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Sample records for boar sperm evaluated

  1. Evaluation of sperm chromatin structure in boar semen

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    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.

  2. 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...... induced AR. A detailed description of the dynamics of sperm viability and acrosomal status of boar sperm following in vitro induction of the AR has to our knowledge not previously been conducted. In the present study, a triple color flow cytometric detection technique was used, which gave simultaneous...... 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...

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

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    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 (pboar sperm.

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

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

    2005-04-15

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

  5. EFFECT OF SEASON ON BOAR SPERM MORPHOLOGY

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    Jan LIPENSKÝ

    2011-01-01

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

  6. Use of heterospermic inseminations and paternity testing to evaluate the relative contributions of common sperm traits and seminal plasma proteins in boar fertility.

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    Flowers, W L; Deller, F; Stewart, K R

    2016-11-01

    The objective of this study was to evaluate relationships between common semen quality estimates including sperm motility, sperm morphology, spontaneous capacitation status and seminal plasma proteins and boar fertility using heterospermic inseminations and subsequent paternity testing. All boars (n=12) used in the study had excellent semen quality (≥70% normal sperm) that resulted in average farrowing rates and litter sizes of 88.9±0.7% and 11.7±0.1 pigs, respectively. Their ejaculates were combined to make heterospermic insemination doses in such a way that each boar was tested against all of his contemporaries. The proportion of piglets sired by each individual was used to separate boars into three fertility groups: High (71.6±4.8%; n=3); Medium (51.6±3.8%; n=6); and Low (25.2%±5.3%; n=3). Ejaculates from High fertility boars had more motile sperm with normal acrosomes that moved faster in a straight-line and were more likely to undergo an acrosome reaction (p≤0.05) compared with their counterparts in the Low fertility group. Ejaculates from High fertility boars contained the greatest concentrations of three seminal plasma proteins (25.9kD/5.9pI; 55.1kD/4.8pI; and 70.1kD/5.2pI; p≤0.05), whereas concentrations of a 19.1kD/6.8pI were highest in semen from Low fertility boars (p≤0.05). Multiple regression analyses indicated that concentrations of the 25.9kD/5.9pI seminal plasma protein explained 66% of the variation observed in the proportion of pigs sired within a litter among boars (p≤0.00001). These results demonstrate that heterospermic inseminations and subsequent paternity testing is an effective technique for defining relationships between common semen quality tests and fertility, especially in situations where reproductive performance of all the boars is high. Motility, normal acrosome morphology, average linear velocity of motile sperm, and the proportion of sperm capable of an acrosome reaction were all positively associated with boar

  7. 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?

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    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

    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 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

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

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

    2016-01-01

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

  9. The nuclear DNA longevity in cryopreserved boar spermatozoa assessed using the Sperm-Sus-Halomax.

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    Alkmin, Diego V; Martinez-Alborcia, Maria J; Parrilla, Inmaculada; Vazquez, Juan M; Martinez, Emilio A; Roca, Jordi

    2013-06-01

    The aim of this experimental study was to evaluate the dynamics of nuclear DNA fragmentation in frozen-thawed (FT) boar spermatozoa incubated over time. Using the Sperm Chromatin Dispersion test (Sperm-Sus-Halomax), this study focused special attention on resolving the hypothesis that the original halo shapes around the sperm head could show dynamic changes over the postthawing incubation time. Twenty FT sperm samples from five boars (four per boar) were incubated at 37 °C during 168 hours and sperm motility (assessed using computer-assisted sperm analysis), viability (evaluated using the LIVE/DEAD Sperm Viability Kit), and nuclear DNA fragmentation were analyzed at 0, 0.5, 2, 4, 6, 24, 48, 72, and 168 hours. The percentages of motile and viable spermatozoa progressively decreased during incubation, with no motile and viable spermatozoa less than 10% in all boars at 24 hours of incubation. Four different halo shapes around the sperm head were considered in the Sperm Chromatin Dispersion test: normal, small, large scattered (typical fragmented nuclear DNA), and absent halo, all of them coexisting at the same time in the boar FT semen samples. Sperm with a large scattered halo did not change during postthaw, consistently showing percentages less than 5% over time in all boars. In contrast, the other three sperm populations showed a dynamic evolution over incubation time, characterized by a gradual reduction of sperm with normal halo, proportional to the increment in the sperm showing a small halo, followed by a switch between the sperm with a small halo and sperm with no halo. These results suggest that three of these four sperm populations, those showing small, large scattered, and absent halo, represent spermatozoa with different degrees of nuclear DNA damage, which should be taken into consideration to indicate the percentage of sperm with fragmented nuclear DNA in boar FT semen samples.

  10. Boar sperm thawing practices: the number of straws does matter.

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    Casas, I; Torner, E; Yeste, M; Bonet, S

    2012-04-15

    The number of straws thawed has been largely neglected in reports of boar sperm cryopreservation. Whereas previous studies confirm the effect of sperm concentration on function and survival of thawed boar spermatozoa, it is still unknown whether, for a same concentration, total number of sperm in the thawing solution affects its mechanics. The present trial sought to define good boar sperm thawing practices by checking if a minimal number of straws as well as the percentage of air volume in the thawing tube should be stated or not to decrease variability from one trial to another. In a first assay, three tubes with different numbers of thawed straws were compared in terms of motility and membrane integrity: control (C, four straws), T1.1 (two straws), and T1.2 (one straw). In a second parallel assay, the sperm motility was evaluated when one straw was thawed in a tube containing 86.67% of air volume (T2.1), and when the tube contained sperm in Beltsville thawing solution (BTS) was 1:3 (v:v) and quality parameters were assessed 4 h after thawing. Results showed the number of straws does affect motility parameters but not the membrane integrity, whereas less air volume in the tube nonsignificantly minimizes data deviation among replicates. In conclusion, it is recommended the use of four straws at 1:3 (v:v) to maintain motility records in boar sperm thawing practices as well as to be provided with vials that fit the sperm volume.

  11. Microscopic analysis of MTT stained boar sperm cells

    OpenAIRE

    B.M. van den Berg

    2015-01-01

    The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few ...

  12. Seasonal variation in sperm characteristics of boars in southern Uruguay

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    Hugo Petrocelli

    2015-01-01

    Full Text Available The objective of this study was to evaluate the effects of season, natural photoperiod, and room temperature at the housing facility on boar semen characteristics in Uruguay (34º66'S; 56º29'W. For this purpose, 117 ejaculates, obtained from eight adult males collected through 12 consecutive months, were assessed for sperm viability, DNA integrity, abnormalities (total, primary, and secondary, ejaculate volume, and sperm concentration. Viability, total and primary abnormalities, volume, and sperm concentration were affected by season. Sperm viability, volume, and sperm concentration were affected by natural photoperiod. In general, autumn and the decreasing photoperiod had a negative impact on most of the semen characteristics, except for volume. Housing temperature did not affect semen characteristics. In boars living in temperate climates, semen quality is negatively affected during autumn and is related to photoperiod changes; however, the effects of temperature changes in housingdo not affect these seminal characteristics. In this scenario, seasonal differences in semen quality may have a negative effect on sow fertilization. Consequently, semen quality control especially during autumn is imperative for the best boar selection to be used for insemination purposes. Seasonal differences in semen quality may have a negative effect on sow reproductive performance. This issue will be addressed in a future investigation.

  13. Effect of Pseudomonas aeruginosa on sperm capacitation and protein phosphorylation of boar spermatozoa.

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    Sepúlveda, Lilian; Bussalleu, Eva; Yeste, Marc; Bonet, Sergi

    2016-05-01

    Several studies have reported the detrimental effects that bacteriospermia causes on boar sperm quality, but little is known about its effects on IVC. Considering that, the present study sought to evaluate the effects of different concentrations of Pseudomonas aeruginosa on different indicators of capacitation status (sperm viability, membrane lipid disorder, sperm motility kinematics, and protein phosphorylation of boar spermatozoa) after IVC. Flow cytometry and computer assisted sperm analysis (CASA) revealed that the presence of P aeruginosa in boar sperm samples, mostly at concentrations greater than 10(6) CFU/mL, is associated with a significant (P sperm membrane integrity and sperm with low membrane lipid disorder, and also with a reduction in sperm motility kinetic parameters when compared with results obtained from the control sample, which presented the typical motility pattern of capacitated-like boar spermatozoa. Moreover, Western blot results also showed significant (P boar sperm, being the most relevant. Indeed, after 3 hours of IVC, phosphotyrosine levels of p32 in the control sample were 3.13 ± 0.81, whereas in the tubes with 10(6) and 10(8) CFU/mL were 1.05 ± 0.20 and 0.36 ± 0.07, respectively. Therefore, the present study provides novel data regarding the effects of bacterial contamination on boar sperm, suggesting that the presence of P aeruginosa affects the fertilizing ability of boar sperm by altering its ability to accomplish IVC.

  14. Acrosin activity is a good predictor of boar sperm freezability.

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    Pinart, Elisabeth; Yeste, Marc; Bonet, Sergi

    2015-06-01

    The main aim of this study was to determine whether acrosin activity could predict boar sperm freezability. For this purpose, we characterized the changes in sperm quality and acrosin activity throughout the cryopreservation procedure of sperm samples from 30 Pietrain boars by analyzing four critical steps: step 1 (extended sperm at 15 °C), step 2 (cooled sperm at 5 °C), step 3 (30 minutes postthaw), and step 4 (240 minutes postthaw). Freezability ejaculate groups were set on the basis of sperm motility and membrane integrity after freeze-thawing. Results obtained highlighted the low predictive value in terms of freezability of sperm motility and kinematics and sperm membrane integrity, as no differences between good and poor freezability ejaculates were seen before cryopreservation. Significant differences (P sperm kinetic parameters, and after thawing for sperm motility and membrane integrity. In contrast, acrosin activity appeared as an indicator of boar sperm freezability because the differences (P sperm kinematics, membrane lipid disorder, intracellular calcium content, acrosome integrity, and acrosin activity throughout the cryopreservation procedure were indicative of a significant damage in spermatozoa during the cooling step in both ejaculate groups. In conclusion, the main finding of our study is that acrosin activity can be used as a reliable predictor of boar sperm freezability because it differs significantly between good and poor freezability ejaculates yet before freeze-thawing procedures took place, i.e., in the refrigeration step at 15 °C.

  15. Phosphatidylinositol 3-kinase pathway regulates sperm viability but not capacitation on boar spermatozoa.

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    Aparicio, I M; Bragado, M J; Gil, M C; Garcia-Herreros, M; Gonzalez-Fernandez, L; Tapia, J A; Garcia-Marin, L J

    2007-08-01

    Phosphatidylinositol 3-kinase (PI3-K) plays an important role in cell survival in somatic cells and recent data pointed out a role for this kinase in sperm capacitation and acrosome reaction (AR). This study was undertaken to evaluate the role of PI3-K pathway on porcine spermatozoa capacitation, AR, and viability using two unrelated PI3-K inhibitors, LY294002 and wortmannin. In boar spermatozoa, we have identified the presence of PDK1, PKB/Akt, and PTEN, three of the main key components of the PI3-K pathway. Incubation of boar sperm in a capacitating medium (TCM) caused a significant increase in the percentage of capacitated (25 +/- 2 to 34 +/- 1% P sperm in basal medium (TBM). Inhibition of PI3-K did affect neither the capacitation status nor AR nor protein p32 tyrosine phosphorylation of boar spermatozoa incubated in TBM or TCM. Boar sperm viability in TBM was significantly decreased by 40 and 20% after pretreatment with LY294002 or wortmannin, respectively. Similar results were observed after incubation of boar spermatozoa in TCM. Treatment of boar spermatozoa with the analog of cAMP, 8Br-cAMP significantly prevented the reduction on sperm viability. Our results provide evidence for an important role of the PI3-K pathway in the regulation of boar sperm viability and suggests that other signaling pathways different from PI3-K must be activated downstream of cAMP to contribute to regulation of sperm viability. Finally, in our conditions the PI3-K pathway seems not related with boar sperm capacitation or AR.

  16. Effects of different concentrations of Pseudomonas aeruginosa on boar sperm quality.

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    Sepúlveda, Lilian; Bussalleu, Eva; Yeste, Marc; Bonet, Sergi

    2014-11-30

    Bacteriospermia in boar ejaculates is a frequent finding that compromises the sperm quality and, consequently, causes economic losses in swine industry. The present study sought to evaluate the effect of different concentrations of Pseudomonas aeruginosa on boar sperm quality over a storing period of 11 days at 15-17 ° C. Ten commercial seminal doses coming from post-pubertal and healthy boars were artificially inoculated with different infective concentrations of P. aeruginosa, ranging from 2 × 10(8) to 2 × 10(4)cfu/mL. Negative controls were non-inoculated doses. Sperm quality, assessed as sperm motility (CASA), sperm viability, acrosome integrity and pH, as well as the bacterial growth, were checked after 0, 1, 2, 4, 7, 9 and 11 days of storage at 15-17 ° C. Results obtained showed significant decreases in the percentages of total and progressive sperm motility, sperm viability and acrosome integrity in the greatest infective concentrations (2 × 10(7) and 2 × 10(8)cfu/mL), when compared to the negative control. In contrast, there was no effect on seminal pH throughout the experiment. Results indicate the presence of P. aeruginosa in boar semen, apart from being a potential source for the spread of infectious diseases and harmful impact on sows, negatively affects the longevity and fertilizing ability of boar sperm when present in high concentrations. Thus, P. aeruginosa causes deleterious effects on boar sperm quality during liquid storage at 15-17 ° C, thus strict hygienic measures must be implemented in boar studs to minimize bacterial concentration of semen doses.

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

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

    2014-07-01

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

  18. Spermometer: electrical characterization of single boar sperm motility

    NARCIS (Netherlands)

    Wagenaar, de Bjorn; Geijs, Daan J.; Boer, de Hans; Bomer, Johan G.; Olthuis, Wouter; Berg, van den Albert; Segerink, Loes I.

    2016-01-01

    Objective: To study single sperm boar motility using electrical impedance measurements in a microfluidic system. Design: Comparison of the optical data and electrical impedance data. Setting: Research laboratory at a university. Animal(s): Boar semen sample were used. Intervention(s): A micr

  19. Classification of Boar Sperm Head Images using Learning Vector Quantization

    NARCIS (Netherlands)

    Biehl, Michael; Pasma, Piter; Pijl, Marten; Sánchez, Lidia; Petkov, Nicolai; Verleysen, Michel

    2006-01-01

    We apply Learning Vector Quantization (LVQ) in automated boar semen quality assessment. The classification of single boar sperm heads into healthy (normal) and non-normal ones is based on grey-scale microscopic images only. Sample data was classified by veterinary experts and is used for training a

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

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    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.

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

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

    1980-01-01

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

  2. Effect of dietary supplementation with amino acids on boar sperm quality and fertility.

    Science.gov (United States)

    Dong, Hong-Jun; Wu, De; Xu, Sheng-Yu; Li, Qiang; Fang, Zheng-Feng; Che, Lian-Qiang; Wu, Cai-Mei; Xu, Xue-Yu; Lin, Yan

    2016-09-01

    The aim of this study was to evaluate the effects of dietary supplementation with amino acids on sperm quality and fertility rates after insemination with boar semen. Twelve Yorkshire boars were paired by age and allocated to one of two dietary treatments composed of total lysine levels of 0.64% (T1) and 0.96% (T2), with the lysine: methionine: threonine: tryptophan: valine ratio in the diets set to 100:27:73:19:69 through the addition of synthetic amino acids. Semen was collected twice weekly (phase 1, 1-12 wk); every other day (phase 2, 13-16 wk); twice weekly (phase 3, 17-26 wk); and daily (phase 4, 27-28 wk). Semen was collected from boars during phase 3 and used to inseminate 64 multiparous sows. Our results showed that sperm concentration and total sperm cells were greater in boars in T2 than in boars in T1 in phases 2 and 4 (Pamino acid concentrations in seminal plasma increased in T2 boars (Pamino acids improves sperm quality, and subsequently increases fertilization capacity and the number of live piglets.

  3. New insights into transduction pathways that regulate boar sperm function.

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

    2016-01-01

    Detailed molecular mechanisms mediating signal transduction cascades that regulate boar sperm function involving Ser/Thr and tyrosine phosphorylation of proteins have been reviewed previously. Therefore, this review will focus in those kinase pathways identified recently (boar spermatozoa that regulate different functional spermatozoa processes. AMP-activated protein kinase (AMPK) is a cell energy sensor kinase that was first identified in mammalian spermatozoa in 2012, and since then it has emerged as an essential regulator of boar sperm function. Signaling pathways leading to AMPK activation in boar sperm are highlighted in this review (PKA, CaMKKα/β, and PKC as well as Ca(2+) and cAMP messengers as upstream regulators). Interestingly, stimuli considered as cell stress (hyperosmotic stress, inhibition of mitochondrial activity, absence of intracellular Ca(2+)) markedly activate AMPK in boar spermatozoa. Moreover, AMPK plays a remarkable and necessary regulatory role in mammalian sperm function, controlling essential boar sperm functional processes such as motility, viability, mitochondrial membrane potential, organization and fluidity of plasma membrane, and outer acrosome membrane integrity. These mentioned processes are all required under fluctuating environment of spermatozoa when transiting through the female reproductive tract to achieve fertilization. An applied role of AMPK in artificial insemination techniques is also suggested as during boar seminal doses preservation at 17 °C, physiological levels of AMPK activity markedly increase (maximum on Day 7) and result essential to maintain the aforementioned fundamental sperm processes. Moreover, regulation of sperm function exerted by the glycogen synthase kinase 3 and Src family kinase pathways is summarized.

  4. Rotation of Boar Semen Doses During Storage Affects Sperm Quality.

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    Schulze, M; Rüdiger, K; Waberski, D

    2015-08-01

    It is common practice to rotate boar semen doses during storage for prevention of sperm sedimentation. In this study, the effect of rotation of boar semen doses during storage on sperm quality was investigated. Manual turning twice daily and automatic rotation five times per hour resulted in the following effects: alkalinization of the BTS-extender, loss of membrane integrity at day 3, and loss of motility and changes in sperm kinematics during a thermoresistance test at day 5. Using a pH-stabilized variant of BTS extender, sperm motility and velocity decreased in continuously rotated samples, whereas membrane integrity and mitochondrial activity remain unaffected. It is concluded that rotation of semen samples adversely affects sperm quality and, therefore, should no longer be recommended for AI practice.

  5. 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.

  6. Microscopic analysis of MTT stained boar sperm cells.

    Science.gov (United States)

    van den Berg, B M

    2015-01-01

    The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few 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.

  7. Addition of cholesterol-loaded cyclodextrins to the thawing extender: effects on boar sperm quality.

    Science.gov (United States)

    Tomás, C; Gómez-Fernández, J; Gómez-Izquierdo, E; Mocé, E; de Mercado, E

    2014-06-01

    The aim of the present study was to evaluate the effect that the addition of cholesterol-loaded cyclodextrins (CLC) to the thawing extender has on the quality of frozen-thawed boar sperm. Pooled semen (n = 5) from three boars was used for the experiments. The semen was cryopreserved with an egg-yolk-based extender, it was diluted after thawing in Beltsville thawing solution (BTS) supplemented with different concentrations of CLC (0, 12.5, 25, 50 or 100 mg/500 × 10(6) sperm), and these samples were incubated at 37°C for 150 min. The following parameters of sperm quality were evaluated 30 and 150 min after incubation: sperm with intact plasma membrane (SIPM; %), sperm with normal acrosomal ridge (NAR; %), total motile sperm (TMS; %), progressively motile sperm (PMS; %) and kinetic parameters. Both SIPM and NAR increased (p < 0.05) when the thawing extender was supplemented with 12.5, 25 and 50 mg CLC/500 × 10(6) sperm. Nevertheless, motility decreased (p < 0.05) when the concentration of CLC exceeded 12.5 mg CLC/500 × 10(6) sperm. In conclusion, our results suggest that the supplementation of thawing extenders with CLC improves sperm viability and reduces acrosome damage after freezing/thawing.

  8. The effects on boar sperm quality of dietary supplementation with omega-3 polyunsaturated fatty acids differ among porcine breeds.

    Science.gov (United States)

    Yeste, Marc; Barrera, Xavier; Coll, David; Bonet, Sergi

    2011-07-01

    The present study was undertaken to shed light on the relationship between boar sperm quality and dietary supplementation with omega-3 polyunsaturated fatty acids, which has been reported inconsistently in the literature. With this aim, such effects were evaluated and compared among three different porcine breeds: Duroc, Large-White, and Pietrain. Animals were randomly separated into two groups and fed either with a control diet or with a diet supplemented with omega-3. Sperm quality of these boar (ejaculate volume, sperm concentration, sperm viability, acrosome and mitochondrial sheath integrity, sperm motility, sperm morphology, and osmotic resistance of spermatozoa) was assessed every week for a 26-week period. Supplementing boar's diet with omega-3 did not affect ejaculate volume, sperm concentration, sperm motility, sperm viability, and acrosome and mitochondrial sheath integrity. In contrast, supplemented diet positively affected both sperm morphology in Large-White and Pietrain breeds and the osmotic resistance of Pietrain spermatozoa. No effects were seen for the same sperm parameters in Duroc breed. These breed-differences in boar fed with the supplemented diet could explain the contradictions in literature and might be related with differences in the composition of plasma membrane among breeds reported by other authors. Because no harmful effects were observed in the three evaluated breeds, but positive effects in Large-White and Pietrain boar, we can conclude that omega-3 fatty acids may be added to boar's diet at the levels used in this study to improve their sperm quality. More research is, however, needed to determine how these fatty acids differently affect the morphology and the osmotic resistance of the spermatozoa in these breeds.

  9. 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 ...

  10. LVQ acrosome integrity assessment of boar sperm cells

    NARCIS (Netherlands)

    Petkov, Nicolai; Alegre, Enrique; Biehl, Michael; Sánchez, Lidia; Tavares, JMRS; Jorge, RMN

    2007-01-01

    We consider images of boar spermatozoa obtained with an optical phase-contrast microscope. Our goal is to automatically classify single sperm cells as acrosome-intact (class 1) or acrosome-reacted (class 2). Such classification is important for the estimation of the fertilization potential of a sper

  11. LVQ acrosome integrity assessment of boar sperm cells

    NARCIS (Netherlands)

    Petkov, Nicolai; Alegre, Enrique; Biehl, Michael; Sánchez, Lidia

    2006-01-01

    We consider images of boar spermatozoa obtained with an optical phase-contrast microscope. Our goal is to automatically classify single sperm cells as acrosome-intact (class 1) or acrosome-reacted (class 2). Such classification is important for the estimation of the fertilization potential of a sper

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

    Science.gov (United States)

    Daigneault, B W; McNamara, K A; Purdy, P H; Krisher, R L; Knox, R V; Rodriguez-Zas, S L; Miller, D J

    2015-05-01

    Due to reduced fertility, cryopreserved semen is seldom used for commercial porcine artificial insemination (AI). Predicting the fertility of individual frozen ejaculates for selection of higher quality semen prior to AI would increase overall success. Our objective was to test novel and traditional laboratory analyses to identify characteristics of cryopreserved spermatozoa that are related to boar fertility. Traditional post-thaw analyses of motility, viability, and acrosome integrity were performed on each ejaculate. In vitro fertilization, cleavage, and blastocyst development were also determined. Finally, spermatozoa-oviduct binding and competitive zona-binding assays were applied to assess sperm adhesion to these two matrices. Fertility of the same ejaculates subjected to laboratory assays was determined for each boar by multi-sire AI and defined as (i) the mean percentage of the litter sired and (ii) the mean number of piglets sired in each litter. Means of each laboratory evaluation were calculated for each boar and those values were applied to multiple linear regression analyses to determine which sperm traits could collectively estimate fertility in the simplest model. The regression model to predict the percent of litter sired by each boar was highly effective (p boar was also effective (p boar spermatozoa can be predicted effectively by including traditional and novel laboratory assays that consider functions of spermatozoa.

  13. Effect of estrogens on boar sperm capacitation in vitro

    Directory of Open Access Journals (Sweden)

    Ded Lukas

    2010-07-01

    Full Text Available Abstract Background Mammalian sperm must undergo a series of controlled molecular processes in the female reproductive tract called capacitation before they are capable of penetrating and fertilizing the egg. Capacitation, as a complex biological process, is influenced by many molecular factors, among which steroidal hormone estrogens play their role. Estrogens, present in a high concentration in the female reproductive tract are generally considered as primarily female hormones. However, there is increasing evidence of their important impact on male reproductive parameters. The purpose of this study is to investigate the effect of three natural estrogens such as estrone (E1, 17beta-estradiol (E2 and estriol (E3 as well as the synthetical one, 17alpha-ethynylestradiol (EE2 on boar sperm capacitation in vitro. Methods Boar sperm were capacitated in vitro in presence of estrogens. Capacitation progress in control and experimental samples was analyzed by flow cytometry with the anti-acrosin monoclonal antibody (ACR.2 at selected times of incubation. Sperm samples were analyzed at 120 min of capacitation by CTC (chlortetracycline assay, immunocytochemistry and flow cytometry with anti-acrosin ACR.2 antibody. Furthermore, sperm samples and capacitating media were analyzed by immunocytochemistry, ELISA with the ACR.2 antibody, and the acrosin activity assay after induced acrosomal reaction (AR. Results Estrogens stimulate sperm capacitation of boar sperm collected from different individuals. The stimulatory effect depends on capacitation time and is highly influenced by differences in the response to estrogens such as E2 by individual animals. Individual estrogens have relatively same effect on capacitation progress. In the boar samples with high estrogen responsiveness, estrogens stimulate the capacitation progress in a concentration-dependent manner. Furthermore, estrogens significantly increase the number of acrosome-reacted sperm after zona

  14. 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-re......-resolution DNA flow cytometry on DAPI-stained sperm cell nuclei (CV...

  15. 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...

  16. Combining reduced glutathione and ascorbic acid has supplementary beneficial effects on boar sperm cryotolerance.

    Science.gov (United States)

    Giaretta, Elisa; Estrada, Efrén; Bucci, Diego; Spinaci, Marcella; Rodríguez-Gil, Joan E; Yeste, Marc

    2015-02-01

    The main aim of this work was to evaluate how supplementing freezing and thawing media with reduced glutathione (GSH) and l-ascorbic acid (AA) affected the quality parameters of frozen-thawed boar spermatozoa. With this purpose, semen samples of 12 ejaculates coming from 12 boars were used. Each ejaculate was split into seven aliquots to which 5 mM of GSH and 100 μM of AA were added separately or together at two different steps of freeze-thawing. Various sperm parameters (levels of free cysteine residues in sperm nucleoproteins, sperm viability, acrosome membrane integrity, intracellular peroxide and superoxide levels [ROS], and total and progressive motility) were evaluated before freezing and at 30 and 240 minutes after thawing. Both GSH and AA significantly improved boar sperm cryotolerance when they were separately added to freezing and thawing media. However, the highest improvement was recorded when both freezing and thawing media were supplemented with 5 mM of GSH plus 100 μM of AA. This improvement was observed in sperm viability and acrosome integrity, sperm motility, and nucleoprotein structure. Although ROS levels were not much increased by freeze-thawing procedures, the addition of GSH and AA to both freezing and thawing extenders significantly decreased intracellular peroxide levels and had no impact on superoxide levels. According to our results, we can conclude that supplementation of freezing and thawing media with both GSH and AA has a combined, beneficial effect on frozen-thawed boar sperm, which is greater than that obtained with the separate addition of either GSH or AA.

  17. Testing an egg yolk supplemented diet on boars to aid in sperm adaptation at 5°C.

    Science.gov (United States)

    Casas, Isabel; Miller-Lux, Yvonne; Osborne, Betty; Bonet, Sergi; Althouse, Gary C

    2015-01-01

    In many species, extended semen can be stored at low temperatures to slow bacterial growth. However, boar semen performs poorly at temperatures below 15 °C and this poses unique challenges, as it is not easy to maintain a constant 15-19 °C during shipment. Some extenders have been formulated with egg yolk for storage at 5 °C but the addition of egg yolk is not applicable in the majority of commercial operations. The purpose of this study was to evaluate if boar dietary supplementation with powdered egg yolk imparts any protective effects on sperm quality when stored at 15 °C and 5 °C for up to 11 days in a conventional extender. Ten boars were fed a commercial diet with the addition of 0.11 Kg of powdered egg yolk for 10 weeks. Ejaculates collected on weeks 4, 6, 8, and 10 were processed for storage at both 15 °C and 5 °C and compared with ejaculates from boars fed a standard diet. Throughout an 11-day storage period, sperm quality was assessed including several motility and morphologic parameters and select plasma membrane properties (fluidity, integrity, and triacylglycerol content). Linear regression models were used to describe effects of treatment, storage day, week and temperature on all sperm parameters. Overall, there were minimal beneficial effects of egg yolk treatment on sperm quality parameters. Sperm from egg yolk supplemented boars did have a slower decline in viability and plasma membrane fluidity than that observed in the control sperm when stored at 5 °C (p sperm from egg yolk fed boars compared to controls at week 10 (p sperm quality or resistance to cold storage when feeding a 10-week dietary supplementation of 0.11 Kg powdered egg yolk to crossbred boars.

