OpenAIRE

    Immler, S.; Birkhead, T. R.

    2007-01-01

    Sperm competition is thought to be a major force driving the evolution of sperm shape and function. However, previous studies investigating the relationship between the risk of sperm competition and sperm morphometry revealed inconclusive results and marked differences between taxonomic groups. In a comparative study of two families of passerines (Fringillidae and Sylviidae) and also across species belonging to different passerine families, we investigated the relative importance of the phylo...

  13. Sperm competition and sperm length influence the rate of mammalian spermatogenesis

    OpenAIRE

    Ramm, Steven A.; Stockley, Paula

    2009-01-01

    Sperm competition typically favours an increased investment in testes, because larger testes can produce more sperm to provide a numerical advantage in competition with rival ejaculates. However, interspecific variation in testis size cannot be equated directly with variation in sperm production rate—which is the trait ultimately selected under sperm competition—because there are also differences between species in the proportion of spermatogenic tissue contained within the testis and in the ...

  14. Development of genetically engineered human sperm immunocontraceptives

    OpenAIRE

    Naz, Rajesh K.

    2009-01-01

    Contraceptive vaccines targeting sperm are an exciting proposition. This review is focused on anti-sperm contraceptive vaccines and genetically engineered human antibodies that can be used as immunocontraceptives. Various methods of vaccinology and antibody engineering have been used to obtain multi-epitope contraceptive vaccines and human single chain variable fragment (scFv) antibodies from immunoinfertile and vasectomized men. Contraceptive vaccines comprised of various sperm antigens, pep...

  15. 添加不同植物油和L-肉碱对公猪精液品质和性欲的影响%Different Vegetable Oils and L-carnitine Influence Libido and Semen Characteristics of Breeding Boars

    Institute of Scientific and Technical Information of China (English)

    白小龙; 吴德; 林燕; 方正锋; 车炼强; 单妹

    2011-01-01

    The study was conducted to investigate the effects of supplementation with polyunsaturated fatty acids of different vegetable oils and L-carnitine on boar libido, semen characteristics, fatty acid composition of sperm and semen antioxidant properties. Thirty adult Pietrain x Duroc hybrid boars with similar age and body condition were randomly divided into 3 groups with 10 replicates and 1 pig in each replicate. The diet of group 1 was supplemented with 1.5% soybean oil; group 2 was supplemented with 1.5% soybean oil +200 mg/kg L-carnitine; group 3 was supplemented with 0. 3% of soybean oil +1.2% flax seed oil +200 mg/kg L-carni-tine. The feeding trial lasted for 16 weeks. The results showed that the semen volume and ejaculation duration of boars in group 3 were significantly higher than those in the other two groups (P 0.05). During 8 to 16 weeks, the sperm docosahexaenoic acid (DHA, n-3) content and activity of superoxide dismutase (SOD) of boars in group 3 were significantly higher than those in the other groups (P <0.05). The malondialdehyde ( MDA) content in semen of boars in group 1 was significantly higher than those in the other groups (P <0.01) during 8 to 16 weeks. The results suggest that dietary polyunsaturated fatty acids and L-camitine could enhance the boar libido, increase the sperm DHA content and semen antioxidant properties, and finally improve the boar semen production, sperm motility and sperm density. [Chinese Journal of Animal Nutrition, 2011, 23(8):1361-1369]%本研究旨在考察富含多不饱和脂肪酸(polyunsaturated fatty acid,PUFA)的不同植物油和L-肉碱的不同添加方式对种公猪精液品质、性欲、精子脂肪酸组成及精液抗氧化性能的影响.试验选用30头年龄、体况相近的成年皮特兰×杜洛克杂交种公猪,随机分为3组,每组设10个重复,每个重复1头猪.试验组公猪分别饲喂添加1.5%大豆油(试验组1)、1.5%大豆油+ 200 mg/kg L-肉碱(试验组2)、0.3%大豆油+1.2%

  16. Artificial insemination with donor sperm (AID): heterogeneity in sperm banking facilities in a single country (Belgium).

    Science.gov (United States)

    Thijssen, A; Dhont, N; Vandormael, E; Cox, A; Klerkx, E; Creemers, E; Ombelet, W

    2014-01-01

    Due to the high inflow of foreign patients seeking cross-border reproductive care in Belgium and the increased number of lesbian couples and single women who call for artificial insemination with donor sperm (AID), Belgian sperm banks nowadays face a shortage in donor sperm. However, since there is no central registration system for sperm donors in Belgium, no figures are currently available supporting this statement. Therefore a study was performed to obtain a detailed overview of the sperm banking facilities in Belgium. Questionnaires were sent to all Belgian centres for assisted reproduction with laboratory facilities (n = 18) to report on their sperm banking methods. The results showed that 82% of the centres rely partially or completely on foreign donor sperm. Moreover, four of the thirteen centres that have their own sperm bank use imported donor sperm in > 95% AID cycles. Our results show that in 63% of the Belgian AID cycles imported Danish donor sperm is being used. Donor recruitment is mainly performed through the centre's website (61%) or by distributing flyers in the centre (46%) and 9 to 180 potential donors have been recruited per centre in 2013. Eventually, 15 to 50% of these candidate donors were accepted. Different criteria for donor acceptance are handled by the centres: donor age limits range from 18-25 to 36-46 years old, and thresholds for sperm normality differ considerably. We can conclude that a wide variation in methods associated with sperm banking is observed in Belgian centres. PMID:25009728

  17. Sperm retention site and its influence on cleavage rate and early development following intracytoplasmic sperm injection

    OpenAIRE

    Yanaihara, Atsushi; Iwasaki, Shinji; Negishi, Momoko; Okai, Takashi

    2006-01-01

    Background: Intracytoplasmic sperm injection (ICSI) has risen to the forefront of reproductive technology. In the present study, the location of the sperm injection was noted, and a prospective study was conducted to evaluate the effect of the sperm retention site on cleavage rates and embryo quality after ICSI.

  18. Effects of sperm viability on fertilization and embryo cleavage following intracytoplasmic sperm injection

    OpenAIRE

    Poe-Zeigler, Robin; Nehchiri, Fariba; Hamacher, Pamela; Boyd, Catherine; Oehninger, Sergio; Muasher, Suheil; Lanzendorf, Susan E.

    1997-01-01

    Purpose: In the human, intracytoplasmic sperm injection is typically performed using “viable” sperm which has been mechanically rendered nonmotile. The purpose of the present study was to determine the ability of nonviable sperm to fertilize human oocytes and the early developmental normalcy of the resulting embryos.

  19. Microdissection testicular sperm extraction: an update

    Institute of Scientific and Technical Information of China (English)

    Ali A Dabaja; Peter N Schlegel

    2013-01-01

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

  20. Regulatory and necrotic volume increase in boar spermatozoa.

    Science.gov (United States)

    Petrunkina, A M; Jebe, E; Töpfer-Petersen, E

    2005-08-01

    Spermatozoa of many species initially respond to hypotonicity as perfect osmometers. Thereafter they undergo a regulatory process resulting in a decrease in cell volume, similar to that reported for somatic cells. Regulatory volume increase (RVI), a complementary process which is assumed to occur following initial shrinkage of sperm volume after exposure to a hypertonic medium, has not yet been described in detail for spermatozoa. In this study, we investigated whether spermatozoa are able to regulate their volume after hypertonic stress and whether this ability is maintained in preserved sperm. Cell volume changes were recorded using electronic cell sizing. Sperm response to the ion channels blockers quinidine, tamoxifen, and dydeoxyforskolin, and to protein kinase/phosphatase inhibitors lavendustin, staurosporine, and vanadate was studied to investigate possible mechanisms of RVI. Annexin V staining was used in combination with propidium iodide to determine whether hypertonic stress may induce apoptosis. Overall protein tyrosine phosphorylation under hypertonic conditions was measured via flow cytometry using antiphosphotyrosine antibody. Spermatozoa exposed to hypertonic stress initially responded with an abundant subpopulation according to the perfect osmometer model and recovered their volume from this shrinkage after 20 min. RVI was inhibited by quinidine and tamoxifen, which indicates the involvement of the important cellular ions sodium and chloride in this process. Volume regulatory ability was essentially maintained during storage of liquid semen. However, the response of the sperm population was heterogeneous. A second population raised, containing spermatozoa with larger volumes, which demonstrated irregularities in the volume response with respect to osmotic challenge, ion channel blockers, and storage. Under hypertonic conditions, both protein kinase inhibitors (PKI) led to increased isotonic volumes and to elevated initial relative volumes and

  1. 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...

  2. Two Types of Assays for Detecting Frog Sperm Chemoattraction

    OpenAIRE

    Burnett, Lindsey A.; Tholl, Nathan; Chandler, Douglas E.

    2011-01-01

    Sperm chemoattraction in invertebrates can be sufficiently robust that one can place a pipette containing the attractive peptide into a sperm suspension and microscopically visualize sperm accumulation around the pipette1. Sperm chemoattraction in vertebrates such as frogs, rodents and humans is more difficult to detect and requires quantitative assays. Such assays are of two major types - assays that quantitate sperm movement to a source of chemoattractant, so-called sperm accumulation assay...

  3. Long sperm fertilize more eggs in a bird

    OpenAIRE

    Bennison, Clair; Hemmings, Nicola; Slate, Jon; Birkhead, Tim

    2015-01-01

    Sperm competition, in which the ejaculates of multiple males compete to fertilize a female's ova, results in strong selection on sperm traits. Although sperm size and swimming velocity are known to independently affect fertilization success in certain species, exploring the relationship between sperm length, swimming velocity and fertilization success still remains a challenge. Here, we use the zebra finch (Taeniopygia guttata), where sperm size influences sperm swimming velocity, to determin...

  4. Chronically infected wild boar can transmit genotype 3 hepatitis E virus to domestic pigs.

    Science.gov (United States)

    Schlosser, Josephine; Vina-Rodriguez, Ariel; Fast, Christine; Groschup, Martin H; Eiden, Martin

    2015-10-22

    Hepatitis E virus (HEV) causes acute hepatitis E in humans in developing countries, but sporadic and autochthonous cases do also occur in industrialized nations. In Europe, food-borne zoonotic transmission of genotype 3 (gt3) has been associated with the consumption of raw and undercooked products from domestic pig and wild boar. As shown recently, naturally acquired HEV gt3 replicates efficiently in experimentally infected wild boar and is transmissible from a wild boar to domestic pigs. Generally, following an acute infection swine suffer from a transient febrile illness and viremia in connection with fecal virus shedding. However, little is known about sub-acute or chronic HEV infections in swine, and how and where HEV survives the immune response. In this paper, we describe the incidental finding of a chronic HEVgt3 infection in two naturally infected European wild boar which were raised and housed at FLI over years. The wild boar displayed fecal HEV RNA excretion and viremia over nearly the whole observation period of more than five months. The animal had mounted a substantial antibody response, yet without initial clearance of the virus by the immune system. Further analysis indicated a subclinical course of HEV with no evidence of chronic hepatitis. Additionally, we could demonstrate that this chronic wild boar infection was still transmissible to domestic pigs, which were housed together with this animal. Sentinel pigs developed fecal virus shedding accompanied by seroconversion. Wild boar should therefore be considered as an important reservoir for transmission of HEV gt3 in Europe. PMID:26344041

  5. Lead, cadmium and organochlorine pesticide residues in hunted red deer and wild boar from northern Italy.

    Science.gov (United States)

    Chiari, Mario; Cortinovis, Cristina; Bertoletti, Marco; Alborali, Loris; Zanoni, Mariagrazia; Ferretti, Enrica; Caloni, Francesca

    2015-01-01

    The objectives of the present study were to assess heavy metal cadmium (Cd), lead (Pb) and organochlorine pesticide concentrations in tissues of red deer (Cervus elaphus) and wild boar (Sus scrofa) from nine hunting areas and to evaluate related risk factors for the host animal. Over a period of 2 years, a total of 1055 and 210 masseters, 424 and 201 livers, 642 and 152 kidneys were collected from wild boar and red deer, respectively, and concentrations of Cd, Pb and organochlorine pesticides were determined. Comparing the two species, Cd concentration in the kidney (3.72 mg/kg), liver (0.67 mg/kg) and muscle (0.02 mg/kg) of wild boar was found to be significantly higher than in the organs of red deer (1.02 mg/kg in the kidneys, 0.07 mg/kg in the liver and 0.006 mg/kg in muscle). Mean Pb concentrations were found to be similar in both animals, with 0.39, 0.52 and 2.60 mg/kg detected in the wild boar kidney, liver and muscle, respectively, and 0.24, 0.21 and 2.04 mg/kg in the respective organs of the red deer. No difference in concentrations were found based on age class, location of tissue sample or contaminant in the case of wild boar. By contrast, a significantly lower Cd concentration was found in the kidney of the young red deer. The search for organochlorine pesticides in both red deer and wild boar produced negative results with values below the limits of detection. Due to the high levels of renal Cd and muscle Pb detected in wild boar and red deer, further research needs to be carried out in an effort to identify the source of contamination and preserve the health of animals and humans. PMID:26365428

  6. Lead and cadmium in red deer and wild boar from different hunting grounds in Croatia

    International Nuclear Information System (INIS)

    The concentration and relations of Cd and Pb as environmental risk factors were studied by atomic absorption spectrophotometry in the liver, kidney and muscle of free ranging wild boar (n = 94) and red deer (n = 45) from hunting grounds in four counties of north-east Croatia. In all four counties, the levels of Cd found in the kidney of red deer ranged from 2.28 to 5.91 mg/kg, and in wild boar from 3.47 to 21.10 mg/kg. The mean renal concentration of Cd was significantly higher in wild boar than in red deer from all four study areas. The mean hepatic (0.11 to 0.49 mg/kg, respectively) and muscle (0.01 to 0.04 mg/kg, respectively) Cd concentrations were similar in both species. The mean renal Cd concentration in wild boar and red deer exceeded 1 mg/kg in all four counties, ranging from 67.0% to 91.4% and from 45.5% to 69.2%, respectively. Also, the hepatic Cd/renal Cd ratio was lower than 1 in all animals. In all four counties, renal Pb concentration ranged from 0.058 to 3.77 mg/kg in red deer and from 0.056 to 11.60 mg/kg in wild boar. Hepatic Pb concentration was similar in both species (0.061 to 0.202 mg/kg in wild boar and 0.077 to 0.108 mg/kg in red deer). Because of the high Cd level in the organs of wild boar and red deer, further research is needed to identify the source of contamination in order to preserve the health of animals and humans.

  7. Single blastocyst transfer after ICSI from ejaculate spermatozoa, percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE)

    OpenAIRE

    Nilsson, Staffan; Waldenström, Urban; Engström, Ann-Britt; Hellberg, Dan

    2007-01-01

    Purpose: To investigate the outcome of IVF following intracytoplasmic sperm injection (ICSI) from ejaculate, percutaneous epididymal sperm aspiration (PESA) and testicular sperm extraction (TESE), with subsequent blastocyst culture and single blastocyst transfer.

  8. A role for a TIMP-3-sensitive, Zn(2+)-dependent metalloprotease in mammalian gamete membrane fusion.

    Science.gov (United States)

    Correa, L M; Cho, C; Myles, D G; Primakoff, P

    2000-09-01

    During fertilization, sperm and egg plasma membranes adhere and then fuse by a mechanism that is not well understood. Zinc metalloproteases are necessary for some intercellular fusion events, for instance, cell-cell fusion in yeast. In this study we tested the effects of class-specific and family-specific protease inhibitors on mouse gamete fusion. Capacitated, acrosome-reacted sperm and zona-free eggs were used in assays designed to define the effects of inhibitors on sperm-egg plasma membrane binding or fusion. Inhibitors of the aspartic, cysteine, and serine protease classes had no effect on sperm-egg binding or fusion. Both a synthetic metalloprotease substrate (succinyl-Ala-Ala-Phe-amidomethylcoumarin) and the zinc chelator 1,10-phenanthroline inhibited sperm-egg fusion but did not decrease sperm-egg binding. The fusion-inhibition effect of phenanthroline was reversible and activity of the inhibitable zinc metalloprotease was shown to be required during a short time window, the first 15 min after insemination. Tissue inhibitor of metalloprotease-3 and Ro 31-9790, specific inhibitors of zinc metalloproteases in the matrixin and adamalysin families, also inhibited sperm-egg fusion but not sperm-egg binding. These data indicate a role in gamete fusion for one or more zinc metalloproteases of the matrixin and/or adamalysin families that act after plasma membrane binding and before sperm-egg membrane fusion. PMID:10964469

  9. The Effects of Different Doses of Ketamine on Quality of Normal Ejaculated Sperm

    Directory of Open Access Journals (Sweden)

    Forouzan Absalan

    2014-07-01

    Full Text Available Background: Ketamine, an injectable anesthetic in human and animal medicine, is also a recreational drug used by young adults. The aim of this study is to evaluate the effects of ketamine on membrane integrity, DNA fragmentation and sperm parameters in humans. Materials and Methods: This prospective study was conducted on 40 males with normal semen samples over one month (August 2012. Subjects were randomly allocated to four groups (Control and case I, II and III whose semen samples were adjusted to different concentrations of ketamine (1, 3, 5 μL for one hour. Sperm analysis was performed for routine parameters, motility and morphology. Evaluation of membrane integrity and DNA fragmentation was done by eosin-Y staining and the sperm chromatin dispersion (SCD test, respectively. The results were analyzed by ANOVA and Tukey’s tests. P≤0.05 was considered statistically significant. Results: Total sperm motility in all case groups were significantly lower compared with the control group. In case group III, progressive motility showed significant difference with case group II. After addition of ketamine, sperm had evidence of coiled tails in all case groups compared to the control group however this observation was not significant. Evaluation of membrane integrity showed the rate of necrospermia increased in all case groups. However, ketamine only significantly affected membrane integrity in case group III. SCD staining showed that in the control group nucleoids with medium halos (63.44 ± 1.2 were significantly different compared to the case groups I (15.44 ± 0.45, II (9.05±1.16 and III (10.55 ± 1.14, respectively. Between case groups, nucleoids with large and medium halos showed significant differences in case groups II and III compared with case group I. Nucleoids with medium halos were significantly different between case groups II and III. Conclusion: Ketamine, through its effect on membrane integrity and DNA fragmentation, decreased

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

    Directory of Open Access Journals (Sweden)

    Oliver Vöcking

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

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

    Science.gov (United States)

    Vöcking, Oliver; Uhl, Gabriele; Michalik, Peter

    2013-01-01

    Storage of sperm inside the female genital tract is an integral phase of reproduction in many animal species. The sperm storage site constitutes the arena for sperm activation, sperm competition and female sperm choice. Consequently, to understand animal mating systems information on the processes that occur from sperm transfer to fertilization is required. Here, we focus on sperm activation in spiders. Male spiders produce sperm whose cell components are coiled within the sperm cell and that are surrounded by a proteinaceous sheath. These inactive and encapsulated sperm are transferred to the female spermathecae where they are stored for later fertilization. We analyzed the ultrastructural changes of sperm cells during residency time in the female genital system of the orb-web spider Argiope bruennichi. We found three clearly distinguishable sperm conditions: encapsulated sperm (secretion sheath present), decapsulated (secretion sheath absent) and uncoiled sperm (cell components uncoiled, presumably activated). After insemination, sperm remain in the encapsulated condition for several days and become decapsulated after variable periods of time. A variable portion of the decapsulated sperm transforms rapidly to the uncoiled condition resulting in a simultaneous occurrence of decapsulated and uncoiled sperm. After oviposition, only decapsulated and uncoiled sperm are left in the spermathecae, strongly suggesting that the activation process is not reversible. Furthermore, we found four different types of secretion in the spermathecae which might play a role in the decapsulation and activation process. PMID:24039790

  12. Cytosine methylation of sperm DNA in horse semen after cryopreservation.

    Science.gov (United States)

    Aurich, Christine; Schreiner, Bettina; Ille, Natascha; Alvarenga, Marco; Scarlet, Dragos

    2016-09-15

    Semen processing may contribute to epigenetic changes in spermatozoa. We have therefore addressed changes in sperm DNA cytosine methylation induced by cryopreservation of stallion semen. The relative amount of 5-methylcytosine relative to the genomic cytosine content of sperm DNA was analyzed by ELISA. In experiment 1, raw semen (n = 6 stallions, one ejaculate each) was shock-frozen. Postthaw semen motility and membrane integrity were completely absent, whereas DNA methylation was similar in raw (0.4 ± 0.2%) and shock-frozen (0.3 ± 0.1%) semen (not significant). In experiment 2, three ejaculates per stallion (n = 6) were included. Semen quality and DNA methylation was assessed before addition of the freezing extender and after freezing-thawing with either Ghent (G) or BotuCrio (BC) extender. Semen motility, morphology, and membrane integrity were significantly reduced by cryopreservation but not influenced by the extender (e.g., total motility: G 69.5 ± 2.0, BC 68.4 ± 2.2%; P < 0.001 vs. centrifugation). Cryopreservation significantly (P < 0.01) increased the level of DNA methylation (before freezing 0.6 ± 0.1%, postthaw G 6.4 ± 3.7, BC 4.4 ± 1.5%; P < 0.01), but no differences between the freezing extenders were seen. The level of DNA methylation was not correlated to semen motility, morphology, or membrane integrity. The results demonstrate that semen processing for cryopreservation increases the DNA methylation level in stallion semen. We conclude that assessment of sperm DNA methylation allows for evaluation of an additional parameter characterizing semen quality. The lower fertility rates of mares after insemination with frozen-thawed semen may at least in part be explained by cytosine methylation of sperm-DNA induced by the cryopreservation procedure. PMID:27242182

  13. Boar taint in entire male pigs: A genomewide association study for direct and indirect genetic effects on androstenone.

    NARCIS (Netherlands)

    Duijvestein, N.; Knol, E.F.; Bijma, P.

    2014-01-01

    Androstenone is one of the compounds causing boar taint of pork and is highly heritable (approximately 0.6). Recently, indirect genetic effects (IGE; also known as associative effects or social genetic effects) were found for androstenone, meaning that pen mates (boars) affect each other’s androsten

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

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

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

  15. Proteolytic release and partial characterization of human sperm-surface glycopeptides

    Directory of Open Access Journals (Sweden)

    Tortorella H.

    1997-01-01

    Full Text Available Sperm-surface glycopeptides were obtained from intact sperm membranes after proteolytic release by different enzymatic treatments such as autoproteolysis, trypsin, papain and pronase. Glycopeptides were isolated, their properties and composition were examined, and their monosaccharide and amino acid constituents were characterized. The monosaccharides identified were fucose, mannose, galactose, N-acetylglucosamine, and N-acetylgalactosamine, which form part of more than one type of oligosaccharide units. Autoproteolytic treatment mainly provided O-glycosidic type oligosaccharides, while a mixture of O- and N-glycosidic oligosaccharides was obtained in variable proportions when treated with trypsin, papain or pronase. The highest degree of peptide cleavage was obtained with pronase. Despite the higher yields reached with trypsin, these glycopeptides contain the lowest percentage of oligosaccharide chains. Proteolytic treatment provides a simple, rapid procedure for the isolation of glycopeptides from the sperm surface

  16. Production of channel catfish with sperm cryopreserved by rapid non-equilibrium cooling.

    Science.gov (United States)

    Cuevas-Uribe, Rafael; Leibo, S P; Daly, Jonathan; Tiersch, Terrence R

    2011-12-01

    This report describes the feasibility of using vitrification for fish sperm. Vitrification can be used to preserve samples in the field and offers an alternative to conventional cryopreservation, although it has not been systematically studied for sperm of aquatic species. The overall goal of the project was to develop streamlined protocols that could be integrated into a standardized approach for vitrification of aquatic species germplasm. The objectives of the present study in channel catfish (Ictalurus punctatus) were to: (1) evaluate the acute toxicity of 5%, 10%, 20% and 30% methanol, N,N-dimethyl acetamide, dimethyl sulfoxide, 1,2-propanediol, and methyl glycol; (2) evaluate a range of devices commonly used for cryopreservation and vitrification of mammalian sperm; (3) compare vitrification with and without cryoprotectants; (4) evaluate the post-thaw membrane integrity of sperm vitrified in different cryoprotectant solutions, and (5) evaluate the ability of vitrified sperm to fertilize eggs. Cryoprotectant concentrations of higher than 20% were found to be toxic to sperm. Methanol and methyl glycol were the least toxic at a concentration of 20% with an exposure time of less than 5 min. We evaluated a method reported for human sperm, using small volumes in loops (15 μl) or cut standard straws (20 μl) with and without cryoprotectants plunged into liquid nitrogen. Cryoprotectant-free vitrification using loops did not yield fertilization (assessed by neurulation), and the fertilization rates observed in two trials using the cut standard straws were low (~2%). In general, fertilization values for vitrification experiments were low and the use of low concentrations of cryoprotectants yielded lower fertilization (propanediol) with a single-step addition. This was reflected in the flow cytometry data from which the highest membrane integrity using loops was for 20% methanol+10% methyl glycol+10% propanediol (~50%). We report the first successful sperm vitrification

  17. 137Cs activity concentration in wild boar meat may still exceed the permitted levels

    Directory of Open Access Journals (Sweden)

    Rachubik J.

    2012-04-01

    Full Text Available The radiocaesium activity concentration may still remain high in natural products such as game meat, wild mushrooms, and forest berries even more than two decades after the Chernobyl accident. The results of regular control studies of game meat conducted in Poland showed wild boars as the most contaminated game animals. It is well documented that some mushrooms, readily consumed by animals, show high ability to accumulate caesium radioisotopes. Bay bolete, one of the most wide-spread mushroom species in Poland, reveals a unique radiocaesium accumulation feature. Moreover, deer truffle, containing also particularly high levels of radiocaesium, could be another radionu-clide source for wild boars. Furthermore, animals consuming deer truffles could digest contaminated soil components. Among 94 wild boar meat samples analysed in 2008–2009, two exceeded the permitted level. Hence, some precautions should be taken in the population with an elevated intake of wild boar meat. Moreover, since each hunted wild boar is examined for the presence of Trichinella larvae, regular measurements of radiocaesium concentrations in these animals may be advisable for enhancing consumer safety.

  18. The role of wild boars in spore dispersal of hypogeous fungi

    Directory of Open Access Journals (Sweden)

    Federica Piattoni

    2013-12-01

    Full Text Available Wild boars (Sus scrofa L. are well-known for soil disturbance in natural and cultivated truffières but their role in spore dispersal is poorly investigated. In the present work we studied the occurrence of hypogeous fungal spores in faecal contents of 14 wild boars randomly hunted in “Parco dei Gessi e Calanchi dell’Abbadessa” Regional Park (North of Italy where truffle production has been previously investigated for three years. Six methods for spore analysis in faeces were compared and the suspension of faeces in ZnSO4 (70% solution resulted to be the most reliable. Hypogeous fungal spores, including Tuber magnatum and Tuber aestivum spores, were detected in 9 animals. This result suggests that the detection of fungal spores in faeces of wild boars may provide information on the presence of hypogeous fungi in an area. However, the poor abundance of spores suggests that the wild boar can be considered an opportunistic mycophagist, ingesting truffles only occasionally, as a seasonal source of food. Considering the magnitude of wild boar movements during seasonal migrations, it is possible to speculate that they play a key role in truffle long distance dispersal.

  19. The Comparison Analysis of the Main Chemical Composition Parameters of Wild Boar Meat and Pork

    Directory of Open Access Journals (Sweden)

    Erika Bodnárné Skobrák

    2011-05-01

    Full Text Available The chemical composition of wild boar meat from wild boar parks with various habitats and different feeding facilities was analysed. Samples were collected from m. serratus anterior – during the winter hunting period. The protein, the total fat, the saturated and unsaturated fatty acid content were measured and also and those elements which have great importance on human alimentation. The results were compared with other author’s data of the same parameters of pork. The aim of the study was to compare the effect of different feeding systems on the ingredients of wild boar meat, and the accidental analogy with the ingredients of pork. There was no difference in protein content. The value of miristic acid (C14:0 in the samples of extensively and intensively fed group was more favourable than that of pork. Pork contains more MUFA (palmitoleic acid – C16:1. The semi-intensively fed wild boar’s meat and pork contained linoleic acid (C18:3 n3 almost on the same level. The greatest difference was detected in the level of arachidonic acid (C20:4 n6. The samples from all wild boar groups contained more of this fatty acid than it was published about domestic pigs. Out of the microelements, the iodine and zinc had higher value in wild boar meat than in pork.