  18. Seminal plasma affects sperm sex sorting in boars.

    Science.gov (United States)

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

    2016-04-01

    Two experiments were conducted in boar semen samples to evaluate how both holding time (24h) and the presence of seminal plasma (SP) before sorting affect sperm sortability and the ability of sex-sorted spermatozoa to tolerate liquid storage. Whole ejaculate samples were divided into three aliquots immediately after collection: one was diluted (1:1, v/v) in Beltsville thawing solution (BTS; 50% SP); the SP of the other two aliquots was removed and the sperm pellets were diluted with BTS + 10% of their own SP (10% SP) or BTS alone (0% SP). The three aliquots of each ejaculate were divided into two portions, one that was processed immediately for sorting and a second that was sorted after 24h storage at 15-17°C. In the first experiment, the ability to exhibit well-defined X- and Y-chromosome-bearing sperm peaks (split) in the cytometry histogram and the subsequent sorting efficiency were assessed (20 ejaculates). In contrast with holding time, the SP proportion influenced the parameters examined, as evidenced by the higher number of ejaculates exhibiting split and better sorting efficiency (P<0.05) in semen samples with 0-10% SP compared with those with 50% SP. In a second experiment, the quality (viability, total and progressive motility) and functionality (plasma membrane fluidity and intracellular generation of reactive oxygen species) of sex-sorted spermatozoa were evaluated after 0, 72 and 120h storage at 15-17°C (10 ejaculates). Holding time and SP proportion did not influence the quality or functionality of stored sex-sorted spermatozoa. In conclusion, a holding time as long as 24h before sorting did not negatively affect sex sorting efficiency or the ability of sorted boar spermatozoa to tolerate long-term liquid storage. A high proportion of SP (50%) in the semen samples before sorting reduced the number of ejaculates to be sorted and negatively influenced the sorting efficiency, but did not affect the ability of sex-sorted spermatozoa to tolerate liquid

  19. Boar sperm quality in relation to presence of sp32-like protein in spermatozoa - preliminary studies

    Directory of Open Access Journals (Sweden)

    Orzołek Aleksandra

    2015-06-01

    Full Text Available The aim of the study was to analyse sperm proteomes of ejaculates from Polish Large White (PLW and Polish Landrace (PL boars and to identify differences which putatively influence semen quality. Spermatozoa protein profiles were analysed by electrophoretic methods followed by selected techniques to evaluate semen quality on the following factors: sperm motility, lipid peroxidation levels (MDA production, ATP content, activities of superoxide dismutase (SOD and catalase (CAT, total antioxidant status (TAS, and total oxidant status (TOS of seminal plasma. A protein with an estimated molecular weight of 30 kDa was found in spermatozoa of selected ejaculates. Mass spectrometry demonstrated that this polypeptide is most similar to proacrosin binding protein (sp32. The presence of the protein was more frequently observed in sperm extracts obtained in spring-summer period. Ejaculates containing sp32-like protein demonstrated significantly higher spermatozoa motility, lower inhibition of MDA production by seminal plasma, and higher SOD activity in seminal plasma. Boar semen which included sp32-like protein also demonstrated lower ATP levels in spermatozoa as well as higher TAS and lower TOS of seminal plasma, though the differences were not statistically significant. Ejaculates from PLW boars, with sp32-like protein present in sperm, were characterised by significantly higher sperm motility, lower ATP content in spermatozoa, and higher TAS of seminal plasma. The diminished parameters of semen quality were observed in ejaculates from PL boars that also contained the discussed protein, but the differences were not statistically significant. These findings suggest that the presence of sp32-like protein in boar spermatozoa could influence semen quality

  20. Effects of centrifugation through three different discontinuous Percoll gradients on boar sperm function.

    Science.gov (United States)

    Matás, C; Vieira, L; García-Vázquez, F A; Avilés-López, K; López-Úbeda, R; Carvajal, J A; Gadea, J

    2011-08-01

    In this study, different combinations of 2-step, discontinuous gradient centrifugation were used, consisting of three different combinations of isotonic Percoll (45/60, 60/75 and 45/90%) that allowed us to select different sperm subpopulations from fertile and normozoospermic boars. Our objective in this study is to evaluate the effects of centrifugation through three different discontinuous Percoll gradients on sperm function parameters (motility, viability, morphology, acrosome status, chromatin condensation, DNA fragmentation, ROS generation, tyrosine phosphorylation and intracellular calcium concentration) and the sperm penetrating capacity in an IVF system. All the Percoll treatments evaluated increased the percentage of spermatozoa with normal morphology, the proportion of un-damaged DNA, normal chromatin condensation, motion parameters measured by CASA and the percentage of capacitated spermatozoa with tyrosine phosphorylated proteins compared to control group. Finally, the in vitro oocyte penetrating capacity of boar spermatozoa was significantly affected by Percoll centrifugation. All the Percoll treatments increased the penetration rates and mean number of sperm per penetrated oocyte. Despite the efficiency of all three of the sperm treatments tested in selecting spermatozoa with improved sperm parameters and capacity to penetrate oocytes in vitro, the optimum performance of this system was demonstrated after preselecting spermatozoa by centrifugation on a discontinuous 45/90 Percoll gradient. The P45/90 treatment leads to obtain a higher percentage of spermatozoa which develop properly the capacitation process as it was shown measuring tyrosine phosphorylation and intracellular calcium concentration.

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

    Science.gov (United States)

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

    2015-08-01

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

  3. 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.

  4. Temperature management during semen processing: Impact on boar sperm quality under laboratory and field conditions.

    Science.gov (United States)

    Schulze, M; Henning, H; Rüdiger, K; Wallner, U; Waberski, D

    2013-12-01

    Freshly collected boar spermatozoa are sensitive to a fast reduction in temperature because of lipid phase transition and phase separation processes. Temperature management during semen processing may determine the quality of stored samples. The aim of this study was to evaluate the influence of isothermic and hypothermic semen processing protocols on boar sperm quality under laboratory and field conditions. In the laboratory study, ejaculates (n = 12) were first diluted (1:1) with Beltsville Thawing Solution (BTS) at 32 °C, then processed either with isothermic (32 °C) or hypothermic (21 °C) BTS, stored at 17 °C, and assessed on days 1, 3, and 6. Temperature curves showed that 150 minutes after the first dilution, semen doses of both groups reached the same temperature. Two-step hypothermic processing resulted in lower sperm motility on days 1 and 6 (P sperm on days 3 and 6 (P boar semen compared with isothermic dilution and that the type of semen extender affects the outcomes.

  5. Effect of seminal plasma and sperm of boars valued by freezability on seminal cryopreservation

    Directory of Open Access Journals (Sweden)

    Francisco Javier Henao Uribe

    2016-07-01

    Full Text Available The aim of this study was to determine the effect of sperm and seminal plasma (SP on the freezability of porcine semen. Semen of eight commercial males from two farms in the central-western region of Colombia (four boars in each farm was frozen and tested to select two males with high freezability (MHF and two with low freezability (MLF, according to the percentage of functionally competent sperm (FCS. Immediately after the collection was completed, the SP and sperm from the males selected were separated by centrifugation to combine the two types of plasma with the two types of sperm, incubate them for three hours and then freeze them. The variables evaluated were: sperm morphology, structural and functional integrity of plasmatic membrane, progressive and total motility, DNA fragmentation, acrosome integrity, capacitated sperm and FCS. The combination of sperm and plasma of MHF recorded the highest value (P<0.01 of acrosome integrity (24.3 ± 0.082 vs 6.076 ± 0.16 when compared to MLF plasma and cells. Membrane structural integrity was higher (P<0.01 with MHF (53.56 ± 0.0395 than with MLF plasma (47.49 ± 0.0419. The differences in porcine semen freezability depend on interactions between seminal plasma and sperm.

  6. The osmotic tolerance of boar spermatozoa and its usefulness as sperm quality parameter.

    Science.gov (United States)

    Yeste, Marc; Briz, Mailo; Pinart, Elisabeth; Sancho, Sílvia; Bussalleu, Eva; Bonet, Sergi

    2010-06-01

    Predicting the fertility outcome of ejaculates is very important in the field of porcine reproduction. The aims of this study were to determine the effects of different osmotic treatments on boar spermatozoa and to correlate them with fertility and prolificacy, assessed as non-return rates within 60 days (NRR(60d)) of the first inseminations, and litter size (LS), respectively. Sperm samples (n=100) from one hundred healthy Piétrain boars were used to assess 48 treatments combining different osmolalities (ranged between 100 and 4000 mOsm kg(-1)), different compounds used to prepare anisotonic solutions, and two different modalities: return and non-return to isotonic conditions. Sperm quality was evaluated before and after applying the treatments on the basis of analyses of sperm viability, motility, morphology and percentages of acrosome-intact spermatozoa. Statistical analyses were performed using a one-way ANOVA and post hoc Tukey's test, linear regression analyses (Pearson correlation and multiple regression) and Jackknife cross-validation. Although three conventional parameters: sperm viability, sperm morphology and the percentages of acrosome-intact spermatozoa were significantly correlated with NRR(60d) and with LS, their respective osmotic tolerance parameters (defined for each parameter and treatment regarding with negative control) presented a higher Pearson coefficient with both fertility and prolificacy in three treatments (150 mOsm kg(-1) with non-return to isotonic conditions, 200 mOsm kg(-1) with return and 500 mOsm kg(-1) using sodium citrate and non-return to isotonic conditions). We conclude that osmotic resistance in sperm viability, sperm morphology and acrosome-intactness in the treatments mentioned above could be assessed along with classical parameters to better predict the fertilising ability of a given ejaculate.

  7. 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...

  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

    2016-03-02

    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.

  9. Administration of flutamide alters sperm ultrastructure, sperm plasma membrane integrity and its stability, and sperm mitochondrial oxidative capability in the boar: in vivo and in vitro approach.

    Science.gov (United States)

    Lydka, M; Piasecka, M; Gaczarzewicz, D; Koziorowski, M; Bilinska, B

    2012-08-01

    Our previous work has shown that an anti-androgen flutamide administered pre- and post-natally induced adverse effects on the epididymal morphology and function of adult boars. The present investigation is aimed to understand the effect of flutamide and its metabolite on changes in sperm plasma membrane integrity and its stability, changes in mitochondrial oxidative capability and frequency of abnormal sperm. In vivo effects of flutamide (50 mg/kg b.w.) on sperm ultrastructure were examined by electron microscopic observations. In vitro effects of 5, 50 and 100 μg/ml hydroxyflutamide, administered for 2 and 24 h, on sperm plasma membrane integrity were measured by LIVE/DEAD Sperm Vitality kit, while those on sperm membrane stability and mitochondrial oxidoreductive activity were investigated using Merocyanine 540 and NADH tests, respectively. The incidence of abnormal spermatozoa increased significantly (p boars compared with controls. In an in vitro approach, low dose of hydroxyflutamide in 2-h incubations appeared less effective in altering the sperm plasma membrane integrity and its stability than two higher doses used (p sperm membrane destabilization and mitochondrial oxidoreductive activity was strengthened after 24 h of hydroxyflutamide administration (p sperm parameters with regard to oxidative capability of mitochondria, plasma membrane changes and sperm ultrastructure provides novel data on the boar sperm sensitivity to anti-androgen action. Results indicate high sensitivity of boar spermatozoa to androgen withdrawal.

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

    Directory of Open Access Journals (Sweden)

    Kondracki S.

    2013-01-01

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

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

    Science.gov (United States)

    Mears, Matthew; Kennelly, Thomas M; Howse, Jonathan R; Tarmey, Drew S; Geoghegan, Mark; Pacey, Allan A

    2014-03-15

    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 layer of the polymer film interacts with the sperm and buffer medium. The VCL of sperm between PS-coated surfaces was significantly reduced compared with PMMA surfaces (P IVF. Controlling the velocity of sperm using the interaction properties of inert polymer coatings could lead to new sperm selection procedures for clinical use or the development of model systems to better understand sperm-surface interactions.

  12. 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

  13. 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...

  14. PROSTAGLANDIN F2α SUPPLEMENTED SEMEN IMPROVES LANDRACE BOARS SPERM MOTILITY

    Directory of Open Access Journals (Sweden)

    NICOLETA IONESCU

    2009-05-01

    Full Text Available This study investigated whether the sperm motility from Landrace boars improveswhen PGF2α (Dinolytic®; 5 mg PGF2α /ml was added to diluted semen. Boars fromone large production unit, were manually collected; semen was either enriched withPGF2α (group 1, n=38, either untreated (group 2, n=32. Total volume of semencollected, percent of motility and number of obtained doses were recorded. Thehighest sperm volume collected from the two groups is corresponding to ejaculatesfrom Landrace boars with PGF2α supplemented semen (267.6 ml. Regardingmotility, the sperm collected from Landrace boars with PGF2α supplemented semenwas higher from the one collected from Landrace boars with untreated semen(81.37% and very significant differences were statistically determined. Theejaculates with highest number of obtained doses is corresponding to the onescollected from boars with PGF2α supplemented semen (25.21. Only boars from thefirst group (with PGF2α supplemented semen showed motility over 70% and even100%. The untreated semen showed motility values around 65-70%.

  15. PROSTAGLANDIN F2α SUPPLEMENTED SEMEN IMPROVES LANDRACE BOARS SPERM MOTILITY

    Directory of Open Access Journals (Sweden)

    IOANA SGURĂ

    2013-07-01

    Full Text Available This study investigated whether the sperm motility from Landrace boars improves when PGF2α (Dinolytic®; 5 mg PGF2α /ml was added to diluted semen. Boars from one large production unit, were manually collected; semen was either enriched with PGF2α (group 1, n=38, either untreated (group 2, n=32. Total volume of semen collected, percent of motility and number of obtained doses were recorded. The highest sperm volume collected from the two groups is corresponding to ejaculates from Landrace boars with PGF2α supplemented semen (267.6 ml. Regarding motility, the sperm collected from Landrace boars with PGF2α supplemented semen was higher from the one collected from Landrace boars with untreated semen (81.37% and very significant differences were statistically determined. The ejaculates with highest number of obtained doses is corresponding to the ones collected from boars with PGF2α supplemented semen (25.21. Only boars from the first group (with PGF2α supplemented semen showed motility over 70% and even 100%. The untreated semen showed motility values around 65-70%.

  16. Fertilidade de sêmen suíno avaliada pelo teste de ligação dos espermatozóides a um substrato sintético Boar semen fertility evaluated by a sperm-binding assay to a synthetic substrate

    Directory of Open Access Journals (Sweden)

    Goreti Ranincheski dos Reis

    2003-11-01

    Full Text Available O objetivo deste trabalho foi avaliar a fertilidade de sêmen suíno pelo teste de ligação de espermatozóides a um substrato sintético. A motilidade (MOT e o porcentual de espermatozóides ligados (PEL foram avaliados após 5, 24, 48 e 72 horas de armazenamento a 17ºC. O PEL foi determinado em soluções contendo 6,25 ou 12,5 milhões de espermatozóides/mL, com ou sem albumina sérica bovina (BSA, preparadas a partir de dois a cinco ejaculados de cada um dos quatro machos. Cinqüenta e oito leitoas foram inseminadas, uma vez, 24 horas após o início do estro. Houve correlação positiva (P = 0,0001; r = 0,33 entre a MOT e o PEL. O PEL foi maior com 12,5 milhões de espermatozóides/mL e na presença de BSA (PThe objective of this work was to evaluate boar semen fertility by a sperm-binding assay to a synthetic substrate. Motility (MOT and percentage of bound sperm (PSB were evaluated after 5, 24, 48 and 72 hours of storage at 17°C. PSB was analyzed in solutions containing 6.25 or 12.5 million of spermatozoa/mL, with or without bovine serum albumin (BSA, processed from two to five ejaculates of four boars. Fifty eight gilts were inseminated, a single time, 24 hours after the beginning of estrus. There was a positive correlation (P = 0.0001; r = 0.33 between MOT and PSB. Higher percentages of PSB were observed with 12.5 million of spermatozoa/mL and in the presence of BSA (P<0.05. After 72 hours, boar 3 showed lower PSB (P<0.05 than the other boars. The cleavage (TC and normal embryo rates did not differ among boars, but boar 3 showed less than 70.0% of TC belonging to the superior quartile while boars 1, 2 and 4 had more than 75.0%. After 24 hours of sperm storage, boars differ in their sperm binding to the synthetic substrate. Binding of swine spermatozoa to the synthetic substrate is higher in the presence of BSA and with the increase of spermatic concentration.

  17. 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.

  18. Boar seminal plasma exosomes: effect on sperm function and protein identification by sequencing.

    Science.gov (United States)

    Piehl, Lidia L; Fischman, M Laura; Hellman, Ulf; Cisale, Humberto; Miranda, Patricia V

    2013-04-15

    Mammalian seminal plasma contains membranous vesicles (exosomes), with a high content of cholesterol and sphingomyelin and a complex protein composition. Their physiological role is uncertain because sperm stabilization and activation effects have been reported. To analyze a putative modulatory role for semen exosomes on sperm activity in the boar, the effects of these vesicles on several sperm functional parameters were examined. Additionally, boar exosome proteins were sequenced and their incorporation into sperm was explored. Boar sperm were incubated under conditions that induce capacitation, manifested as increased tyrosine phosphorylation, cholesterol loss and greater fluidity in apical membranes, and the ability to undergo the lysophosphatidylcholine-induced acrosome reaction. After establishing this cluster of capacitation-dependent functional parameters, the effect produced by exosomes when present during or after sperm capacitation was analyzed. Exosomes inhibited the capacitation-dependent cholesterol efflux and fluidity increase in apical membranes, and the disappearance of a 14-kD phosphorylated polypeptide. In contrast, the acrosome reaction (spontaneous and lysophosphatidylcholine-induced) was not affected, and sperm binding to the oocyte zona pellucida was reduced only when vesicles were present during gamete coincubation. Liposomes with a lipid composition similar to that present in exosomes mimicked these effects, except the one on zona pellucida binding. Interaction between exosomes and sperm was confirmed by transfer of aminopeptidase activity. In addition, the major exosome protein, identified as actin, appeared to associate with sperm after coincubation. Exosome composition had a predominance for structural proteins (actin, plastin, ezrin, and condensin), enzymes, and several porcine seminal plasma-specific polypeptides (e.g., spermadhesins). Transfer of proteins from exosome to sperm and their ability to block cholesterol efflux supports a

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

    OpenAIRE

    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-st...

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

    Science.gov (United States)

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

    2015-03-01

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

  1. Methods for Improving In Vitro and In Vivo Boar Sperm Fertility.

    Science.gov (United States)

    Funahashi, H

    2015-07-01

    Fertility of boar spermatozoa is changed after ejaculation in vivo and in vitro. During processing for in vitro fertilization (IVF), although spermatozoa are induced capacitation, resulting in a high penetration rate, persistent obstacle of polyspermic penetration is still observed with a high incidence. For artificial insemination (AI), we still need a large number of spermatozoa and lose a majority of those in the female reproductive tract. Fertility of cryopreserved boar spermatozoa is still injured through freezing and thawing process. In the present brief review, factors affecting fertility of boar sperm during IVF, AI and cryopreservation are discussed in the context of discovering methodologies to improve it.

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

    Science.gov (United States)

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

    2016-04-01

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

  3. 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

  4. Effect of addition of coconut water (Cocos nucifera) to the freezing media on post-thaw viability of boar sperm.

    Science.gov (United States)

    Bottini-Luzardo, María; Centurión-Castro, Fernando; Alfaro-Gamboa, Militza; Aké-López, Ricardo; Herrera-Camacho, José

    2013-01-01

    The aims of this experiment were to evaluate the addition of coconut water in natura to the freezing media, compare the effect of deionized water vs filtered water of coconut over the post-thaw seminal characteristics, and evaluate the effect of the deionized water and in natura coconut water on the seminal characteristics of boar sperm at different post-thaw times. Thirty-four ejaculates were used divided in three aliquots which received one of the following treatments (T): T1, LEY (bidistilled water, lactose, and egg yolk) and LEYGO (LEY + glycerol and Orvus ET paste); T2, LEY(A) (coconut deionized water, lactose, and egg yolk)-LEYGO(A); and T3, LEY(B) (in natura coconut water, lactose, and egg yolk)-LEYGO(B). Samples of boar semen were frozen according to the Westendorf method, thawed at 38°C, and evaluated at three incubation times (0, 30, and 60 min). Seminal characteristics assessed were motility (Mot), acrosomal integrity (AInt), membrane integrity (MInt), and mitochondrial activity (MAct). T1 showed a higher percentage of viable sperm than T3 (Mot 36.5 vs 5.4 %, AInt 61.8 vs 41.2 %, MInt 50.4 vs 41.3 %, and MAct 56.9 vs 50.5 %). T2 kept a higher percentage of viable sperm at all incubation times. In natura coconut water showed a detrimental effect over the viability of the frozen-thawed boar semen. Deionized coconut water improved the boar semen viability post-thaw, outperforming results of in natura coconut water.

  5. Changes in exposed membrane proteins during in vitro capacitation of boar sperm

    Energy Technology Data Exchange (ETDEWEB)

    Berger, T. (Univ. of California, Davis (USA))

    1990-11-01

    Exposed plasma membrane proteins were labeled with {sup 125}I before and after incubation of boar sperm under capacitating conditions. Labeled protein profiles were compared to the ability of the sperm to penetrate zona-free hamster ova. Quantitatively, the labeled sperm membrane proteins were primarily low Mr prior to capacitation. The majority of the labeled seminal plasma protein was also low Mr. After capacitation, two new proteins (64,000 Mr and 78,000 Mr) were labeled. Sperm did not exhibit these exposed membrane proteins when incubated under noncapacitating conditions. Appearance of these proteins was not correlated to the percentage of acrosome-reacted sperm. Although the 64,000 Mr protein was not consistently observed, the relative labeling of the 78,000 Mr protein was highly correlated with the ability of sperm to fuse with zona-free hamster ova. The 78,000 Mr protein may be a sperm protein involved in fusion with the egg plasma membrane.

  6. 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).

  7. Effects of freezing/thawing on motile sperm subpopulations of boar and donkey ejaculates.

    Science.gov (United States)

    Flores, E; Taberner, E; Rivera, M M; Peña, A; Rigau, T; Miró, J; Rodríguez-Gil, J E

    2008-10-01

    The main aim of this study is to assess the influence of freeze/thawing on motile sperm subpopulations in ejaculates from two phylogenetically different mammalian species, boar and donkey. Our results indicate that, whereas boar and donkey sperm respond very differently in their mean motion characteristics to freezing/thawing, this process did not change the existence of a 4-subpopulations structure in the ejaculates in either species when these subpopulations were defined by taking values of curvilinear velocity (VCL) as reference. Moreover, the freezing/thawing-linked changes in mean sperm-motion characteristics in both boar and donkey semen were especially due to changes in the proportion among each concrete subpopulation. In this way, the freezing/thawing-induced mean increase in motion characteristics observed in boar sperm was a result of the decrease in the percentage of sperm in Subpopulation 1 (from 53.9%+/-4.7% to 31.2%+/-3.9% after thawing) and a concomitant increase of sperm from Subpopulations 3 (from 13.3%+/-2.5% to 32.6%+/-3.9% after thawing) and 4 (from 3.4%+/-0.9% to 8.0%+/-1.1% after thawing). On the contrary, changes in mean motility of frozen/thawed donkey sperm were linked to an increase in the percentage of sperm in Subpopulation 1 (from 31.5%+/-4.3% to 58.8%+/-4.9% after thawing) and a concomitant decrease of sperm from Subpopulations 3 (from 32.4%+/-3.2% to 6.6%+/-1.8% after thawing) and 4 (from 12.2%+/-2.5% to 7.3%+/-1.9% after thawing). In conclusion, our results seem to indicate that motility changes induced by the freezing/thawing protocol are linked to concomitant changes in both the specific parameters and, more importantly, to the specific percentage of each of the motile sperm subpopulations. These changes did not affect the overall proportion of motile sperm present in both boar and donkey, which is conserved despite the detrimental effect caused by freezing/thawing in both species. Finally, the presence of some kind of motile sperm

  8. Sperm quality and fertility of boar seminal doses after 2 days of storage: does the type of extender really matter?

    Science.gov (United States)

    Pinart, Elisabeth; Yeste, Marc; Prieto-Martínez, Noelia; Reixach, Josep; Bonet, Sergi

    2015-06-01

    The present approach was designed to evaluate the extender effects on sperm quality and fertility of short-term refrigerated seminal doses from Landrace boars lodged in husbandry-controlled conditions. For this purpose, we analyzed the sperm quality of seminal doses diluted in short-term (Beltsville Thawing Solution) and extra-long-term (Duragen) extenders from Days 0 to 2 of storage at 17 °C during an 8-month period. Pregnancy rates and litter size were evaluated from double inseminations within an interval of 12 hours (36 and 48 hours of refrigeration) of multiparous females using seminal doses diluted in each extender type. Sperm quality was assessed from the analyses of sperm motility and kinetics, sperm viability, expressed as plasma and acrosome membrane integrity, membrane lipid disorder, intracellular calcium levels, and acrosin activity. Results indicated significant differences between the extenders in the sperm quality of seminal doses. Therefore, the seminal doses diluted in Duragen had higher percentages of progressive motile spermatozoa and membrane-intact spermatozoa than those diluted in Beltsville Thawing Solution throughout all the experimental months. Nevertheless, despite these differences in preserving the sperm quality, pregnancy rates (>90%) and litter sizes (>10 piglets born per litter) were similar between the extenders. Our results had great relevance from a practical point of view because they reported lack of an extender effect on the reproductive performance of seminal doses during short-tem storage.

  9. 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.

  10. Quality Control of Boar Sperm Processing : Implications from European AI Centres and Two Spermatology Reference Laboratories

    NARCIS (Netherlands)

    Riesenbeck, A; Schulze, M; Rüdiger, K; Henning, H; Waberski, D

    2015-01-01

    In recent years, increased automatization has resulted in a higher efficiency of boar semen processing in AI laboratories. Sophisticated laboratory management and efficient quality control programmes are needed for current tendencies in major pork-producing countries to reduce the sperm number per A

  11. Successful laparoscopic insemination with a very low number of flow cytometrically sorted boar sperm in field conditions.

    Science.gov (United States)

    del Olmo, David; Parrilla, Inmaculada; Sanchez-Osorio, Jonatan; Gomis, Jesus; Angel, Miguel A; Tarantini, Tatiana; Gil, Maria A; Cuello, Cristina; Vazquez, Jose L; Roca, Jordi; Vaquez, Juan M; Martinez, Emilio A

    2014-01-15

    The aim of this study was to develop a useful procedure for laparoscopic insemination (LI) with sex-sorted boar spermatozoa that yields adequate fertility results in farm conditions. In experiment 1, we evaluated the effects of single (oviducts) and double (oviducts and tips of the uterine horns) LI with X-sorted sperm on the reproductive performance of sows. Sows (N = 109) were inseminated once as follows: (1) single LI with 0.5 × 10(6) unsorted sperm per oviduct; (2) single LI with 0.5 × 10(6) sex-sorted sperm per oviduct; or (3) double LI with 0.5 × 10(6) sex-sorted sperm per oviduct and 0.5 × 10(6) sex-sorted sperm per uterine horn. The farrowing rates were lower (P sperm (43.2% and 61.9% for the single and double insemination groups, respectively) than in sows from the unsorted group (91.3%). Within the sex-sorted groups, the farrowing rate tended (P = 0.09) to be greater in sows inseminated using double LI. There were no differences in the litter size among groups. In experiment 2, we evaluated the effect of the number of sex-sorted sperm on the reproductive performance of sows when using double LI. Sows (N = 109) were inseminated with sex-sorted sperm once using double LI with: (1) 0.5 × 10(6) sperm per oviduct and 1 × 10(6) sperm per uterine horn; or (2) 1 × 10(6) sperm per oviduct and 2 × 10(6) sperm per uterine horn. Similarly high pregnancy (90%) and farrowing (80%) rates were achieved in both groups. The sows inseminated with the highest number of sperm tended (P = 0.09) to have more piglets (10.8 ± 0.7 vs. 9.2 ± 0.6). A high female proportion (number of female births divided by the total of all births ≥0.92) was obtained in both experiments using X-sorted sperm. Our results indicate that the double LI procedure, using between 3 and 6 × 10(6) sex-sorted sperm per sow produces adequate fertility at the farm level, making sperm-sexing technology potentially applicable in elite breeding units.

  12. 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.

  13. Hexavalent chromium affects sperm motility by influencing protein tyrosine phosphorylation in the midpiece of boar spermatozoa.

    Science.gov (United States)

    Zhen, Linqing; Wang, Lirui; Fu, Jieli; Li, Yuhua; Zhao, Na; Li, Xinhong

    2016-01-01

    Hexavalent chromium reportedly induces reproductive toxicity and further inhibits male fertility in mammals. In this study, we investigated the molecular mechanism by which hexavalent chromium affects motility signaling in boar spermatozoa in vitro. The results indicated that Cr(VI) decreased sperm motility, protein phosphorylation, mitochondrial membrane potential (ΔΨm) and metabolic enzyme activity starting at 4μmol/mL following incubation for 1.5h. Notably, all parameters were potently inhibited by 10μmol/mL Cr, while supplementation with the dibutyryl-cAMP (dbcAMP) and the 3-isobutyl-1-methylxanthine (IBMX) prevented the inhibition of protein phosphorylation. Interestingly, high concentrations of Cr (>10μmol/mL) increased the tyrosine phosphorylation of some high-molecular-weight proteins in the principle piece but decreased that in the middle piece associated with an extreme reduction of sperm motility. These results suggest that chromium affects boar sperm motility by impairing tyrosine phosphorylation in the midpiece of sperm by blocking the cAMP/PKA pathway in boar sperm in vitro.

  14. 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

    A flow cytometric method has been developed for rapid determination of sperm concentration in semen from various mammalian species.* All cells containing DNA are stained with SYBR-14 or propidium iodide (PI) and sperm concentration is determined in relation to an internal standard of fluorescent...... microspheres ( beads). Satisfactory staining can be achieved within 2-3 min and the following flow cytometric analysis on the FACSCount AF System rapidly provides the user with a precise and accurate assessment of the sperm concentration. In this study, the FACSCount AF System and Sperm Counting Reagent ( BD...... Biosciences) was compared with microscopic counting using a 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...

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

    Science.gov (United States)

    Kasimanickam, Vanmathy; Kastelic, John

    2016-09-06

    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 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.

  16. 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 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

  17. Effect of magnetized extender on sperm membrane integrity and development of oocytes in vitro fertilized with liquid storage boar semen.

    Science.gov (United States)

    Lee, Sang-Hee; Park, Choon-Keun

    2015-03-01

    The objective of this study was to evaluate the effect of a magnetized extender on sperm membrane damage and development of oocytes in vitro fertilized with liquid storage boar semen. Before semen dilution, extender was flowed through a neodymium magnet (0, 2000, 4000 and 6000G) for 5min and collected semen was preserved for 168h at 18°C. In results, plasma membrane integrity with live sperm was significantly higher in semen treated with extenders magnetized at 4000G than sperm treated with extenders magnetized at 0G during semen preservation for 120-168h (psperm was significantly lower in semen treated with extenders magnetized at 2000G than other groups during semen preservation for 168h. The ability of semen to achieve successful in vitro fertilization was also not significantly different among the groups during preservation. However, when the semen was preserved for 168h, the blastocyst formation rates were significantly higher at 6000G compared to 0 and 2000G (psperm membrane from damage, and improve the ability of rates of in vitro blastocyst development and magnetized semen diluter is beneficial for long liquid preservation of boar semen.

  18. Effects of bovine serum albumin on boar sperm quality during liquid storage at 17°C.

    Science.gov (United States)

    Zhang, X-G; Yan, G-J; Hong, J-Y; Su, Z-Z; Yang, G-S; Li, Q-W; Hu, J-H

    2015-04-01

    This study aimed to investigate the effects of bovine serum albumin (BSA) on boar sperm quality during liquid storage at 17°C. Boar semen samples were collected and diluted with Modena containing different concentrations (0, 1, 2, 3, 4, 5 and 6 g/l) of BSA, and sperm motility, plasma membrane integrity, acrosome integrity, total antioxidative capacity (T-AOC) activity and malondialdehyde (MDA) content were measured and analysed. The results showed that Modena supplemented with 3, 4 and 5 g/l BSA could improve boar sperm motility, effective survival time and plasma membrane integrity (p sperm acrosome integrity and T-AOC activity among these three groups (p > 0.05). The semen sample diluted with Modena containing 4 g/l BSA could achieve optimum effect, and sperm survival time was 7.5 days. After 7 days preservation, sperm motility, plasma membrane integrity and acrosome integrity were 54%, 49% and 78%, respectively. T-AOC activity and MDA content were 1.03 U/ml and 17.5 nmol/ml, respectively. In conclusion, Modena supplemented with BSA reduced the oxidative stress and improved the sperm quality of boar semen during liquid storage at 17°C, and 4 g/l BSA was the optimum concentration. Further studies are required to obtain more concrete results on the determination of antioxidant capacities of BSA in liquid preserved boar semen.

  19. Effects of different concentrations of enterotoxigenic and verotoxigenic E. coli on boar sperm quality.

    Science.gov (United States)

    Bussalleu, E; Yeste, M; Sepúlveda, L; Torner, E; Pinart, E; Bonet, S

    2011-09-01

    The presence of bacteria in boar semen causes economic losses in artificial insemination (AI) centers, as a consequence of alterations on boar sperm quality. For this reason, the effects of different concentrations of enterotoxigenic Escherichia coli (ETEC) and verotoxigenic E. coli (VTEC) on boar sperm quality were determined in this study, by conducting two experiments. The first one consisted of assessing these effects on boar sperm quality after incubating the inoculated doses at 37°C for a 96-h period, whereas the second inoculated doses were stored at 15°C during 11 days. In both experiments, the infective concentrations ranged from 10(8)cfu mL(-1) to 10(2)cfu mL(-1); the negative control being a non-inoculated dose. Twenty-four hours after inoculation, we checked by PCR for the presence of bacteria in all tubes. Sperm quality (sperm motility, sperm viability and sperm morphology) was assessed at 24h, 48h, 72h and 96h after inoculations in the first experiment (37°C), and after 3, 5, 7, 9 and 11 days in the second (15°C). Whereas no changes were observed in sperm morphology in both experiments, the percentages of progressive motile spermatozoa dramatically diminished after 24h of incubation at 37°C, the effect being more detrimental at the highest infective concentration of microbes. Moreover, a significant decrease in the percentage of viable spermatozoa in the tube inoculated with the highest concentration (10(8)cfu mL(-1)) was detected after 24h of incubating contaminated doses at 37°C. After 48h of incubation, the presence of infective concentrations of ETEC and VTEC from 10(8)cfu mL(-1) to 10(3)cfu mL(-1) resulted in a significant diminution in the percentage of viable spermatozoa. These results suggest that ETEC and VTEC PCR analyses should be done in doses destined for AI to minimize the use of doses with diminished sperm quality due to the presence of bacteria and to avoid the potential spread of infective diseases.

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

    Science.gov (United States)

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

  1. Acrosin-binding protein (ACRBP) and triosephosphate isomerase (TPI) are good markers to predict boar sperm freezing capacity.

    Science.gov (United States)

    Vilagran, Ingrid; Castillo, Judit; Bonet, Sergi; Sancho, Sílvia; Yeste, Marc; Estanyol, Josep M; Oliva, Rafael

    2013-09-15

    Sperm cryopreservation is the most efficient method for storing boar sperm samples for a long time. However, one of the inconveniences of this method is the large variation between and within boars in the cryopreservation success of their sperm. The aim of the present work was thus to find reliable and useful predictive biomarkers of the good and poor capacity to withstand the freeze-thawing process in boar ejaculates. To find these biomarkers, the amount of proteins present in the total proteome in sperm cells were compared between good freezability ejaculates (GFE) and poor freezability ejaculates (PFE) using the two-dimensional difference gel electrophoresis technique. Samples were classified as GFE and PFE using progressive motility and viability of the sperm at 30 and 240 minutes after thawing, and the proteomes from each group, before starting cryopreservation protocols, were compared. Because two proteins, acrosin binding protein (ACRBP) and triosephosphate isomerase (TPI), presented the highest significant differences between GFE and PFE groups in two-dimensional difference gel electrophoresis assessment, Western blot analyses for ACRBP and TPI were also performed for validation. ACRBP normalized content was significantly lower in PFE than in GFE (P sperm viability and motility was confirmed using Pearson's linear correlation. In conclusion, ACRBP and TPI can be used as markers of boar sperm freezability before starting the cryopreservation procedure, thereby avoiding unnecessary costs involved in this practice.

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

    Science.gov (United States)

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

    2011-01-01

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

  3. Cluster analysis reveals a binary effect of storage on boar sperm motility function.

    Science.gov (United States)

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

    2014-06-01

    Storage of liquid-preserved boar spermatozoa is associated with a loss of fertilising ability of the preserved spermatozoa, which standard semen parameters barely reflect. Monitoring responses to molecular effectors of sperm function (e.g. bicarbonate) has proven to be a more sensitive approach to investigating storage effects. Bicarbonate not only initiates capacitation in spermatozoa, but also induces motility activation. This occurs at ejaculation, but also happens throughout passage through the oviduct. In the present study we tested whether the specific response of boar sperm subpopulations to bicarbonate, as assessed by motility activation, is altered with the duration of storage in vitro. Three ejaculates from each of seven boars were diluted in Beltsville thawing solution and stored at 17°C. Only minor changes in the parameters of diluted semen were revealed over a period of 72h storage. For assessment of bicarbonate responses, subsamples of diluted spermatozoa were centrifuged through a discontinuous Percoll gradient after 12, 24 and 72h storage. Subsequently, spermatozoa were incubated in two Ca2+-free variants of Tyrode's medium either without (TyrControl) or with (TyrBic) 15mM bicarbonate, and computer-aided sperm analysis motility measurements were made. Cluster analysis of imaging data from motile spermatozoa revealed the presence of five major sperm subpopulations with distinct motility characteristics, differing between TyrBic and TyrControl at any given time (Psperm motility function descriptors to storage: although the quantitative descriptor (percentage of motile spermatozoa) declines in washed semen samples, the qualitative descriptor (percentage of spermatozoa stimulated into fast linear motion by bicarbonate) is sustained independent of the duration of storage.

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

    Directory of Open Access Journals (Sweden)

    A. V. RUSU

    2013-07-01

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

  5. A Review of New Technologies that may Become Useful for in vitro Production of Boar Sperm.

    Science.gov (United States)

    Gadella, B M; Ferraz, M A

    2015-07-01

    Making sperm cells outside the original testicular environment in a culture dish has been considered for a long time as impossible due to the very complicated process of spermatogenesis and sperm maturation, which altogether, encompasses a 2-month period. However, new approaches in complex three-dimensional co-cell cultures, micro-perfusion and micro-fluidics technologies, new knowledge in the functioning, culturing and differentiation of spermatogonial stem cells (SSC) and their precursor cells have revolutionized this field. Furthermore, the use of better molecular markers as well as stimulatory factors has led to successful in vitro culture of stem cells either derived from germ line stem cells, from induced pluripotent stem cells (iPSC) or from embryonic stem cells (ESC). These stem cells when placed into small seminiferous tubule fragments are able to become SSC. The SSC beyond self-renewal can also be induced into haploid sperm-like cells under in vitro conditions. In mouse, this in vitro produced sperm can be injected into a mature oocyte and allow post-fertilization development into an early embryo in vitro. After transferring such obtained embryos into the uterus of a recipient mouse, they can further develop into healthy offspring. Recently, a similar approach has been performed with combining selected cells from testicular cell suspensions followed by a complete in vitro culture of seminiferous cords producing sperm-like cells. However, most of the techniques developed are laborious, time-consuming and have low efficiency, placing questionable that it will become useful used for setting up an efficient in vitro sperm production system for the boar. The benefits and drawbacks as well as the likeliness of in vitro pig sperm production to become applied in assisted technologies for swine reproduction are critically discussed. In this contribution, also the process of sperm production in the testis and sperm maturation is reviewed.

  6. Assessment of sperm quality traits in relation to fertility in boar semen

    Directory of Open Access Journals (Sweden)

    Zilinskas Henrikas

    2009-12-01

    Full Text Available Abstract Background Several studies have been published where sperm plasma membrane integrity correlated to fertility. In this study we describe a simple fluorometer-based assay where we monitored the fluorescence intensity of artificially membrane-ruptured spermatozoa with a fixed time staining with fluorescent DNA dyes. Methods Membrane-impermeant fluorescent dyes Hoechst 33258 (H258 and propidium iodide (PI were used to measure the fluorescence of the nucleus in artificially membrane ruptured spermatozoa and membrane-permeant dye Hoechst 33342 (H342 was used to measure fluorescence of intact spermatozoa. The concentration of spermatozoa in insemination doses varied from 31.2 × 106/ml to 50 × 106/ml and the average value was 35 × 106/ml. Each boar was represented by three consecutive ejaculates, collected at weekly intervals. Nonreturn rate within 60 days of first insemination (NR % and litter size (total number of piglets born of multiparous farrowings were used as fertility measures. Results Sperm fluorescence intensity of H258 and H342, but not the fluorescence intensity of PI-stained spermatozoa correlated significantly with the litter size of multiparous farrowings, values being r = - 0.68 (P Conclusions The increase in fluorescence values of membrane-ruptured H258 and unruptured H342-stained spermatozoa in boar AI doses can be associated with smaller litter size after AI. This finding indicates that the fluorescence properties of the sperm nucleus could be used to select for AI doses with greater fertilizing potential.

  7. EFFECT OF PROTEINE CONTENT IN BOAR SEMINAL PLASMA ON THE SPERM MOTILITY IN DILUTED SEMEN STORED FOR 3 DAYS

    Directory of Open Access Journals (Sweden)

    Jelena Apić

    2015-02-01

    Full Text Available Recently, it was frequently demonstrated that fertility of sows after artificially inseminated is lower than after mating. This is associated with a reduced fertilization capacity of overdiluted insemination doses. The aim of this study was to investigate the sperm motility in the semen samples, forming from the ejaculates with high or low protein content, stored in vitro on 17oC for 3 days. Progressive motility was significantly higher (p<0.01 in the ejaculates with high, compared to the ejaculates with low protein content (82% vs. 76%. After 3 days of storage, in the1:4 dilution proportion, the average progressive motility was significantly (p<0.01 decreased in relation to this value in native semen from the boars with high (82% to64%, as well from the boars with low protein content in seminal plasma (76% to48%. However, the average diluted semen progressive motility was significantly greater (p<0.01 in the boars with high (64%, compared to the boars with low protein content in seminal plasma (48%. The number of good diluted semen samples (≥65% progressive motility, was also significantly (p<0.01 greater in the boars with high (41%, compared to the boars with low protein content in seminal plasma (12%. These results show that seminal plasma proteins play an important role in maintaining the sperm progressive motility of diluted semen in vitro stored for 3 days.

  8. New strategies of boar sperm cryopreservation: development of novel freezing and thawing methods with a focus on the roles of seminal plasma.

    Science.gov (United States)

    Okazaki, Tetsuji; Shimada, Masayuki

    2012-09-01

    Cryopreservation of boar spermatozoa offers an effective means of long-term storage of important genetic material. Many researchers have investigated how to improve reproductive performance by artificial insemination (AI) using cryopreserved boar spermatozoa. Recently, we and other groups reported that high conception rates (70-80%) can be achieved by AI with frozen-thawed boar spermatozoa using a modified temperature program during freezing, or a novel cryopreservation extender to improve sperm quality (including sperm survivability, motility, membrane status and fertilization ability) after thawing, or a novel sperm infusion method, deep intra uterine insemination. However, these techniques have not yet been used for commercial pig production. The variation in sperm freezability among boars or among ejaculations in an identical boar is one of the main reasons for this problem. In our previous study, it was revealed that some components of seminal plasma have a negative effect on the freezability of boar sperm. One of these factors is bacteria-released endotoxin (lipopolysaccharide: LPS). LPS binds to Toll-like receptor-4 (TLR-4) expressed on the sperm surface, resulting in induction of apoptosis. On the other hand, seminal plasma suppresses cryo-capacitation induced by thawing stress. On the basis of these findings, we designed a novel protocol of AI using frozen-thawed boar sperm.

  9. Influence of chamber type integrated with computer-assisted semen analysis (CASA) system on the results of boar semen evaluation.

    Science.gov (United States)

    Gączarzewicz, D

    2015-01-01

    The objective of the study was to evaluate the effect of different types of chambers used in computer-assisted semen analysis (CASA) on boar sperm concentration and motility parameters. CASA measurements were performed on 45 ejaculates by comparing three commonly used chambers: Leja chamber (LJ), Makler chamber (MK) and microscopic slide-coverslip (SL). Concentration results obtained with CASA were verified by manual counting on a Bürker hemocytometer (BH). No significant differences were found between the concentrations determined with BH vs. LJ and SL, whereas higher (p0.05). The results obtained show that CASA assessment of boar semen should account for the effect of counting chamber on the results of sperm motility and concentration, which confirms the need for further study on standardizing the automatic analysis of boar semen.

  10. 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

  11. Effect of dietary selenium and vitamin E on the ultrastructure and ATP concentration of boar spermatozoa, and the efficacy of added sodium selenite in extended semen on sperm motility.

    Science.gov (United States)

    Marin-Guzman, J; Mahan, D C; Whitmoyer, R

    2000-06-01

    Three experiments evaluated the effects of dietary Se and vitamin E on the ultrastructure of spermatozoa, ATP concentration of spermatozoa, and the effects of adding sodium selenite to semen extenders on subsequent sperm motility. The experiment was a 2 x 2 arrangement of treatments in a randomized complete block design. A total of 10 mature boars were fed from weaning to 18 mo of age diets fortified with two levels of supplemental Se (0 or .5 ppm) or vitamin E (0 or 220 IU/kg diet). The nonfortified diets contained .06 ppm Se and 4.4 IU vitamin E/kg. In Exp. 1, the spermatozoa from all boars were examined by electron microscopy. Vitamin E had no effect on structural abnormalities in the spermatozoa. When the low-Se diet was fed the acrosome or nuclei of the spermatozoa was unaffected, but the mitochondria in the tail midpiece were more oval with wider gaps between organelles. The plasma membrane connection to the tail midpiece was not tightly bound as when boars were fed Se. Immature spermatozoa with cytoplasmic droplets were more numerous when boars were fed the low-Se diet, but the occurrence of midpiece abnormalities occurred in boars fed diets with or without Se or vitamin E. Our results suggest that Se may enhance spermatozoa maturation in the epididymis and may reduce the number of sperm with cytoplasmic droplets. In Exp. 2, the concentration of ATP in the spermatozoa was evaluated in the semen of all treatment boars. When the low-Se diet was fed, ATP concentration was lower (P spermatozoa. Overall, these results suggest that low Se-diets fed to boars resulted in abnormal spermatozoal mitochondria, a lower ATP concentration in the spermatozoa, and a loose apposition of the plasma membrane to the helical coil of the tail midpiece, but no effect from inadequate vitamin E was demonstrated. Adding sodium selenite to the semen extender reduced sperm cell motility.

  12. Comparison of post-thaw DNA integrity of boar spermatozoa assessed with the neutral comet assay and Sperm-Sus Halomax test kit.

    Science.gov (United States)

    Fraser, L; Parda, A; Filipowicz, K; Strzeżek, J

    2010-10-01

    In this study, we tested the hypothesis whether the neutral Comet assay (NCA) and the Sperm-Sus-Halomax (SSH) test kit could provide similar measurements of post-thaw DNA fragmentation of boar spermatozoa. Whole ejaculates or sperm-rich fractions of boar semen were frozen in an extender containing lactose, lipoprotein fractions isolated from ostrich egg yolk (LPFo), glycerol (lactose-LPFo-G) or in a standard boar semen extender (K3), without the addition of cryoprotective substances. In all boars, both the NCA and SSH test showed similar levels of post-thaw sperm DNA fragmentation in samples of the same ejaculates, regardless of the ejaculate collection procedure and extender. Yet, the levels of post-thaw sperm DNA damage, detected by the NCA and SSH test, were more accentuated in spermatozoa frozen in the absence of cryoprotective substances. Both the NCA and SSH detected variations among individual boars in terms of post-thaw sperm DNA fragmentation. Agreement between the measurements of the NCA and SSH was confirmed by scatter plots of differences, suggesting that the DNA integrity tests could detect the same sperm populations, which were susceptible to cryo-induced DNA damage. The findings of this study indicate that the NCA and the SSH test are effective in detecting similar levels of sperm DNA fragmentation and reinforce their importance in the assessment of frozen-thawed boar semen quality.

  13. Elevated dietary intake of Zn-methionate is associated with increased sperm DNA fragmentation in the boar.

    Science.gov (United States)

    García-Contreras, Adelfa; De Loera, Yasmin; García-Artiga, Carlos; Palomo, Antonio; Guevara, Jesús A; Herrera-Haro, José; López-Fernández, Carmen; Johnston, Steve; Gosálvez, Jaime

    2011-05-01

    Boars fed on ration of 200 ppm Zn methionate showed a significant increase (P Zn methionate. There was a positive correlation (R2 = 0.207; P = 0.002) between % sperm DNA fragmentation (SDF) and the concentration of Zn in spermatozoa. Increased Zn in the diet also resulted in a non-proportional increase in Zn concentration in the testis and spermatozoa but not in the epididymis; Zn in sperm accumulated at levels up to 50 times higher than that found in the seminal plasma and 10-13 times that found in the epididymis and testis, respectively. These results show that supplementation of dietary Zn at a concentration of 200 ppm had an adverse effect on boar sperm DNA quality and may be related to the ability of spermatozoa to accumulate Zn during spermiogenesis.

  14. EFFECT OF THE ADDITION OF SEMINAL PLASMA, VITAMIN E AND INCUBATION TIME ON POST-THAWED SPERM VIABILITY IN BOAR SEMEN

    OpenAIRE

    A. G. C. Pech- Sansores; F. G. Centurión- Castro; J. C. Rodríguez-Buenfil; J. C. Segura-Correa; J. R. Aké-Lopez

    2011-01-01

    The objective of the study was to evaluate the effect of seminal plasma (SP), vitamin E (VE), and incubation time on sperm viability of post-thawed boar semen. Thirty six ejaculates were used and allocated to four treatments: T1, semen + BTS (Belstville Thawing Solution) + 10% SP; T2, semen + BTS + 200¿g/ml VE; T3, semen + BTS + 10% SP + 200¿g/ml VE; T4, semen + BTS (control). Motility (MOT), intact acrosomes (IA), membrane integrity (MI) and mitochondrial activity (MA) were evaluated, at 0 a...

  15. Calcium regulates motility and protein phosphorylation by changing cAMP and ATP concentrations in boar sperm in vitro.

    Science.gov (United States)

    Li, Xinhong; Wang, Lirui; Li, Yuhua; Zhao, Na; Zhen, Linqing; Fu, Jieli; Yang, Qiangzhen

    2016-09-01

    Considering the importance of calcium (Ca(2+)) in regulating sperm capacitation, hyperactivation and acrosome reaction, little is known about the molecular mechanism of action of this ion in this process. In the present study, assessment of the molecular mechanism from the perspective of energy metabolism occurred. Sperm motility variables were determined using computer-assisted sperm analysis (CASA) and the phosphorylation of PKA substrates, tyrosine residues and AMP-activated protein kinase (AMPK) were analyzed by Western blot. Moreover, intracellular sperm-specific glyceraldehyde 3-phosphatedehydrogenase (GAPDH) activity, 3'-5'-cyclic adenosine monophosphate (cAMP) and adenosine 5'-triphosphate (ATP) concentrations were assessed in boar sperm treated with Ca(2+). Results of the present study indicated that, under greater extracellular Ca(2+)concentrations (≥3.0mM), sperm motility and protein phosphorylation were inhibited. Interestingly, these changes were correlated with that of GAPDH activity, AMPK phosphorylation, cAMP and ATP concentrations. The negative effects of Ca(2+) on these intracellular processes were attenuated by addition of the calmodulin (CaM) inhibitor W7 and the inhibitor of calmodulin-dependent protein kinase (CaMK), KN-93. In the presence of greater extracellular Ca(2+), however, the phosphorylation pathway was suppressed by H-89. Taken together, these results suggested that Ca(2+) had a dual role in regulating boar sperm motility and protein phosphorylation due to the changes of cAMP and ATP concentrations, in response to cAMP-mediated signal transduction and the Ca(2+) signaling cascade. The present study provided some novel insights into the molecular mechanism underlying the effects of Ca(2+) on boar sperm as well as the involvement of energy metabolism in this mechanism.

  16. Expression of α-gustducin and α-transducin, G proteins coupled with taste receptors, in boar sperm.

    Science.gov (United States)

    Spinaci, M; Bucci, D; Mazzoni, M; Giaretta, E; Bernardini, C; Vallorani, C; Tamanini, C; Clavenzani, P; Galeati, G

    2014-07-01

    During the transit in the female genital tract, spermatozoa are exposed to an environment that varies in composition from the vagina to the oviduct. Because G proteins, α-gustducin and α-transducin, are accepted as markers of chemosensitive cells, this study was aimed at assessing whether these proteins are expressed in boar germ cells. Ejaculated sperm extracts were analyzed by Western blot, and indirect immunofluorescence was performed on testis sections, smears of epididymal and ejaculated spermatozoa, sperm cells after in vitro induction of capacitation and acrosome reaction (IVAR), and in sperm cells bound to zona pellucida during IVF. Based on immunoblot results, both G proteins are present in boar sperm. In the testicular tissue sections, α-gustducin and α-transducin positivity was recorded in the germinal cells near the tubular lumen, whereas no positive signal was evident in spermatogonia located in the outer region of the seminiferous tubules. α-Gustducin expression in epididymal and ejaculated spermatozoa was mainly detectable in both the acrosome and the principal piece of the tail, whereas α-transducin was confined to the acrosome and the midpiece. No changes after in vitro induction of capacitation and IVAR were observed, except for the disappearance of acrosomal positivity in reacted spermatozoa. In sperm bound to zona pellucida, the G protein signal was congruent with that observed in IVAR cells. To the best of our knowledge, this is the first description of α-transducin in mammalian sperm and the first description of α-gustducin in boar sperm. Further studies are needed to clarify the possible role of these G proteins in sperm physiology.

  17. 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 sperm with high intracellular H2O2 level were significantly lower in the 6000G + BSA group than under other treatments (P boar sperm.

  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.

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

    Science.gov (United States)

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

    2011-06-01

    The presented work focuses on electrophoretic and zymographic characterization of boar sperm proteins isolated by various extraction methods and on comparison of the protein profiles obtained from ejaculated and in vitro capacitated spermatozoa. Sperm proteins of ejaculated and in vitro capacitated boar sperms were isolated with the following agents: 1% v/v Triton X-100, 1% v/v Triton X-114, 2% v/v acetic acid, 1% m/v sodium dodecyl sulphate (SDS), 30 mM N-octyl-β-D-glucopyranoside (OBG), rehydration buffer (RHB) for isoelectric focusing and finally by the freezing-thawing approach. The extracts were characterized in terms of 1-DE, 2-DE protein profiles, 1-DE glycoprotein staining and proteinase and hyaluronidase substrate zymographic profiles. The results have shown quantitative and qualitative differences in 1-DE protein and glycoprotein profiles with respect to the employed isolation approach. These differences were seen even more clearly in 2-DE protein profiles, where it was possible to distinguish the presence/absence, changes in relative abundance and pI/M(r) shifts of various protein spots. Proteinase and hyaluronidase zymograms supported the prediction that various isolation protocols result in various profiles of enzymatically active molecules.

  1. Effect of the holding time at 15 °C prior to cryopreservation, the thawing rate and the post-thaw incubation temperature on the boar sperm quality after cryopreservation.

    Science.gov (United States)

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

    2014-01-30

    The aim of the present study was to evaluate the effect of the holding time at 15 °C prior to cryopreservation (2, 4 and 8h), thawing rate (37 °C for 20s or 70 °C for 8s) and post-thaw incubation temperature (15 °C or 37 °C) on the post-thaw boar sperm quality. These are important time periods in the freezing-thawing process which have been less studied. Sperm-rich ejaculate fractions from three healthy boars were collected once a week for five consecutive weeks and were cryopreserved with the lactose-egg yolk extender (LEY). Sperm quality was determined by assessing the motility, the acrosome status, and the sperm plasma membrane integrity at 30, 150 and 240 min of incubation. The results show that with the holding time at 15 °C prior to cryopreservation there was not a clear effect until at least 24h of holding time. The thawing rate and the post-thaw incubation temperature, however, had a marked effect on sperm quality. When the samples were thawed at 70 °C for 8s, the sperm viability, motility and some kinetic variables (VCL, VSL, VAP and ALH) were greater than with results observed when the samples were thawed at 37 °C for 20s. In addition after thawing the sperm samples incubated at 15 °C had a sustained sperm quality for longer, up to 4h post-thawing.

  2. The activity of paraoxonase type 1 (PON-1) in boar seminal plasma and its relationship with sperm quality, functionality, and in vivo fertility.

    Science.gov (United States)

    Barranco, I; Tvarijonaviciute, A; Perez-Patiño, C; Alkmin, D V; Ceron, J J; Martinez, E A; Rodriguez-Martinez, H; Roca, J

    2015-03-01

    Paraoxonase 1 (PON-1) is a hydrolytic enzyme present in body fluids, capable of protecting cells against oxidative stress. The hypothesis was hereby to test that PON-1, present in seminal plasma (SP), acts protecting boar spermatozoa when showing a reasonable high activity in the ejaculate. SP-PON-1 activity differed (p boars (from 0.10 to 0.29 IU/mL). Intra-boar variability was also observed (p boars. SP-PON-1 activity differed among ejaculate portions, showing the spermatozoa-peak portion of spermatozoa-rich ejaculate fraction the highest levels (0.35 ± 0.03 IU/mL, ranging from 0.12 to 0.69) and the post-sperm ejaculate fraction the lowest levels (0.12 ± 0.01 IU/mL, ranging from 0.03 to 0.21). SP-PON-1 activity was positively correlated with the percentage of spermatozoa with rapid and progressive movement (p boars with highest farrowing rates. In conclusion, SP-PON-1 activity differed among boars and ejaculate fractions/portions. SP-PON-1 activity was positively correlated with sperm quality and functionality of liquid-stored semen samples and it evidenced a positive association with in vivo fertility.

  3. Boar sperm changes after sorting and encapsulation in barium alginate membranes.

    Science.gov (United States)

    Spinaci, M; Bucci, D; Chlapanidas, T; Vallorani, C; Perteghella, S; Communod, R; Vigo, D; Tamanini, C; Galeati, G; Faustini, M; Torre, M L

    2013-09-15

    A routine use of boar-sexed semen is limited by the long sorting time necessary to obtain an adequate number of sexed spermatozoa for artificial insemination and by the high susceptibility of spermatozoa of this species to damages induced by sorting procedure and subsequent cryopreservation. The aim of this work was to study the impact of encapsulation in barium alginate membrane on sorted boar spermatozoa by evaluating membrane integrity, chlortetracycline staining patterns, protein tyrosine phosphorylation, and Hsp70 immunolocalization during storage over 72 hours in liquid or encapsulated form. The encapsulation procedure significantly (P < 0.05) decreased the overall membrane integrity of control unsorted semen (81.8 vs. 57.4, CTR vs. CPS), but did not negatively affect the overall viability and the chlortetracycline staining patterns of sorted encapsulated cells. Moreover, encapsulation significantly decreased (P < 0.05) the overall phosphotyrosin A pattern cell percentage in unsorted (98.4 vs. 92.6, CTR vs. CPS) but not in sorted semen (64.0 vs. 74.2; SORT CTR vs. SORT CPS). As for Hsp70, the overall percentage of cells displaying the different patterns was significantly influenced (P < 0.05) by treatment but not by storage time. The sorting procedure seems to induce the major changes, whereas encapsulation tends to exert a protective effect on sorted semen by increasing the percentage of spermatozoa displaying the T pattern (2.8 vs. 24.3; SORT CTR vs. SORT CPS). In conclusion, our data confirm that the damaging impact of the encapsulation in barium alginate capsules seems to be limited when compared with that of the sorting procedure and, moreover, the association of the two procedures does not result in an algebraic sum of the negative effects. These results suggest the possibility of a future utilization of the encapsulation technology in order to store sorted spermatozoa and permit their controlled release in the female genital tract.

  4. 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 (Psperm 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.

  5. Effect of number of motile frozen-thawed boar sperm and number of fixed-time inseminations on fertility in estrous-synchronized gilts

    Science.gov (United States)

    There are advantages for use of frozen-thawed boar sperm (FTS) as a tool for preservation and transfer of valuable genetic material, despite its practical limitations. We hypothesized that increasing the number of motile FTS and number of timed artificial inseminations (AI) would improve pregnancy r...

  6. 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 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 < 0.0001) and an effect of time of evaluation (P < 0.0001) on MOT and ACR. Thawing at 70 °C for 20 seconds reduced (P < 0.05) MOT, VIA, and ACR compared to other treatments. Experiment 3 tested the effects of storage temperature and time after thawing using 20 ejaculates. Samples were thawed, diluted, and allotted to storage at 17 °C, 26 °C, or 37 °C with evaluation at 2, 6, 12, and 24 hours. There was a storage temperature and time effect and an interaction for MOT and VIA (P < 0.0001). Storage at 17 °C and 26 °C increased (P < 0.05) MOT over all times (38.5%) compared to 37 °C (26%), whereas MOT was reduced at intervals. Viability was also greatest with 17 °C and 26 °C compared to 37 °C and was also affected by time and decreased with time. These results indicate that FTS can be held at 17 °C or 26 °C for up to 2 hours before use and would allow for preparation of multiple doses. These data suggest in vitro fertility of FTS is affected by extenders, thawing, and storage.

  7. Direct contact between boar spermatozoa and porcine oviductal epithelial cell (OEC) cultures is needed for optimal sperm survival in vitro.

    Science.gov (United States)

    Yeste, M; Lloyd, R E; Badia, E; Briz, M; Bonet, S; Holt, W V

    2009-07-01

    Oviductal epithelial cell (OEC) co-culture prolongs sperm viability and motility in vitro in a number of species including humans and horses. This study has sought to determine the effects of homologous OEC co-culture on boar sperm function. To determine whether the effects on spermatozoa were specifically caused by co-culture with or by OEC secretions, or by both factors together, a number of co-culture and cell-conditioned medium (CM) experiments were conducted. Firstly, Percoll-washed spermatozoa were co-cultured with OECs and pig kidney epithelial (LLC-PK1) cells, and in medium without cells. Secondly, Percoll-washed spermatozoa were incubated with CM derived from both OECs and LLC-PK1 cells and in unconditioned medium. A number of sperm function parameters were assessed after 5, 30, 60, 90, 120, and 180 min, and 24h of co-culturing or incubation with CM. Of all the sperm function parameters investigated, the percentage (%) viability data yielded the most interesting results. OECs (mean+/-S.E.M.; 31.2+/-1.10) were better than LLC-PK1 cells (24.3+/-0.93) at prolonging the viability of unbound spermatozoa after 24h of co-culturing (Pcells (53.5+/-1.43; Psperm function parameters, e.g., capacitation and motility, were also influenced by OEC co-culturing and incubation with CM, although to a lesser degree. In conclusion, porcine homologous OEC co-culture and CM incubation specifically affect sperm function. However, we propose that it is OEC co-culturing, rather than OEC-CM, that has the greater influence.