  20. Predominance of sperm motion in corners.

    Science.gov (United States)

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

    2016-01-01

    Sperm migration through the female tract is crucial to fertilization, but the role of the complex and confined structure of the fallopian tube in sperm guidance remains unknown. Here, by confocal imaging microchannels head-on, we distinguish corner- vs. wall- vs. bulk-swimming bull sperm in confined geometries. Corner-swimming dominates with local areal concentrations as high as 200-fold that of the bulk. The relative degree of corner-swimming is strongest in small channels, decreases with increasing channel size, and plateaus for channels above 200 μm. Corner-swimming remains predominant across the physiologically-relevant range of viscosity and pH. Together, boundary-following sperm account for over 95% of the sperm distribution in small rectangular channels, which is similar to the percentage of wall swimmers in circular channels of similar size. We also demonstrate that wall-swimming sperm travel closer to walls in smaller channels (~100 μm), where the opposite wall is within the hydrodynamic interaction length-scale. The corner accumulation effect is more than the superposition of the influence of two walls, and over 5-fold stronger than that of a single wall. These findings suggest that folds and corners are dominant in sperm migration in the narrow (sub-mm) lumen of the fallopian tube and microchannel-based sperm selection devices. PMID:27211846

  1. Preservation of mouse sperm by convective drying and storing in 3-O-methyl-D-glucose.

    Science.gov (United States)

    Liu, Jie; Lee, Gloria Y; Lawitts, Joel A; Toner, Mehmet; Biggers, John D

    2012-01-01

    With the fast advancement in the genetics and bio-medical fields, the vast number of valuable transgenic and rare genetic mouse models need to be preserved. Preservation of mouse sperm by convective drying and subsequent storing at above freezing temperatures could dramatically reduce the cost and facilitate shipping. Mouse sperm were convectively dried under nitrogen gas in the Na-EGTA solution containing 100 mmol/L 3-O-methyl-D-glucose and stored in LiCl sorption jars (Relative Humidity, RH, 12%) at 4°C and 22°C for up to one year. The functionality of these sperm samples after storage was tested by intracytoplasmic injection into mouse oocytes. The percentages of blastocysts produced from sperm stored at 4°C for 1, 2, 3, 6, and 12 months were 62.6%, 53.4%, 39.6%, 33.3%, and 30.4%, respectively, while those stored at 22°C for 1, 2, and 3 months were 28.8%, 26.6%, and 12.2%, respectively. Transfer of 38 two- to four-cell embryos from sperm stored at 4°C for 1 year produced two live pups while 59 two- to four-cell embryos from sperm stored at 22°C for 3 months also produced two live pups. Although all the pups looked healthy at 3 weeks of age, normality of offspring produced using convectively dried sperm needs further investigation. The percentages of blastocyst from sperm stored in the higher relative humidity conditions of NaBr and MgCl(2) jars and driest condition of P(2)O(5) jars at 4°C and 22°C were all lower. A simple method of mouse sperm preservation is demonstrated. Three-O-methyl-D-glucose, a metabolically inactive derivative of glucose, offers significant protection for dried mouse sperm at above freezing temperatures without the need for poration of cell membrane. PMID:22272261

  2. Characterization of Mammalian ADAM2 and Its Absence from Human Sperm.

    Directory of Open Access Journals (Sweden)

    Heejin Choi

    Full Text Available The members of the ADAM (a disintegrin and metalloprotease family are membrane-anchored multi-domain proteins that play prominent roles in male reproduction. ADAM2, which was one of the first identified ADAMs, is the best studied ADAM in reproduction. In the male germ cells of mice, ADAM2 and other ADAMs form complexes that contribute to sperm-sperm adhesion, sperm-egg interactions, and the migration of sperm in the female reproductive tract. Here, we generated specific antibodies against mouse and human ADAM2, and investigated various features of ADAM2 in mice, monkeys and humans. We found that the cytoplasmic domain of ADAM2 might enable the differential association of this protein with other ADAMs in mice. Western blot analysis with the anti-human ADAM2 antibodies showed that ADAM2 is present in the testis and sperm of monkeys. Monkey ADAM2 was found to associate with chaperone proteins in testis. In humans, we identified ADAM2 as a 100-kDa protein in the testis, but failed to detect it in sperm. This is surprising given the results in mice and monkeys, but it is consistent with the failure of ADAM2 identification in the previous proteomic analyses of human sperm. These findings suggest that the reproductive functions of ADAM2 differ between humans and mice. Our protein analysis showed the presence of potential ADAM2 complexes involving yet-unknown proteins in human testis. Taken together, our results provide new information regarding the characteristics of ADAM2 in mammalian species, including humans.

  3. Characterization of Mammalian ADAM2 and Its Absence from Human Sperm

    Science.gov (United States)

    Choi, Heejin; Jin, Sora; Kwon, Jun Tae; Kim, Jihye; Jeong, Juri; Kim, Jaehwan; Jeon, Suyeon; Park, Zee Yong; Jung, Kang-Jin; Park, Kwangsung; Cho, Chunghee

    2016-01-01

    The members of the ADAM (a disintegrin and metalloprotease) family are membrane-anchored multi-domain proteins that play prominent roles in male reproduction. ADAM2, which was one of the first identified ADAMs, is the best studied ADAM in reproduction. In the male germ cells of mice, ADAM2 and other ADAMs form complexes that contribute to sperm-sperm adhesion, sperm-egg interactions, and the migration of sperm in the female reproductive tract. Here, we generated specific antibodies against mouse and human ADAM2, and investigated various features of ADAM2 in mice, monkeys and humans. We found that the cytoplasmic domain of ADAM2 might enable the differential association of this protein with other ADAMs in mice. Western blot analysis with the anti-human ADAM2 antibodies showed that ADAM2 is present in the testis and sperm of monkeys. Monkey ADAM2 was found to associate with chaperone proteins in testis. In humans, we identified ADAM2 as a 100-kDa protein in the testis, but failed to detect it in sperm. This is surprising given the results in mice and monkeys, but it is consistent with the failure of ADAM2 identification in the previous proteomic analyses of human sperm. These findings suggest that the reproductive functions of ADAM2 differ between humans and mice. Our protein analysis showed the presence of potential ADAM2 complexes involving yet-unknown proteins in human testis. Taken together, our results provide new information regarding the characteristics of ADAM2 in mammalian species, including humans. PMID:27341348

  4. Assessment of in vitro sperm characteristics and their importance in the prediction of conception rate in a bovine timed-AI program.

    Science.gov (United States)

    Oliveira, Letícia Zoccolaro; de Arruda, Rubens Paes; de Andrade, André Furugen Cesar; Celeghini, Eneiva Carla Carvalho; Reeb, Pablo Daniel; Martins, João Paulo Nascimento; dos Santos, Ricarda Maria; Beletti, Marcelo Emílio; Peres, Rogério Fonseca Guimarães; Monteiro, Fabio Morato; Hossepian de Lima, Vera Fernanda Martins

    2013-03-01

    The aims of this study were to assess in vivo fertility and in vitro sperm characteristics of different sires and to identify sperm variables important for the prediction of conception rate. Multiparous Nelore cows (n = 191) from a commercial farm underwent the same timed artificial insemination (timed-AI) protocol. Three batches of frozen semen from three Angus bulls were used (n = 9). A routine semen thawing protocol was performed in the laboratory to mimic field conditions. The following in vitro sperm analyses were performed: Computer Assisted Semen Analysis (CASA), Thermal Resistance Test (TRT), Hyposmotic Swelling Test (HOST), assessment of plasma and acrosomal membrane integrity, assessment of sperm plasma membrane stability and of lipid peroxidation by flow cytometry and assessment of sperm morphometry and chromatin structure by Toluidine Blue staining. For statistical analyses, Partial Least Squares (PLS) regression was used to explore the importance of various sperm variables in the prediction of conception rate. The following in vitro sperm variables were determined to be important predictors of conception rate: total motility (TM), progressive motility (PM), TM after 2 h of thermal incubation (TM_2 h), PM after 2 h of thermal incubation (PM_2 h), Beat Cross Frequency after 2 h of thermal incubation (BCF_2 h), percentage of rapidly moving cells after 2 h of thermal incubation (RAP_2 h), intact plasma membrane evaluated by HOST, intact plasma and acrosomal membranes evaluated by flow cytometry, intact plasma membrane suffering lipid peroxidation, major defects, total defects, morphometric width/length ratio, Fourier_0 and Fourier_2 and Chromatin Heterogeneity. We concluded that PLS regression is a suitable statistical method to identify in vitro sperm characteristics that have an important relationship with in vivo bull fertility. PMID:23428291

  5. Sperm protein 17 is expressed in the sperm fibrous sheath

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    Albani Elena

    2009-07-01

    Full Text Available Abstract Background Sperm protein 17 (Sp17 is a highly conserved mammalian protein characterized in rabbit, mouse, monkey, baboon, macaque, human testis and spermatozoa. mRNA encoding Sp17 has been detected in a range of murine and human somatic tissues. It was also recognized in two myeloma cell lines and in neoplastic cells from patients with multiple myeloma and ovarian carcinoma. These data all indicate that Sp17 is widely distributed in humans, expressed not only in germinal cells and in a variety of somatic tissues, but also in neoplastic cells of unrelated origin. Methods Sp17 expression was analyzed by immunocytochemistry and transmission electron microscopy on spermatozoa. Results Here, we demonstrate the ultrastructural localization of human Sp17 throughout the spermatozoa flagellar fibrous sheath, and its presence in spermatozoa during in vitro states from their ejaculation to the oocyte fertilization. Conclusion These findings suggest a possible role of Sp17 in regulating sperm maturation, capacitation, acrosomal reaction and interactions with the oocyte zona pellucida during the fertilization process. Further, the high degree of sequence conservation throughout its N-terminal half, and the presence of an A-kinase anchoring protein (AKAP-binding motif within this region, suggest that Sp17 might play a regulatory role in a protein kinase A-independent AKAP complex in both germinal and somatic cells.

  6. Sperm Whales Suffer the Bends

    Institute of Scientific and Technical Information of China (English)

    Michael; Hopkin; 席芳

    2005-01-01

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

  7. Liquid storage of equine semen: Assessing the effect of d-penicillamine on longevity of ejaculated and epididymal stallion sperm.

    Science.gov (United States)

    Brogan, P T; Beitsma, M; Henning, H; Gadella, B M; Stout, T A E

    2015-08-01

    Short-term storage of equine sperm at 5°C in an extender containing milk and/or egg yolk components is common practice in the equine breeding industry. Sperm motility, viability, DNA integrity and, consequently, fertilizing ability decline over time, partly due to reactive oxygen species (ROS) generation. We investigated whether adding the anti-oxidant d-penicillamine to a commercial milk/egg yolk extender delayed the decrease in semen quality. Semen was recovered on four consecutive days from eight 3-year old Warmblood stallions. On day 5, seven of the stallions were castrated and sperm recovered from the caudae epididymides. Ejaculated samples were split, and one portion was centrifuged and re-suspended to reduce seminal plasma content. All samples were diluted to 50millionsperm/ml and divided into two portions, one of which was supplemented with 0.5mM d-penicillamine. After 48h, 96h, 144h and 192h storage, sperm motility was assessed by computer-assisted semen analysis (CASA), viability by SYBR14/PI staining, and DNA integrity using the sperm chromatin structure assay (SCSA). d-Penicillamine had no effect on motility of ejaculated sperm (P>0.05) but reduced total and progressive motility of epididymal sperm. Sperm chromatin integrity was not influenced by storage time, seminal plasma or d-penicillamine. In short, adding d-penicillamine to a commercial semen extender was neither beneficial nor detrimental to the maintenance of quality in ejaculated semen stored at 5°C. The negative effect on motility of epididymal sperm may reflect differences in (membrane) physiology of spermatozoa that have not been exposed to seminal plasma. PMID:26130601

  8. Ultrastructure of fresh and post thawed sperm of pejerrey Odontesthes bonariensis (Atheriniformes

    Directory of Open Access Journals (Sweden)

    Ángela Gárriz

    2013-01-01

    Full Text Available In the present study it was showed for the first time the ultrastructural morphology of O. bonariensis sperm using electron microscopy techniques. Different kinds of abnormalities were described in fresh and post thawed sperm caused by crogenic protocols. Pejerrey spermatozoon is uniflagellated and is differentiated into three parts: a small roundish head (~1.80µm in length and 1.67µm in width, a midpiece or transitional region (~1.11µm in length and 1.56µm in width, and a long tail or flagellum (~29.08µm. Samples of fresh and post thawed sperm showed evidence of morphological anomalies affecting various intracellular compartments. Spermatozoa with swollen, ruptured, or absent membranes in the head showing excess of cytoplasm, and with alteration of the spatial orientation of the mitochondria were observed. A swollen flagellum was observed containing cytoplasmic vesicles, distributed along the whole length or concentrated in a restricted part of the tail. It was also found a high level of abnormalities (60% in frozen sperm when compared with normal sperm (18% reflecting the damage provoked by cryopreservation procedures.

  9. Thermodynamics of the interaction between bovine binder of sperm BSP1 and low-density lipoprotein from hen's egg yolk

    Energy Technology Data Exchange (ETDEWEB)

    Lusignan, Marie-France; Manjunath, Puttaswamy [Maisonneuve-Rosemont Hospital Research Center and Department of Medicine, Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada); Lafleur, Michel, E-mail: michel.lafleur@umontreal.ca [Department of Chemistry, Centre for Self-Assembled Chemical Structures (CSACS), Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada)

    2011-03-20

    Research highlights: {yields} Isothermal titration calorimetry revealed Binder-of-SPerm BSP1 protein has high affinity for egg yolk low density lipoproteins. {yields} It is estimated that 104 BSP1 molecules could bind one LDL particle. {yields} BSP1 has 2 phosphocholine binding sites and the BSP1/LDL ratio corresponds to 1.6 phosphatidylcholine per bound BSP1. {yields} The strong interaction between LDL particles and BSP1 is proposed to be important for sperm protection by egg yolk extender. - Abstract: Egg yolk is used in extender to protect sperm from cold shock and freezing during preservation. It is the low-density lipoprotein (LDL) fraction of egg yolk that protects sperm. Even though essential for sperm capacitation, the major proteins from bull seminal plasma, the Binder of SPerm (BSP) proteins, are detrimental for sperm preservation because they induce a continual phospholipids and cholesterol efflux from sperm membranes. The BSP proteins were proposed to bind to egg yolk LDL, preventing the sperm membrane damage. We characterized the binding between the BSP proteins and the LDL by isothermal titration calorimetry, providing the thermodynamics and quantitative description of this putative association. The association between BSP1 (major BSP proteins) and LDL is characterized by an affinity constant (K{sub a}) of 3.4 {+-} 0.4 {mu}M{sup -1}. A protein/LDL ratio of 104 {+-} 5 was determined indicating that 104 molecules of BSP1 would bind to one LDL particle. This stoichiometry leads to proposing that the association involves 1.6 {+-} 0.1 phosphatidylcholines (PC) per BSP protein. This finding is satisfactorily consistent with the fact that each BSP1 protein has 2 binding sites for choline group. In conclusion, the formation of a high affinity complex between BSP1 and LDL is proposed to be important for the protection of sperm by egg yolk extender.

  10. Epizootiology of trichinellosis in pigs and wild boars in Western Romania, 1998-2011.

    Science.gov (United States)

    Borza, Claudia; Neghina, Adriana M; Dumitrascu, Victor; Tirnea, Livius; Calma, Crenguta L; Neghina, Raul

    2012-08-01

    This article reports for the first time data regarding trichinellosis in sacrificed animals (domestic pigs and wild boars) inspected during the period from January 1998 to November 2011 in the largest of the Romanian counties. The data were collected from the Veterinary Public Health Department of Timis County. A total of 5,586,431 domestic pigs on farms, 609,325 pigs in private households, and 823 wild boars were examined during the study period. The results indicated that 681 domestic pigs (0.01%) on farms and 407 pigs in private households (0.07%) were found to be infected with Trichinella. Regarding wild boars, infection was detected in only 4 animals (0.5%). Although no infected animals have recently been identified during routine examinations (2009-2011), human outbreaks continue to occur yearly, and the main factors for these include improper handling by the population. PMID:22651387

  11. Effects of soybean isoflavones on reproductive parameters in Chinese mini-pig boars

    Directory of Open Access Journals (Sweden)

    Yuan Xiao-xue

    2012-10-01

    Full Text Available Abstract Background Soybean isoflavones are structurally similar to mammalian estrogens and therefore may act as estrogen agonists or antagonists. However, it has not been determined if they have any negative effects on reproductive parameters in male livestock. Therefore, the objective of this study was to evaluate the effects of soybean isoflavones on male reproduction using Chinese mini-pig boars as a model. Fifty Xiang boars were randomly divided into five groups and fed diets containing 0, 125, 250, or 500 ppm soybean isoflavones or 0.5 ppm diethylstilbestrol for 60 days. Results Dietary supplementation with 250 ppm of soy isoflavones markedly increased the testis index (P P P P P P P P P P P Conclusions The results of this study indicate that consumption of soy isoflavones at dietary levels up to 250 ppm did not adversely affect reproductive parameters in Chinese mini-pig boars whereas higher levels of soy isoflavones may adversely affect male reproduction.