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

    Directory of Open Access Journals (Sweden)

    Dorota BANASZEWSKA

    2010-06-01

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

  9. Oligomycin A-induced inhibition of mitochondrial ATP-synthase activity suppresses boar sperm motility and in vitro capacitation achievement without modifying overall sperm energy levels.

    Science.gov (United States)

    Ramió-Lluch, Laura; Yeste, Marc; Fernández-Novell, Josep M; Estrada, Efrén; Rocha, Luiz; Cebrián-Pérez, José A; Muiño-Blanco, Teresa; Concha, Ilona I; Ramírez, Alfredo; Rodríguez-Gil, Joan E

    2014-01-01

    Incubation of boar spermatozoa in a capacitation medium with oligomycin A, a specific inhibitor of the F0 component of the mitochondrial ATP synthase, induced an immediate and almost complete immobilisation of cells. Oligomycin A also inhibited the ability of spermatozoa to achieve feasible in vitro capacitation (IVC), as measured through IVC-compatible changes in motility patterns, tyrosine phosphorylation levels of the acrosomal p32 protein, membrane fluidity and the ability of spermatozoa to achieve subsequent, progesterone-induced in vitro acrosome exocytosis (IVAE). Both inhibitory effects were caused without changes in the rhythm of O2 consumption, intracellular ATP levels or mitochondrial membrane potential (MMP). IVAE was accompanied by a fast and intense peak in O2 consumption and ATP levels in control spermatozoa. Oligomycin A also inhibited progesterone-induced IVAE as well as the concomitant peaks of O2 consumption and ATP levels. The effect of oligomycin on IVAE was also accompanied by concomitant alterations in the IVAE-induced changes on intracellular Ca(2+) levels and MMP. Our results suggest that the oligomycin A-sensitive mitochondrial ATP-synthase activity is instrumental in the achievement of an adequate boar sperm motion pattern, IVC and IVAE. However, this effect seems not to be linked to changes in the overall maintenance of adequate energy levels in stages other than IVAE.

  10. Toxigenic diversity of two different RAPD groups of Stachybotrys chartarum isolates analyzed by potential for trichothecene production and for boar sperm cell motility inhibition

    DEFF Research Database (Denmark)

    Peltola, J.; Niessen, L.; Nielsen, Kristian Fog

    2002-01-01

    Thirty-one isolates of Stachybotrys chartarum from indoor and outdoor environments were analyzed for the presence of the trichodiene synthase (Tri5) gene, trichothecenes, boar sperm cell motility inhibition, and randomly amplified polymorphic DNA banding patterns (RAPDs). Twenty-two S. chartarum...... satratoxins or trichodermol. Nineteen S. chartarum isolates, distributed among the Tri5 gene negative and positive groups, inhibited boar spermatozoan motility at concentrations of less than or equal to60 mug of crude cell extract/mL. The inhibition of motility was independent of satratoxins or atranones...

  11. Morphometry of boar sperm head and flagellum in semen backflow after insemination.

    Science.gov (United States)

    García-Vázquez, Francisco Alberto; Hernández-Caravaca, Iván; Yánez-Quintana, Wellington; Matás, Carmen; Soriano-Úbeda, Cristina; Izquierdo-Rico, María José

    2015-09-01

    Once deposited in the female reproductive system, sperm begin their competition and undergo a selection to reach the site of fertilization. Little is known about the special characteristics of sperm that reach the oviduct and are able to fertilize, with even less information on the role of sperm dimension and shape in transport and fertilization. Here, we examine whether sperm morphometry could be involved in their journey within the uterus. For this purpose, sperm head dimension (length, width, area, and perimeter) and shape (shape factor, ellipticity, elongation, and regularity), and flagellum length were analyzed in the backflow at different times after insemination (0-15, 16-30, and 31-60 minutes). Sperm morphometry in the backflow was also analyzed taking into account the site of semen deposition (cervical vs. intrauterine). Finally, flagellum length was measured at the uterotubal junction. Sperm analyzed in the backflow were small (head and flagellum) with different head shapes compared with sperm observed in the dose before insemination. The site of deposition influenced head morphometry and tail size both being smaller in the backflow after cervical insemination compared with intrauterine insemination. Mean tail length of sperm collected in the backflow was smaller than that in the insemination dose and at the uterotubal junction. Overall, our results suggest that sperm size may be involved in sperm transport either because of environment or through sperm selection and competence on their way to encounter the female gamete.

  12. 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.

  13. Statistical Approach to Boar Semen Evaluation Using Intracellular Intensity Distribution of Head Images

    NARCIS (Netherlands)

    Sánchez, L.; Petkov, N.; Alegre, E.

    2006-01-01

    We propose a method for the classification of boar sperm heads based on their intracellular intensity distributions observed in microscopic images. The image pre-processing comprises segmentation of cell heads and normalization for brightness, contrast and size. Next, we define a model distribution

  14. 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.

  15. 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.

  16. The effect of numbers of frozen-thawed boar sperm and addition of prostaglandin F2α at insemination on fertility in pigs.

    Science.gov (United States)

    Knox, Robert V; Yantis, Brandon M

    2014-12-30

    Frozen-thawed boar sperm (FTS) has reduced fertility compared to liquid semen. Exogenous prostaglandin administered at insemination has been reported to improve cases of low fertility. This experiment tested the effect of number of FTS and addition of prostaglandin (PGF2α) on fertility. The experiment was performed in replicates using weaned sows (n=24) and synchronized gilts (n=94). All females were induced into estrus using PG600® at weaning or following estrus synchronization. At estrus, females received 0.5, 1.0, or 2 billion motile FTS (n=9 boars) with 0 or 5mg of PGF2α added into each AI dose at insemination. Inseminations occurred at 24 and 36h after onset of estrus and ovulation was monitored by ultrasound. Pregnancy and litter size were determined for sows at farrowing and d 50 of gestation for gilts at slaughter. There was no effect of PGF2α and no interaction with dose of FTS or parity on fertility (P>0.10). Pregnancy rate was affected by FTS dose (P0.10) but was influenced by boar (Ppigs than 0.5×10(9) sperm (6.9±0.9). Litter size was also affected by parity (P=0.001) and boar (P<0.01). These results indicate that AI using 2.0×10(9) FTS can result in acceptable pregnancy rates and litter sizes but with no measurable benefit for addition of prostaglandin.

  17. Type-1 cannabinoid receptors reduce membrane fluidity of capacitated boar sperm by impairing their activation by bicarbonate.

    Directory of Open Access Journals (Sweden)

    Barbara Barboni

    Full Text Available BACKGROUND: Mammalian spermatozoa acquire their full fertilizing ability (so called capacitation within the female genital tract, where they are progressively exposed to inverse gradients of inhibiting and stimulating molecules. METHODOLOGY/PRINCIPAL FINDINGS: In the present research, the effect on this process of anandamide, an endocannabinoid that can either activate or inhibit cannabinoid receptors depending on its concentration, and bicarbonate, an oviductal activatory molecule, was assessed, in order to study the role exerted by the type 1 cannabinoid receptor (CB1R in the process of lipid membrane remodeling crucial to complete capacitation. To this aim, boar sperm were incubated in vitro under capacitating conditions (stimulated by bicarbonate in the presence or in the absence of methanandamide (Met-AEA, a non-hydrolysable analogue of anandamide. The CB1R involvement was studied by using the specific inhibitor (SR141716 or mimicking its activation by adding a permeable cAMP analogue (8Br-cAMP. By an immunocytochemistry approach it was shown that the Met-AEA inhibits the bicarbonate-dependent translocation of CB1R from the post-equatorial to equatorial region of sperm head. In addition it was found that Met-AEA is able to prevent the bicarbonate-induced increase in membrane disorder and the cholesterol extraction, both preliminary to capacitation, acting through a CB1R-cAMP mediated pathway, as indicated by MC540 and filipin staining, EPR spectroscopy and biochemical analysis on whole membranes (CB1R activity and on membrane enriched fraction (C/P content and anisotropy. CONCLUSIONS/SIGNIFICANCE: Altogether, these data demonstrate that the endocannabinoid system strongly inhibits the process of sperm capacitation, acting as membrane stabilizing agent, thus increasing the basic knowledge on capacitation-related signaling and potentially opening new perspectives in diagnostics and therapeutics of male infertility.

  18. Quality of boar spermatozoa from the sperm-peak portion of the ejaculate after simplified freezing in MiniFlatpacks compared to the remaining spermatozoa of the sperm-rich fraction.

    Science.gov (United States)

    Siqueira, A P; Wallgren, M; Hossain, M S; Johannisson, A; Sanz, L; Calvete, J J; Rodríguez-Martínez, H

    2011-04-15

    Boar sperm viability post-thaw differs depending on the ejaculate fraction used, with spermatozoa present in the first 10 mL of the sperm-rich fraction (SRF) (portion 1, P1, sperm-peak portion) displaying the best cryosurvival in vitro compared with that of spermatozoa from the rest of the ejaculate (portion 2 of the SRF plus the post-spermatic fraction), even when using simplified freezing routines. This viability apparently relates to the specific profile of seminal plasma in P1 (i.e., glycoprotein and bicarbonate concentrations, and pH). However, spermatozoa from P1 have not been compared with spermatozoa from the rest of the SRF (SRF-P1, usually 30-40 mL of the SRF), which is routinely used for freezing. We compared P1 with SRF-P1 in terms of sperm kinematics (using the QualiSperm™ system), while membrane integrity (SYBR-14/PI), acrosome integrity (FITC PNA/PI), and sperm membrane stability (Annexin-V) were explored using flow cytometry. As well, total protein concentration and the proteomics of the seminal plasma (SP) of both portions of the SRF were studied using two-dimensional electrophoresis (2DE), mass fingerprinting (MALDI-TOF), and collision-induced dissociation tandem mass spectrometry (CID-MS/MS) on selected peptides. The SRF portions were collected weekly from four mature boars (4-5 replicates per boar, sperm concentration: P1, 1.86 ± 0.20; SRF-P1, 1.25 ± 0.14 × 10(9) spz/mL) and processed using a quick freezing method in MiniFlatPacks. Post-thaw sperm motility reached 50%, without differences between SRF portions, but with clear inter-boar variation. Neither plasma membrane nor acrosome integrity differed (ns) between fractions. These results indicate that there are no differences in cryosurvival after quick freezing of boar spermatozoa derived from either of the two SRF portions. While P1 and SRF-P1 clearly differed in relative total protein contents, as expected, they displayed very similar protein profiles as assessed using 2DE and mass

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

    NARCIS (Netherlands)

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

    2005-01-01

    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

  20. Effects of Three Different Diluents on Quality of Boar Semen Stored at 17℃

    Institute of Scientific and Technical Information of China (English)

    Hu Shan; Zhang Xiao-gang; Han Cong; Wei Shuai-yi; Xie Dong-qi; Du Ren-rang; Hu Jian-hong

    2015-01-01

    To investigate the effects of different diluents on the quality of the boar semen stored at 17℃, and assess the relationship between sperm motility and the relative levels of enzymes, three commercial diluents (DiluentⅠ, DiluentⅡand DiluentⅢ) and three boar breed semens (Yorkshire, Landrace and Duroc) were utilized. The sperm motility, effective survival time, survival index, catalase (CAT), the total anti-oxidative capacity (T-AOC) and malondialdehyde (MDA) levels were evaluated. The results showed that there were significant interaction effects between diluents and breeds on the boar sperm motility (P0.05). All of the parameters varied significantly with the increase of the storage time (P<0.001). The survival time increased 12.9% in Yorkshire boar semen diluted with DiluentⅢ than with DiluentⅡ, while the survival time increased 6.6% in Landrace boar semen diluted with DiluentⅡ than with DiluentⅢ. Both CAT and T-AOC levels were significantly positive correlated with sperm motility in all the three boar breeds (P<0.001), while MDA levels were significantly negative correlated with sperm motility (P<0.001). These results indicated that DiluentⅢ and DiluentⅡwere the optimal commercial diluents for Yorkshire and Landrace boar semen stored at 17℃, respectively.

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

    Science.gov (United States)

    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 cr...

  2. A genome-wide association study reveals a novel candidate gene for sperm motility in pigs

    NARCIS (Netherlands)

    Diniz, D.B.; Lopes, M.S.; Broekhuijse, M.L.W.J.; Lopes, P.S.; Harlizius, B.; Guimaraes, S.E.F.; Duijvesteijn, N.; Knol, E.F.; Silva, F.F.

    2014-01-01

    Sperm motility is one of the most widely used parameters in order to evaluate boar semen quality. However, this trait can only be measured after puberty. Thus, the use of genomic information appears as an appealing alternative to evaluate and improve selection for boar fertility traits earlier in li

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

    Science.gov (United States)

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

    2012-06-01

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

  4. Effect of conventional and controlled freezing method on the post thaw characteristics of boar spermatozoa.

    Science.gov (United States)

    Baishya, S K; Biswas, R K; Kadirvel, G; Deka, B C; Kumar, Suresh; Sinha, S; Dutta, D J; Saikia, G K

    2014-10-01

    The objective of the present study was to evaluate the effectiveness of conventional, and controlled freezing method adopting three freezing rates 20°C, 40°C and 60°C/min for cryopreservation of boar semen. Sixty sperm-rich fractions of ejaculates from six boars were utilized for freezing of semen with different freezing methods in lactose-egg yolk glycerol extender using 0.5 ml straws. Semen samples were evaluated for sperm motility, live sperm, acrosome integrity, plasma membrane integrity (PMI) by carboxyfluorescein diacetate plus propidium iodide (PI) staining, mitochondrial membrane potential (MMP) by combined JC-1 plus PI staining and lipid peroxidation (LPO) by BODIPY (581/591)-C11 probe after equilibration and after freezing. The results revealed that the post thaw sperm motility, live sperm, live intact acrosome and plasma membrane integrity were significantly (p0.05) mean values of live sperm with high MMP as compared to conventional freezing. However, the post thaw sperm LPO did not influence by difference in freezing methods. No significant difference on the post thaw sperm qualities was recorded among the three controlled freezing rates. All the sperm parameters assessed declined significantly (pboar semen with controlled freezing methods conferred better post thaw sperm quality as compared to conventional method, and the freezing rates of either 20, 40 or 60°C/min could provide better freezability of boar semen.

  5. 实时精子分离系统对猪精子冷冻品质的影响%Effect of real-time sperm separation system on the quality of boar sperm frozen

    Institute of Scientific and Technical Information of China (English)

    杨海; 李青旺; 胡建红; 张树山

    2011-01-01

    【Objective】 The study was to explore the effect of real-time sperm separation system on the quality of boar semen frozen.【Method】 Original semen was separated with real-time sperm separation system and tubule-froze with liquid nitrogen.The original semen was looked as control group,and the effect of real-time sperm separation system on the quality of boar semen frozen was detected.【Result】 After the separated semen was thawed,the sperm motility,the rate of spermatozoa with intact acrosome,the rate of spermatozoa with normal plasma membrane,the rate of spermatozoa with normal chromatin of the separated group were higher than those of the control group(P0.01);For the rate of spermatozoa with normal morphology,the separated group was better than that of the control group(P0.01),but it was not affected by the freeze-thaw process(P0.01);The spermatozoa density was not significantly affected by the separation process(P0.01).【Conclusion】 The real-time sperm separation system can significantly improve the quality of boar frozen semen.%【目的】探索实时精子分离系统对猪精子冷冻品质的影响。【方法】采用实时精子分离系统对精液进行分离优选,液氮冷冻法制作细管冻精,以未分离精液为对照,检测实时精子分离系统对猪精子冷冻品质的影响。【结果】分离优选精子解冻后,精子活率、顶体完整率、质膜完整率、正常染色质率均明显高于对照组(P〈0.01);分离组精子的正常形态率也显著高于对照组,且未受冷冻-解冻过程的影响(P〉0.01);分离过程对精子密度也未产生显著影响(P〉0.01)。【结论】实时精子分离系统可以显著改善猪冷冻精液的品质。

  6. 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.

  7. Fluorescent multiple staining and CASA system to assess boar sperm viability and membranes integrity in short and long-term extenders

    Directory of Open Access Journals (Sweden)

    F. Cremonesi

    2013-03-01

    Full Text Available The aim of this study was to assess the effect on boar spermatozoa quality of in vitro storage in short and long-term extenders by fluorescent multiple staining (FMS and computer assisted semen analyzer (CASA. Fresh ejaculates from three healthy, sexually mature boars were diluted with equal volumes of six short-term or three long-term commercial extenders and stored at 19°C for 6 days (short-term or 12 days (long-term. The integrity of spermatozoa membranes was analyzed by FMS using propidium iodide, 5,5’,6,6’-tetrachloro-1,1’,3,3’ tetraethylbenzimidazolyl-carbocyanine iodide (JC-1 and fluorescein isothiocyanate-conjugated peanut agglutinin (PNA. The results obtained from this staining were compared with spermatozoa motility assessed by CASA. Our study showed that the number of viable spermatozoa with non-reacted acrosomes and intact mitochondria was positively correlated with the rate of motile spermatozoa (r2>0.9 irrespective of the extender used. In all extenders the number of motile spermatozoa significantly decreased as preservation period increased (P<0.05. FMS test is a potent indicator of sperm motility because it analyses mitochondrial integrity independently from observable alterations in motility. The best performing extenders were BTS for short-term storage and TRI-x-Cell for long-term storage.

  8. Fluorescent multiple staining and CASA system to assess boar sperm viability and membranes integrity in short and long-term extenders.

    Science.gov (United States)

    Lange-Consiglio, A; Meucci, A; Cremonesi, F

    2013-01-01

    The aim of this study was to assess the effect on boar spermatozoa quality of in vitro storage in short and long-term extenders by fluorescent multiple staining (FMS) and computer assisted semen analyzer (CASA). Fresh ejaculates from three healthy, sexually mature boars were diluted with equal volumes of six short-term or three long-term commercial extenders and stored at 19°C for 6 days (short-term) or 12 days (long-term). The integrity of spermatozoa membranes was analyzed by FMS using propidium iodide, 5,5',6,6'-tetrachloro-1,1',3,3' tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) and fluorescein isothiocyanate-conjugated peanut agglutinin (PNA). The results obtained from this staining were compared with spermatozoa motility assessed by CASA. Our study showed that the number of viable spermatozoa with non-reacted acrosomes and intact mitochondria was positively correlated with the rate of motile spermatozoa (r(2)>0.9) irrespective of the extender used. In all extenders the number of motile spermatozoa significantly decreased as preservation period increased (P<0.05). FMS test is a potent indicator of sperm motility because it analyses mitochondrial integrity independently from observable alterations in motility. The best performing extenders were BTS for short-term storage and TRI-x-Cell for long-term storage.

  9. Fennel (Foeniculum vulgare) provides antioxidant protection for boar semen cryopreservation.

    Science.gov (United States)

    Malo, C; Gil, L; Cano, R; González, N; Luño, V

    2012-05-01

    Boar semen is extremely vulnerable to cold shock and it is also sensitive to peroxidation due to the high content of unsaturated fatty acids in the plasma membrane. Antioxidants exert a protective effect on the plasma membrane of frozen boar sperm. Fennel has been shown to contain antioxidant substances. Therefore, this study was performed to evaluate the effect of different concentrations of fennel added to the freezing extender on boar semen quality and lipid peroxidation after thawing. Semen collected from four boars was cryopreserved in lactose-egg-yolk extender or in the same extender with varying concentration of fennel essences: low (LF); medium (MF); high (HF). Analysis of data clearly indicated that higher concentrations of fennel produced significant improvement in total motility. Moreover, when fennel was included in the extender, a dose-dependent tendency to increase sperm viability was observed. In contrast, the addition of fennel had no effect on acrosome integrity or hypoosmotic swelling test (HOST) compared with the control. Malondialdehyde (MDA) formation decreased significantly in fennel groups, yielding similar results for MF and HF. Fennel seems a new antioxidant for use in sperm cryopreservation, but its particular effects on sperm physiology must be further studied, especially the causes of motility stimulation and its effect on lipoxidation.

  10. Effects of Chinese Herb Feed Additives on Sperm Quality and Reproductive Hormone of Boars%中草药复方制剂对公猪精液品质和生殖激素的影响

    Institute of Scientific and Technical Information of China (English)

    严迪华; 常争艳; 李锐; 肖湘; 陈志军; 李文平

    2012-01-01

    为了研究中草药复方制剂对公猪精液品质和生殖激素的影响,对27头精液品质相近的成年大约克种公猪饲喂不同剂量的中草药复方制剂,测定精液指标和血清生殖激素水平。结果表明:与对照组相比,2%低剂量组(试验Ⅰ组)的采精量、采精持续时间、精子密度,血清中促卵泡刺激素(FSH)、促黄体生成素(LH)和睾酮(T)含量差异显著(P〈0.05);与对照组相比,3%高剂量组(试验Ⅱ组)的采精持续时间、精子活力和血液中LH、T含量显著提高,采精量和血液中FSH含量差异极显著(P〈0.01);高、低剂量组的精子畸形率与对照组相比均有所降低。说明日粮中添加一定剂量中草药复方制剂可显著提高种公猪的精液品质,促进生殖激素的分泌。%In order to research the effects of Chinese herb feed additives on the sperm quality and repro- ductive hormone of boars, 27 Large White boars with similar quality of semen were fed different dose of Chinese herb feed additives and their semen index and level of reproductive hormone in serum were evalu- ated. The experiment results showed that compared with control group, the length of semen, collection time, fresh semen volume, sperm density, and the content of FSH, LH and T in the serum of pigs in the 2% low dose group were improved significantly ( P 〈 0.05), the serum of pigs in the 3 % high dose group had significant difference compared with the control group; fresh semen volume and the content of FSH in the serum of pigs in the high dose group had very significant difference compared with the control group( P 〈 0.01 ). And abnormal sperm rate of pigs in low dose group and high dose group compared with the control group were reduced. In conclution, Chinese herb feed additives was very important for boars to keep a good sperm quality.

  11. Effect of Salvia miltiorrhiza polysaccharides on boar spermatozoa during freezing-thawing.

    Science.gov (United States)

    Shen, Tao; Jiang, Zhong-Liang; Liu, Hong; Li, Qing-Wang

    2015-08-01

    Salvia miltiorrhiza polysaccharides (SMPs) were extracted from S. miltiorrhiza in this study. The aim of the present study was to evaluate the effect of SMP on the motility of boar sperm, including the antioxidant effect of SMP on boar sperm and the effect of SMP on the in vivo fertilizing ability of frozen-thawed boar sperm. Fifty ejaculates from 5 Swagger boars were collected and diluted with an extender, which contained 3% glycerol (v/v) with five concentrations of SMP (0.2, 0.4, 0.6, 0.8, and 1.0mg/mL). The semen was frozen in 0.25mL straws at 1.0×10(9) cells/mL. Sixty gilts were inseminated using fresh semen, frozen semen with 0.4mg/mL of SMP and frozen semen without SMP. The results indicate that the addition of SMP to the extender results in a higher percentage of motile sperm post-thaw (Pboar sperm from peroxidative damage and increase sperm motility and litter size during the process of freezing-thawing. The optimal concentration of SMP for the frozen extenders in this study was determined to be 0.4mg/mL.

  12. Sage (Salvia officinalis) and fennel (Foeniculum vulgare) improve cryopreserved boar epididymal semen quality study.

    Science.gov (United States)

    Monton, A; Gil, L; Malo, C; Olaciregui, M; Gonzalez, N; de Blas, I

    2015-01-01

    The aim of this study was to evaluate the effect of fennel and sage extracts and the influence of the egg yolk source (fresh or pasteurized) on the success of freezing boar epididymal spermatozoa. In experiment 1, epididymal sperm was recovered by flushing and cryopreserved in a lactose-egg yolk solution supplemented with various concentrations (10, 5 and 2.5 g/L) of sage or fennel. Sperm quality was evaluated (motility, viability, HOST and acrosome integrity) at 0 h and 2 h after thawing. Fennel 10 g/L and sage 5 g/L and control (no extracts) were selected for experiment 2 which also compared fresh or pasteurized egg yolk in the freezing extender and measured DNA integrity of the frozen sperm. Results showed that the interaction between fennel and sage antioxidants with fresh egg yolk significantly improved post thaw sperm quality and protected boar epididymal spermatozoa from cryopreservation damage as a result of oxidative stress.

  13. Evaluation of a Commercial ELISA for the Detection of Antibodies to Sarcoptes scabiei in Wild Boar (Sus scrofa).

    Science.gov (United States)

    Haas, Chloé; Rossi, Sophie; Meier, Roman; Ryser-Degiorgis, Marie-Pierre

    2015-07-01

    Sarcoptic mange occurs in free-ranging wild boar (Sus scrofa) but has been poorly described in this species. We evaluated the performance of a commercial indirect enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of sarcoptic mange in domestic swine when applied to wild boar sera. We tested 96 sera from wild boar in populations without mange history ("truly noninfected") collected in Switzerland between December 2012 and February 2014, and 141 sera from free-ranging wild boar presenting mange-like lesions, including 50 live animals captured and sampled multiple times in France between May and August 2006 and three cases submitted to necropsy in Switzerland between April 2010 and February 2014. Mite infestation was confirmed by skin scraping in 20 of them ("truly infected"). We defined sensitivity of the test as the proportion of truly infected that were found ELISA-positive, and specificity as the proportion of truly noninfected that were found negative. Sensitivity and specificity were 75% and 80%, respectively. Success of antibody detection increased with the chronicity of lesions, and seroconversion was documented in 19 of 27 wild boar sampled multiple times that were initially negative or doubtful. In conclusion, the evaluated ELISA has been successfully applied to wild boar sera. It appears to be unreliable for early detection in individual animals but may represent a useful tool for population surveys.

  14. Status of sperm morphology assessment: an evaluation of methodology and clinical value

    NARCIS (Netherlands)

    Hoven, L. van den; Hendriks, J.C.M.; Verbeet, J.G.M.; Westphal, J.R.; Wetzels, A.M.M.

    2015-01-01

    OBJECTIVE: To characterize methodological changes in sperm morphology assessment and to correlate sperm morphology with clinical outcome. DESIGN: In this observational study, sperm morphology profiles of patients were analyzed. The percentages of morphologically normal spermatozoa were evaluated wit

  15. 高温季节糖萜素对种公猪精液质量的影响%Effect of saccharicterpenin on the quality of boar sperm in high temperature season

    Institute of Scientific and Technical Information of China (English)

    褚晓红; 胡锦平; 徐如海; 路伏增; 戴丽荷; 姜红进

    2012-01-01

    研究了浙江高温季节(7~9月)时,种公猪饲料中添加400 g·t-1糖萜素对其精液质量的影响,结果表明,在高温炎热季节里,4头种公猪饲料中添加糖萜素可以提高采精量、精子密度与活力,降低精液pH,但差异不显著(P>0.05),并显著提高睾酮含量(P<0.05),对种公猪的精液质量有一定的促进作用.%This study was conducted to study the effect of saccharicterpenin on the quality of boar sperm in high temperature season (from July to September, 2011) in Zhejiang province, adding 400 g·t-1 saccharicterpenin in a diet of boar. The result indicated that there was an increase in semen volume, density, motility and pH value, but no significant difference (P > 0. 05 ). Meanwhile, there was significant increase ( P < 0. 05 ) in testosterone level. In summary, saccharicterpenin can increase the quality of boar sperm in high temperature season.

  16. 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.

  17. Perspectives on cryopreservation, quality control, and artificial insemination of frozen-thawed boar sperm in a national repository

    Science.gov (United States)

    The National Animal Germplasm Program (NAGP) is a repository for agricultural germplasm (sperm, eggs, embryos, blood, tissue, DNA) that has been collected, cryopreserved, and cataloged for the purpose of creating a living compilation of genetic resources. The repository is multipurpose and can be u...

  18. Effects of dietary supplementation with an organic source of selenium on characteristics of semen quality and in vitro fertility in boars.

    Science.gov (United States)

    Speight, S M; Estienne, M J; Harper, A F; Crawford, R J; Knight, J W; Whitaker, B D

    2012-03-01

    Semen characteristics in boars fed organic or inorganic sources of Se were assessed in 3 experiments. Crossbred boars were randomly assigned at weaning to 1 of 3 dietary treatments: I) basal diets with no supplemental Se (control), II) basal diets with 0.3 mg/kg of supplemental Se from an organic source (Sel-Plex, Alltech Inc., Nicholasville, KY), and III) basal diets supplemented with 0.3 mg/kg of supplemental Se from sodium selenite (Premium Selenium 270, North American Nutrition Co. Inc., Lewisburg, OH). For Exp. 1, semen was collected from boars (n = 10/dietary treatment) on 5 consecutive days at 15 mo of age. Effects of treatment × day were detected for the proportions of progressively motile (P = 0.02) and rapidly moving (P = 0.03) spermatozoa, and measures of sperm velocity, including path velocity of the smoothed cell path (P = 0.05) and average velocity measured in a straight line from the beginning to the end of the track (P = 0.05). Negative effects of day of semen collection on sperm motility were least pronounced in boars fed Sel-Plex. Experiment 2 was conducted when boars were 17 mo of age, and semen was collected (n = 10 boars/dietary treatment), diluted in commercially available extenders, and stored at 18°C for 9 d. Effects of treatment × day were detected for percentages of motile (P = 0.01) and static (P = 0.01) spermatozoa, amplitude of lateral head displacement (P = 0.02), frequency with which the sperm track crossed the sperm path (P = 0.04), straightness (P = 0.01), and average size of all sperm heads (P = 0.03). In general, sperm cells from boars fed Sel-Plex were better able to maintain motility during liquid storage compared with boars fed sodium selenite. For Exp. 3, semen was collected from boars (n = 6/dietary treatment) at 23 mo of age, and spermatozoa were evaluated at d 1 and 8 after semen collection using in vitro fertilization procedures. There was a tendency for an effect (P = 0.11) of dietary treatment on fertilization rate

  19. Effects of (-)-epigallocatechin gallate on the motility and penetrability of frozen-thawed boar spermatozoa incubated in the fertilization medium.

    Science.gov (United States)

    Kaedei, Y; Naito, M; Naoi, H; Sato, Y; Taniguchi, M; Tanihara, F; Kikuchi, K; Nagai, T; Otoi, T

    2012-12-01

    Epigallocatechin gallate (EGCG) is the major polyphenol in green tea (Camellia sinensis) and is known for its antioxidant effects. The objective of the present study was to examine the effects of EGCG during in vitro fertilization (IVF) on the sperm quality and penetrability into oocytes. In the first experiment, the effects of concentration and incubation period of EGCG on the motility and penetrability of spermatozoa were examined. When frozen-thawed spermatozoa were incubated in IVF medium supplemented with 0 (control), 1, 50 and 100 μm EGCG for 1, 3 and 5 h, supplementation with 50 and 100 μm EGCG improved motility of the spermatozoa (p sperm penetration rates. In the second experiment, the effects of supplementation of EGCG in IVF medium on penetrability of sperm from different boars and development of fertilized oocytes were evaluated. When frozen-thawed spermatozoa from six boars were co-incubated with IVM oocytes in IVF medium supplemented with 50 μm EGCG, the effect of EGCG on sperm penetration and development of oocytes after fertilization was found to vary with individual boar. Our results indicate that motility and penetrability of boar spermatozoa are improved by co-incubation with 50 μm EGCG, but the effects vary with individual boars.