  12. Evidence for Chlamydiaceae and Parachlamydiaceae in a wild boar (Sus scrofa population in Italy

    Directory of Open Access Journals (Sweden)

    Antonietta Di Francesco

    2013-03-01

    Full Text Available Conjunctival swabs from 44 free-living wild boars culled during a demographic control programme applied in a Regional Park located in the Northern Italy were examined by 16S rRNA encoding gene nested PCR. In total, 22 (50% wild boars were PCR positive. Sequencing of the amplicons identified Chlamydia suis and Chlamydia pecorum in 12 and 5 samples, respectively. For one sample found PCR positive, the nucleotide sequence could not be determined. Four conjunctival samples showed ≥ 92% sequence similarities to 16S rRNA sequences from Chlamydia-like organisms, as did large intestine, uterus, and vaginal swabs from the same four animals. Amoeba DNA was found in one Chlamydia-like organism positive conjunctival swab. To our knowledge, this is the first detection of members of the Parachlamydiaceae family in wild boars, confirming a large animal host range for Chlamydia-like organisms.

  13. Caries, Periodontal Disease, Supernumerary Teeth and Other Dental Disorders in Swedish Wild Boar (Sus scrofa).

    Science.gov (United States)

    Malmsten, A; Dalin, A-M; Pettersson, A

    2015-07-01

    Between January and December 2013, the dental and periodontal health of 99 Swedish wild boars (Sus scrofa) was investigated. Sampling occurred in conjunction with routine hunting at six large estates in the southern and middle parts of Sweden. All six of the estates use supplemental feeding. The weight of the animals, their sex and their dates of death were noted. Age was estimated using tooth eruption and tooth replacement patterns. The oral cavity was inspected and abnormalities were recorded on a dental chart modified for wild boars. The findings included supernumerary teeth, absence of teeth, mild class II malocclusion, severe tooth wear, periodontitis, calculus, caries, tooth fractures and the presence of enamel defects. Swedish wild boars suffer from different dental lesions and the impact of supplemental feeding on dental and periodontal health is still to be investigated. PMID:25979683

  14. Sparganosis in wild boar (Sus scrofa) - Implications for veterinarians, hunters, and consumers.

    Science.gov (United States)

    Kołodziej-Sobocińska, Marta; Miniuk, Mariusz; Ruczyńska, Iwona; Tokarska, Małgorzata

    2016-08-30

    From February to March 2016 we found plerocercoids of Spirometra sp. in four wild boar hunted in Białowieża Primeval Forest, north-eastern Poland. Plerocercoids were located subcutaneously and in muscle tissue. A sequence of a nuclear 18S rRNA gene was used for genetic specification of the samples. The analyzed gene fragment showed 100% identity with the Spirometra erinacei sequence. Thus, the emerge of human sparganosis due to consumption of undercooked or smoked wild boar meat is likely in the areas where wild boar is an approved food source, especially in the absence of routine guidelines for vets. It has become a priority to inform the public about possibilities and consequences of this zoonosis. PMID:27523946

  15. Attitudes of Swiss consumers towards meat from entire or immunocastrated boars: a representative survey.

    Science.gov (United States)

    Huber-Eicher, B; Spring, P

    2008-12-01

    Male piglets are castrated in order to prevent boar taint in pork. The surgical intervention is currently done without anaesthesia. Growing public concern about the welfare issue of this procedure forces the meat industry to evaluate alternative methods. The acceptance of such methods was studied in Switzerland within a large representative survey on the image of Swiss meat. Five questions were aimed at our subject. It was found that only a small part of the population has actually experienced boar taint. Nevertheless, the majority would not buy products made from tainted meat even if the absence of any perceivable boar taint and identical quality with current products could be guaranteed. The acceptance of meat from immunocastrated animals was low. Among the proposed four alternative methods, the production of entire males (with two options regarding processing of the tainted meat), immunocastration and castration with anaesthesia, only the last one seems to be acceptable to the interviewees. PMID:18433811

  16. Avaliação da suplementação de vitamina A nas características seminais em reprodutores suínos Assessment of vitamin A supplementation on boar semen characteristics

    Directory of Open Access Journals (Sweden)

    Simone Maria Massami Kitamura Martins

    2009-08-01

    Full Text Available Avaliou-se o efeito da suplementação de vitamina A na dieta sobre as características seminais (volume, motilidade, pH, vigor e concentração espermática, número total de espermatozoides, percentual de espermatozoides vivos e anormalidades morfológicas de reprodutores suínos no período de um ano (junho de 2004 a junho de 2005. Utilizaram-se dez reprodutores híbridos com 328,5 ± 2,12 dias de idade e 191,0 ± 12,0 kg distribuídos em duas rações, uma controle, com 10.000 UI de vit. A por kg de ração por dia, e outra com 16.000 UI de vit. A, fornecidas na quantidade de 2,5 kg/dia. O delineamento foi inteiramente casualizado com medidas repetidas no tempo. Não houve efeito significativo da suplementação de vitamina A na dieta nem de interação com o tempo nas características seminais estudadas. O tempo teve efeito significativo no pH, na concentração espermática, no número total de espermatozoides, no percentual de espermatozoides vivos, nas anormalidades morfológicas, na motilidade e no vigor espermático. Apesar da não-significância da suplementação de vitamina A, foi possível detectar diferenças numéricas no aumento da motilidade e de espermatozoides vivos, bem como na diminuição das anormalidades morfológicas, resultados que indicam ação da vitamina A.The objective of this study was to evaluate the effect of vitamin A feed supplementation on the following boar semen characteristics: volume, motility, vigor, seminal pH, spermatic concentration, total number of spermatozoa, percentage of living sperm cells and morphologic abnormalities. The experiment was carried out with 10 hybrid boars (328.5 ± 2.12 days of age and 191.0 ± 12.0 kg live weight. The boars were separated into two treatments and fed with two vitamin A levels: control, 10.000 UI/kg feed daily; and vitamin A, 16.000 UI. The boars received 2.5 kg feed daily. A randomized complete design was used with replication measurements in time. There was no

  17. Protein and carbohydrate intake influence sperm number and fertility in male cockroaches, but not sperm viability

    Science.gov (United States)

    Bunning, Harriet; Rapkin, James; Belcher, Laurence; Archer, C. Ruth; Jensen, Kim; Hunt, John

    2015-01-01

    It is commonly assumed that because males produce many, tiny sperm, they are cheap to produce. Recent work, however, suggests that sperm production is not cost-free. If sperm are costly to produce, sperm number and/or viability should be influenced by diet, and this has been documented in numerous species. Yet few studies have examined the exact nutrients responsible for mediating these effects. Here, we quantify the effects of protein (P) and carbohydrate (C) intake on sperm number and viability in the cockroach Nauphoeta cinerea, as well as the consequences for male fertility. We found the intake of P and C influenced sperm number, being maximized at a high intake of diets with a P : C ratio of 1 : 2, but not sperm viability. The nutritional landscapes for male fertility and sperm number were closely aligned, suggesting that sperm number is the major determinant of male fertility in N. cinerea. Under dietary choice, males regulate nutrient intake at a P : C ratio of 1 : 4.95, which is midway between the ratios needed to maximize sperm production and pre-copulatory attractiveness in this species. This raises the possibility that males regulate nutrient intake to balance the trade-off between pre- and post-copulatory traits in this species. PMID:25608881

  18. Sperm characterization and identification of sperm sub-populations in ejaculates from pampas deer (Ozotoceros bezoarticus).

    Science.gov (United States)

    Beracochea, F; Gil, J; Sestelo, A; Garde, J J; Santiago-Moreno, J; Fumagalli, F; Ungerfeld, R

    2014-10-01

    Pampas deer (Ozotoceros bezoarticus) is a native endangered species. Knowledge of the basic spermiogram characteristics and the morphometric descriptors is necessary to effectively develop sperm cryopreservation. In other species, sperm sub-population is related to sperm cryo-resistance. The objective was to provide a general description of the sperm, including sperm head morphometric descriptors, its repeatability, and the existence of sperm sub-populations. Sperm were obtained from adult males by electroejaculation during the breeding season. The motility score was 3.4 ± 0.2 (mean ± SEM) and progressive motility was 59.4 ± 3.7%. Ejaculated volume was 413.9 ± 51.0 μl, the total number of sperm ejaculated was 321.2 ± 55.4 × 10(6). Also, 63.3 ± 3.1% of the sperm were morphologically abnormal and 23.7 ± 2.3% had acrosome damage. The sperm head length was 7.6 ± 0.01 μm, width 4.4 ± 0.01 μm, area 28.1 ± 0.07 μm(2) and the perimeter was 21.9 ± 0.04 μm. There was a positive relationship among morphometric descriptors and the motility score, overall motility and progressive motility. Also length (P=0.011), width (P=0.003), area (P=0.006) and perimeter (P=0.009) of sperm head obtained in two different collections were positively related. Overall, the low concentration, volume, overall quality and abnormal morphology, and wide variation of these variables may be a limitation for the development of sperm cryopreserved banks. There were three sperm sub-populations with different morphometric characteristics. The morphometric descriptors are maintained similarly among different collections. PMID:25104472

  19. Wild boar (Sus scrofa) - reservoir host of Toxoplasma gondii, Neospora caninum and Anaplasma phagocytophilum in Slovakia.

    Science.gov (United States)

    Reiterová, Katarína; Špilovská, Silvia; Blaňarová, Lucia; Derdáková, Markéta; Čobádiová, Andrea; Hisira, Vladimír

    2016-06-01

    In Central Europe the wild boar population is permanently growing and consequently Cf foodborne infections. In this study serological and molecular detection of Toxoplasma gondii and Neospora caninum in wild boars was evaluated. Moreover, same samples were screened for the presence and genetic variability of tick-borne bacterium Anaplasma phagocytophilum. Blood samples collected from 113 wild boars from Southern Slovakia were examined for antibodies to T. gondii by indirect and to N. caninum by competitive ELISA. The presence of parasitic DNA in blood samples was determined by standard or real time PCR techniques. Antibodies against T. gondii and N. caninum were detected in 45 (39.8%) and 38 (33.6%) animals, respectively. Females were more frequently infected for both pathogens than males. The high seropositivity against both coccidia indicates a permanent occurrence of these pathogens in the studied locality. T. gondii DNA was confirmed in five seropositive boars (4.4%) and N. caninum in 23 blood samples (20.4%). Three out of 23 N. caninum PCR positive animals did not show seropositivity. Three out of 113 blood samples of wild boars were positive for A. phagocytophilum (2.7%). The obtained A. phagocytophilum sequences were 100% identical with GenBankTM isolates from Slovak dog (KC985242); German horse (JF893938) or wild boar (EF143810) and red deer (EF143808) from Poland. Coinfections of T. gondii with N. caninum and N. caninum with A. phagocytophilum were detected in single cases. Results suggest a potential zoonotic risk of toxoplasmosis transmission to humans and the spread of neosporosis to farm animals. PMID:27078648

  20. Sperm dumping as a defense against meiotic drive

    OpenAIRE

    Price, Tom; Lewis, Zenobia; Wedell, Nina

    2009-01-01

    Sperm from Drosophila simulans that carry a sex-ratio distorter is preferentially lost from females' sperm-storage organs. This suggests that sperm dumping is a major factor affecting sperm competition in this species, and may have evolved in response to sex-ratio distorters.

  1. Effect of different thawing temperatures on the viability, in vitro fertilizing capacity and chromatin condensation of frozen boar semen packaged in 5 ml straws.

    Science.gov (United States)

    Córdova-Izquierdo, A; Oliva, J H; Lleó, B; García-Artiga, C; Corcuera, B D; Pérez-Gutiérrez, J F

    2006-03-01

    The effect of two different thawing temperatures on frozen boar semen viability, in vitro fertilizing capacity and chromatin condensation and stability was studied. Freeze-thaw motility, normal apical ridge (NAR), in vitro fertilizing (IVF) capacity and chromatin condensation and stability were evaluated after thawing at 42 degrees C, 40s and 50 degrees C, 40s. Chromatin condensation degree was determined by flow cytometry, using propidium iodide as fluorochrome intercalating agent, and chromatin stability was evaluated by the same procedure after inducing sperm chromatin decondensation with ethylene diamine tetraacetic acid (EDTA) and sodium dodecyl sulfate (SDS). The results showed that thawing straws at 42 degrees C, 40s significantly reduced motility compared to straws thawed at 50 degrees C, 40s. NAR, penetration, monospermy and polyspermy were not different between the two groups of samples thawed at different temperatures. Chromatin was significantly more compact when thawing was performed at 50 degrees C, but its stability did not show any difference relative to thawing at 42 degrees C. It is suggested that the interactions involved in chromatin overcondensation had a non-covalent nature. PMID:15975744

  2. The Trumorph® system: The new universal technique for the observation and analysis of the morphology of living sperm. [corrected].

    Science.gov (United States)

    Soler, C; García-Molina, A; Contell, J; Silvestre, M A; Sancho, M

    2015-07-01

    Evaluation of sperm morphology is a fundamental component of semen analysis, but its real significance has been obscured by a plethora of techniques that involve fixation and staining procedures that induce artefacts. Here we describe Trumorph℗®, a new method for sperm morphology assessment that is based upon examination of wet preparations of living spermatozoa immobilized by a short 60°C shock using negative phase contrast microscopy. We have observed samples from five animals of the following species: bull, boar, goat and rabbit. In every case, all the components of the sperm head and tail were perfectly defined, including the acrosome and midpiece (in all its length, including cytoplasmic droplets). A range of morphological forms was observed, similar to those found by conventional fixed and stained preparations, but other forms were found, distinguishable only by the optics used. The ease of preparation makes it a robust method applicable for analysis of living unmodified spermatozoa in a range of situations. Subsequent studies on well-characterized samples are required to describe the morphology of potentially fertilizing spermatozoa. PMID:26003741

  3. Worldwide phylogeography of wild boar reveals multiple centers of pig domestication.

    Science.gov (United States)

    Larson, Greger; Dobney, Keith; Albarella, Umberto; Fang, Meiying; Matisoo-Smith, Elizabeth; Robins, Judith; Lowden, Stewart; Finlayson, Heather; Brand, Tina; Willerslev, Eske; Rowley-Conwy, Peter; Andersson, Leif; Cooper, Alan

    2005-03-11

    Mitochondrial DNA (mtDNA) sequences from 686 wild and domestic pig specimens place the origin of wild boar in island Southeast Asia (ISEA), where they dispersed across Eurasia. Previous morphological and genetic evidence suggested pig domestication took place in a limited number of locations (principally the Near East and Far East). In contrast, new genetic data reveal multiple centers of domestication across Eurasia and that European, rather than Near Eastern, wild boar are the principal source of modern European domestic pigs. PMID:15761152

  4. Do intensive drive hunts affect wild boar () spatial behaviour in Italy? Some evidences and management implications

    OpenAIRE

    Scillitani, Laura; Monaco, Andrea; Toso, Silvano

    2009-01-01

    Abstract Wild boar have been increasing in numbers all over Western Europe in the last 30 years. The species is a major pest for agriculture, but it has a high value as a game species, and in Italy, as in several other countries, it is traditionally hunted in drive hunts by hunting teams with several dogs. This hunting method can have disruptive effects on the demography and spatial behaviour of wild boar, especially family groups. We conducted a 2-year study (2003 and 2004) to det...

  5. Suspended sediments limit coral sperm availability

    Science.gov (United States)

    Ricardo, Gerard F.; Jones, Ross J.; Clode, Peta L.; Humanes, Adriana; Negri, Andrew P.

    2015-01-01

    Suspended sediment from dredging activities and natural resuspension events represent a risk to the reproductive processes of coral, and therefore the ongoing maintenance of reefal populations. To investigate the underlying mechanisms that could reduce the fertilisation success in turbid water, we conducted several experiments exposing gametes of the corals Acropora tenuis and A. millepora to two sediment types. Sperm limitation was identified in the presence of siliciclastic sediment (230 and ~700 mg L−1), with 2–37 fold more sperm required to achieve maximum fertilisation rates, when compared with sediment-free treatments. This effect was more pronounced at sub-optimum sperm concentrations. Considerable (>45%) decreases in sperm concentration at the water’s surface was recorded in the presence of siliciclastic sediment and a >20% decrease for carbonate sediment. Electron microscopy then confirmed sediment entangled sperm and we propose entrapment and sinking is the primary mechanism reducing sperm available to the egg. Longer exposure to suspended sediments and gamete aging further decreased fertilisation success when compared with a shorter exposure. Collectively, these findings demonstrate that high concentrations of suspended sediments effectively remove sperm from the water’s surface during coral spawning events, reducing the window for fertilisation with potential subsequent flow-on effects for recruitment. PMID:26659008

  6. The stochastic dance of circling sperm cells: sperm chemotaxis in the plane

    International Nuclear Information System (INIS)

    Biological systems such as single cells must function in the presence of fluctuations. It has been shown in a two-dimensional experimental setup that sea urchin sperm cells move toward a source of chemoattractant along planar trochoidal swimming paths, i.e. drifting circles. In these experiments, a pronounced variability of the swimming paths is observed. We present a theoretical description of sperm chemotaxis in two dimensions which takes fluctuations into account. We derive a coarse-grained theory of stochastic sperm swimming paths in a concentration field of chemoattractant. Fluctuations enter as multiplicative noise in the equations for the sperm swimming path. We discuss the stochastic properties of sperm swimming and predict a concentration-dependence of the effective diffusion constant of sperm swimming which could be tested in experiments.

  7. The stochastic dance of circling sperm cells: sperm chemotaxis in the plane

    Energy Technology Data Exchange (ETDEWEB)

    Friedrich, B M; Juelicher, F [Max Planck Institute for the Physics of Complex Systems, Noethnitzer Strasse 38, 01187 Dresden (Germany)], E-mail: ben@pks.mpg.de, E-mail: julicher@pks.mpg.de

    2008-12-15

    Biological systems such as single cells must function in the presence of fluctuations. It has been shown in a two-dimensional experimental setup that sea urchin sperm cells move toward a source of chemoattractant along planar trochoidal swimming paths, i.e. drifting circles. In these experiments, a pronounced variability of the swimming paths is observed. We present a theoretical description of sperm chemotaxis in two dimensions which takes fluctuations into account. We derive a coarse-grained theory of stochastic sperm swimming paths in a concentration field of chemoattractant. Fluctuations enter as multiplicative noise in the equations for the sperm swimming path. We discuss the stochastic properties of sperm swimming and predict a concentration-dependence of the effective diffusion constant of sperm swimming which could be tested in experiments.

  8. Polyspermy in birds: sperm numbers and embryo survival

    OpenAIRE

    Hemmings, N.; Birkhead, T. R.

    2015-01-01

    Polyspermy is a major puzzle in reproductive biology. In some taxa, multiple sperm enter the ovum as part of the normal fertilization process, whereas in others, penetration of the ovum by more than one sperm is lethal. In birds, several sperm typically enter the germinal disc, yet only one fuses with the female pronucleus. It is unclear whether supernumerary sperm play an essential role in the avian fertilization process and, if they do, how females regulate the progression of sperm through ...

  9. Female presence influences sperm velocity in the guppy

    OpenAIRE

    Gasparini, Clelia; Alfredo V Peretti; Pilastro, Andrea

    2009-01-01

    As sperm production is costly, males are expected to strategically allocate resources to sperm production according to mating opportunities. While sperm number adjustments have been reported in several taxa, only a few studies investigated whether sperm quality shows adaptive plasticity as well. We tested this prediction in the guppy, Poecilia reticulata. A total of 46 males were initially stripped of all retrievable sperm before being randomly allocated to one of two treatments simulating di...

  10. Progress in Oral Vaccination against Tuberculosis in Its Main Wildlife Reservoir in Iberia, the Eurasian Wild Boar

    Directory of Open Access Journals (Sweden)

    Beatriz Beltrán-Beck

    2012-01-01

    Full Text Available Eurasian wild boar (Sus scrofa is the main wildlife reservoir for tuberculosis (TB in Iberia. This review summarizes the current knowledge on wild boar vaccination including aspects of bait design, delivery and field deployment success; wild boar response to vaccination with Bacillus Calmette-Guérin (BCG and inactivated Mycobacterium bovis; and wild boar vaccination biosafety issues as well as prospects on future research. Oral vaccination with BCG in captive wild boar has shown to be safe with significant levels of protection against challenge with virulent M. bovis. An oral vaccination with a new heat-killed M. bovis vaccine conferred a protection similar to BCG. The study of host-pathogen interactions identified biomarkers of resistance/susceptibility to tuberculosis in wild boar such as complement component 3 (C3 and methylmalonyl coenzyme A mutase (MUT that were used for vaccine development. Finally, specific delivery systems were developed for bait-containing vaccines to target different age groups. Ongoing research includes laboratory experiments combining live and heat-killed vaccines and the first field trial for TB control in wild boar.

  11. Mass-Specific Metabolic Rate and Sperm Competition Determine Sperm Size in Marsupial Mammals

    OpenAIRE

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

    2011-01-01

    Two complementary hypotheses have been proposed to explain variation in sperm size. The first proposes that post-copulatory sexual selection favors an increase in sperm size because it enhances sperm swimming speed, which is an important determinant of fertilization success in competitive contexts. The second hypothesis proposes that mass-specific metabolic rate acts as a constraint, because large animals with low mass-specific metabolic rates will not be able to process resources at the rate...

  12. Outcomes with intracytoplasmic sperm injection of cryopreserved sperm from men with spinal cord injury

    OpenAIRE

    Bechoua, Shaliha; Berki-Morin, Yasmine; Michel, Frédéric; Girod, Sophie; Sagot, Paul; Fauque, Patricia

    2013-01-01

    Background Erectile dysfunction, ejaculatory dysfunction and poor semen quality are the main causes of infertility in men with spinal cord injury (SCI). Different sperm retrieval techniques such as penile vibratory stimulation (PVS), electro-ejaculation (EEJ) or surgical sperm retrieval (SSR) associated or not with sperm cryopreservation can be offered to these patients to preserve their fertility. If fatherhood cannot be achieved naturally, assisted reproductive techniques can be offered to ...

  13. Ovarian fluid mediates the temporal decline in sperm viability in a fish with sperm storage.

    Directory of Open Access Journals (Sweden)

    Clelia Gasparini

    Full Text Available A loss of sperm viability and functionality during sperm transfer and storage within the female reproductive tract can have important fitness implications by disrupting fertilization and impairing offspring development and survival. Consequently, mechanisms that mitigate the temporal decline in sperm function are likely to be important targets of selection. In many species, ovarian fluid is known to regulate and maintain sperm quality. In this paper, we use the guppy Poecilia reticulata, a highly polyandrous freshwater fish exhibiting internal fertilization and sperm storage, to determine whether ovarian fluid (OF influences the decline in sperm viability (the proportion of live sperm in the ejaculate over time and whether any observed effects depend on male sexual ornamentation. To address these questions we used a paired experimental design in which ejaculates from individual males were tested in vitro both in presence and absence of OF. Our results revealed that the temporal decline in sperm viability was significantly reduced in the presence of OF compared to a saline control. This finding raises the intriguing possibility that OF may play a role in mediating the decline in sperm quality due to the deleterious effects of sperm ageing, although other possible explanations for this observation are discussed. Interestingly, we also show that the age-related decline in sperm viability was contingent on male sexual ornamentation; males with relatively high levels of iridescence (indicating higher sexual attractiveness exhibited a more pronounced decline in sperm viability over time than their less ornamented counterparts. This latter finding offers possible insights into the functional basis for the previously observed trade-off between these key components of pre- and postcopulatory sexual selection.

  14. Long sperm fertilize more eggs in a bird.

    Science.gov (United States)

    Bennison, Clair; Hemmings, Nicola; Slate, Jon; Birkhead, Tim

    2015-01-22

    Sperm competition, in which the ejaculates of multiple males compete to fertilize a female's ova, results in strong selection on sperm traits. Although sperm size and swimming velocity are known to independently affect fertilization success in certain species, exploring the relationship between sperm length, swimming velocity and fertilization success still remains a challenge. Here, we use the zebra finch (Taeniopygia guttata), where sperm size influences sperm swimming velocity, to determine the effect of sperm total length on fertilization success. Sperm competition experiments, in which pairs of males whose sperm differed only in length and swimming speed, revealed that males producing long sperm were more successful in terms of (i) the number of sperm reaching the ova and (ii) fertilizing those ova. Our results reveal that although sperm length is the main factor determining the outcome of sperm competition, complex interactions between male and female reproductive traits may also be important. The mechanisms underlying these interactions are poorly understood, but we suggest that differences in sperm storage and utilization by females may contribute to the outcome of sperm competition. PMID:25621327

  15. Lactic acid is a sperm motility inactivation factor in the sperm storage tubules

    Science.gov (United States)

    Matsuzaki, Mei; Mizushima, Shusei; Hiyama, Gen; Hirohashi, Noritaka; Shiba, Kogiku; Inaba, Kazuo; Suzuki, Tomohiro; Dohra, Hideo; Ohnishi, Toshiyuki; Sato, Yoshikatsu; Kohsaka, Tetsuya; Ichikawa, Yoshinobu; Atsumi, Yusuke; Yoshimura, Takashi; Sasanami, Tomohiro

    2015-01-01

    Although successful fertilization depends on timely encounters between sperm and egg, the decoupling of mating and fertilization often confers reproductive advantages to internally fertilizing animals. In several vertebrate groups, postcopulatory sperm viability is prolonged by storage in specialized organs within the female reproductive tract. In birds, ejaculated sperm can be stored in a quiescent state within oviductal sperm storage tubules (SSTs), thereby retaining fertilizability for up to 15 weeks at body temperature (41 °C); however, the mechanism by which motile sperm become quiescent within SSTs is unknown. Here, we show that low oxygen and high lactic acid concentrations are established in quail SSTs. Flagellar quiescence was induced by lactic acid in the concentration range found in SSTs through flagellar dynein ATPase inactivation following cytoplasmic acidification (sperm morphology under hypoxic and high temperature conditions indicates that a combination of these factors enables sperm cells to survive during the ovulation cycles. Our findings suggested a novel physiological role for lactic acid in promoting sperm quiescence in SSTs and opened up a new opportunity for technological improvement in prolonging sperm longevity at ambient or body temperature. PMID:26619826

  16. Motile sperm organelle morphology examination (MSOME) and sperm head vacuoles: state of the art in 2013.

    Science.gov (United States)

    Perdrix, Anne; Rives, Nathalie

    2013-01-01

    BACKGROUND Approximately 10 years after the first publication introducing the motile sperm organelle morphology examination (MSOME), many questions remained about sperm vacuoles: frequency, size, localization, mode of occurrence, biological significance and impact on male fertility potential. Many studies have tried to characterize sperm vacuoles, to determine the sperm abnormalities possibly associated with vacuoles, to test the diagnostic value of MSOME for male infertility or to question the benefits of intracytoplasmic morphologically selected sperm injection (IMSI). METHODS We searched PubMed for articles in the English language published in 2001-2012 regarding human sperm head vacuoles, MSOME and IMSI. RESULTS A bibliographic analysis revealed consensus for the following findings: (i) sperm vacuoles appeared frequently, often multiple and preferentially anterior; (ii) sperm vacuoles and sperm chromatin immaturity have been associated, particularly in the case of large vacuoles; (iii) teratozoospermia was a preferred indication of MSOME and IMSI. CONCLUSION The high-magnification system appears to be a powerful method to improve our understanding of human spermatozoa. However, its clinical use remains unclear in the fields of male infertility diagnosis and assisted reproduction techniques (ARTs). PMID:23825157

  17. Sperm Proteases that May Be Involved in the Initiation of Sperm Motility in the Newt, Cynops pyrrhogaster

    OpenAIRE

    Misato Yokoe; Makoto Sano; Honami Shibata; Daisuke Shibata; Eriko Takayama-Watanabe; Kazuo Inaba; Akihiko Watanabe

    2014-01-01

    A protease of sperm in the newt Cynops pyrrhogaster that is released after the acrosome reaction (AR) is proposed to lyse the sheet structure on the outer surface of egg jelly and release sperm motility-initiating substance (SMIS). Here, we found that protease activity in the sperm head was potent to widely digest substrates beneath the sperm. The protease activity measured by fluorescein thiocarbamoyl-casein digestion was detected in the supernatant of the sperm after the AR and the activi...

  18. Nuclear reconstitution of demembranated Orychophragmus violaceus sperm in Xenopus laevis egg extracts

    Institute of Scientific and Technical Information of China (English)

    LU; Ping(卢萍); REN; Min(任民); ZHAI; Zhonghe(翟中和)

    2002-01-01

    The cell-free extracts from animal Xenopus laevis egg could induce chromatin decon- densation and pronuclear formation from demembranated plant (Orychophragmus violaceus)sperm. The demembranated Orychophragmus violaceus sperm began to swell in 30 min incubation, and then were gradually decondensed. The reassembly of nuclear envelope in the reconstituted nuclei had been visualized by means of electron microscopy and fluorescent microscopy. Membrane vesicles fused to form the double envelope around the periphery of the decondensed chromatin. The morphology of the newly formed nucleus, with a double membrane, was similar to those nuclei after fertilization. Transmission electron microscope micrograph of the whole mount prepared nuclear matrix-lamina showed the reconstituted nucleus to be filled with a dense network.

  19. Successive electrical responses to insemination and concurrent sperm entries in the polyspermic egg of the ctenophore Beroe ovata.