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

    Science.gov (United States)

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

    2015-10-01

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

  1. Advances in Boar Semen Cryopreservation

    Directory of Open Access Journals (Sweden)

    Heriberto Rodriguez-Martinez

    2011-01-01

    Full Text Available The present paper highlights aspects of the cryopreservation of boar semen, a species with particular large, fractionated ejaculates, and a cumbersome cryotechnology that had prevented its commercial application. With the dramatic increase of use of liquid pig semen for artificial breeding over the past decade, developments on cryopreservation alongside the routine use of stud boar semen for AI had been promoted. Recent advances in our laboratory, accommodating the best use of portions of the sperm-rich fraction of the ejaculate for cryopreservation of the sperm-peak portion (P1 and parallel use of the rest of the collected ejaculated spermatozoa, appears as a suitable commercial alternative.

  2. Effects of selenium-enriched probiotics on sperm quality of stock boars%富硒益生菌对种公猪精液品质的影响

    Institute of Scientific and Technical Information of China (English)

    苏惠龙; 李儒曙; 贺湘仁; 陈锦珍; 韩卓宙; 刘宇; 张健騑

    2009-01-01

    在成年长白种公猪的饲料中,以0.3 mg/kg(以硒浓度计算)添加富硒益生菌,研究了富硒益牛菌对种公猪精液品质的影响.结果显示,试验组种公猪精液中的精子活率、顶体完整率、还原型谷胱甘肽过氧化物酶的活性均比对照组高,但精液中的精子畸形率比对照组低,说明富硒益生菌能明显改善种公猪的精液品质.%Selenium-enriched probiotics was added into Changbai stock boars' feedstuff according to 0.3 mg/kg in order to investigate the effect of selenium-enriched probiotics on sperm quality in stock boats. The results showed that the sperm motility rate, acrosome integrity rate and GSH-Px active in trial group were better than the control group, while the teratospermia rate in trial group was lower than that in the control group. So we can conclude that selenium-enriched probioties supplementation could significantly enhance sperm quality in stock boars.

  3. Sperm MTT Viability Assay: A New Method for Evaluation of Human Sperm Viability

    OpenAIRE

    Nasr-Esfahani, Mohammad Hossein; Aboutorabi, Roshanak; Esfandiari, Ebrahim; Mardani, Mohammad

    2002-01-01

    Purpose: MTT (3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay is commonly used as a cell proliferation assay. The objective of this study was to evaluate the capability of MTT assay to discriminate between viable and nonviable sperms and compare its efficiency with E&N (eosin and nigrosin) and HOST (hypo-osmotic swelling test).

  4. Digital holographic microscopy for the evaluation of human sperm structure

    CERN Document Server

    Coppola, Gianluca; Wilding, Martin; Ferraro, Pietro; Esposito, Giusy; Di Matteo, Loredana; Dale, Roberta; Coppola, Giuseppe; Dale, Brian

    2013-01-01

    The morphology of the sperm head has often been correlated with the outcome of in vitro fertilization (IVF), and has been shown to be the sole parameter in semen of value in predicting the success of intracytoplasmic sperm injection (ICSI) and intracytoplasmic morphologically selected sperm injection (IMSI). In this paper, we have studied whether Digital Holographic (DH) microscopy may be useful to obtain quantitative data on human sperm head structure and compared this technique to high power digitally enhanced Nomarski microscope. The main advantage of DH is that a high resolution 3-D quantitative sample imaging may be obtained thorugh numerical refocusing at different object planes without any mechanical scanning. We show that DH can furnish useful information on the dimensions and structure of human spermatozoo, that cannot be revealed by conventional phase contrast microscopy. In fact, in this paper DH has been used to evaluate volume and indicate precise location of vacuoles, thus suggesting its use as ...

  5. Determination of sperm acrosin activity for evaluation of male fertility

    Institute of Scientific and Technical Information of China (English)

    Yun-HeCUI; Rui-LanZHAO; QiangWANG; Zi-YingZHANG

    2000-01-01

    Aim: To investigate a simple method for assaying acrosin activity for the evaluation of male fertility. Methods: The acrosin activity of 7.5 × 106 sperm without seminal plasma and acrosin activity inhibitors was assayed using N-α-benzoyl-DL-arginine-p-nitroanilide (BAPNA) and detergent (Triton X-100) as substrate. Results: The acrosin activity of 60 normal fertile men (35±10μIU/106 sperm ) was higher than that of 168 infertile men ( 16±8μIU/106 sperm) (P < 0.01 ). It was indicated that there was a significant positive correlation between the acrosin activity and the sperm motility ( r≥0. 6534, P < 0.01 ) and a significant negative correlation between the sperm malformed rate and the WBC number ( r≤-0. 5426, P < 0.01 ). The temperature and time of incubation and the sperm concentration could influence the assay results. Conclusion: Acrosin activity is an important index for the evaluation of male fertility. The approach developed by the authors is a simple method for the determination of acrosin activity.

  6. Impact of genetic selection on management of boar replacement.

    Science.gov (United States)

    Robinson, J A B; Buhr, M M

    2005-01-15

    Boars in an artificial insemination centre have been selected for their superior genetic potential, with 'superior' being defined as having traits the customer wants transmitted to his herd. The ability to meet the customers' needs depends on the heritability of the trait, the geneticist's success in devising a selection scheme for the trait in balance with other economically important traits, and the boar's ability to produce sperm that can fertilise oocytes. Genetic evaluation research over the past 20 years has greatly increased the number of traits for which a boar can be selected: currently in the Canadian national program, these include age at 100 kg, backfat at 100 kg, feed efficiency, lean yield and litter size. In the near future, traits that are very likely to be added to this selection list include piglet survival, marbling, loin eye area and structure traits. In Canada, sires are ranked on two estimated breeding value (EBV) indices; one, focused on development of terminal sire lines, is based on the growth and yield traits and another, primarily focused on maternal line development, de-emphasises these traits and incorporates litter size. Boars that are in Canadian AI centres because of their excellent growth traits are typically in the top 5-10% of the national population for terminal sire line index, but they may be only average or substandard for litter size. Conversely, boars selected to be in the top 5-10% for conveying such reproductive traits as litter size may only be in the top 33% for growth traits. The more offspring from a superior boar in either of these indices, the faster the population average for the trait improves. The original sire gets knocked out of the elite group, is culled and replaced by a higher ranked young boar from the now improved general population. Although genetic superiority should govern an AI centre's selection and culling of boars, decision-making in real life is seldom that simple. Selection criteria may be

  7. Evaluation of DNA fragmentation in llama (Lama glama) sperm using the sperm chromatin dispersion test.

    Science.gov (United States)

    Carretero, M I; Lombardo, D; Arraztoa, C C; Giuliano, S M; Gambarotta, M C; Neild, D M

    2012-03-01

    The integrity of sperm chromatin is now viewed as an important factor in male fertility and in early embryonic development. The objectives of this study were: (1) adapt the simple and inexpensive sperm chromatin dispersion (SCD) test to evaluate DNA fragmentation in llama sperm and establish the halo patterns observed in this species, (2) determine an effective and reliable positive control for this technique and (3) evaluate correlation between the SCD test and the toluidine blue (TB) stain. To adapt the SCD test, three different mercaptoethanol (ME) concentrations were assayed (2.5%, 5% and 10% ME). To determine an effective positive control, three treatments (incubation at 100 °C for 30 min, incubation with 0.3 M NaOH for 30 min at room temperature and exposure to UV light for 2h) were assayed. The concentration selected to use in the SCD test was 5% ME, because it produced the largest halo while still conserving the structure of the core. Four DNA dispersion patterns were clearly observed: (I) nuclei with large DNA dispersion halos; (II) nuclei with medium halos; (III) nuclei with very small halos and (IV) nuclei with no halo. All treatments used as positive controls were effective in producing DNA fragmentation. A high correlation (r=0.84, P=0.03) was observed between spermatozoa without halos and TB positive cells. To conclude, SCD patterns in llama sperm have been established as well as a repeatable positive control for the assay. The SCD test and TB stain are simple and inexpensive techniques that can be used to evaluate DNA damage in llama sperm.

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

    Directory of Open Access Journals (Sweden)

    Thais Rose dos Santos Hamilton

    2016-01-01

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

  9. Surveillance strategies for Classical Swine Fever in wild boar – a comprehensive evaluation study to ensure powerful surveillance

    Science.gov (United States)

    Schulz, Katja; Peyre, Marisa; Staubach, Christoph; Schauer, Birgit; Schulz, Jana; Calba, Clémentine; Häsler, Barbara; Conraths, Franz J.

    2017-01-01

    Surveillance of Classical Swine Fever (CSF) should not only focus on livestock, but must also include wild boar. To prevent disease transmission into commercial pig herds, it is therefore vital to have knowledge about the disease status in wild boar. In the present study, we performed a comprehensive evaluation of alternative surveillance strategies for Classical Swine Fever (CSF) in wild boar and compared them with the currently implemented conventional approach. The evaluation protocol was designed using the EVA tool, a decision support tool to help in the development of an economic and epidemiological evaluation protocol for surveillance. To evaluate the effectiveness of the surveillance strategies, we investigated their sensitivity and timeliness. Acceptability was analysed and finally, the cost-effectiveness of the surveillance strategies was determined. We developed 69 surveillance strategies for comparative evaluation between the existing approach and the novel proposed strategies. Sampling only within sub-adults resulted in a better acceptability and timeliness than the currently implemented strategy. Strategies that were completely based on passive surveillance performance did not achieve the desired detection probability of 95%. In conclusion, the results of the study suggest that risk-based approaches can be an option to design more effective CSF surveillance strategies in wild boar. PMID:28266576

  10. The increase in phosphorylation levels of serine residues of protein HSP70 during holding time at 17°C is concomitant with a higher cryotolerance of boar spermatozoa.

    Science.gov (United States)

    Yeste, Marc; Estrada, Efrén; Rivera Del Álamo, Maria-Montserat; Bonet, Sergi; Rigau, Teresa; Rodríguez-Gil, Joan-Enric

    2014-01-01

    Boar-sperm cryopreservation is not usually performed immediately after semen collection, but rather a holding time (HT) of 4 h-30 h at 17°C is spent before starting this procedure. Taking this into account, the aim of this study was to go further in-depth into the mechanisms underlying the improving effects of HT at 17°C on boar-sperm cryotolerance by evaluating the effects of two different HTs (3 h and 24 h) on overall boar-sperm function and survival before and after cryopreservation. Given that phospho/dephosphorylation mechanisms are of utmost importance in the overall regulation of sperm function, the phosphorylation levels of serine residues (pSer) in 30 different sperm proteins after a 3 h- or 24 h-HT period were also assessed. We found that a HT of 24 h contributed to a higher sperm resistance to freeze-thawing procedures, whereas mini-array protein analyses showed that a HT of 24 h induced a significant (Psperm cryotolerance was significantly correlated with pSer levels in HSP70. In addition, from all the parameters evaluated before freeze-thawing, only pSer levels in HSP70 resulted to be able to predict sperm cryotolerance. In conclusion, our results suggest that boar spermatozoa modulate its function during HT, at least partially, by changes in pSer levels of proteins like HSP70, and this is related to a higher cryotolerance.

  11. 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.

  12. Evaluation on Sperm Acrosome Integrity of Infertile Men with Varicocele

    Institute of Scientific and Technical Information of China (English)

    P.Tzvetkova; Wei-jie ZHU; Jing LI; D.Tzvetkov

    2007-01-01

    Objective To determine the sperm acrosome integrity of samples from infertile men with varicocele.Methods Forty-nine infertile men with varicocele were divided into three groups according to the grade of varicocele. Group A (grade Ⅰ), B (grade Ⅱ), and C (grade Ⅲ) consisted of 15, 18, and 16 cases, respectively. Besides, 15 semen samples from normospermic donors were used as the control. The acrosome integrity of sperm was examined with fluorescein-labeled Pisum sativum agglutinin. Acrosomal ultrastructure was observed with transmission electron microscopy.Results In three varicocele groups, most samples had high sperm abnormal morphology rates. There were significant differences in acrosome integrity rates between each varicocele group and the control (P<0.01). Group C had the lowest acrosome integrity rate among the three groups. Ultrastructural observation showed that acrosome malformations revealed acrosomal membranes defects, swelling, hypoplasia, and dissolution of the matrix.Conclusions Infertile men with varicocele had low level of acrosome integrity. Severe varicocele for infertile men might be associated with severe acrosomal defects. Evaluating sperm acrosome should aid the understanding of the sperm structural state and benefit the treatment for infertile men.

  13. Evaluation of different strategies to mask boar taint in cooked sausage.

    Science.gov (United States)

    Martínez, B; Rubio, B; Viera, C; Linares, M B; Egea, M; Panella-Riera, N; Garrido, M D

    2016-06-01

    The use of smoking and/or spices was evaluated for their ability to mask boar taint in frankfurters manufactured from entire pigs with high levels of androstenone. Five frankfurter types were considered: control, smoked, flavouring+smoked, spicy and spicy+smoked. A trained panel in androstenone perception carried out a sensory profile on the different sausages. The highest scores for androstenone perception (odour, flavour and aftertaste) were found in frankfurters that included no masking strategy which indicated the effectiveness of the evaluated strategies. Regarding masking strategies, the contribution of spices and smoking to sensory perception of frankfurters was detected by the panellists. Smoking was the best strategy to mask androstenone odour, while the use of spices masked androstenone odour to a greater extent than androstenone flavour. Only the combined use of spices and smoking was able to eliminate the perception of androstenone. The application of this strategy in frankfurters could be an alternative in the commercialization of entire pigs.

  14. Suitability and effectiveness of single layer centrifugation using Androcoll-P in the cryopreservation protocol for boar spermatozoa.

    Science.gov (United States)

    Martinez-Alborcia, Maria J; Morrell, Jane M; Gil, Maria A; Barranco, Isabel; Maside, Carolina; Alkmin, Diego V; Parrilla, Inmaculada; Martinez, Emilio A; Roca, Jordi

    2013-08-01

    The goal of the present experiment was to evaluate the suitability and effectiveness of single layer centrifugation (SLC), using the pig-specific colloid Androcoll-P, as a routine procedure for selecting boar spermatozoa for cryopreservation. The study focuses special attention on the effectiveness of SLC for processing a whole sperm rich ejaculate fraction and the fertilizing ability of frozen-thawed (FT) sperm selected using SLC prior to freezing. Thirteen sperm rich ejaculate fractions (one per boar) were split into three aliquots. Two aliquots of 15 and 150mL were SLC-processed (500×g for 20min) using 15 and 150mL (v/v) of Androcoll-P-Large and Androcoll-P-XL, respectively. The third aliquot remained un-processed as a control. The percentages of spermatozoa that were morphologically normal and showed rapid and progressive motility (assessed by CASA) spermatozoa were higher (Psperm chromatin dispersion test) were lower (Psperm motility (both total motility and rapid progressive motility), viability and intact nuclear DNA were higher (Psperm (679 in vitro matured oocytes inseminated with a viable sperm:oocyte ratio of 300:1 and coincubated for 6h), measured as the percentage of penetrated oocytes and the mean number of swollen sperm heads and/or male pronuclei in penetrated oocytes. However, there was no effect of SLC-processing on the in vitro ability of putative zygotes to develop to blastocysts. Overall these results indicate that SLC-processing of boar ejaculates using Androcoll-P improves the quality and fertilizing ability of cryosurvival boar sperm. However, efforts should be made to ensure continued high recovery yields before considering the inclusion of SLC as a routine procedure in the cryopreservation protocol of boar ejaculates.

  15. 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

    Directory of Open Access Journals (Sweden)

    Müller Karin

    2009-11-01

    Full Text Available 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 detergents Pluronic F-127, sodium cholate, CHAPS, Tween 20, Triton X-100 and Brij 96V. Different patterns of membrane disintegration were observed by light and electron microscopy. In accordance with microscopic data, different amounts of lipids and proteins were released from the cells by the different detergents. The biochemical methods to assay the phosphorus and cholesterol contents as well as 31P NMR to determine the phospholipids were not influenced by the presence of detergents since comparable amounts of lipids were detected in the organic extracts from whole cell suspensions after exposure to each detergent. However, matrix-assisted laser desorption and ionization time-of-flight mass spectrometry applied to identify phospholipids was essentially disturbed by the presence of detergents which exerted particular suppression effects on signal intensities. After separation of the membrane fractions released by detergents on a sucrose gradient only Triton X-100 and sodium cholate produced sharp turbid DRM bands. Only membrane solubilisation by Triton X-100 leads to an enrichment of cholesterol, sphingomyelin, phosphatidylinositol and phosphatidylethanolamine in a visible DRM band accompanied by a selective accumulation of proteins. Conclusion The boar sperm membranes are solubilised to a different extent by the used detergents. Particularly, the very unique DRMs isolated after Triton X-100 exposure are interesting candidates for further studies regarding the

  16. Evaluation on the Morphology and Membrane Integrity of Immotile Human Sperm

    Institute of Scientific and Technical Information of China (English)

    Wei-Jie ZHU; Jing LI

    2006-01-01

    Objective To determine the membrane integrity in the head and tail regions of individual spermatozoon, and observe sperm morphology for samples with totally immotile sperm.Methods Ten infertile men with immotile sperm were enrolled into this study (group A).The membrane integrity in the head and tail regions of individual spermatozoon of immotile sperm was examined by using the combined hypo-osmotic swelling-eosin Y exclusion test (HOS-EY test). Sperm morphology was observed by light, scanning and transmission electron microscopy. Ten semen samples from normospermic donors were used as the control (group B).Results The percentage of sperm with intact both head and tail membranes in group A was significantly lower than that in group B (P<0.01), whereas the value of sperm with defective head membrane but intact tail membrane in group A was significantly higher than that in group B (P<0.01) Abnormal sperm morphology in group A had a high incidence, and immotile sperm with viability and normal morphology could be observed in some cases. Most sperm had multiple ultrastructural defects.Conclussion Some immotile sperm had intact tail membrane but defective head membrane. Immotile sperm with viability and normal morphology could exist in some cases though abnormal sperm were in a great proportion. Carefully evaluating immotile sperm membrane integrity and morphology should benefit the treatment of patients with immotile sperm.

  17. Handling of boar spermatozoa during and after flow cytometric sex-sorting process to improve their in vitro fertilizing ability.

    Science.gov (United States)

    del Olmo, D; Parrilla, I; Gil, M A; Maside, C; Tarantini, T; Angel, M A; Roca, J; Martinez, E A; Vazquez, J M

    2013-09-01

    The objective of this study was to develop an adequate sperm handling protocol in order to obtain a sex-sorted sperm population with an optimal fertilizing ability. For this purpose, different aspects of the sorting procedure were examined. The effects of the high dilution rates (experiment 1), type of collection medium used (experiment 2), and sheath fluid composition (experiment 3) on sorted boar sperm quality and function were evaluated. Sperm quality was assessed by motility and viability tests, whereas sperm function was evaluated by an in vitro fertilization assay which determined the penetration and polyspermy rates as well as the mean number of sperm penetrating each oocyte. In experiment 1, the results obtained indicated that the high dilution rates did not cause a decrease either in the sperm quality parameters evaluated or the in vitro fertilization ability of spermatozoa. In experiment 2, although sperm quality was not affected, fertilizing ability was compromised after sorting, regardless of the collection medium that was used. In the experiment 3, all groups displayed adequate sperm quality values, but higher in vitro fertility parameters were obtained for spermatozoa sorted in presence of EDTA in the sheath fluid and egg yolk (EY) in the collection media when compared with those sorted in absence of these protective agents. No differences in penetration rates between unsorted highly diluted (control) and sorted sperm in the presence of EDTA and EY were observed. In conclusion, fertilizing ability was compromised in sex-sorted sperm. The addition of EDTA to sheath fluid and EY to collection medium improved boar sperm fertilizing ability, and both agents should be included as essential media components in future studies.

  18. Changes in the distribution and molecular mass of boar sperm acrosome-associated 1 proteins during the acrosome reaction; their validity as indicators for occurrence of the true acrosome reaction.

    Science.gov (United States)

    Ogura, Yukari; Takagishi, Yuki; Harayama, Hiroshi

    2016-09-01

    The aims of this study were to investigate changes in the distribution and molecular mass of boar sperm acrosome-associated 1 (SPACA1) proteins during the acrosome reaction and to discuss validity of SPACA1 proteins as indicators for occurrence of the true acrosome reaction. Boar ejaculated spermatozoa were used for induction of the extracellular Ca(2+)-dependent acrosome reaction (true acrosome reaction) or acrosomal damages (false acrosome reaction) and then subjected to double staining with the anti-SPACA1 protein antibody and FITC-PNA and Western blotting. Extracellular Ca(2+)-dependently acrosome-reacted spermatozoa were characterized by appearance of SPACA1 proteins in the postacrosomal region (; these spermatozoa were classified into SP-3&AR pattern of double staining). However, SPACA1 proteins were not observed in the postacrosomal region of frozen-thawed spermatozoa with severely damaged acrosomes (; these spermatozoa were classified into SP-2&AR pattern). Moreover, the spermatozoa in which acrosomes were severely damaged by incubation with cyclodextrins and without CaCl2 were classified into either SP-2&AR or SP-3&AR pattern. Although SPACA1 proteins were detected mainly as 36-42kDa proteins in the spermatozoa with intact acrosomes, small types of SPACA1 proteins (15-28kDa) increased in extracellular Ca(2+)-dependently acrosome-reacted spermatozoa as well as frozen-thawed spermatozoa with damaged acrosomes. These results show the increase of boar spermatozoa classified into SP-3&AR pattern after incubation in the medium with CaCl2 and without cyclodextrins indicates occurrence of the true acrosome reaction. Moreover, we suggest the increase of small types of SPACA1 proteins is a valid indicator for occurrence of the acrosomal disintegration arising from the true and false acrosome reactions.

  19. Evaluation of Different Methods of Sperm Immunization in Mice

    Directory of Open Access Journals (Sweden)

    H Moatamed

    2005-07-01

    Full Text Available Introduction: Antifertility effects of naturally occuring antisperm antibody (ASA in infertile couples and studies on experimental immunization of various animals with sperm antigens represent ASA as an immunocontraceptive target. The effects of different factors on sperm immunogenecity and ASA production have been studied and different results have been reported. In this study, whole sperm immunization was evaluated. Methods: In this experimental study, whole mice sperm with different adjuvants i.e. complete Freund’s adjuvant (CFA, incomplete Freund’s adjuvant (ICFA, cholera toxin subunit-β (CTS-β were administrated to mice by different routes; Intramuscular (IM, Subcutaneous (SC, Intranasal (IN, Intra peritoneal (IP, Intrarectal (IR, Intravaginal (IVA and oral. Control groups were inoculated with phosphate buffer saline (PBS plus corresponding adjuvant. Immunization was carried out on day 0,7,14,28 and ASA titers were detected by indirect immunofluorescence (IFA technique. The results were compared between control and experimental groups by Mann Whitney and Fisher exact tests. Results: The number of positive mice for ASA in IM, IN and SC experimental and control groups were significantly different (P=0.01, P=0.01, P=0.04 respectively. However, there were no significant differences between the IR, IVA, and oral experimental and control groups. No differences were observed between ASA in vaginal washing of all groups. Due to high mortality, the IP group was excluded from the study. Conclusion: It can be concluded that whole sperm antigen can induce immune response in female mice by IM, SC and IN routes, but not through IAV, IR and oral administration routes.

  20. [Classical Swine Fever in wild boar in Rhineland-Palatinate: evaluation of the official control measures from 2005-2011].

    Science.gov (United States)

    Romelt, Maria; Klingelhefer, Irene; Konig, Astrid; Braun, Bettina; Reiner, Gerald

    2015-01-01

    The present study describes the control strategy for fighting Classical Swine Fever in wild boar in Rhineland-Palatinate from 2005 to 2011 and evaluates its effectiveness. The official control measures were based on the following three main pillars:--Serological and virological monitoring: By means of serological monitoring Classical Swine Fever outbreaks could be detected very early. Increasing antibody prevalences indicated an imminent Classical Swine Fever outbreak. This could be confirmed by the virological investigations. The geographical evaluations of the virological investigations showed that the outbreaks occurred only in localized areas and a spreading of the virus had not taken place yet or could be prevented.--Oral immunization: After virological detection of Classical Swine Fever Virus oral immunization was started immediately. This oral immunization achieved antibody prevalence rates of 57% on an average. The analysis of the distribution of the antibodies in the vaccination areas concerning the different age groups in the vaccination areas showed that 41% of the young animals, 66% of animals from one to two years and 77% of the adult animals were immunized.--Hunting measures: For the reduction of the wild boar population an all-year, intensive hunt with special attention to the young animals and the female animals was carried out. The hunting bag increased on more than 80 000 wild boar per hunting season. Out of the total 108,772 hunted wild boar were 47% of young animals, 40% of animals from one to two years and 13% of adult animals. Concerning the gender distribution on an average 53% female and 47% male animals were shot. in summary, the current control strategy was effective because there had been no further proof of Classical Swine Fever in wild boar in Rhineland-Palatinate since 2009. Nevertheless, the fight strategy can be optimized even further. For an optimum monitoring the development of a marker vaccine which allows a differentiation of

  1. Influence of Boar and Semen Parameters on Motility and Acrosome Integrity in Liquid Boar Semen Stored for Five Days

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    Sehested E

    2002-03-01

    Full Text Available Ninety ejaculates from a total of 76 AI boars were extended in Beltsville Thawing Solution (BTS. Boar identity, breed, weight of the ejaculate and sperm concentration were registered. Motility and acrosome integrity were assessed after storage at 16–18°C for 6, 30, 54, 78, and 102 h. Storage time had a significant influence on both motility (p

  2. Glycolipid migration from the apical to the equatorial subdomains of the sperm head plasma membrane precedes the acrosome reaction: evidence for a primary capacitation event in boar spermatozoa

    NARCIS (Netherlands)

    Gadella, B.M.; Lopes-Cardozo, M.; Colenbrander, B.; van Goldie, L.M.G.; Gadella, Th.W.J.

    1995-01-01

    In order to extend the static information of immunolabelling sulphogalactolipids in fixed boar spermatozoa, a fluorescent sulphogalactolipid analogue, galactose(3-sulphate)-b1-1¢[( N-lissamine rhodaminyl)-12-aminododecanoyl]-sphingosine, was incorporated into plasma membranes of living spermatozoa a

  3. 季节、品种对种公猪精液量及精子活力的影响%Effects of Season and Va-riety on the Semen Vol-ume and Sperm Motility of Breeding Boar

    Institute of Scientific and Technical Information of China (English)

    姚建军

    2014-01-01

    The research aimed to study the effects of season and variety on the semen volume and sperm motility of adult boar. [Method] The se-men volume and sperm motility of 21 Landrace pigs and 26 Yorkshire pigs in different seasons were observed and de-termined. And the statistical analysis was made on these quality indices of semen. [Result] The effects of different seasons on the semen volume of adult boar reached significant level (P0.05). The semen volume of Landrace pigs in spring was the most, and that in autumn, winter and summer was decreased in succession. The semen volume of York-shire pigs in autumn was the most, and that in summer, spring and winter was decreased in succession. The effects of season and variety on the sperm motility of adult boar were not significant (P>0.05), but the sperm motility of Landrace pigs and Yorkshire pigs reached the maximum values in spring and reached the minimum values in summer. Further-more, the sperm motility of Landrace pigs in four seasons was obviously higher than that of Yorkshire pigs. [Conclusion] The results laid the foundation for im-proving the reproductive efficiency and production level of pig farms.%[目的]研究季节、品种两大因素对成年公猪精液量及精子活力的影响效应。[方法]对21头长白猪和26头大白猪在一年内不同季节的精液量和精子活力进行观察和测定,并对这些精液品质进行统计学分析。[结果]季节对成年公猪精液量的影响达到显著水平(P0.05),其中长白猪的精液量在春季最多,秋、冬、夏依次减少,而大白猪的精液量在秋季最多,夏、春、冬三季则依次减少;季节和品种对成年公猪精子活力的影响均不显著(P>0.05),但是长白猪和大白猪的精子活力均在春季最高,而夏季最低,此外一年四季长白猪的精子活力明显高于大白猪。[结论]结果为提高猪场的繁殖效率和生产水平奠定基础。

  4. Evaluation of Sperm Parameters of Infertile Men with Retrograde Ejaculation

    Institute of Scientific and Technical Information of China (English)

    Hong-xing ZHONG; Wei-jie ZHU; Jing LI

    2006-01-01

    Objective To investigate sperm parameters of infertile men with retrograde ejaculation.Methods Twelve infertile men with retrograde ejaculation (group A) were enrolled into this study. Sperm samples were obtained from the postejaculation urine. After sperm recovery and washing procedure, sperm parameters were assessed. Twelve semen samples from normospermic donors were used as the control (group B).Results In all retrograde cases, motile sperm with forward movement were observed in the medium. Motility of group A was significantly lower than that of group B (P<0. 01).In group A, sperm motility ranged from 11% to 56%, sperm with intact both head and tail membranes was 42.2 ± 12.3%, sperm count ranged (13-85)×106/ml, and the sperm survival time was highly shortened. Sperm with normal morphology and intact acrosome were observed in retrograde specimens.Conclusion Sperm parameters recovered from retrograde specimens were highly variable between subjects. The toxicity of urine caused deleterious to sperm functions.Motile sperm could be collected by sperm recovery procedure. Sperm parameters could meet the requirement for the use of assisted reproductive techniques for treating infertile men with retrograde ejaculation.

  5. Supplementing oregano essential oil to boar diet with strengthened fish oil: Effects on semen antioxidant status and semen quality parameters.

    Science.gov (United States)

    Liu, Q; Duan, R J; Zhou, Y F; Wei, H K; Peng, J; Li, J L

    2017-02-22

    Previous research has shown benefits of dietary fish oil supplementation on semen quality of boars. However, little is known about how antioxidant protects lipid peroxidation on spermatozoa from n-3 polyunsaturated fatty acid (PUFA) addition. This study evaluated the effect of oregano essential oil (OEO) supplementation on semen antioxidant status and semen quality in boars fed a diet enriched with fish oil. Thirty-four mature boars of proven fertility, received daily 2.5 kg basal diet top-dressed with 45 g soybean oil and 15 g fish oil to meet the n-3 PUFA requirement of spermatozoa, randomly allocated to one of four groups supplemented with 100 mg α-tocopheryl acetate kg(-1) (control), or 250 or 500 or 750 mg OEO kg(-1) for 16 weeks. Semen was collected at weeks 0, 8, 12 and 16 for measurements of sperm production, motion characteristics, sperm α-tocopherol content, antioxidant enzyme activities, reactive oxygen species (ROS), DNA damage (8-hydroxydeoxyguanosine, 8-OHdG), lipoperoxidation (malondialdehyde, MDA) and seminal total antioxidant capacity (TAC). Sperm production and motion characteristics were similar (p > .05) among groups throughout the experimental week 16, but increased (p oil has a positive effect on antioxidant capacity in boar when used fish oil.