    Science.gov (United States)

    Goudeau, M; Goudeau, H

    1993-04-01

    In the ctenophore Beroe ovata, action potentials and sperm-induced egg electrical responses were recorded, in unfertilized and in vitro inseminated eggs, respectively. The unfertilized egg had a resting membrane potential of -65.6 +/- 1.3 mV and a membrane resistance of 1.45 +/- 0.09 M omega. When depolarized beyond a threshold of about -20 mV, the membrane displayed an action potential with a fast phase and then a peak reaching 38 +/- 0.6 mV, followed by a positive declining plateau which lasted about 20 sec and ended by a biphasic repolarization of the membrane. The action potential was mainly Ca(2+)-dependent and to a lesser extent, Na(+)-dependent. Upon one or several inseminations, monospermic eggs generated one fertilization potential and polyspermic eggs generated several sperm induced egg electrical responses. These sperm-induced egg electrical responses were composed of a voltage-gated action potential elicited by sperm-egg contact, resembling the action potential triggered by a depolarizing current, followed by a true sperm-gated depolarizing plateau which lasted about 60 sec and was mainly Na(+)-dependent. The number of electrical responses displayed by each egg corresponded to the number of male pronuclei detected in its cortex. An electrical response occurred for all egg membrane potentials ranging from -78 to +16 mV. In addition, the histogram giving the number of eggs in each class of egg characterized by a number of sperm entries indicated that each sperm entry constituted an independent process. However, when we compared the effectiveness of the in vitro inseminations during electrophysiological measurements and the in vivo inseminations under simulated natural conditions, the results showed two predominant classes of fertilized eggs, with one and two male pronuclei in the cortex respectively. Consequently, although our overall results exhibit features characteristic of polyspermy, they do not yet preclude the possibility that the eggs of the

  20. Mammalian Sperm Motility: Observation and Theory

    Science.gov (United States)

    Gaffney, E. A.; Gadêlha, H.; Smith, D. J.; Blake, J. R.; Kirkman-Brown, J. C.

    2011-01-01

    Mammalian spermatozoa motility is a subject of growing importance because of rising human infertility and the possibility of improving animal breeding. We highlight opportunities for fluid and continuum dynamics to provide novel insights concerning the mechanics of these specialized cells, especially during their remarkable journey to the egg. The biological structure of the motile sperm appendage, the flagellum, is described and placed in the context of the mechanics underlying the migration of mammalian sperm through the numerous environments of the female reproductive tract. This process demands certain specific changes to flagellar movement and motility for which further mechanical insight would be valuable, although this requires improved modeling capabilities, particularly to increase our understanding of sperm progression in vivo. We summarize current theoretical studies, highlighting the synergistic combination of imaging and theory in exploring sperm motility, and discuss the challenges for future observational and theoretical studies in understanding the underlying mechanics.

  1. 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. PMID:26925808

  2. Mammalian Sperm Motility: Observation and Theory

    KAUST Repository

    Gaffney, E.A.

    2011-01-21

    Mammalian spermatozoa motility is a subject of growing importance because of rising human infertility and the possibility of improving animal breeding. We highlight opportunities for fluid and continuum dynamics to provide novel insights concerning the mechanics of these specialized cells, especially during their remarkable journey to the egg. The biological structure of the motile sperm appendage, the flagellum, is described and placed in the context of the mechanics underlying the migration of mammalian sperm through the numerous environments of the female reproductive tract. This process demands certain specific changes to flagellar movement and motility for which further mechanical insight would be valuable, although this requires improved modeling capabilities, particularly to increase our understanding of sperm progression in vivo. We summarize current theoretical studies, highlighting the synergistic combination of imaging and theory in exploring sperm motility, and discuss the challenges for future observational and theoretical studies in understanding the underlying mechanics. © 2011 by Annual Reviews. All rights reserved.

  3. 精子脂质过氧化反应与致炎细胞因子%Sperm lipid peroxidation and pro-inflammatory cytokines

    Institute of Scientific and Technical Information of China (English)

    P.Martinez; F.Proverbio; M.I.Camejo

    2007-01-01

    Aim:To investigate if interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), interferon-gamma (IFN-γ) or tumor necrosis factor-alpha (TNF-α) are able to stimulate the level of lipid peroxidation of sperm membranes, either alone or in the presence of leukocytes. Methods: Semen samples from normozoospermic donors were prepared by density gradient. The sperms were exposed to the indicated cytokines, at physiological and infection-inflammation concentrations, in the absence or presence of leukocytes. Lipid peroxidation of the sperm membranes was determined by measuring malondialdehyde (MDA) and 4-hydroxialkenals (HAE) formation. Results: TNF-α, IL-8 and IFN-γ increased the level of sperm membrane lipid peroxidation when tested at physiological concentrations. At infectioninflammation concentrations, only IL-8 was able to produce a higher effect. When assayed in the presence of leucocytes,IL-8 and TNF-α showed a higher effect at infection-inflammation concentrations than at physiological concentrations.Finally, IL-8 showed a higher effect in the presence of leukocytes than in their absence at both physiological and infection-inflammation concentrations. TNF-α also showed a higher effect when assayed in the presence of leukocytes than in their absence, but only at infection-inflammation concentrations. There was no effect of IL-6 or IL- 10 in any of the tested conditions. Conclusion: Several pro-inflammatory cytokines at physiological concentrations increase the level of lipid peroxidation of sperm membranes, which could be important for the sperm fecundation process. However, infection-inflammation concentrations of some cytokines, such as IL-8 and TNF-α, either alone or in the presence of leukocytes, could drive the lipid peroxidation of the spermatozoa plasma membrane to levels that can affect the sperm fertility capacity.

  4. Effects of pH during liquid storage of goat semen on sperm viability and fertilizing potential.

    Science.gov (United States)

    Liu, Chang-He; Dong, Hai-Bo; Ma, Dong-Li; Li, You-Wei; Han, Dong; Luo, Ming-Jiu; Chang, Zhong-Le; Tan, Jing-He

    2016-01-01

    A specific problem in goat semen preservation is the detrimental effect of seminal plasma on sperm viability in extenders containing yolk or milk. Thus, the use of chemically defined extenders will have obvious advantages. Although previous studies indicate that the initial pH of an extender is crucial to sustain high sperm motility, changes in extender pH during long-term semen storage have not been observed. Monitoring extender pH at different times of semen storage and modeling its variation according to nonlinear models is thus important for protocol optimization for long-term liquid semen preservation. The present results showed that during long-term liquid storage of goat semen, both sperm motility and semen pH decreased gradually, and a strong correlation was observed between the two. Whereas increasing the initial extender pH from 6.04 to 6.25 or storage with stabilized pH improved, storage with artificially lowered pH impaired sperm motility. Extender renewal improved sperm motility by maintaining a stable pH. Sperm coating with chicken (Gallus gallus) egg yolk improved motility by increasing tolerance to pH decline. A new extender (n-mZAP) with a higher buffering capacity was formulated, and n-mZAP maintained higher sperm motility, membrane integrity and acrosome intactness than the currently used mZAP extender did. Goat semen liquid-stored for 12 d in n-mZAP produced pregnancy and kidding rates similar to those obtained with freshly collected semen following artificial insemination. In conclusion, maintenance of a stable pH during liquid semen storage dramatically improved sperm viability and fertilizing potential. PMID:26612188

  5. Changes in intracellular Ca2+ cf sperm fore-and-aft acrosome reaction in the swimming crab Portunus trituberculatus

    OpenAIRE

    ZHU Dong-Fa; Shen, Li-ping; Shen, Jian-Ming; Wu, Qiong

    2008-01-01

    The acrosome reaction process of sperm in the swimming crab Portunus trituberculatus ( Miers ) consists of four consecutive stages: (1) swelling of the apical cap; (2) eversion of the acrosomal vesicle; (3) forward elongation of the perforatorium and reversal of the acrosomal vesicle membrane; and (4) shedding of the acrosomal vesicle membrane and formation of the acrosomal filament. In the present study, Laser Scanning Confocal Microscope (LSCM) was used to image spermatozoa stained with Flu...

  6. Epidemiological studies on Trichinellosis among swine, wild boars and humans in Poland

    OpenAIRE

    Ramisz A.; Szymborski J.; Balicka-Ramisz A.

    2001-01-01

    The introduction of the digestion method (the magnetic stirrer type] resulted directly in the reduction of Trichinella sp. infection among people in Poland.Pork and its products are still the main cause of human trichinellosis in Poland. However, epidemics caused by eating wild boar meat suggested that this way of the transmission of Trichinella sp. larvae to humans might be of considerable importance.

  7. Overview of the current measuring results on radioactive contamination of wild boars

    International Nuclear Information System (INIS)

    In order to clarify the reasons for the continuously high contamination of wild boars when the soil is of low contamination, a study was initiated and sample of wild boars were analysed. From 2132 wild boars samples examined in the year 2001 in the Palatine forest, 9.4 % had more than 600 Bq/kg radiocesium (not allowed to be marketed). Wild boars take up radiocesium via their fodder. Elaphomyces granulatus play a special role, because their fruit body growing underground is a preferred delicacy. With a peak value of about 122 000 Bq/kg in the year 2002, elaphomyces granulatus in the Bavarian forest were contaminated by one order of magnitude higher than edible mushrooms. Also large thick forest areas are of great significance: the more animals move to agricultural areas on search for food and feed low contaminated fodder, the less is the radiocesium content in their muscle flesh. Breeding animals fed exclusively with agricultural products have only very low contamination levels which can be compared with that of beef, pork or chicken. (N.C.)

  8. Evaluation of different soil parameters and wild boar (Sus scrofa [L.] grassland damage

    Directory of Open Access Journals (Sweden)

    Žiga Laznik

    2014-10-01

    Full Text Available Presented in this paper are the correlations between different soil parameters [presence of grubs, earthworms, pH, content of P2O5, K2O and organic matter (OM in soil] and wild boar (Sus scrofa [L.] damage to grasslands. The soil samples and damage assessments were performed at six locations in the Kočevje region, which is a densely wooded part of South East Slovenia. A significant positive correlation was discovered between the extent of damage due to wild boar rooting in grasslands and the number of grubs (r=0.73, the weight of grubs (r=0.69 and the content of P2O5 (r=0.87 in the soil. The quantity and weight of grubs in soil were significantly influenced by soil pH, the content of CaCl2 (r=0.71/0.72, P2O5 (r=0.90/0.91, and OM (r=0.74/0.77; while the quantity and weight of earthworms in soil were influenced by the content of K2O (r=0.81/-0.84. A moderate yet insignificant correlation (r=0.48/0.56 was discovered between the number and weight of earthworms in soil and the extent of grassland damage. Grubs represent a more important source of protein for wild boars than earthworms; consequently, reducing the quantity of grubs in soil could minimise the extent of damage caused by boars.

  9. Genomics and Systems Biology of Boar Taint and Meat Quality in Pigs

    DEFF Research Database (Denmark)

    Drag, Markus; Kogelman, Lisette; Meinert, Lene;

    objective of the PhD project is to unravel the underlying mechanisms of BT at the genomic, transcriptomic, metabolomic and phenotypic levels as well as its connection with sensory meat quality (SMQ). Male pigs with different genetic merit of BT will be produced by crossbreeding Duroc boars with Landrace X...

  10. Trichinella infection in wild boars and synanthropic rats in northwest Vietnam.

    Science.gov (United States)

    Thi, N Vu; Nguyen, V D; Praet, N; Claes, L; Gabriël, S; Huyen, N T; Dorny, P

    2014-02-24

    Trichinellosis is an emerging parasitic zoonosis in North Vietnam. In this survey, hunted and farm-bred wild boars as well as synanthropic rats were sampled in two provinces of northwest Vietnam where outbreaks of trichinellosis have recently occurred. Evidence of Trichinella infection was studied by parasitological, serological and molecular methods. The results showed relatively low prevalence of Trichinella spiralis in hunted wild boars (2/62 (3.2%; 95% CI: 0.8- 4.8)) and rats (23/820 (2.8%; 95% CI: 13.7-32.3)). Parasite burdens in the muscle tissues were between 0.1 and 0.03 larvae/g, and 0.1 and 7 larvae/g in wild boars and rats, respectively. Seroprevalence in farm-bred wild boars was negative. The findings of Trichinella-infected rats in 7 of the 20 districts of Dien Bien and Son La provinces suggest that the parasite is circulating in these regions. These results indicate that the local population and health centers should be made aware of the risks of eating raw or undercooked meat dishes prepared from wild animals. PMID:24360291

  11. The influence of sex mature of wild boar (Sus scrofa) to reproduction in the Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Ježek, M.; Štípek, K.; Červený, Jaroslav

    Prague : Czech University of Life Sciences, 2009. s. 132. ISBN 978-80-213-1961-5. [European Congress of Conservation Biology /2./. 01.09.2009-05.09.2009, Prague] Institutional research plan: CEZ:AV0Z60930519 Keywords : wild boar * Czech Republic Subject RIV: EH - Ecology, Behaviour http://www.eccb2009.org/uploads/book_of_abstracts_errata.pdf

  12. Worldwide Phylogeography of Wild Boar Reveals Multiple Centers of Pig Domestication

    DEFF Research Database (Denmark)

    Larson, Greger; Dobney, Keith; Albarella, Umberto;

    2005-01-01

    Mitochondrial DNA (mtDNA) sequences from 686 wild and domestic pig specimens place the origin of wild boar in island Southeast Asia (ISEA), where they dispersed across Eurasia. Previous morphological and genetic evidence suggested pig domestication took place in a limited number of locations (pri...

  13. Activity of Cs-137 in red deer and wild boar in Slovakia

    International Nuclear Information System (INIS)

    Results of monitoring the activity of radiocesium in game animals from various parts of Slovakia are presented. Samples of game flesh were collected by veterinary officials during hunting seasons 1988-1994. More than 80 % of samples came from following districts of Slovakia: Ziar nad Hronom, Prievidza, Martin, Rimavska Sobota, Senica, Banska Bystrica, Roznava, Poprad and Spisska Nova Ves. All measurements were carried out using gamma spectrometric system equipped with 4 high purity germanium detectors. Presented results were obtained using statistical evaluation for left-censored log-normal distribution of data sets. Overall activities of Cs-137 found in red deer and wild boards in Slovakia are considerably lower, than activities reported in game animals from some parts of Northern Moravia, Southern Bohemia and Austria. While the mean activities in red deer show a decreasing tendency, mean activities of wild boar are low, but with higher occurrence of extreme values, and hence, higher variance. The observed difference could be explained by the feeding habits of wild boar: grubbing in the ground for worms, larvae, roots, etc. can lead to presence of up to 20 % of contaminated soil in their stomach. At the same time wild boars often graze farmlands, where the activity of the Cs-137 in the top soil layer is reduced by ploughing and radiocesium on clay particles. Fraction of farmlands in the home range of the wild boars and the time of shooting could contribute to observed variations in radiocesium activity. (J.K.) 2 tabs., 3 refs

  14. Effect of boar contact and housing conditions on estrus expression in weaned sows

    NARCIS (Netherlands)

    Langendijk, P.; Soede, N.M.; Kemp, B.

    2000-01-01

    Our objective was to study the effects of housing conditions and the amount of boar contact in a protocol for estrus detection on estrus detection rate, timing of onset of estrus, duration of estrus, and timing of ovulation. After weaning, 130 multiparous sows were assigned to three treatments: HI,

  15. Effect of alpha-lipoic acid on boar spermatozoa quality during freezing-thawing.

    Science.gov (United States)

    Shen, Tao; Jiang, Zhong-Liang; Li, Cong-Jun; Hu, Xiao-Chen; Li, Qing-Wang

    2016-04-01

    Alpha-lipoic acid (ALA) is known to be a natural antioxidant. The aim of the present study was to evaluate the cryoprotective effect of ALA on the motility of boar spermatozoa and its antioxidant effect on boar spermatozoa during freezing-thawing. Different concentrations (2.0, 4.0, 6.0, 8.0 or 10.0 mg/ml) of ALA were added to the extender used to freeze boar semen, and the effects on the quality and endogenous antioxidant enzyme activities of frozen-thawed spermatozoa were assessed. The results indicated that the addition of ALA to the extender resulted in a higher percentage of motile spermatozoa post-thaw (P < 0.05). The activities of superoxide dismutase, lactate dehydrogenase, glutamic-oxaloacetic transaminase and catalase improved after adding ALA to the extender (P < 0.05). Artificial insemination results showed that pregnancy rate and litter size were significantly higher at 6.0 mg/ml in the ALA group than in the control group (P < 0.05). In conclusion, ALA conferred a cryoprotective capacity to the extender used for boar semen during the process of freezing-thawing, and the optimal concentration of ALA for the frozen extender was 6.0 mg/ml. PMID:26099848

  16. Morphometrical Analysis of Reproduction Traits for the Wild Boar (Sus scrofa L. in Croatia

    Directory of Open Access Journals (Sweden)

    Nikica Šprem

    2011-09-01

    Full Text Available The wild boar (Sus scrofa L. is native game in Croatia, whose population have tendency of increasing as well throughout the Europe. The wild boar is a natural inhabitant of Europe, Asia, and North Africa and is phylogenetically the ancestor of the domestic pig. Because of its phylogenetic and economic importance, this species is an interesting model for studying testis function. Therefore, the present study was performed to investigate the testis morphometry, and gonadosomatic index (GSI for 77 individuals. The mean live body weight was 75.03 kg, testis weight was 0.355 kg and with a gonadosomatic index (GSI of approximately 0.40%. The mean circumference for the left and right testes were not significant, but a significant and positive correlation was observed between testis weight and body weight (r = 0.88, p<0.05. A high reproductive contribution of juveniles is a likely consequence of a high hunting pressure rather than a species specific life history pattern characterizing wild boar. Generally, beside female seasonal reproductive activity knowledge of male reproduction cycle in wild boar is very important for established better management of free-ranging population.

  17. Unexpected but welcome. Artificially selected traits may increase fitness in wild boar.

    Science.gov (United States)

    Fulgione, Domenico; Rippa, Daniela; Buglione, Maria; Trapanese, Martina; Petrelli, Simona; Maselli, Valeria

    2016-07-01

    Artificial selection affects phenotypes differently by natural selection. Domestic traits, which pass into the wild, are usually negatively selected. Yet, exceptionally, this axiom may fail to apply if genes, from the domestic animals, increase fertility in the wild. We studied a rare case of a wild boar population under the framework of Wright's interdemic selection model, which could explain gene flow between wild boar and pig, both considered as demes. We analysed the MC1R gene and microsatellite neutral loci in 62 pregnant wild boars as markers of hybridization, and we correlated nucleotide mutations on MC1R (which are common in domestic breeds) to litter size, as an evaluation of fitness in wild sow. Regardless of body size and phyletic effects, wild boar sows bearing nonsynonymous MC1R mutations produced larger litters. This directly suggests that artificially selected traits reaching wild populations, through interdemic gene flow, could bypass natural selection if and only if they increase the fitness in the wild. PMID:27330553

  18. Methyl-parathion decreases sperm function and fertilization capacity after targeting spermatocytes and maturing spermatozoa

    Energy Technology Data Exchange (ETDEWEB)

    Pina-Guzman, Belem; Sanchez-Gutierrez, M.; Marchetti, Francesco; Hernandez-Ochoa, I.; Solis-Heredia, M.J .; Quintanilla-Vega, B.

    2009-05-03

    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. Oxidative damage has been involved in the genotoxic and reproductive effects of OP. The purpose of this study was to determine the effects of Me-Pa on spermatozoa function and 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. DNA damage was evaluated by nick translation (NT-positive cells) and SCSA (percentDFI); lipoperoxidation (LPO) by malondialdehyde production; sperm function by spontaneous- and induced-acrosome reactions (AR); mitochondrial membrane potential (MMP) by using the JC-1 flurochrome; and, fertilization ability by an in vitro assay and in vivo mating. Results showed alterations in DNA integrity (percentDFI and NT-positive cells) at 7 and 28 dpt, in addition to decreased sperm quality and a decrease in induced-AR; reduced MMP and LPO was observed only at 7 dpt. We found negative correlations between LPO and all sperm alterations. Altered sperm functional parameters were associated with reduced fertilization rates at both times, evaluated either in vitro or in vivo. 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 ofthe detrimental effects of Me-Pa in male germ cells.

  19. Influence of semen collection method on sperm cryoresistance in small ruminants.

    Science.gov (United States)

    Jiménez-Rabadán, P; Soler, A J; Ramón, M; García-Álvarez, O; Maroto-Morales, A; Iniesta-Cuerda, M; Fernández-Santos, M R; Montoro, V; Pérez-Guzmán, M D; Garde, J J

    2016-04-01

    Semen collection for cryopreservation is a key step for small ruminant conservation programs. While in these species semen is mainly collected via artificial vagina (AV), electroejaculation (EE) provides a viable alternative for untrained males. Herein we investigated the effect of semen collection method on post-thaw sperm quality by comparing two small ruminant species, sheep and goats. Semen from Blanca-Celtibérica bucks and Manchega rams was collected by AV and EE on the same day and cryopreserved using a standard protocol. At thawing, sperm motion parameters were evaluated by CASA, whereas membrane stability (YO-PRO-1), sperm viability (propidium iodide, PI) and mitochondrial activity (Mitotracker Deep Red) were analyzed using flow cytometry. The semen collection method negatively affected post-thaw sperm quality in bucks but not in rams. Thus, in bucks, post-thaw sperm motility was higher for samples collected by AV as compared to those obtained via EE. Similarly, post-thaw sperm parameters evaluated by flow cytometry were worse for buck samples collected by EE than those collected by AV in the same species, or than ram samples regardless of collection method. These results suggest that ovine and caprine spermatozoa have a different response to the cryopreservation process depending upon the semen collection method used. We hypothesize that the EE procedure may lead to changes in the composition of the ejaculate in bucks that would make spermatozoa more susceptible to the cryopreservation process, whereas this procedure would have had no effect on ram spermatozoa. This assumption requires further investigation. PMID:26917359

  20. Regulation of Sperm Capacitation by the 26S Proteasome: An Emerging New Paradigm in Spermatology.

    Science.gov (United States)

    Kerns, Karl; Morales, Patricio; Sutovsky, Peter

    2016-05-01

    The ubiquitin proteasome system (UPS) participates in many biological processes ranging from cell cycle and antigen processing to cellular defense and signaling. Work of the last decade has made it evident that the UPS is involved in many sperm-related processes leading up to and as part of fertilization. The current knowledge of UPS involvement and changes during sperm capacitation are reviewed together with a list of known proteasome-associated sperm proteins and a discussion of the relationships between these proteins and the proteasome. Proteasomal inhibitors such as MG-132 and epoxomicin significantly alter capacitation and prevent acrosome reaction. The 26S proteasome degrades AKAP3, an A-kinase anchoring protein, partially regulating the release of protein-kinase A (PKA), a vital component necessary for the steps leading up to capacitation. Further, changes occur in 20S core subunit localization and abundance throughout capacitation. Proteasome-interacting valosine-containing protein (VCP) undergoes tyrosine phosphorylation; however, its physiological roles in capacitation and fertilization remain unknown. The E1-type ubiquitin-activating enzyme (UBA1) inhibitor PYR-41 also alters acrosomal membrane remodeling during capacitation. Furthermore, after capacitation, the acrosomal proteasomes facilitate the degradation of zona pellucida glycoproteins leading up to fertilization. Methods to modulate the sperm proteasome activity during sperm storage and capacitation may translate to increased reproductive efficiency in livestock animals. Human male infertility diagnostics may benefit from incorporation of research outcomes built upon relationships between UPS and capacitation. Altogether, the studies reviewed here support the involvement of UPS in sperm capacitation and present opportunities for new discoveries. PMID:27053366

  1. Methyl-parathion decreases sperm function and fertilization capacity after targeting spermatocytes and maturing spermatozoa

    International Nuclear Information System (INIS)

    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.

  2. Sperm DNA oxidative damage and DNA adducts.

    Science.gov (United States)

    Jeng, Hueiwang Anna; Pan, Chih-Hong; Chao, Mu-Rong; Lin, Wen-Yi

    2015-12-01

    The objective of this study was to investigate DNA damage and adducts in sperm from coke oven workers who have been exposed to polycyclic aromatic hydrocarbons. A longitudinal study was conducted with repeated measurements during spermatogenesis. Coke-oven workers (n=112) from a coke-oven plant served the PAH-exposed group, while administrators and security personnel (n=67) served the control. Routine semen parameters (concentration, motility, vitality, and morphology) were analyzed simultaneously; the assessment of sperm DNA integrity endpoints included DNA fragmentation, bulky DNA adducts, and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dGuo). The degree of sperm DNA fragmentation was measured using the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and sperm chromatin structure assay (SCSA). The PAH-exposed group had a significant increase in bulky DNA adducts and 8-oxo-dGuo compared to the control subjects (Ps=0.002 and 0.045, respectively). Coke oven workers' percentages of DNA fragmentation and denaturation from the PAH-exposed group were not significantly different from those of the control subjects (Ps=0.232 and 0.245, respectively). Routine semen parameters and DNA integrity endpoints were not correlated. Concentrations of 8-oxo-dGuo were positively correlated with percentages of DNA fragmentation measured by both TUNEL and SCSA (Ps=0.045 and 0.034, respectively). However, the concentrations of 8-oxo-dGuo and percentages of DNA fragmentation did not correlate with concentrations of bulky DNA adducts. In summary, coke oven workers with chronic exposure to PAHs experienced decreased sperm DNA integrity. Oxidative stress could contribute to the degree of DNA fragmentation. Bulky DNA adducts may be independent of the formation of DNA fragmentation and oxidative adducts in sperm. Monitoring sperm DNA integrity is recommended as a part of the process of assessing the impact of occupational and environmental toxins on sperm

  3. CHROMOSOMAL ABNORMALITIES IN PATIENTS WITH SPERM DISORDERS

    OpenAIRE

    L. Y. Pylyp; L. A. Spinenko; V. D. Zukin; N. M. Bilko

    2013-01-01

    Chromosomal abnormalities are among the most common genetic causes of spermatogenic disruptions. Carriers of chromosomal abnormalities are at increased risk of infertility, miscarriage or birth of a child with unbalanced karyotype due to the production of unbalanced gametes. The natural selection against chromosomally abnormal sperm usually prevents fertilization with sperm barring in cases of serious chromosomal abnormalities. However, assisted reproductive technologies in general and intrac...

  4. Role of the epididymis in sperm competition

    Institute of Scientific and Technical Information of China (English)

    Russell C. Jones; Jean-Louis Dacheux; Brett Nixon; Heath W. Ecroyd

    2007-01-01

    Although it is generally understood that the testes recruited kidney ducts for reproductive function during the evolution of vertebrates, little is understood of the biological significance of the adaptation. In the context of the evolution of the mammalian epididymis, this report provides evidence that a major role of the epididymis is to enhance a male's chance of achieving paternity in a competitive mating system. A unique example of sperm cooperation in monotremes is used as evidence that the epididymis produces sperm competition proteins to form groups of 100 sperm into bundles that have a forward motility nearly thrice that of individual spermatozoa. As it required 3-h incubation in vitro under capacitation conditions to release motile sperm from the bundles, it is suggested that the monotremes provide an example of capacitation that is quite different from capacitation in higher mammals. It is Suggested that variation between species in the intensity of sperm competition could explain the variation that occurs between species in the amount of post-testicular sperm maturation and storage in the epididymis, an explanation of why the human epididymis does not play as important a role in reproduction as the epididymis of most mammals.

  5. Lifestyle influences human sperm functional quality

    Institute of Scientific and Technical Information of China (English)

    Mnica Ferreira; Joana Vieira Silva; Vladimiro Silva; Antnio Barros; Margarida Fardilha

    2012-01-01

    Objective:To investigate the impact of acute lifestyle changes on human sperm functional quality.Methods:In the academic festivities week, young and apparently healthy male students who voluntarily submit themselves to acute lifestyle alterations(among the potentially important variations are increase in alcohol, caffeine, and tobacco consumption and circadian rhythm shifts) were used as a model system.Sperm samples were obtained before and after the academic week and compared by traditional semen analysis(n=54) and also tested for cleavedPolyADP-ribose polymerase(PARP) protein, an apoptotic marker(n=35).Results:Acute lifestyle changes that occurred during the academic week festivities(the study model) resulted both in a significant reduction in sperm quality, assessed by basic semen analysis(decrease in sperm concentration, total number of spermatozoa, progressive and non-progressive motility and increase in sperm morphological abnormalities) and by an increase in the expression of the apoptotic marker, cleavedPARP, in the ejaculate.Conclusions:Acute lifestyle changes have clear deleterious effects on sperm quality.We propose cleavedPARP as a novel molecular marker, valuable for assessing spermquality in parallel with the basic semen analysis method.