  6. The activity of N-acetyl-β-hexosaminidase in boar seminal plasma is linked with semen quality and its suitability for cryopreservation.

    Science.gov (United States)

    Wysocki, Paweł; Orzołek, Aleksandra; Strzeżek, Jerzy; Koziorowska-Gilun, Magdalena; Zasiadczyk, Łukasz; Kordan, Władysław

    2015-04-15

    The determination of sperm cryotolerance is an important step in the process of developing optimal techniques for the storage of boar semen. The objective of this study was to determine individual proteome variations in boar seminal plasma and spermatozoa and establish their influence on the cryotolerance of ejaculate. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed the presence of protein with estimated molecular weight of 90 kDa in sperm extracts from ejaculates of selected boars. In all cases, dialysis performed at the initial stage of cryopreservation effectively removed the protein from sperm cells. The protein had an affinity for Zn(2+) ions. Mass spectrometry revealed similarities between the discussed protein and the β subunit of N-acetyl-β-hexosaminidase (β-HEX). Seminal plasma β-HEX was purified 252-fold with approximately 27% recovery and specific activity of 1800 U/mg of protein. Enzyme activity in fresh seminal plasma was correlated with superoxide dismutase activity (r = -0.42, P 20,000 U/L) levels of β-HEX activity in seminal plasma. In plasma with high β-HEX activity, spermatozoa were characterized by lower plasma membrane integrity (84.7%, P spermatozoa/h) were reported in ejaculates with high seminal plasma β-HEX activity. The results of this study indicate that β-HEX activity in seminal plasma is a useful indicator in preliminary evaluations of boar sperm cryotolerance.

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

    Science.gov (United States)

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

    2011-10-01

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

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

    Directory of Open Access Journals (Sweden)

    V. HAREA

    2009-05-01

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

  9. Boar and season effects on some parameters of semen fertilizing potential

    Directory of Open Access Journals (Sweden)

    Apić Jelena

    2016-01-01

    Full Text Available In order to determine the more accurate fertile potential of sperm, it seems that the conventional parameters of boar semen quality (the ejaculate volume, sperm concentra­tion, progressive motility, percentage of live sperm and of those with intact acrosomal mor­phology are insufficient. Since recently, there have been numerous studies proving that protein concentration in sperm plasma has high positive correlation with boar fertile po­tential. The research objective was to determine the effect of boars as well as the season on the variation of protein content in the sperm plasma. For the research there were used spermal fractions of 2 boars with high (V-boar and 2 boars with low (N-boar protein con­tent in spermal plasma. The ejaculates of boars were taken once a week, for a month, during one year (4 × 12 = 48 ejaculates per boar. For protein analysis in the spermal plas­ma, the samples were prepared by centrifugation. The ejaculate volume, protein concen­tration and progressive motility varied considerably (p 0.01 protein content in sperm plasma (V-boars: 4 to 4.5% in warm and cold season; N-boars: 2.3 do 2.6% in warm and 2.3 to 2.5% in cold season. The obtained results showed that measurement of protein con­tent in boar sperm plasma could be a useful method for their ranking, based on fertile po­tential of fresh semen.

  10. The Differences in Sperm Concentration and Semen Volume in Different Segments in one Boar Ejaculation%公猪射精过程不同阶段精液精子密度和精液量的变化

    Institute of Scientific and Technical Information of China (English)

    幸宇云; 吴延博; 杨明; 李平华; 李凯; 周利华

    2011-01-01

    There is one to several short pauses during the boar' s whole ejaculation. The semen of the whole ejaculation was divided into three segments: concentrated semen ( segment 1) , the semen of the first e jaculate with the exception of the concentrated semen ( segment 2 ) , the rest part ( segment 3 ). The differences in sperm concentration and semen volume in these three different segments were studied. The results showed that the volume of the concentrated semen was significantly lower than that of the other two segments (P<0.01) , whereas the sperm concentration and sperm number were significantly higher than those of the other two segments ( P < 0.01). The concentrated semen occupied only about 17% of the whole semen volume whereas about 71% of the total sperm number. The first ejaculate accounted for about 70% of semen volume and 95% of sperms of the whole ejaculation.%公猪在一次完整射精过程中有1至数次不等的短暂停顿,将公猪一次完整射精的精液分成三段:浓精(S1)、第一次射精中除浓精外的精液(S2)及其余部分精液(S3),研究三段精液精子密度、精液量的变化规律.结果表明:浓精精液量显著低于其它两段精液(P<0.01),而精子密度及总精子数显著高于其它两段精液(P<0.01);浓精量只占总精液量的约17%,而精子数占总精子数的约71%;公猪完整射精过程中的第一次射精精液量占总精液量的约70%、精子数占总精子数的约95%.

  11. Sperm morphological and morphometric evaluation in captive collared peccaries (Pecari tajacu

    Directory of Open Access Journals (Sweden)

    Patrícia C. Sousa

    2013-07-01

    Full Text Available The aim of this study was to compare different staining methods for the evaluation of sperm morphology by light microscopy and also to describe the morphometry of the entire sperm in collared peccaries (Pecari tajacu. Semen from 10 males was obtained by electroejaculation and evaluated for sperm motility, vigor, and concentration. Semen smears were prepared through three different staining methods: Bengal rose, brome-phenol blue, and eosin-nigrosin. Smears were evaluated under light microscopy and sperm morphologic alterations were determined in percentage. In addition, sperm morphometric analysis was conducted by light microscopy coupled to image analyzer software. The smears stained with Bengal Rose provide the best results for the visualization of the sperm tail, midpiece, and head. The use of eosin-nigrosin stain did not allow an adequate impregnation, and some sperm presented a few contrasts with the background. A higher incidence of bent coiled tails was verified in the use of brome-phenol blue staining (P<0.05. Through morphometric evaluation, it was observed that the tail occupies the greatest proportion (89% of the sperm which presents a discretely elongated head. According to the results, the use of the Bengal Rose stain is recommended for the morphologic evaluation of the collared peccary sperm.

  12. 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.

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

    Science.gov (United States)

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

    2009-10-01

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

  14. Comparative Study of LDL Extracted from Five Avian Species on the Cryopreservation of Boar Sperm%5种禽类卵黄低密度脂蛋白对猪精子冷冻效果的影响

    Institute of Scientific and Technical Information of China (English)

    吕瑞凯; 胡建宏; 王红; 程亮; 江中良; 李青旺; 姚俊; 张鹏飞

    2011-01-01

    为了确定具有最佳抗冷冻效果的禽类卵黄低密度脂蛋白(LDL)及其添加质量分数,在猪精液冷冻稀释液中分别添加质量分数为6%、7%、8%、9%和10%的鸡、鸭、鹌鹑、鸽子和鸵鸟的卵黄LDL,分析不同禽类的LDL对猪精子的冷冻保存效果.结果表明,稀释液中添加质量分数为9%的鸡、鸭、鹌鹑、鸽子卵黄LDL以及质量分数8%的鸵鸟卵黄LDL时,冷冻-解冻后精子活率最高,分别达到42.33%、35.63%、31.47%、47.33%和36.40%.以5种禽类LDL最佳质量分数配制冷冻稀释液冷冻精子,发现质量分数为9%的鸽蛋LDL冻存猪精子时解冻后精子活率达到47.33%,顶体完整性达到62.57%,质膜完整性达到48.13%,均显著优于其他处理组(P<0.05).说明鸽子卵黄LDL对猪精子具有良好的冷冻保护性能,可提高猪精子抵抗低温打击的能力.%Low density lipolipid (LDL) can protect sperm from freezing damage during the sperm frozen-thawed process. In order to distinguish the efficiency of LDL extracted from different avian species, to confirm which one and how much of it has the best anti-freezing effect, LDL extracted from eggs of hen, duck, quail, pigeon and ostrich with the mass fraction of 6%, 7%, 8%, 9% and 10% was added into the cryopreservation diluents of boar sperm separately. The result showed that after the frozen-thawed process, LDL extracted from eggs of hen, duck, quail and pigeon with concentration of 9% and ostrich LDL with concentration of 8% could protect sperm with highest livability of 42. 33%, 35. 63% , 31. 47%, 47. 33% and 36. 40%, respectively. They were significantly better than that of other concentrations (P<0. 05). Among the cryopreservation diluents and cryopreservation sperm prepared with the five avian species with the optimum concentration, it was found that the 9% pigeon LDL group showed 47. 33% sperm livability, 62. 57% acrosome integrity and 48. 13% membrane integrity after

  15. Separating Subcellular Proteins in Uncapacitation and Capacitation of Boar Sperm Cells and Identifying the Tyrosine Phosphoprotein%猪精子获能前后细胞亚组分蛋白分离及酪氨酸磷酸化蛋白的鉴定

    Institute of Scientific and Technical Information of China (English)

    胡启蒙; 张媛媛; 陈振亮; 王亮亮; 李新红

    2012-01-01

    The aim of this study was to separate subsets proteins in uncapacitation and capacitation of boar sperm cells and to identify the protein tyrosine phosphorylation, and to establish the theoretical foundation for the mammalian sperm fertilization biology research. In vitro , the boar sperms were cultivated for capacitation and separated subsets proteins. The tyrosine phosphoryla-ted proteins in uncapacitation and capacitation sperms were identified by using SDS-PAGE and Western blot. In capacitation boar sperm, the degree of tyrosine phosphorylation on 126, 108, 79 ku proteins were significantly higher than them on uncapacition boar sperm. The vigor indexes and the results of SDS-PAGE of uncapacitation and capacitation sperms were different,all the vigor indexes of capacitation sperms were significantly improved and these changes were related with tyrosine phosphorylation. The molecular weight of tyrosine phosphorylated proteins on membrane were approximately 25, 47 and 50 ku and on cytoplasm is about 47 ku. The degree of tyrosine phosphoproteins of 25 and 47 ku in capacitation sperm were significantly higher than them in the uncapacitation sperm(P<0. 05). This phenomenon also existed in cell nucleus. In sperm nucleus,these proteins' molecular weight were 23, 37 and 42-50 ku. The degree of tyrosine phos-phrylated 23 ku protein in nucleus of capacitation sperms are higher. The results suggests that the main tyrosine phosphorylated proteins were on the nucleus and membrane proteins, and a few of them are plasma proteins.%本研究对猪精子获能前后细胞亚组分蛋白进行分离以及对酪氨酸磷酸化蛋白进行鉴定,旨在为哺乳动物精子受精生物学研究奠定理论基础.利用动物精子体外获能培养、细胞亚组分分离技术及蛋白免疫印迹的方法,分离猪精子细胞亚组分蛋白及酪氨酸磷酸化蛋白鉴定.结果表明,猪精子经过获能培养后各项活力指标均得到显著提高,且与精子蛋白发生

  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. Evaluation of possible mitigation measures to prevent introduction and spread of African swine fever virus through wild boar

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    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.

  18. The effect of glycerol concentrations on the post-thaw in vitro characteristics of cryopreserved sex-sorted boar spermatozoa.

    Science.gov (United States)

    Parrilla, I; del Olmo, D; Caballero, I; Tarantini, T; Cuello, C; Gil, M A; Roca, J; Martinez, E A; Vazquez, J M

    2012-12-01

    The objective of this study was to optimize protocols for the cryopreservation of sex-sorted boar spermatozoa. In the experiment 1, we evaluated the effects of a standard boar sperm cryopreservation procedure (3% final glycerol concentration) on the in vitro characteristics of sex-sorted sperm frozen at low sperm concentrations (20 × 10(6) sperm/ml; S20 group). Non-sorted spermatozoa frozen at 1000 × 10(6) (C1000 group) and 20 × 10(6) (C20 group) sperm/ml were used as the freezing control groups. In experiment 2, the effects of different final glycerol concentrations (0.16%, 0.5%, 1.0%, 2.0% and 3.0%) on post-thaw quality of the S20 and C20 groups were evaluated. In both experiments, the samples were evaluated prior to freezing (5°C) and at 30, 90 and 150 min after thawing. Experiment 1 indicated that freezing sperm at low concentrations decreased (p sperm were sorted or not. However, the sperm membrane integrity was not affected at any evaluation step. Inexperiment 2, significant effects on the TM and PM because of increased glycerol concentrations in the S20 and C20 groups were observed only at 90 and 150 min after thawing. The samples frozen in 3% glycerol showed lower (p < 0.05) TM and PM values when compared to those frozen in the presence of 0.5% and 1% glycerol. In both experiments, non-sorted control samples displayed higher percentages of spermatozoa with damaged DNA than sorted spermatozoa. In conclusion, the optimization of cryopreservation conditions by decreasing the glycerol concentrations can improve post-thaw motility of sex-sorted spermatozoa frozen at low concentrations.

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

    Science.gov (United States)

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

    2016-11-01

    Cholesterol (cholesterol-loaded cyclodextrins: CLC) treatment of dromedary camel sperm prior to freezing enhances cryosurvival. The present study first validated the efficacy of a heterologous zona-free goat oocyte assay (n=115 oocytes) to evaluate camel sperm function in vitro (Experiment 1: n=6 bulls), then examined the effects of CLC treatment (1.5mg/mL CLC; CLC+) versus no treatment (0 CLC) of fresh (Experiment 2: n=4 bulls) and frozen-thawed (Experiment 3: n=5 bulls) camel sperm to penetrate, de-condense and form pro-nuclei in in vitro-matured goat oocytes. Finally, the ability of fresh 0 CLC and CLC+ sperm to fertilize in vivo was studied by artificially inseminating super-ovulated females (n=7-9 per treatment) and examining embryo production (Experiment 4: n=4-5 bulls/treatment). Camel spermatozoa penetrated (60%) and formed pro-nuclei (33%) in goat oocytes demonstrating the utility of this heterologous system for assessing sperm function in vitro. For fresh spermatozoa, 0 CLC-treated sperm performed better than their CLC+ counterparts for all parameters measured (Pdromedary camel sperm improves sperm motility it fails to confer an advantage to them in terms of improved in vitro sperm-oocyte interaction or in vivo fertilization under the conditions tested.

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

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    Maria J Martinez-Alborcia

    Full Text Available In the present study, the impact of non-functional spermatozoa on the cryopreservation success of functional boar spermatozoa was evaluated. Fifteen sperm-rich ejaculate fractions collected from five fertile boars were frozen with different proportions of induced non-functional sperm (0--native semen sample-, 25, 50 and 75% non-functional spermatozoa. After thawing, the recovery of motile and viable spermatozoa was assessed, and the functional of the spermatozoa was evaluated from plasma membrane fluidity and intracellular reactive oxygen species (ROS generation upon exposure to capacitation conditions. In addition, the lipid peroxidation of the plasma membrane was assessed by the indirect measurement of malondialdehyde (MDA generation. The normalized (with respect to a native semen sample sperm motility (assessed by CASA and viability (cytometrically assessed after staining with Hoechst 33342, propidium iodide and fluorescein-conjugated peanut agglutinin decreased (p<0.01 as the proportion of functional spermatozoa in the semen samples before freezing decreased, irrespective of the semen donor. However, the magnitude of the effect differed (p<0.01 among boars. Moreover, semen samples with the largest non-functional sperm subpopulation before freezing showed the highest (p<0.01 levels of MDA after thawing. The thawed viable spermatozoa of semen samples with a high proportion of non-functional spermatozoa before freezing were also functionally different from those of samples with a low proportion of non-functional spermatozoa. These differences consisted of higher (p<0.01 levels of intracellular ROS generation (assessed with 5-(and-6 chloromethyl-20,70-dichlorodihydrofluorescein diacetate acetyl ester; CM-H(2DCFDA and increased (p<0.01 membrane fluidity (assessed with Merocyanine 540. These findings indicate that non-functional spermatozoa in the semen samples before freezing negatively influence the freezability of functional spermatozoa.

  1. COOLING CURVES OF THE BOAR SEMEN DILUTED IN ACP®103 EXTENDER ADDED OF POWDERED EGG YOLK IN FIXED CONCENTRATION

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    Tatyane Bandeira Barros

    2016-10-01

    Full Text Available The conservation of boar semen at lower temperatures might contribute to the further expansion of artificial insemination in this species. Egg yolk cryoprotectant properties have already been extensively tested on sperm cryopreservation of several species. This study aimed to test different temperature curves for the conservation of boar semen diluted with coconut milk powdered (ACP®-103 add 7% egg yolk and to verify which one better maintains sperm viability. For this, 36 ejaculates were diluted and stored at 17, 10 and 5 °C. Daily analysis of vigor and motility were performed, and on days D0, D2, and D4 semen was evaluated regarding vitality, morphology, and osmotic resistance. For the statistical analysis we performed the tests of Kruska-Wallis with Dunns post-test (nonparametric data and ANOVA and Tukey test (parametric data. The storage temperature of 10 °C was the best one   to maintain spermatic motility at appropriate levels to be used in an artificial insemination program. Analyses of viability, morphology, and hypoosmotic test did not show statistical difference among the treatments. In conclusion, the best temperature curve was 10 °C with diluted semen previously kept at 17 °C to maintain the viability of sperm cells in pigs for a longer period. Keywords: boar semen; coconut water powder; conservation; egg yolk.

  2. Trehalose in glycerol-free freezing extender enhances post-thaw survival of boar spermatozoa.

    Science.gov (United States)

    Athurupana, Rukmali; Takahashi, Daisen; Ioki, Sumire; Funahashi, Hiroaki

    2015-01-01

    Cryopreservation of boar semen is still considered suboptimal due to lower fertility as compared with fresh samples when glycerol, a permeating cryoprotectant, is used. Trehalose is a non-permeable cryoprotectant and nonreducing disaccharide known to stabilize proteins and biologic membranes. The aim of this study was to evaluate the cryosurvival and in vitro penetrability of boar spermatozoa when glycerol was replaced with trehalose in a freezing extender. Ejaculated Berkshire semen samples were diluted in egg yolk-based freezing extender containing glycerol (100 mM) or trehalose (0, 50, 100, 150, 200 and 250 mM) and cryopreserved using a straw freezing procedure. Thawed samples were analyzed for motility, viability, mitochondrial membrane potential (MMP), and acrosome integrity. In experiment 2, penetrability of spermatozoa cryopreserved with 100 mM glycerol or trehalose was examined. Replacement of cryoprotectant glycerol (100 mM) with trehalose had no effect on sperm viability, but replacing it with 100 mM trehalose improved motility, MMP and acrosome integrity significantly. Sperm motility and MMP were considerably higher in 100 mM trehalose, whereas the acrosome integrity was substantially higher in 100-250 mM trehalose. The in vitro penetration rate was also significantly higher in spermatozoa cryopreserved with trehalose (61.3%) than in those cryopreserved with glycerol (43.6%). In conclusion, 100 mM non-permeable trehalose can be used to replace glycerol, a permeating cryoprotectant, for maintenance of better post-thaw quality of boar spermatozoa.

  3. Antioxidant effect of lemon balm (Melissa officinalis) and mate tea (Ilex paraguensys) on quality, lipid peroxidation and DNA oxidation of cryopreserved boar epididymal spermatozoa.

    Science.gov (United States)

    Luño, V; Gil, L; Olaciregui, M; Jerez, R A; de Blas, I; Hozbor, F

    2015-11-01

    In this study, we investigated the protective ability of the addition of two antioxidant herb extracts, mate tea and lemon balm, on boar epididymal frozen-thawed spermatozoa quality. Testes from mature boars were collected at local slaughterhouse, and sperm samples from epididymis were recovered by flushing. Spermatozoa were cryopreserved in lactose-egg yolk buffer supplemented with various concentrations of lemon balm and mate tea (0, 2.5, 5 and 10 g l(-1) ) using the straw-freezing procedure. Motion parameters, acrosome and plasma membrane integrity, lipoperoxidation levels and DNA oxidative damage (8-hydroxy-2'-deoxyguanosine base lesion) were evaluated. There were no differences among experimental groups with regard to motility characteristics, viability, acrosome and plasma membrane integrity; however, the highest concentration of lemon balm produced significant (P sperm lipid membrane peroxidation, and only mate tea protected DNA against oxidative damage during cryopreservation at 120 min post-thawing (P sperm oxidised DNA and malondialdehyde generation; thus, mate tea is a potential candidate such as antioxidant compound on boar sperm cryopreservation medium.

  4. Direct binding of boar ejaculate and epididymal spermatozoa to porcine epididymal epithelial cells is also needed to maintain sperm survival in in vitro co-culture.

    Science.gov (United States)

    Yeste, Marc; Castillo-Martín, Míriam; Bonet, Sergi; Briz, Maria Dolors

    2012-04-01

    The aim of the present study was to compare the influence of cultured epididymal epithelial cells (EEC) from corpus, caput or cauda, oviductal epithelial cells (OEC) and non-reproductive epithelial cells (LLC-PK1) on function and survival of epididymal and ejaculated spermatozoa, in the latter case to determine whether such influence differed between morphologically normal and abnormal spermatozoa. For this purpose, either spermatozoa were directly co-cultured with EEC from caput, corpus, or cauda, OEC and LLC-PK1 cells (experiment 1) or a membrane-diffusible insert was included in these co-cultures (experiment 2). EEC cultured from the three epididymal regions did not differently affect the sperm parameters. Morphologically normal spermatozoa presented a higher ability to bind EEC, OEC, and LLC-PK1 than abnormal spermatozoa with cytoplasmic droplets or with tail/head malformations. Epididymal spermatozoa were more able to bind EEC during the first 24 h of co-culture, while ejaculated spermatozoa presented a higher capacity to bind OEC between 30 min and 3 h of co-incubation. In all cases, the ability to bind to epithelial cells was higher when they were co-cultured with EEC and OEC than with LLC-PK1. After 2 h of co-culture, the viability of epididymal spermatozoa was better maintained when they bound EEC than when they bound OEC. Conversely, the viability of ejaculated spermatozoa was better maintained when bound OEC than when bound EEC after 24 and 48 h of co-culture. Our work, apart from corroborating the involvement of morphologically normal spermatozoa in the formation of sperm reservoir, highlights the importance of direct contact spermatozoa-EEC in maintaining the sperm survival in in vitro co-culture, and also suggests that a specific binding between EEC and epididymal spermatozoa exists.

  5. Effects of commercial selenium products on glutathione peroxidase activity and semen quality in stud boars

    Science.gov (United States)

    The aim of this study was to determine how dietary supplementation of inorganic and organic selenium affects selenium concentration and glutathione peroxidase activity in blood and sperm of sexually mature stud boars. Twenty-four boars of the Large White, Landrace, Pietrain, and Duroc breeds of opt...

  6. Validation of the polysemen admixture on viability and acrosomal morphology of boar spermatozoa

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    Ogbuewu IP

    2007-07-01

    Full Text Available Semen were collected using artificial vagina (AV, from 5 large white boars aged 2-2.5 years twice a week for 16 weeks in each of the two seasons, early rainy (ER and late rainy (LR seasons, to determine the effects of multiple semen pool admixture on the viability and acrosomal morphology. The semen qualities studied were sperm motility, live sperm and sperm concentration, while the acrosomal parameters includes normal apical ridge (NAR, damaged apical ridge (DAR, missing apical ridge (MAR and loose apical ridge (LAC. There were no significant (P>0.05 seasonal effects. Three-boar semen admixture gave the highest percentage NAR, motility, live sperm concentration and least DAR and LAC, although these were not significantly (P>0.05 different from the 2-boar semen admixture. The result of this study suggests that 3-boar semen admixture is most suitable for use in artificial insemination program.

  7. Dietary n-6:n-3 ratio and Vitamin E improve motility characteristics in association with membrane properties of boar spermatozoa.

    Science.gov (United States)

    Liu, Qing; Zhou, Yuan-Fei; Duan, Run-Jia; Wei, Hong-Kui; Peng, Jian; Jiang, Si-Wen

    2017-01-01

    This study was aimed to evaluate the effects of dietary n-6:n-3 ratio and Vitamin E on the membrane properties and motility characteristics of spermatozoa in boars. Forty Duroc boars were randomly distributed in a 2 × 2 factorial design with two n-6:n-3 ratios (14.4 and 6.6) and two Vitamin E levels (200 and 400 mg kg-1 ). During 16 weeks of treatment, fresh semen was collected at weeks 0, 8, 12, and 16 for measurements of motility characteristics, contents of fatty acids, membrane properties (membrane fluidity and membrane integrity), and lipid peroxidation of the spermatozoa. The semen was diluted in Beltsville Thawing Solution (BTS) extender and stored at 17°C, and the sperm motility was assessed at 12, 36, 72, and 120 h of storage. The 6.6 n-6:n-3 dietary ratio increased the contents of n-3 polyunsaturated fatty acids (PUFAs) and docosahexaenoic acid (DHA) and improved the membrane integrity and membrane fluidity of the spermatozoa, resulting in notably increased total motility, sperm progressive motility, and velocity parameters of fresh semen. Feeding diet with Vitamin E (400 mg kg-1 ) prevented sperm lipid peroxidation, and resulted in higher total motility and sperm progressive motility in fresh and liquid stored semen. In conclusion, the adjustment of n-6:n-3 ratio (6.6) and supply of Vitamin E (400 mg kg-1 ) successfully improved sperm motility characteristics and thus may be beneficial to the fertility of boars, which might be due to the modification of the physical and functional properties of spermatozoa membrane in response to dietary supplementation.

  8. Dietary n-6:n-3 ratio and Vitamin E improve motility characteristics in association with membrane properties of boar spermatozoa

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    Qing Liu

    2017-01-01

    Full Text Available This study was aimed to evaluate the effects of dietary n-6:n-3 ratio and Vitamin E on the membrane properties and motility characteristics of spermatozoa in boars. Forty Duroc boars were randomly distributed in a 2 × 2 factorial design with two n-6:n-3 ratios (14.4 and 6.6 and two Vitamin E levels (200 and 400 mg kg−1 . During 16 weeks of treatment, fresh semen was collected at weeks 0, 8, 12, and 16 for measurements of motility characteristics, contents of fatty acids, membrane properties (membrane fluidity and membrane integrity, and lipid peroxidation of the spermatozoa. The semen was diluted in Beltsville Thawing Solution (BTS extender and stored at 17°C, and the sperm motility was assessed at 12, 36, 72, and 120 h of storage. The 6.6 n-6:n-3 dietary ratio increased the contents of n-3 polyunsaturated fatty acids (PUFAs and docosahexaenoic acid (DHA and improved the membrane integrity and membrane fluidity of the spermatozoa, resulting in notably increased total motility, sperm progressive motility, and velocity parameters of fresh semen. Feeding diet with Vitamin E (400 mg kg−1 prevented sperm lipid peroxidation, and resulted in higher total motility and sperm progressive motility in fresh and liquid stored semen. In conclusion, the adjustment of n-6:n-3 ratio (6.6 and supply of Vitamin E (400 mg kg−1 successfully improved sperm motility characteristics and thus may be beneficial to the fertility of boars, which might be due to the modification of the physical and functional properties of spermatozoa membrane in response to dietary supplementation.

  9. Cryopreservation of Iberian pig spermatozoa. Comparison of different freezing extenders based on post-thaw sperm quality.

    Science.gov (United States)

    De Mercado, Eduardo; Rodríguez, Ana; Gómez, Emilio; Sanz, Elena

    2010-03-01

    The aim of this study was to evaluate the cryoprotective effect of different freezing extenders against cryopreservation injuries on Iberian boar sperm. The sperm-rich fraction was collected and pooled from six sexually mature Iberian boars, and was frozen in different extenders containing glucose, lactose or fructose as sugar source and including Orvus ES Paste only in the freezing extender-2 (Glucose; Lactose and Fructose) or in both freezing extenders (Glucose2; Lactose2 and Fructose2). During the cryopreservation process, the supernatant was removed after the centrifugation step, then was extended with freezing extender-1 for the equilibration period and with freezing extender-2 immediately before freezing. Post-thaw sperm characteristics, such as plasma membrane integrity (SYBR-14/PI), mitochondrial function (Rhodamine 123) and acrosome integrity (NAR), were monitored. Overall sperm motility and the individual kinematic parameters of motile spermatozoa (assessed by the computer-aided sperm analysis system Sperm Class Analyzer [SCA]) were recorded in the different experimental treatments. Measurements were taken at 30 and 150 min post-thaw. The state of the acrosome after thawing did not show significant differences between the freezing extenders studied. Freezing-thawing caused a significant decrease (Psperm motility and kinematic parameters were concurrent with reduced sperm characteristics. It can be suggested that in the Iberian pig, the beneficial effects of Orvus ES Paste during the freezing process of spermatozoa is time dependent. The analysis of different sperm characteristics such as motility, plasma membrane integrity and mitochondrial function, determined that the extenders studied in the present experiment affected the quality of frozen-thawed semen in Iberian boar.

  10. Acrosin inhibitor detection along the boar epididymis.

    Science.gov (United States)

    Maňásková-Postlerová, Pavla; Cozlová, Nina; Dorosh, Andriy; Šulc, Miroslav; Guyonnet, Benoit; Jonáková, Věra

    2016-01-01

    Epididymal sperm maturation represents a key step in the reproduction process. Spermatozoa are exposed to epididymal fluid components representing the natural environment essential for their post-testicular maturation. Changes in sperm membrane proteins are influenced by proteolytic, glycosylation and deglycosylation enzymes present in the epididymal fluid. Accordingly, the occurrence of inhibitors of these enzymes in the epididymis is very important for the regulation of sperm membrane protein processing. In the present study, we monitored acrosin inhibitor distribution in boar epididymal fluid and in spermatozoa from different segments of the organ. Using specific polyclonal antibody we registered increasing signal of the acrosin inhibitor (AI) from caput to cauda epididymis. Mass spectroscopy examination of the immunoprecipitated acrosin inhibitor (12 kDa) unequivocally identified sperm-associated acrosin inhibitor (SAAI) in the epididymal tissue. Lectin staining showed N-glycosylation in AI from boar epididymis. Protein detection of AI was supported by the results of semi-quantitative RT-PCR showing the presence of mRNA specifically coding for SAAI and similarly increasing throughout the epididymal duct, from its proximal to distal part. Additionally, the immunofluorescence technique showed the AI localization in the secretory tissue of caput, corpus and cauda epididymis, and in the acrosome region and midpiece of the sperm.

  11. Field data analysis of boar semen quality.

    Science.gov (United States)

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

    2011-09-01

    This contribution provides an overview of approaches to correlate sow fertility data with boar semen quality characteristics. Large data sets of fertility data and ejaculate data are more suitable to analyse effects of semen quality characteristics on field fertility. Variation in fertility in sows is large. The effect of semen factors is relatively small and therefore impossible to find in smaller data sets. Large data sets allow for statistical corrections on both sow- and boar-related parameters. Remaining sow fertility variation can then be assigned to semen quality parameters, which is of huge interest to AI (artificial insemination) companies. Previous studies of Varkens KI Nederland to find the contribution to field fertility of (i) the number of sperm cells in an insemination dose, (ii) the sperm motility and morphological defects and (iii) the age of semen at the moment of insemination are discussed in context of the possibility to apply such knowledge to select boars on the basis of their sperm parameters for AI purposes.

  12. Evaluation of the expression of sperm proteins in normozoospermic and asthenozoospermic men using monoclonal antibodies

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    Jana Capkova

    2016-01-01

    Full Text Available Recent studies have shown that infertility affects estimated 15% of all couples. Male infertility is the primary or contributory cause in 60% of these cases. Consequently, the application of assisted reproduction is increasing. These methods could benefit from an extended evaluation of sperm quality. For this reason, we analyzed sperm proteins from 30 men with normal spermiograms and 30 men with asthenozoospermia. Ejaculates of both groups were tested by flow cytometry (FCM and fluorescence with a set of well-characterized anti-human sperm Hs-monoclonal antibodies (MoAbs, which were generated in our laboratory. No statistically significant differences were found between normospermics and asthenospermics in the expression of the sperm surface protein clusterin, evaluated with Hs-3 MoAb, and semenogelin, evaluated with Hs-9 MoAb. However, FCM revealed quantitative differences in the acrosomal proteins between normozoospermic and asthenozoospermic men, namely, in glyceraldehyde-3-phosphate dehydrogenase, evaluated with Hs-8 MoAb, valosin-containing protein, evaluated with Hs-14 MoAb, and ATP synthase (cAMP-dependent protein kinase II, PRKAR2A, evaluated with MoAb Hs-36. Asthenozoospermic men displayed a highly reduced expression of intra-acrosomal proteins, with a likely decrease in sperm quality, and thus a negative impact on successful reproduction. Asthenozoospermia seems to be a complex disorder involving intra-acrosomal proteins.

  13. Osmolarity of Coconut Water (Cocos nucifera Based Diluents and their Effect Over Viability of Frozen Boar Semen

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    Bottini Luzardo

    2010-01-01

    Full Text Available Problem statement: Boar sperm cells are sensitive to the freezing process, which compromises viability of frozen-thawed sperm. In a constant search for minimizing or suppressing sperm cell damage caused by the temperature and osmolarity changes during the freezing process, crioprotective and antioxidant substances have been added to the freezing media, such as coconut water, in order to increase the viability of frozen-thawed swine semen. The addition of any substance to the freezing diluent, directly affects osmolarity of the media, which can have positive or negative effects over the sperm cell. Approach: There are no published studies currently that indicate the effect of adding coconut water over the osmolarity of freezing media and their effect over viability of sperm cells, therefore, the objective of the present study was to evaluate the effect of coconut water based diluents osmolarity over the Motility (Mot, Acrosome Integrity (AI, Membrane Integrity (MI and Mitochondrial Activity (MA of thawed boar sperm cells. The treatments used were control T1 (LEY with bidistilled water + LEYGO with an osmolarity range of 296-368 mOsmol Kg-1, T2 (LEY and deionized coconut water + LEYGO between 381 and 480 mOsmol Kg-1 and T3 (LEY and in natura coconut water + LEYGO between 519 and 1041 mOsmol Kg-1. The Westendorf modified method was the freezing method used. The obtained data were statistically analyzed by GLM, using the SAS software (SAS, 2000. Results: A significant difference was observed on T2 compared to T1 regarding Mot 41.9 Vs 36.9% and MI 58.0 Vs 50.2. T3 had a detrimental effect on all studied variables. Conclusion: Under our study conditions, the osmolarity range of T2, due to the non ionic solutes content, contributed to improve the viability of frozen-thawed sperm cells.

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

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    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.

  15. Intracellular calcium movements of boar spermatozoa during 'in vitro' capacitation and subsequent acrosome exocytosis follow a multiple-storage place, extracellular calcium-dependent model.

    Science.gov (United States)

    Yeste, M; Fernández-Novell, J M; Ramió-Lluch, L; Estrada, E; Rocha, L G; Cebrián-Pérez, J A; Muiño-Blanco, T; Concha, I I; Ramírez, A; Rodríguez-Gil, J E

    2015-07-01

    This work analysed intracellular calcium stores of boar spermatozoa subjected to 'in vitro' capacitation (IVC) and subsequent progesterone-induced acrosome exocytosis (IVAE). Intracellular calcium was analysed through two calcium markers with different physico-chemical properties, Fluo-3 and Rhod-5N. Indicative parameters of IVC and IVAE were also evaluated. Fluo-3 was located at both the midpiece and the whole head. Rhod-5N was present at the sperm head. This distribution did not change in any of the assayed conditions. Induction of IVC was concomitant with an increase in both head and midpiece Ca(2+) signals. Additionally, while IVC induction was concurrent with a significant (p sperm membrane permeability, no significant changes were observed in O2 consumption and ATP levels. Incubation of boar spermatozoa in the absence of calcium showed a loss of both Ca(2+) labellings concomitantly with the sperm's inability to achieve IVC. The absence of extracellular calcium also induced a severe decrease in the percentage of spermatozoa exhibiting high mitochondrial membrane potential (hMMP). The IVAE was accompanied by a fast increase in both Ca(2+) signalling in control spermatozoa. These peaks were either not detected or much lessened in the absence of calcium. Remarkably, Fluo-3 marking at the midpiece increased after progesterone addition to sperm cells incubated in a medium without Ca(2+) . The simultaneous addition of progesterone with the calcium chelant EGTA inhibited IVAE, and this was accompanied by a significant (p boar spermatozoa present different calcium deposits with a dynamic equilibrium among them and with the extracellular environment. Additionally, the modulation role of the intracellular calcium in spermatozoa function seems to rely on its precise localization in boar spermatozoa.

  16. Genetic parameters for male fertility and its relationship to skatole and androstenone in Danish Landrace boars

    DEFF Research Database (Denmark)

    Strathe, Anders Bjerring; Velander, I.H.; Mark, Thomas;

    2013-01-01

    ). Hence, the objective of this investigation was to study the genetic association between direct measures of male fertility and the boar taint compounds in Danish Landrace pigs. Concentrations of skatole and androstenone in the back fat were available for approximately 6,000 and 1,000 Landrace boars......, respectively. The litter size traits, such as total number born, live piglets at d 5, and piglet survival until d 5 on relatives of the slaughter boars, were extracted from the Danish Landrace breeding program, yielding 35,715 records. Semen volume, sperm concentration, subjective sperm quality score......, and total number of sperm were available from 95,267 ejaculates. These ejaculates were collected between 2005 and 2012 and originated from 3,145 Landrace boars from 12 AI stations in Denmark. The traits were analyzed using single and multitrait animal models including univariate random regression models...

  17. Evaluation of sperm quality snakes Erythrolamprus poecilogyrus sublineatus (Cope, 1860 (Serpentes, Dipsadidae

    Directory of Open Access Journals (Sweden)

    A. C. Silva

    Full Text Available Abstract Erythrolamprus poecilogyrus sublineatus (Cope, 1860, is a species widely distributed in the Pampa Domain, occurring in Rio Grande do Sul, Argentina and Uruguay, mainlyin the pampa region. In the coastal region of southern Brazil this is serpent is considered one of the most abundant. The purpose of the present study is to describe the techniques of sperm evaluation in vitro for E. poecilogyrus sublineatus in the coastal plain of Rio Grande do Sul, Brazil. After laparatomy the efferent vases were collected and the semen was diluted in 1ml Beltsville Thawing Solution. The characteristics of motility, membrane integrity, mitochondria, acrosome, DNA, cell viability and cellular functionality were evaluated. Fluorescent probes were used for the evaluation of sperm structure in epifluorescence microscope. With the techniques described, it was possible to identify intact and injured cells, enabling the determination of cell characteristics for the spring season (October and November. It was observed in the analyses that 80% of sperm cells were mobile and that 84.1 ± 8.0% of sperm membranes were intact. The standards found were of 48 ± 13.8% of intact acrosome, 73.6 ± 6.0 of perfect DNA and of 91.8 ± 4.0 of functional mitochondria. Thus, these values from the sperm analysis can be used as standards for the species Erythrolamprus poecilogyrus sublineatus.

  18. Evaluation of sperm quality snakes Erythrolamprus poecilogyrus sublineatus (Cope, 1860) (Serpentes, Dipsadidae).

    Science.gov (United States)

    Silva, A C; Varela, A S; Cardoso, T F; Silva, E F; Loebmann, D; Corcini, C D

    2017-01-12

    Erythrolamprus poecilogyrus sublineatus (Cope, 1860), is a species widely distributed in the Pampa Domain, occurring in Rio Grande do Sul, Argentina and Uruguay, mainlyin the pampa region. In the coastal region of southern Brazil this is serpent is considered one of the most abundant. The purpose of the present study is to describe the techniques of sperm evaluation in vitro for E. poecilogyrus sublineatus in the coastal plain of Rio Grande do Sul, Brazil. After laparatomy the efferent vases were collected and the semen was diluted in 1ml Beltsville Thawing Solution. The characteristics of motility, membrane integrity, mitochondria, acrosome, DNA, cell viability and cellular functionality were evaluated. Fluorescent probes were used for the evaluation of sperm structure in epifluorescence microscope. With the techniques described, it was possible to identify intact and injured cells, enabling the determination of cell characteristics for the spring season (October and November). It was observed in the analyses that 80% of sperm cells were mobile and that 84.1 ± 8.0% of sperm membranes were intact. The standards found were of 48 ± 13.8% of intact acrosome, 73.6 ± 6.0 of perfect DNA and of 91.8 ± 4.0 of functional mitochondria. Thus, these values from the sperm analysis can be used as standards for the species Erythrolamprus poecilogyrus sublineatus.

  19. Nanomedicine and mammalian sperm: Lessons from the porcine model.

    Science.gov (United States)

    Barkalina, Natalia; Jones, Celine; Coward, Kevin

    2016-01-01

    Biomedical nanotechnology allows us to engineer versatile nanosized platforms that are comparable in size to biological molecules and intracellular organelles. These platforms can be loaded with large amounts of biological cargo, administered systemically and act at a distance, target specific cell populations, undergo intracellular internalization via endogenous uptake mechanisms, and act as contrast agents or release cargo for therapeutic purposes. Over recent years, nanomaterials have been increasingly viewed as favorable candidates for intragamete delivery. Particularly in the case of sperm, nanomaterial-based approaches have been shown to improve the efficacy of existing techniques such as sperm-mediated gene transfer, loading sperm with exogenous proteins, and tagging sperm for subsequent sex- or function-based sorting. In this short review, we provide an outline of the current state of nanotechnology for biomedical applications in reproductive biology and present highlights from a series of our studies evaluating the use of specialized silica nanoparticles in boar sperm as a potential delivery vehicle into mammalian gametes. The encouraging data obtained already from the porcine model in our laboratory have formed the basis for ethical approval of similar experiments in human sperm, thereby bringing us a step closer toward the potential use of this novel technology in the clinical environment.

  20. Effect of photoperiod on sexual activity of boar

    Directory of Open Access Journals (Sweden)

    Radomir Savić

    2015-08-01

    Full Text Available The main objective of this study was to assess the effect of photoperiod on sexual activity of three breeds of boars: Swedish Landrace (n=34, Large White (n=38, and Duroc (n=32. Boar sexual activity was analysed based on the libido index and intensity of ejaculation. The libido index was calculated as the ratio between the duration of ejaculation and time of preparation until ejaculation. The intensity of ejaculation was the volume of ejaculate (mL secreted in the unit of time (min. The effect of photoperiod was analysed as the effect of duration of daylight (12 h within photoperiod intervals (increasing and decreasing. Impact assessment was carried out by applying the General Linear Model procedure. Libido and intensity of ejaculation varied under the impact of photoperiod and the breed of boars. With the increase in age, the boar libido weakened, while the volume of ejaculate and intensity of ejaculation increased. Boars manifested better libido when the daylight lasted longer than 12 h in both photoperiod intervals. Different from libido, the volume of ejaculate and intensity of ejaculation were highest when the daylight was shorter than 12 h, but only in the decreasing photoperiod interval. Swedish Landrace boars manifested best libido, while in the production of sperm the Duroc boars were inferior compared with Swedish Landrace and Large White. The phenotypic relationship among libido, ejaculate volume, and ejaculation intensity ranges from very low to high; however, the coefficients were positive, which indicates the possibility of simultaneous improvement of these traits.

  1. Evaluation of pathogenesis caused in cattle and guinea pig by a Mycobacterium bovis strain isolated from wild boar

    Directory of Open Access Journals (Sweden)

    Di Rienzo Julio

    2011-07-01

    Full Text Available Abstract Background In many regions of the world, wild mammals act as reservoir of Mycobacterium bovis, a situation that prevents the eradication of bovine tuberculosis. In order to observe whether a strain isolated from a wild boar, previously tested as highly virulent in a mice model, is also virulent in cattle, we performed cattle experimental inoculation with this strain Results Groups of Friesian calves were either infected with the wild boar strain M. bovis 04-303 or with the bovine strain NCTC10772 as a control. We found that antigen-specific IFN-γ release in whole blood samples occurred earlier in animals infected with M. bovis 04-303. Both M. bovis strains resulted in a positive skin test, with animals infected with the wild boar isolate showing a stronger response. These results and the presence of more severe organ lesions, with granuloma and pneumonic areas in cattle demonstrate that the wild boar isolate is more virulent than the NCTC10772 strain. Additionally, we tested the infectivity of the M. bovis strains in guinea pigs and found that M. bovis 04-303 had the highest pathogenicity. Conclusions M. bovis strains isolated from wild boars may be pathogenic for cattle, producing TB lesions.

  2. Impact of porcine circovirus type 2 (PCV2) vaccination on boar semen quality and quantity using two different vaccines.

    Science.gov (United States)

    Caspari, K; Henning, H; Schreiber, F; Maass, P; Gössl, R; Schaller, C; Waberski, D

    2014-09-01

    Porcine circovirus type-2 (PCV2) is widespread in domestic pig populations. It can be shed with boar semen, but the role boars have in epidemiology is still unclear. Vaccinating boars against PCV2 can reduce disease and virus load in semen, but may have unwanted side effects, that is, impairment of spermatogenesis. Therefore, the aim of this study was to investigate the effect and impact of two different PCV2 vaccines on boar semen quality and quantity. Healthy normospermic Large White boars in three groups of 12 each were vaccinated with either Circovac, Ingelvac CircoFLEX, or received NaCl. Eight ejaculates were collected starting 1 week after vaccination and assessed for quantitative traits. In general, sperm quantity and quality parameters did not change due to the vaccination (P > 0.05). Only DNA integrity between the Circovac and control group was P vaccination, fever period, and impaired sperm quality could be observed. The results indicate that both vaccines did not have a major impact on sperm quality or quantity. Therefore, vaccination of boars against PCV2 seems to be feasible. However, one boar treated with the oil-based vaccine showed a temporarily impaired semen quality after elevated body temperature after vaccination. Thus, possible systemic reactions and the subsequent impact on sperm quality should be taken into account when choosing a PCV2 vaccine for boars.

  3. No effect of the plant growth regulator, chlormequat, on boar fertility

    DEFF Research Database (Denmark)

    Sørensen, M.T.; Poulsen, Mette Erecius; Leffers, H.;

    2009-01-01

    weeks of age and two boar littermates continued on the same treatment as the dam until maturity and delivery of semen for in vitro fertilization (IVF) and in vivo fertilization. Semen volume, sperm concentration and fraction of live sperms were not (P >= 0.46) detrimentally affected by chlormequat...

  4. Evaluating the function of calcium antagonist on the Cd-induced stress in sperm of Russian sturgeon, Acipenser gueldenstaedtii

    Energy Technology Data Exchange (ETDEWEB)

    Li Zhihua, E-mail: zhihuali06@yahoo.com [University of South Bohemia in Ceske Budejovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Zatisi 728/II, 389 25 Vodnany (Czech Republic); Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Jingzhou 434000 (China); Li Ping [University of South Bohemia in Ceske Budejovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Zatisi 728/II, 389 25 Vodnany (Czech Republic); Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Jingzhou 434000 (China); Rodina, Marek; Randak, Tomas [University of South Bohemia in Ceske Budejovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Zatisi 728/II, 389 25 Vodnany (Czech Republic)

    2010-11-15

    In the current study, the sperm of Russian sturgeon (Acipenser gueldenstaedtii) was used to evaluate the roles of Verapamil (VRP), a calcium channel blocker, against cadmium (Cd)-induced stress. Sturgeon sperm were exposed for 2 h at 50 {mu}g/L VRP, 5.0 {mu}g/L Cd, the mixture of 50 {mu}g/L VRP + 5.0 {mu}g/L Cd, 50 {mu}g/L Cd and the mixture of 50 {mu}g/L VRP + 50 {mu}g/L Cd. After exposure, the sperm motility parameters (motility and velocity), oxidative stress levels (lipid peroxidation [LPO] and carbonyl protein [CP]) and antioxidant enzyme activities (superoxide dismutase [SOD], glutathione reductase [GR], glutathione peroxidase [GPx]) were measured in sturgeon sperm. Compared to the control, Cd-induced stress was apparent as reflected by depressed motility parameters, induced oxidative stress and inhibited antioxidant enzyme activities at both Cd concentrations. In the presence of VRP, Cd-induced stress was reduced in sturgeon sperm, especially all the measured parameters in the sperm exposed at 5.0 {mu}g/L Cd returned to control levels, expect for the sperm motility. The present results indicate that VRP can reduce the Cd-induced stress in sturgeon sperm and suggest that using of sperm in vitro assays may provide a novel and efficient means for evaluating the effects of residual metals in the aquatic environment of sturgeon.

  5. Evaluating the function of calcium antagonist on the Cd-induced stress in sperm of Russian sturgeon, Acipenser gueldenstaedtii.

    Science.gov (United States)

    Li, Zhi-Hua; Li, Ping; Rodina, Marek; Randak, Tomas

    2010-11-15

    In the current study, the sperm of Russian sturgeon (Acipenser gueldenstaedtii) was used to evaluate the roles of Verapamil (VRP), a calcium channel blocker, against cadmium (Cd)-induced stress. Sturgeon sperm were exposed for 2h at 50μg/L VRP, 5.0μg/L Cd, the mixture of 50μg/L VRP+5.0μg/L Cd, 50μg/L Cd and the mixture of 50μg/L VRP+50μg/L Cd. After exposure, the sperm motility parameters (motility and velocity), oxidative stress levels (lipid peroxidation [LPO] and carbonyl protein [CP]) and antioxidant enzyme activities (superoxide dismutase [SOD], glutathione reductase [GR], glutathione peroxidase [GPx]) were measured in sturgeon sperm. Compared to the control, Cd-induced stress was apparent as reflected by depressed motility parameters, induced oxidative stress and inhibited antioxidant enzyme activities at both Cd concentrations. In the presence of VRP, Cd-induced stress was reduced in sturgeon sperm, especially all the measured parameters in the sperm exposed at 5.0μg/L Cd returned to control levels, expect for the sperm motility. The present results indicate that VRP can reduce the Cd-induced stress in sturgeon sperm and suggest that using of sperm in vitro assays may provide a novel and efficient means for evaluating the effects of residual metals in the aquatic environment of sturgeon.

  6. Hydroxyflutamide alters the characteristics of live boar spermatozoa.

    Science.gov (United States)

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

    2014-10-15

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

  7. EVALUATION OF CHROMOMYCIN A3 ASSAY IN HUMAN SPERM AFTER SIMULATED OVERNIGHT SHIPMENT

    Science.gov (United States)

    EVALUATION OF CHROMOMYCIN A3ASSAY IN HUMAN SPERM AFTER SIMULATED OVERNIGHT SHIPMENT. SC Jeffay1, R Morris Buus1, LF Strader1, AF Olshan2, DP Evenson3, SD Perreault1. 1US EPA/ORD, RTP, NC;2UNC-CH, Chapel Hill, NC;3SDSU, Brookings, SD.Semen collection kits that allow ...

  8. OBJECTIVE EVALUATION OF HYPERACTIVATED MOTILITY IN RAT SPERMATOZA USING COMPUTER-ASSISTED SPERM ANALYSIS (CASA)

    Science.gov (United States)

    Objective evaluation of hyperactivated motility in rat spermatozoa using computer-assisted sperm analysis.Cancel AM, Lobdell D, Mendola P, Perreault SD.Toxicology Program, University of North Carolina, Chapel Hill, NC 27599, USA.The aim of this study was t...

  9. 猪精子微胶囊的制备及其对精子质量的影响%Preparation of Boar Sperm Microcapsule and Its Effect on Sperm Quality

    Institute of Scientific and Technical Information of China (English)

    张海涛; 马志远; 孙天宇; 王国鑫; 李聪慧; 魏国生; 李井春

    2016-01-01

    为了探讨猪精子微胶囊制作参数,优化猪精子微胶囊制作工艺,提高微胶囊及微胶囊化保存的猪精液质量.将猪精液进行稀释变成1×108个/mL,利用海藻酸钠和钡离子反应制成猪精子微胶囊,检测海藻酸钠和钡离子不同反应时间对胶囊的参数及猪精子质量的影响,以确定最优的微胶囊反应时间的参数.结果表明:5、20、60 min反应时间组之间在胶囊强度分别为23.5、35.5、66.0 g/个,壁厚分别为557.5、1 009.0、1 156.5μm,重量分别为80.0、112.0、135.0 mg以及缓释率(24 h时分别为9.40×104、6.00×104、4.50×104)sperm/个胶囊,以上指标存在显著差异(P<0.05).其中5 min反应组的强度最差;而60 min反应组缓释率最低,不利于精子的释放.此外,20 min和5 min反应时间组在微胶囊化保存后猪精子的活力显著高于60 min反应组,而在精子畸形率上都明显低于60 min反应时间组(P<0.05).综上可见,钡离子-海藻酸钠微胶囊反应时间在20 min时达到最佳,微胶囊保存猪精子质量最好.

  10. Preservation Effect of Different Extenders on Tibetan Boar Semen at 4℃%不同稀释液对藏猪精液4℃保存效果的影响

    Institute of Scientific and Technical Information of China (English)

    陈晓英; 马金英; 宋天增; 唐建华; 赵彦玲; 任子利; 朱彦宾; 王良润

    2016-01-01

    This study was conducted to investigate the preservation effect of different extenders on Tibetan Boar semen at 4℃and to provide scientific basis for reasonable and high-efficient utilization of excellent Tibetan Boar breeder semen. Five different extenders commonly used for preservation of Tibetan Boar semen in room temperature were selected. The sperm motility, sperm deformity rate,and the integrity rate of sperm acrosome and plasma membrane preserved by the five extenders at the 5th day of preservation period at 4℃were statistically evaluated and compared. The results revealed that the NO.2 extenders exhibited the best preservation effect with the sperm motility of 0.54 at the 5th day of preservation period, and the sperm motility, sperm deformity rate and the integrity rate of sperm acrosome and plasma membrane preserved with NO.2 extenders were higher than those of the semen preserved with the other four extenders (P<0.05). This study preliminary selected the appropriate extenders for Tibetan Boar semen used at 4℃and laid the foundation for the production and application of Tibetan Boar semen in the future.%为探讨不同精液稀释液对4℃保存藏猪精液的效果,保证优良藏猪种公猪精液的合理、高效利用,该试验选用5种常用的猪精液常温保存稀释液,并在藏猪精液4℃保存至第5天时检查藏猪精子活力、精子畸形率、精子顶体完整率、精子质膜完整率等。结果表明,在精液保存过程中,2号液对藏猪精液的保存效果最佳,精子活力在保存至第5天时仍然保持在0.54,且藏猪精子活力、精子畸形率、精子顶体完整率、精子质膜完整率均高于其余4组(P<0.05)。该试验研究初步筛选了4℃保存藏猪精液的稀释液,为今后藏猪精液的生产应用奠定了基础。

  11. Lectin histochemistry of the boar bulbourethral glands

    Directory of Open Access Journals (Sweden)

    E Badia

    2009-06-01

    Full Text Available The present study describes, for the first time, the glycosidic content of boar bulbourethral glands using lectin histochemistry. Fourteen horseradish peroxidase- or digoxigeninlabelled lectins with different carbohydrate specificities were used in samples obtained from 3 healthy Landrace boars. The results obtained indicate that endpiece and duct cells synthesize and secrete mainly O-glycoproteins with a- and b-D-N-acetylgalactosamine, b-D-galactose-b(1®3-D-Nacetylgalactosamine, D-N-acetylglucosamine and neuraminic acid residues. Glycoproteins secreted by bulbourethral glands have a role in the protection and lubrication of the urethra. In addition, they may be also involved in the regulation of the sperm metabolic activity and in the maintenance of the structural integrity of acrosomal and plasma membranes.

  12. Reduced glutathione and procaine hydrochloride protect the nucleoprotein structure of boar spermatozoa during freeze-thawing by stabilising disulfide bonds.

    Science.gov (United States)

    Yeste, Marc; Flores, Eva; Estrada, Efrén; Bonet, Sergi; Rigau, Teresa; Rodríguez-Gil, Joan E

    2013-01-01

    One important change the head of boar spermatozoa during freeze-thawing is the destabilisation of its nucleoprotein structure due to a disruption of disulfide bonds. With the aim of better understanding these changes in frozen-thawed spermatozoa, two agents, namely reduced glutathione (GSH) and procaine hydrochloride (ProHCl), were added at different concentrations to the freezing media at different concentrations and combinations over the range 1-2mM. Then, 30 and 240 min after thawing, cysteine-free residue levels of boar sperm nucleoproteins, DNA fragmentation and other sperm functional parameters were evaluated. Both GSH and ProHCl, at final concentrations of 2mM, induced a significant (Psperm head disulfide bonds 30 and 240 min after thawing compared with the frozen-thawed control. This effect was accompanied by a significant (Psperm peroxide levels, motility patterns and plasma membrane integrity. In conclusion, the results show that both GSH and ProHCl have a stabilising effect on the nucleoprotein structure of frozen-thawed spermatozoa, although only GSH exerts an appreciable effect on sperm viability.

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

    Science.gov (United States)

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

    2016-01-01

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

  14. The increase in phosphorylation levels of serine residues of protein HSP70 during holding time at 17°C is concomitant with a higher cryotolerance of boar spermatozoa.

    Directory of Open Access Journals (Sweden)

    Marc Yeste

    Full Text Available Boar-sperm cryopreservation is not usually performed immediately after semen collection, but rather a holding time (HT of 4 h-30 h at 17°C is spent before starting this procedure. Taking this into account, the aim of this study was to go further in-depth into the mechanisms underlying the improving effects of HT at 17°C on boar-sperm cryotolerance by evaluating the effects of two different HTs (3 h and 24 h on overall boar-sperm function and survival before and after cryopreservation. Given that phospho/dephosphorylation mechanisms are of utmost importance in the overall regulation of sperm function, the phosphorylation levels of serine residues (pSer in 30 different sperm proteins after a 3 h- or 24 h-HT period were also assessed. We found that a HT of 24 h contributed to a higher sperm resistance to freeze-thawing procedures, whereas mini-array protein analyses showed that a HT of 24 h induced a significant (P<0.05 increase in pSer (from 100.0±1.8 arbitrary units in HT 3 h to 150.2±5.1 arbitrary units in HT 24 h of HSP70 and, to a lesser extent, in protein kinases GSK3 and total TRK and in the cell-cycle regulatory protein CDC2/CDK1. In the case of HSP70, this increase was confirmed through immunoprecipation analyses. Principal component and multiple regression analyses indicated that a component explaining a percentage of variance higher than 50% in sperm cryotolerance was significantly correlated with pSer levels in HSP70. In addition, from all the parameters evaluated before freeze-thawing, only pSer levels in HSP70 resulted to be able to predict sperm cryotolerance. In conclusion, our results suggest that boar spermatozoa modulate its function during HT, at least partially, by changes in pSer levels of proteins like HSP70, and this is related to a higher cryotolerance.

  15. Using test station and on-farm data for the genetic evaluation of Pietrain boars used on Landrace sows for growth performance.

    Science.gov (United States)

    Dufrasne, M; Rustin, M; Jaspart, V; Wavreile, J; Gengler, N

    2011-12-01

    The aim of this study was to develop a new genetic evaluation model to estimate the genetic merit of boars for growth based on 1) performance of their crossbred progeny fattened in the test station and 2) their own performance or those of relatives from the on-farm testing system. The model was a bivariate random regression animal model with linear splines and was applied to Piétrain boars from the Walloon Region of Belgium mated with Landrace sows. Data contained 1) 12,610 BW records from the test station collected on 1,435 crossbred pigs from Piétrain boars and Landrace sows, and 2) 52,993 BW records from the on-farm testing system collected on 50,670 pigs with a breed composition of at least 40% Piétrain or Landrace. Since 2007, 56 Piétrain boars have been tested in the station. Data used to estimate variance components and breeding values were standardized for the age to take into account heterogeneity of variances and then pre-adjusted at 210 d of age to put all records on the same scale. Body weight records from the test station and from the on-farm testing system were considered as 2 different traits. The heterosis effect was modeled as fixed regression on the heterozygosity coefficient. As all test station animals were similarly crossbred, smaller variation in heterozygosity caused the sampling error of the regression estimate at 210 d to be larger in the test station than in on-farm data with estimates of 28.35 ± 14.55 kg and 9.02 ± 0.67 kg, respectively. Therefore, the most likely reason for the large differences in estimates was sampling. Heritability estimates ranged from 0.37 to 0.60 at 210 and 75 d, respectively, for test station BW and from 0.42 to 0.60 at 210 d and 175 d, respectively, for on-farm BW. Genetic correlation decreased when the age interval between records increased, and were greater between ages for test station than for on-farm data. Genetic correlations between test station and on-farm BW at the same age were high: 0.90 at 175 d

  16. Use of refractometry as a new management tool in AI boar centers for quality assurance of extender preparations.

    Science.gov (United States)

    Schulze, M; Rüdiger, K; Jung, M; Grossfeld, R

    2015-01-01

    A study was performed to see if refractometry can be used as a new quality control tool for boar semen extenders. For this the refractive index and osmolality of BTS extender concentrations (EC) were recorded in 10%-steps from 50% to 150% and 200% of the correct amount. Twelve boar ejaculates were evaluated for semen quality. The refractive index for the correctly prepared extender was 4.6±0.0°Bx, corresponded to 316±16mOsmkg(-1), and correlated highly with osmolality (r=0.99; P<0.001). Total sperm motility with 100% EC differed significantly from ≤70% EC (P<0.001) and 200% EC (P<0.001) on day 1 (d1) and d4, respectively. The percentage of motile spermatozoa in a thermoresistance test on d2 showed a significant drop using ≤70% EC (P=0.047) and ≥140% EC (P=0.004). Secondary apical ridge defects were significantly higher using 50% EC (P<0.001) and ≥150% EC (P=0.032) compared to 100% EC, respectively. An increased number of coiled tails were observed using ≤60% EC (P<0.001). Percentages of spermatozoa with intact membranes on d2 resulted in a significant decrease using 50% EC (P<0.001) and ≥150% EC (P=0.005), respectively. The mean percentage of PI negative spermatozoa with active mitochondria on d2 showed a significant difference using ≤60% EC (P=0.016) and ≥140% EC (P<0.001) compared to 100% EC, respectively. Boar sperm quality is affected by inexact extender preparation. The refractive-index is an indicator of osmolality and may be used to verify semen extender preparation. The sensitivity is sufficient to detect deviations from correct extender preparation before negative effects on sperm quality occur.