  6. Sperm competitiveness in frogs: slow and steady wins the race.

    Science.gov (United States)

    Dziminski, Martin A; Roberts, J Dale; Beveridge, Maxine; Simmons, Leigh W

    2009-11-22

    When sperm compete to fertilize available ova, selection is expected to favour ejaculate traits that contribute to a male's fertilization success. While there is much evidence to show that selection favours increased numbers of sperm, only a handful of empirical studies have examined how variation in sperm form and function contributes to competitive fertilization success. Here, we examine selection acting on sperm form and function in the externally fertilizing myobatrachid frog, Crinia georgiana. Using in vitro fertilization techniques and controlling for variation in the number of sperm contributed by males in competitive situations, we show that males with a greater proportion of motile sperm, and motile sperm with slower swimming velocities, have an advantage when competing for fertilizations. Sperm morphology and the degree of genetic similarity between putative sires and the female had no influence on competitive fertilization success. These unusual patterns of selection might explain why frog sperm typically exhibit relatively slow swimming speeds and sustained longevity. PMID:19710059

  7. Sperm ultrastructure of the spider crab Maja brachydactyla (Decapoda: Brachyura).

    Science.gov (United States)

    Simeó, Carles G; Kurtz, Kathryn; Rotllant, Guiomar; Chiva, Manel; Ribes, Enric

    2010-04-01

    This study describes the morphology of the sperm cell of Maja brachydactyla, with emphasis on localizing actin and tubulin. The spermatozoon of M. brachydactyla is similar in appearance and organization to other brachyuran spermatozoa. The spermatozoon is a globular cell composed of a central acrosome, which is surrounded by a thin layer of cytoplasm and a cup-shaped nucleus with four radiating lateral arms. The acrosome is a subspheroidal vesicle composed of three concentric zones surrounded by a capsule. The acrosome is apically covered by an operculum. The perforatorium penetrates the center of the acrosome and has granular material partially composed of actin. The cytoplasm contains one centriole in the subacrosomal region. A cytoplasmic ring encircles the acrosome in the subapical region of the cell and contains the structures-organelles complex (SO-complex), which is composed of a membrane system, mitochondria with few cristae, and microtubules. In the nucleus, slightly condensed chromatin extends along the lateral arms, in which no microtubules have been observed. Chromatin fibers aggregate in certain areas and are often associated with the SO-complex. During the acrosomal reaction, the acrosome could provide support for the penetration of the sperm nucleus, the SO-complex could serve as an anchor point for chromatin, and the lateral arms could play an important role triggering the acrosomal reaction, while slightly decondensed chromatin may be necessary for the deformation of the nucleus. PMID:19885919

  8. Association of heat shock protein 90 with motility of post-thawed sperm in bulls.

    Science.gov (United States)

    Zhang, Xiao-Gang; Hu, Shan; Han, Cong; Zhu, Qing-Chao; Yan, Guan-Jie; Hu, Jian-Hong

    2015-04-01

    The correlation between the 90 kDa heat-shock protein (HSP90) and sperm quality following the process of freezing-thawing in bulls has not been studied clearly. Therefore, the objective of the present was to clarify the relationship between HSP90 level and semen parameters during the process of cryopreservation in bulls. Semen samples from 5 Holstein bulls were obtained by artificial vagina. Characteristics of these semen at three stages (fresh, after equilibration and frozen-thawed), including motility, plasma membrane integrity and acrosome integrity were evaluated. The mRNA expression level of HSP90 at the three stages was evaluated by using quantitative Real-Time PCR. Meanwhile, the protein level of HSP90 expression at the three stages was detected according to Western blot. The results showed that sperm parameters evaluated in fresh semen was the highest in the three groups. Sperm parameters in semen after equilibration were lower than those in fresh semen (P>0.05) and higher than those in post-thawed semen (PSperm parameters in frozen-thawed semen were the lowest among the three groups (PHSP90 expression is proportional to sperm quality. HSP90 expression level in fresh semen was significantly higher than that in frozen-thawed semen (PHSP90 expression were observed between fresh semen and semen after equilibration (P>0.05). Results in this study suggest that HSP90 level in bull spermatozoa was gradually declined following the process of freezing-thawing, and might be associated with sperm motility, plasma membrane integrity and acrosome integrity. PMID:25578982

  9. A patch-clamp study on human sperm Cl-channel reassembled into giant liposome

    Institute of Scientific and Technical Information of China (English)

    Jun-Ping BAI; Yu-Liang SHI

    2001-01-01

    Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl-channels in human sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembled into liposome bilayer, and the liposomes were fused into giant liposomes with a diameter more than 10 μm by dehydrationrehydration procedure. The giant liposomes were used to study the Cl-channel activities by patch-clamp technique.Results: By patch clamping the giant liposome in an asymmetric NMDG (N-methyl-D-glucamine)-Cl (bath 100//pipette 200 mmol/L) solution system, three kinds of single-channel events with unit conductances of (74.1 ± 8.3) pS,( 117.0 ± 5.7) pS and ( 144.7 ±+ 4.5 ) pS, respectively, were detected. Their activities were voltage-dependent and all manner. By constructing the open and close dwell time distribution histograms and then fitting them with exponential function, two time constants were obtained in both the open and the close states. The burst activity and conductance substate of the channels were observed. Conclusion: There exist three kinds of Cl-channels with different conductance in human sperm membrane at least.

  10. Sperm selection in assisted reproductive technology: An update%辅助生殖中的精子优选研究进展

    Institute of Scientific and Technical Information of China (English)

    宋岳强; 沙艳伟

    2012-01-01

    精子优选是辅助生殖技术中的重要环节.精子活力和形态等常规参数是评估精子的重要内容,近年来精子超微结构、DNA完整性、凋亡和膜成熟度等特征逐渐受到关注,针对这些特征筛选精子成为精子优选技术发展的方向.本文介绍了近年来发展起来的电泳法、电动电位法、透明质酸结合法、膜联蛋白V选择法、形态选择性单精子注射技术(IMSI)和微流控芯片法的技术原理,除微流控芯片法外其它技术均已应用于IVF或ICSI治疗中,同时对各种技术的优缺点和应用效果等方面做一综述.%Sperm selection plays an important role in assisted reproductive technology. In recent years, sperm evaluation is not limited to the assessment of sperm motility and morphology, but involves more other sperm characteristics such as sperm infrastructure, DNA integrity, apoptosis and membrane. Assessment based on these characteristics is becoming the aim of sperm selection. This article gives an overview on several newly developed techniques for sperm selection according to different technical principles, such as electrophoretic separation, zeta potential, HA binding, Annexin V binding, intracytoplasmic morphologically selected sperm injection (IMSI) and microfluidic sperm sorter, which have all been applied to IVF or ICSI with the exception of microfluidic sperm sorter. It also introduces the advantages, disadvantages and application effects of these techniques.

  11. Comparison of efficacy of two techniques for testicular sperm retrieval in nonobstructive azoospermia: multifocal testicular sperm extraction versus multifocal testicular sperm aspiration.

    Science.gov (United States)

    Hauser, Ron; Yogev, Leah; Paz, Gedalia; Yavetz, Haim; Azem, Fuad; Lessing, Joseph B; Botchan, Amnon

    2006-01-01

    To compare the efficacy of 2 sperm-retrieval procedures, testicular sperm extraction (TESE) and testicular sperm aspiration (TESA), during the same procedure using the same subjects as their own controls. The presence of mature testicular sperm cells and motility were evaluated in 87 men with nonobstructive azoospermia (NOA) by means of multifocal TESE and multifocal TESA, which were performed during the same procedure using the same subjects as their own controls. Sperm cells were recovered by TESE in 54 cases, but by TESA in only 36 cases. There were significantly more cases (n = 20) in which sperm cells were recovered by TESE only, compared with 2 cases in whom cells were recovered by TESA only (McNemar's test, P < .001). The mean number of locations in each testis in which sperm cells were detected was significantly higher in the TESE group. In significantly more cases (n = 27), motility was observed in TESE material only, compared with 3 cases in which motility was present in material extracted by TESA only (McNemar's test, P < .001). Mean number of locations in each testis with motile sperm cells was significantly higher in the TESE group. The TESE procedure yielded significantly more sperm cells, as was also reflected by the difference in number of straws with cryopreserved sperm. This comparative prospective clinical study revealed that multifocal TESE is more efficient than multifocal TESA for sperm detection and recovery in men with NOA and should be the procedure of choice for sperm retrieval for them. PMID:16400074

  12. Severity of bovine tuberculosis is associated with co-infection with common pathogens in wild boar.

    Science.gov (United States)

    Risco, David; Serrano, Emmanuel; Fernández-Llario, Pedro; Cuesta, Jesús M; Gonçalves, Pilar; García-Jiménez, Waldo L; Martínez, Remigio; Cerrato, Rosario; Velarde, Roser; Gómez, Luis; Segalés, Joaquím; Hermoso de Mendoza, Javier

    2014-01-01

    Co-infections with parasites or viruses drive tuberculosis dynamics in humans, but little is known about their effects in other non-human hosts. This work aims to investigate the relationship between Mycobacterium bovis infection and other pathogens in wild boar (Sus scrofa), a recognized reservoir of bovine tuberculosis (bTB) in Mediterranean ecosystems. For this purpose, it has been assessed whether contacts with common concomitant pathogens are associated with the development of severe bTB lesions in 165 wild boar from mid-western Spain. The presence of bTB lesions affecting only one anatomic location (cervical lymph nodes), or more severe patterns affecting more than one location (mainly cervical lymph nodes and lungs), was assessed in infected animals. In addition, the existence of contacts with other pathogens such as porcine circovirus type 2 (PCV2), Aujeszky's disease virus (ADV), swine influenza virus, porcine reproductive and respiratory syndrome virus, Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae, Haemophilus parasuis and Metastrongylus spp, was evaluated by means of serological, microbiological and parasitological techniques. The existence of contacts with a structured community of pathogens in wild boar infected by M. bovis was statistically investigated by null models. Association between this community of pathogens and bTB severity was examined using a Partial Least Squares regression approach. Results showed that adult wild boar infected by M. bovis had contacted with some specific, non-random pathogen combinations. Contact with PCV2, ADV and infection by Metastrongylus spp, was positively correlated to tuberculosis severity. Therefore, measures against these concomitant pathogens such as vaccination or deworming, might be useful in tuberculosis control programmes in the wild boar. However, given the unexpected consequences of altering any community of organisms, further research should evaluate the impact of such measures under

  13. Severity of bovine tuberculosis is associated with co-infection with common pathogens in wild boar.

    Directory of Open Access Journals (Sweden)

    David Risco

    Full Text Available Co-infections with parasites or viruses drive tuberculosis dynamics in humans, but little is known about their effects in other non-human hosts. This work aims to investigate the relationship between Mycobacterium bovis infection and other pathogens in wild boar (Sus scrofa, a recognized reservoir of bovine tuberculosis (bTB in Mediterranean ecosystems. For this purpose, it has been assessed whether contacts with common concomitant pathogens are associated with the development of severe bTB lesions in 165 wild boar from mid-western Spain. The presence of bTB lesions affecting only one anatomic location (cervical lymph nodes, or more severe patterns affecting more than one location (mainly cervical lymph nodes and lungs, was assessed in infected animals. In addition, the existence of contacts with other pathogens such as porcine circovirus type 2 (PCV2, Aujeszky's disease virus (ADV, swine influenza virus, porcine reproductive and respiratory syndrome virus, Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae, Haemophilus parasuis and Metastrongylus spp, was evaluated by means of serological, microbiological and parasitological techniques. The existence of contacts with a structured community of pathogens in wild boar infected by M. bovis was statistically investigated by null models. Association between this community of pathogens and bTB severity was examined using a Partial Least Squares regression approach. Results showed that adult wild boar infected by M. bovis had contacted with some specific, non-random pathogen combinations. Contact with PCV2, ADV and infection by Metastrongylus spp, was positively correlated to tuberculosis severity. Therefore, measures against these concomitant pathogens such as vaccination or deworming, might be useful in tuberculosis control programmes in the wild boar. However, given the unexpected consequences of altering any community of organisms, further research should evaluate the impact of such measures

  14. Variation in sperm morphometry and sperm competition among barn swallow (Hirundo rustica) populations

    Czech Academy of Sciences Publication Activity Database

    Laskemoen, T.; Albrecht, Tomáš; Bonisoli-Alquati, A.; Cepák, J.; De Lope, F.; Hermosell, I. G.; Johannessen, L. E.; Kleven, O.; Marzal, A.; Mousseau, T. A.; Moller, A. P.; Robertson, R. J.; Rudolfsen, G.; Saino, N.; Vortman, Y.; Lifjeld, J. T.

    2013-01-01

    Roč. 67, č. 2 (2013), s. 301-309. ISSN 0340-5443 R&D Projects: GA ČR(CZ) GAP506/12/2472 Institutional support: RVO:68081766 Keywords : Barn swallow * Extrapair paternity * Hirundo rustica * Sperm competition * Sperm size Subject RIV: EG - Zoology Impact factor: 3.049, year: 2013

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

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past two decades, Denmark has developed into an important destination for fertility travelers in need of donor sperm. A variety of different clinics providing sperm with different modes of clinical care have developed, and an increasing number of their customers come from abroad. One o...

  16. SMART USE OF COMPUTER-AIDED SPERM ANALYSIS (CASA) TO CHARACTERIZE SPERM MOTION

    Science.gov (United States)

    Computer-aided sperm analysis (CASA) has evolved over the past fifteen years to provide an objective, practical means of measuring and characterizing the velocity and parttern of sperm motion. CASA instruments use video frame-grabber boards to capture multiple images of spermato...

  17. MONTHLY VARIATION IN SPERM MOTILITY IN COMMON CARP ASSESSED USING COMPUTER-ASSISTED SPERM ANALYSIS (CASA)

    Science.gov (United States)

    Sperm motility variables from the milt of the common carp Cyprinus carpio were assessed using a computer-assisted sperm analysis (CASA) system across several months (March-August 1992) known to encompass the natural spawning period. Two-year-old pond-raised males obtained each mo...

  18. Prudent sperm use by leaf-cutter ant queens

    DEFF Research Database (Denmark)

    Den Boer, Susanne Petronella A; Baer, Boris; Dreier, Stéphanie Agnès Jeanine;

    2009-01-01

    In many species, females store sperm between copulation and egg fertilization, but the consequences of sperm storage and patterns of sperm use for female life history and reproductive success have not been investigated in great detail. In hymenopteran insect societies (ants, bees, wasps), reprodu......In many species, females store sperm between copulation and egg fertilization, but the consequences of sperm storage and patterns of sperm use for female life history and reproductive success have not been investigated in great detail. In hymenopteran insect societies (ants, bees, wasps...

  19. The Semen pH Affects Sperm Motility and Capacitation

    OpenAIRE

    Ji Zhou; Li Chen; Jie Li; Hongjun Li; Zhiwei Hong; Min Xie; Shengrong Chen; Bing Yao

    2015-01-01

    As the chemical environment of semen can have a profound effect on sperm quality, we examined the effect of pH on the motility, viability and capacitation of human sperm. The sperm in this study was collected from healthy males to avoid interference from other factors. The spermatozoa cultured in sperm nutrition solution at pH 5.2, 6.2, 7.2 and 8.2 were analyzed for sperm total motility, progressive motility (PR), hypo-osmotic swelling (HOS) rate, and sperm penetration. Our results showed tha...

  20. Cross-Reactivity of Porcine Immunoglobulin A Antibodies with Fecal Immunoglobulins of Wild Boar (Sus scrofa) and Other Animal Species

    Science.gov (United States)

    Seo, Sang won; Yoo, Sung J.; Sunwoo, Sunyoung; Hyun, Bang hun

    2016-01-01

    Fecal samples obtained from wild boar habitats are useful for the surveillance of diseases in wild boar populations; however, it is difficult to determine the species of origin of feces collected in natural habitats. In this study, a fecal IgA ELISA was evaluated as a method for identifying the porcine species from fecal samples. Both domestic pigs (Sus scrofa domestica) and wild boars (Sus scrofa coreanus) showed significantly higher levels of fecal IgA than other animal species. Additionally, age dependent changes in the level of Ig A in wild boars and domestic pigs were identified; Titers of Ig A were highest in suckling period and lowest in weanling period. PMID:27340389