  17. Evaluation of methods to determine sperm density for the european eel, anguilla anguilla

    DEFF Research Database (Denmark)

    Sørensen, Sune Riis; Gallego, V.; Pérez, L.;

    2013-01-01

    , computer-assisted sperm analysis (CASA) and flow cytometry (FCM), using Neubauer Improved haemocytometer as benchmark. Initially, relationships between spermatocrit, haemocytometer counts and sperm motility were analysed, as well as the effect of sperm dilution on haemocytometer counts. Furthermore...

  18. Evaluation of effects of 1,3-dinitrobenzene on sperm motility of hamster using computer assisted semen analysis (CASA)

    Institute of Scientific and Technical Information of China (English)

    L.D.C. Peiris; H.D.M. Moore

    2001-01-01

    Aim: To evaluate the effects of 1,3-dinitrobenznene (mDNB) on sperm motility of hamster and to correlate the results with the fertility. Methods: Adult male hamsters were gavaged with one of the 3 dose regimes of mDNB (1.5 mg daily for 4 weeks, 1.5 mg one day a week for 4 weeks and 1.0 mg 3 days a week for 4 weeks). Computer assisted semen analysis (CASA) was used to analyse the sperm motility parameters, curvilinear velocity (VCL) and straight line velocity (VSL) of sperm in distal corpus epididymides and distal cauda epididymides. In vitro fertilisation was carded out only for 1.5 mg mDNB daily group to determine the sperm fertilising capacity. Results: There was a significant reduction in sperm velocity parameters at weeks 3 and 4 after treatment, which was correlated with a de cline in sperm fertility. Conclusion: Sperm velocity parameters may be used to determine the effect of a toxic insult on the sperm function.

  19. Effects of low-density lipoproteins extracted from different avian yolks on boar spermatozoa quality following freezing-thawing.

    Science.gov (United States)

    Wang, Peng; Wang, Yan-Feng; Wang, Chun-Wei; Bu, Shu-Hai; Hu, Jian-Hong; Li, Qing-Wang; Pang, Wei-Jun; Yang, Gong-She

    2014-05-01

    Low-density lipoproteins (LDL) is known to protect boar sperm during freezing-thawing, but little information is known about the effects of LDL extracted from different avian egg yolks on post-thaw boar semen quality. The purpose of this study was to compare and analyze the effects of LDL at various concentrations and different species on boar sperm quality after freezing-thawing. LDL extracted from the yolk of hen egg, duck egg, quail egg, pigeon egg or ostrich egg was added to the extender at the concentrations of 0.06, 0.07, 0.08, 0.09 and 0.1 g/ml, respectively, and their effects on frozen-thawed boar sperm quality were assessed. According to all measured parameters, the results showed that sperm motility, acrosome integrity and plasma membrane integrity were 43.20%, 52.57% and 48.13%, respectively, after being frozen-thawed with 0.09 g/ml LDL extracted from pigeon egg yolk. All these quality parameters were higher than that of other groups (P boar sperm among all of the groups supplemented with LDL from five kinds of avian egg in extender. The optimum concentration of LDL extracted from pigeon egg in boar semen freezing extender was 0.09 g/ml.

  20. Evaluation of ELISA coupled with Western blot as a surveillance tool for Trichinella infection in wild boar (Sus scrofa).

    Science.gov (United States)

    Cuttell, Leigh; Gómez-Morales, Maria Angeles; Cookson, Beth; Adams, Peter J; Reid, Simon A; Vanderlinde, Paul B; Jackson, Louise A; Gray, C; Traub, Rebecca J

    2014-01-31

    Trichinella surveillance in wildlife relies on muscle digestion of large samples which are logistically difficult to store and transport in remote and tropical regions as well as labour-intensive to process. Serological methods such as enzyme-linked immunosorbent assays (ELISAs) offer rapid, cost-effective alternatives for surveillance but should be paired with additional tests because of the high false-positive rates encountered in wildlife. We investigated the utility of ELISAs coupled with Western blot (WB) in providing evidence of Trichinella exposure or infection in wild boar. Serum samples were collected from 673 wild boar from a high- and low-risk region for Trichinella introduction within mainland Australia, which is considered Trichinella-free. Sera were examined using both an 'in-house' and a commercially available indirect-ELISA that used excretory-secretory (E/S) antigens. Cut-off values for positive results were determined using sera from the low-risk population. All wild boar from the high-risk region (352) and 139/321 (43.3%) of the wild boar from the low-risk region were tested by artificial digestion. Testing by Western blot using E/S antigens, and a Trichinella-specific real-time PCR was also carried out on all ELISA-positive samples. The two ELISAs correctly classified all positive controls as well as one naturally infected wild boar from Gabba Island in the Torres Strait. In both the high- and low-risk populations, the ELISA results showed substantial agreement (k-value=0.66) that increased to very good (k-value=0.82) when WB-positive only samples were compared. The results of testing sera collected from the Australian mainland showed the Trichinella seroprevalence was 3.5% (95% C.I. 0.0-8.0) and 2.3% (95% C.I. 0.0-5.6) using the in-house and commercial ELISA coupled with WB respectively. These estimates were significantly higher (PTrichinella DNA in any mainland animals, but did reveal the presence of a second larvae-positive wild boar on Gabba

  1. Evaluation of the proliferative activity of immunocompetent cells in the jejunal and iliac lymph nodes of prepubertal female wild boars diagnosed with mixed mycotoxicosis

    Directory of Open Access Journals (Sweden)

    Zielonka Łukasz

    2015-06-01

    Full Text Available The study evaluated the proliferative activity of immunocompetent cells in the jejunal and iliac lymph nodes of prepubertal female wild boars exposed to deoxynivalenol and zearalenone in naturally contaminated feed. The evaluation was performed with the use of the MTT assay and 2 mitogens: lipopolysaccharide (LPS and concanavalin A. Intensified proliferative processes in T and B lymphocytes were revealed. The mitogenic activity of LPS was more expressed in the lymphocytes of both iliac and jejunal lymph nodes in comparison with the control group. Proliferative activity was higher in iliac lymph nodes than in jejunal lymph nodes. A reverse trend was observed in the percentage of live cells, which was higher in jejunal lymph nodes during the evaluation of lymphocyte proliferation.

  2. Lean meat prediction with HGP, CGM and CSB-Image-Meater, with prediction accuracy evaluated for different proportions of gilts, boars and castrated boars in the pig population.

    Science.gov (United States)

    Engel, B; Lambooij, E; Buist, W G; Vereijken, P

    2012-02-01

    Prediction equations for the percentage lean meat in pig carcasses in The Netherlands were derived for the Hennessy Grading Probe 7, Capteur Gras/Maigre--Sydel and CSB-Image-Meater. Because castrated males are expected to vanish from the Dutch pig population in the near future, accuracy of prediction was evaluated for different scenarios representing a wide range of different proportions for entire males, castrated males and females in the Dutch pig population. The prediction equations for the instruments are in compliance with the EC regulations for prediction accuracy for the different scenarios. So, these equations will remain valid when castrated males are (gradually) removed from the Dutch slaughter population. Results of this study are of interest for researchers from countries or areas contemplating the use of one of the aforementioned instruments. The statistical approach for evaluation of prediction accuracy is of particular interest when changes in proportions of important subpopulations in the target population are foreseen.

  3. 纳米化聚酰胺-胺型树枝状聚合物对猪精子介导基因转移效率的影响%Positive effects of PAMAM-D on the efficiency of boar sperm-mediated gene transfer

    Institute of Scientific and Technical Information of China (English)

    吴斌; 戴建军; 张廷宇; 张树山; 吴彩凤; 顾晓龙; 陈会兰; 张德福; 马恒东

    2013-01-01

    To optimize production of transgenic pig by sperm-mediated gene transfer, the present study was designed to examine the effects of treatment with Nano-PAMAM-D on exogenous DNA uptake of boar spermatozoa and gene expression in in vitro fertilized eggs. Exogenous DNA treated with Nano-PAMAM-D was incubated with boar spermatozoa,the exogenous DNA uptake was detected by in situ hybridization, and exogenous gene expression was examined by fluorescence microscope and PCR analysis. The results showed that positive rate of sperm treated with Nano-PAMAM-D and DNA according to 0. 5 μg linear plasmid plus PAMAM-D at the ratio of 20 : 1 was the highest group (P<0. 05). The positive rates were significantly increased for incubation 90 or 120 min compared to incubation 30 or 60 min(19. 94%, 18. 57% vs 8. 50%, 13. 87%; P< 0. 05). The complexes formed between exogenous DNA and Nano-PAMAM-D were internalized (23.93% vs 7. 25%, 13. 25%,P<0. 05)on boar spermatozoa at a higher efficiency compared to methods using DNA or lipofectin alone. This study suggested that boar spermatozoa using Nano-PAMAM-D as transfectant could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. In conclusion,Nano-PAMAM-D vector could be used to efficiently introduce a exogenous gene into embryo via spermatozoa.%为优化猪精子介导的基因转移(SMGT)技术,提高转基因效率,本试验利用纳米化聚酰胺-胺型树枝状聚合物(PAMAM-D)为载体介导猪精子转染外源DNA,经原位杂交检测其阳性率,体外受精(IVF)分析外源基因的表达.结果显示,0.5 μg线状质粒DNA中加入PAMAM-D(质量比20∶1)阳性率最高(P<0.05);孵育时间为90、120min时,阳性率显著高于30、60 min组(19.94%、18.57%与8.50%、13.87%;P<0.05);以此条件处理精子,阳性率显著高于无载体和脂质体处理组(23.93%与7.25%0、13.25%;P<0.05).各处

  4. Evaluation of the effect of cooling and of the addition of collagenase on llama sperm DNA using toluidine blue.

    Science.gov (United States)

    Carretero, M I; Giuliano, S M; Casaretto, C I; Gambarotta, M C; Neild, D M

    2012-05-01

    The effect cryopreservation has on sperm chromatin condensation has been studied in many species but not in South American camelids. The objectives of this study were to evaluate with toluidine blue (TB) the effects of cooling and of adding collagenase on llama sperm DNA condensation. The optimum incubation time (30 s, 1.5 and 3 min) with a reducing agent (dithiothreitol) was also determined. When comparing cooled samples with the raw ejaculate, a significant increase in sperm showing a high degree of decondensation (TB positive) was observed (P = 0.005). A positive correlation was observed, both in raw and cooled semen, between sperm head morphological abnormalities observed in TB-stained cells and TB-positive sperm (highly decondensed DNA), but not with TB-intermediate spermatozoa (moderately decondensed DNA). No significant differences (P > 0.05) were observed in samples incubated with or without 0.1% collagenase. In cooled semen, but not in raw, a significant increase (P = 0.000) in reacted sperm (TB positive) was observed using 3-min incubation with 1% dithiothreitol (DTT). To conclude, cooling would seem to produce an increase in llama sperm chromatin decondensation. Also, 0.1% collagenase in H-TALP-BSA could be added to raw semen to aid its manipulation as it would not seem to increase DNA decondensation.

  5. Genetic correlations between male reproductive traits and growth traits in growth performance tested Duroc, Landrace and Yorkshire breed boars.

    Science.gov (United States)

    Chang, Hsiu-Luan; Lai, Yung-Yu; Wu, Ming-Che; Sasaki, Osamu

    2017-02-09

    Male-related traits at 180-225 days of age for 6464 grow-finish performance tested boars were measured from 2000 to 2016. Heritability estimates and genetic correlations among average daily gain, feed efficiency, back fat thickness, teat counts, mounting libido, leg locomotion, penile length, sperm motility, sperm concentration and total sperm counts were estimated by VCE software using a multiple traits animal model in each breed. Growth-tested boars had heritability estimates of male reproductive traits in 0.34-0.56 of teat counts, 0.12-0.20 of libido, 0.08-0.12 of locomotion, 0.17-0.58 of penile length, 0.04-0.21 of sperm motility and concentration, 0.17-0.30 of total sperm counts. Total sperm counts were genetically positively correlated with penile length in all breeds. Boars with higher total sperm counts had genetically better libido and locomotion. Genetic correlation between feed efficiency and sperm motility and feed efficiency and sperm concentration were positive in Duroc and negative in Landrace and Yorkshire. Sperm motility and concentration were genetically negatively correlated with average daily gain in Yorkshire. Male reproductive traits of imported breeds could be improved with care in the change of growth traits, especially in Yorkshire.

  6. Genetic parameters for male fertility and its relationship to skatole and androstenone in Danish Landrace boars.

    Science.gov (United States)

    Strathe, A B; Velander, I H; Mark, T; Ostersen, T; Hansen, C; Kadarmideen, H N

    2013-10-01

    Concerns have been raised regarding selection against the boar taint compounds, androstenone and skatole, due to potential unfavorable genetic correlations with important male fertility traits (i.e., selection of boars with low levels of these boar taint compounds might also reduce male fertility). Hence, the objective of this investigation was to study the genetic association between direct measures of male fertility and the boar taint compounds in Danish Landrace pigs. Concentrations of skatole and androstenone in the back fat were available for approximately 6,000 and 1,000 Landrace boars, respectively. The litter size traits, such as total number born, live piglets at d 5, and piglet survival until d 5 on relatives of the slaughter boars, were extracted from the Danish Landrace breeding program, yielding 35,715 records. Semen volume, sperm concentration, subjective sperm quality score, and total number of sperm were available from 95,267 ejaculates. These ejaculates were collected between 2005 and 2012 and originated from 3,145 Landrace boars from 12 AI stations in Denmark. The traits were analyzed using single and multitrait animal models including univariate random regression models. Skatole and androstenone concentrations were moderate to highly heritable (i.e., 0.33 and 0.59, respectively). The genetic correlation between the two compounds was moderate (0.40). Genetic variance of sperm production per ejaculate increased during the productive life of the boar, resulting in heritability estimates increasing from 0.18 to 0.31. Genetic correlations between sperm production per ejaculate at different ages were high and generally larger than 0.8, indicating that later genetic merit can be predicted from records at an early age. The heritability (based on service-sire genetic component) of both total number of piglets born and survival to d 5 were 0.02, and the correlation between these effects and the additive genetic effect on boar taint ranged from 0.05 to -0

  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. Interactions of egg yolk lipoprotein fraction with boar spermatozoa assessed with a fluorescent membrane probe.

    Directory of Open Access Journals (Sweden)

    Łukasz Zasiadczyk

    2010-08-01

    Full Text Available The interactions of a fluorescent membrane probe, 1-anilinonaphthalene-8-sulfonic acid (1,8-ANS, with boar spermatozoa were followed through the use of lipoprotein fraction of ostrich egg yolk (LPFo. Semen samples, extended in Kortowo 3 (K3 extender, were supplemented with 2% or 5% LPFo and stored for 3h at 16 degrees C. Additionally, cold shock-treated spermatozoa (1h at 4 degrees C were stored in K3 extender supplemented with LPFo for 3h at 16 degrees C. In each boar, the fluorescent enhancement of ANS was observed in K3-extended semen supplemented with LPFo, prior to storage. Following storage, there was a significant increase in LPFo-ANS fluorescence, particularly in the sperm membrane overlying the head and midpiece regions. There were significant differences among the boars with respect to the sperm populations defined by the LPFo-ANS fluorescence. Sperm viability was not significantly affected during the storage period. Furthermore, the proportions of spermatozoa defined by the different patterns of LPFo-ANS fluorescence were low and remained unchanged after storage of cold shock-treated spermatozoa with 2% or 5% LPFo, suggesting irreversible damage to the sperm membrane architecture. These findings indicate that the ANS fluorescent probe could be used to shed more light on the nature of the interactions between LPFo and sperm membrane following semen preservation. Such valuable information could contribute to the development of an optimal protocol for cryopreservation of boar semen.

  9. Guangxi Donor Sperm Enterprises and Institutions boars 4 disease virus survey in 2007-2013%2007-2013年广西供精单位种公猪4种疫病带毒情况调查

    Institute of Scientific and Technical Information of China (English)

    覃芳芸; 马琳; 胡丽萍; 杨荣; 冯淑萍; 胡巧云; 黄胜斌; 熊毅

    2014-01-01

    为了解广西供精单位种公猪主要疫病带毒情况,对开展公猪疫病控制与净化工作提供科学数据。通过对2007-2013年广西158个供精单位5611份公猪精液和血清进行猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪伪狂犬病毒(PRV)和布鲁氏菌抗体的检测,检出 CSFV、PRRSV的阳性数及阳性率分别为21份(0.37%)、186份(3.31%),检出CSFVP+RRSV混合阳性样品4份,阳性率0.07%,未检出PRV和布鲁氏菌抗体阳性样品。因此,得出公猪精液传播病毒仍是母猪发生繁殖障碍疫病的重要原因之一的结论,建议加强种公猪的疫病控制与净化工作,加强公猪精液的管理,从源头上控制病原的传播。%Objective:This study was aimed to understand the diseases of boar from insemination units in Guangxi,that provides a scientific data for disease control in boar.Method:5611 boar semen and serum samples were collected from 158 insemination units in Guangxi province from Year 2007 to 2013,and tested for 3 viruses and 1 antibody including classical swine fever virus (CSFV),porcine reproductive and respiratory syndrome virus (PRRSV),pseudorabies virus (PRV) and antibody of Brucella.ResuIt:The positive rate of CSFV、PRRSV 、PRV and antibody of Brucella were 0.37%、3.31%、0% and 0% respectively.2 samples were tested as mixed infection with CSFV and PRRSV,the positive rate was 0.07%.ConcIusion:The results indicated that the virus spread by semen was still an important fector for breeding disease of sows.To control the disease from source,one suggestion is further strengthen disease control and manggenment in boar and semen.

  10. Boar taint detection using parasitoid biosensors

    Science.gov (United States)

    To evaluate the potential for a non-stinging wasp to be used as a biosensor in the pig industry, we trained wasps to 3 individual chemicals associated with boar taint. Training consisted of presenting the odors to hungry wasps while they were feeding on sugar. This associates the chemical with a fo...

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

    Science.gov (United States)

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

    2015-01-01

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

  12. New insights about the evaluation of human sperm quality: the aromatase example.

    Directory of Open Access Journals (Sweden)

    A Saad

    2010-01-01

    Full Text Available Male contribution to the couple's infertility is at first evaluated by the routine examination of semen parameters upon optical microscopy providing valuable information for a rational initial diagnosis and for a clinical management of infertility. But the different forms of infertility defined according to the WHO criteria especially teratozoospermia are not always related to the chromatin structure or to the fertilization capacity. New investigations at the molecular level (transcript and protein could be developed in order to understand the nature of sperm malformation responsible of human infertility and thus to evaluate the sperm quality. The profile analysis of spermatozoal transcripts could be considered as a fingerprint of the past spermatogenic events. The selection of representative transcripts of normal spermatozoa remains complex because a differential expression (increased, decreased or not modified levels of specific transcripts has been revealed between immotile and motile sperm fractions issued from normozoospermic donors. Microarrays tests or real-time quantitative PCR could be helpful for the identification of factors involved in the male infertility. Differences in the expression of specific transcripts have been reported between normal and abnormal semen samples. With the aromatase example, we have noted a negative strong correlation between the amount of transcript and the percentage of abnormal forms especially in presence of head defects. Immunocytochemical procedures using fluorescent probes associated with either confocal microscopy or flow cytometry can be also helpful to proceed with further investigations about the localization of proteins in the compartmentalized spermatozoa or the acrosome reaction. The dual location of aromatase both in the equatorial segment, the mid-piece and the tail could explain the double role of this enzyme in acrosome reaction and motility.

  13. Apoptotic-like changes of boar spermatozoa in freezing media supplemented with different antioxidants.

    Science.gov (United States)

    Trzcińska, M; Bryła, M

    2015-01-01

    This study evaluated the effect of supplementing the freezing extender with exogenous anti-oxidants on apoptotic-like changes in post-thaw boar spermatozoa. A total of 36 ejaculates were resuspended in standard lactose-egg yolk-glycerol extender supplemented with antioxidant to final concentrations of 0 (as control), 2.5mM GSH (group I), 5.0 mM GSH (group II), 150 IU/mL SOD (group III), 300 IU/mL SOD (group IV), 200 IU/mL CAT (group V), 400 IU/mL CAT (group VI), 150 IU/mL SOD+200 IU/mL CAT (group VII), 300 IU/mL SOD+400 IU/mL CAT (group VIII). Sperm motility and apoptotic-like changes were determined before and after freeze-thawing. The various markers of apoptotic-like changes were measured: plasma membrane permeability by YO-PRO-1/PI assay, phosphatidylserine (PS) translocation across the plasma membrane using fluorescein-labeled Annexin-V, mitochondrial transmembrane potential detected by JC-1, and DNA fragmentation evaluated by TUNEL assay. The highest percentage of progressive motile sperm was noticed in group II (PM% 64.2±15.4) compared with control (PM% 36.8±5.5). The supplementation of 400 IU/mL CAT (group VI) revealed significant (Psperm survival compared with the control. Evaluation by TUNEL assay revealed that cryopreservation and thawing did not induce DNA fragmentation in boar spermatozoa.

  14. Evaluation of an animal protein-free semen extender for cryopreservation of epididymal sperm from North American bison (Bison bison).

    Science.gov (United States)

    Krishnakumar, S; Whiteside, D P; Elkin, B; Thundathil, J C

    2011-07-15

    The objective was to evaluate the suitability of an animal protein-free semen extender for cryopreservation of epididymal sperm from the two subspecies of North American bison: plains (Bison bison bison) and wood (Bison bison athabascae) bison. Both cauda epididymides (from six plains and five wood bison) were minced and incubated in Sp-TALPH buffer for approximately 2 h at 37 °C to release actively motile sperm. Sperm suspensions were filtered, centrifuged and the sperm pellet from each bull was divided into two fractions and diluted either in egg yolk containing extender, Triladyl, or in an animal protein-free extender, Andromed, and equilibrated for 20 min at 37 °C. Thereafter, samples were chilled and cryopreserved. Frozen-thawed sperm were evaluated for motility (computer assisted sperm analysis), viability (SYBR 14 and propidium iodide), acrosome integrity (FITC conjugated PSA), cryocapacitation (tyrosine phosphorylation of sperm proteins as a biomarker), and fertilizing ability (in a heterologous IVF system). There was no significant difference for progressive motility, viability, and acrosome integrity between the two extenders for plains bison (36.8 ± 9.0, 60.5 ± 17.4, and 77.3 ± 4.6%; overall mean ± SD) as well as for wood bison (11.7 ± 8.1, 13.7 ± 5.6, and 73.4 ± 4.2%). Levels of tyrosine phosphorylation did not differ for sperm preserved in the two extenders for both subspecies, although an inter-bull variability in the response to tyrosine phosphorylation between extenders was suggested for plains bison. Fertilization percent did not differ significantly between extenders for plains bison (84.16 ± 9.92%, overall mean ± SD) and for wood bison (59.53 ± 19.99%). In conclusion, in the absence of significant difference between extenders in post-thaw sperm characteristics, we inferred that Andromed (animal protein-free) was suitable for cryopreservation of epididymal sperm from North American bison.

  15. Semen evaluation in four autochthonous wild raptor species using computer-aided sperm analyzer.

    Science.gov (United States)

    Dogliero, Andrea; Rota, Ada; Lofiego, Renato; Mauthe von Degerfeld, Mitzy; Quaranta, Giuseppe

    2016-04-01

    At least 10 percent of the approximately 300 species of the order Falconiformes are listed as being globally threatened. The present work describes the seminal characteristics of three diurnal and one nocturnal raptor species. Semen was collected from clinically healthy Accipiter nisus (n = 1), Falco subbuteo (n = 6), and Falco tinnunculus (n = 5) adult males that were housed at the 'Centro Animali Non Convenzionali' of the Department of Veterinary Sciences of the University of Turin. The semen was collected after a period of recovery and before their release as well as from seven Bubo bubo males bred in captivity as part of a raptor conservation project. All the potential semen donors were trained in semen collection during the breeding season via a ritualized procedure. Ejaculation was achieved using a massaging technique. Each sample was evaluated for volume, degree of contamination, and spermatozoa concentration. The semen motility and kinetic parameters were assessed on diluted semen (modified tyroides albumin lactate pyruvate, pH 7.5, temperature 37.5 °C) using a computer-aided sperm analyzer. Semen collection was successful in all the diurnal species and in five B bubo individuals. The sperm motility and sperm kinetic parameters were very variable both among and within species. In contrast with previous studies that involved raptors bred in captivity and imprinted on humans, we worked with wild birds and attempted to overcome the problem of poor semen quality, which is strongly influenced by stress, by adopting a ritualized procedure that has never been reported for semen collection purposes.

  16. 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.

  17. The influence of short-term exposure to tropical sunlight on boar seminal characteristics

    Science.gov (United States)

    Egbunike, G. N.; Dede, T. I.

    1980-06-01

    The seminal characteristics of 4 Large White boars exposed to direct tropical sunlight 45 min daily for three days were compared to those of their mates that were maintained under shade in the barn. During the period of exposure, both respiratory rate and rectal temperature increased significantly by 276.84 and 5.13% respectively in the exposed over the unexposed boars, thus indicating a high degree of hyperthermia. Although libido, as judged from the reaction time, was unaffected, the ejaculation time appeared to be longer for the stressed than unstressed animals. Gel mass, semen volume and pH appeared to be stable inspite of the treatment, unlike sperm motility and concentration which deteriorated. Also, the dehydrogenase activity of the semen was inferior in the stressed animals. Sperm output per ejaculate dropped drastically only in the week following exposure from 58.22 to 28.42 billion sperm as compared to corresponding values of 54.83 and 47.87 by the unexposed boars. Similarly, the frequency of sperm abnormality was higher in the stressed boars in this period after which the animals appeared to have recovered.

  18. Mitochondrial function and reactive oxygen species action in relation to boar motility.

    Science.gov (United States)

    Flow cytometric assays of viable boar sperm were developed to measure reactive oxygen species (ROS) formation (oxidization of hydroethidine to ethidium), membrane lipid peroxidation (oxidation of lipophilic probe C11-BODIPY581/591), and mitochondrial inner transmembrane potential (aggregation of mit...

  19. Automatic classification of the acrosome status of boar spermatozoa using digital image processing and LVQ

    NARCIS (Netherlands)

    Alegre, Enrique; Biehl, Michael; Petkov, Nicolai; Sanchez, Lidia

    2008-01-01

    We consider images of boar spermatozoa obtained with ail optical phase-contrast microscope. Our goal is to automatically classify single sperm cells as acrosome-intact (class 1) or acrosome-damaged (class 2). Such classification is important for the estimation of the fertilization potential of a spe

  20. Evaluation of human sperm chromatin status after selection using a modified Diff-Quik stain indicates embryo quality and pregnancy outcomes following in vitro fertilization

    OpenAIRE

    Tavares, R. S.; A.F. Silva; Lourenço, B.; Almeida-Santos, T; Sousa, A. P.; Ramalho-Santos, J.

    2013-01-01

    Sperm chromatin/DNA damage can be measured by a variety of assays. However, it has been reported that these tests may lose prognostic value in Assisted Reproductive Technology (ART) cycles when assessed in post-prepared samples, possibly due to the normalizing effect promoted by sperm preparation procedures. We have recently implemented a modified version of the Diff-Quik staining assay that allows for the evaluation of human sperm chromatin status in native samples, together with st...

  1. Enhanced binding of zona pellucida proteins to the acrosomal region of intact boar spermatozoa in response to fertilizing conditions: a flow cytometric study

    NARCIS (Netherlands)

    Harkema, W.; Harrison, R.A.P.; Miller, N.G.A.; Topper, E.K.; Woelders, H.

    1998-01-01

    In this investigation we sought to determine whether sperm capacitation in vitro is accompanied by changes in the functional presence of zona binding sites on the plasma membrane of boar spermatozoa. During sperm incubation at 39°C in various modifications of a Tyrode's-based in vitro fertilization

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

    Science.gov (United States)

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

    2015-01-01

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

  3. 刃天青还原分析法在用接收器特性法评估雄猪精液中的诊断意义%Determination of the diagnostic value of the resazurin reduction assay for evaluating boar semen by receiver operating characteristic analysis

    Institute of Scientific and Technical Information of China (English)

    Petra Zrim(s)ek; Marjan Kosec; Janez Kunc; Janko Mrkun

    2006-01-01

    Aim: To assess that metabolic status of spermatozoa could provide a useful tool for evaluation of semen quality.Methods: The accuracy of the spectrophotometric application of the resazurin reduction assay was assessed using receiver operating characteristic (ROC) analysis. Results: Areas under ROC curves (AUC) for motile sperm concentration and sperm index (SI) (sperm concentration multiplied by the square root of percentage sperm motility multiplied by the percentage normal sperm morphology) were 0.922. The best discrimination between poor and good semen samples according to the SI was achieved at a cut-off point of A610 = 0.209, where high sensitivity (94.1%) and specificity (91.7%) were calculated. The assay was less accurate when motile sperm concentration was used as the criterion value, yielding sensitivity of 88.2% and specificity of 87.5%, respectively. Likelihood ratios (LR) indicate that absorbances lower than 0.209 were at least 11.3 times as likely to be found in good semen samples than those in poor according to the SI, whereas in the case of motile sperm concentration, the LR was calculated to be 7.06. Conclusion:These results show that the resazurin reduction assay combined with spectrophotometry is an accurate method of assessing the quality of boar semen.%目的:确定精子代谢状态可以作为一个有用的精液质量分析工具.方法:用接收器特性法(ROC)评估刃天青还原法联合分光光度计法的精确度.结果:活动精子密度和精子指数(SI)(精子密度×精子活力百分比的平方根×形态正常精子百分比)的ROC曲线下面积为0.992.参考SI值,采集到的精液样本的质量高低区分点的吸光度为0.209(610nm波长),该点的敏感性和特异性值分别为94.1%和91.7%.如果以活动精子密度为标准值,这种分析方法的精确度就会降低,敏感性和特异性值分别为88.2%和87.5%.似然比(LR)表明在质量良好的精液样本中测得的吸光度低于0.209,至少相当

  4. An evaluation of soybean lecithin as an alternative to avian egg yolk in the cryopreservation of fish sperm.

    Science.gov (United States)

    Yildiz, Cengiz; Bozkurt, Yusuf; Yavas, Ilker

    2013-08-01

    Plant-derived lecithin has been used as a more sanitary alternative to avian egg yolk in livestock sperm cryopreservation protocols but its efficacy for cryopreserving fish sperm has not previously been tested comparatively. Here various concentrations of soybean lecithin were evaluated for the cryopreservation of carp (Cyprinus carpio) sperm. Sexually mature fish were induced to spermiation and ovulation with ovopel. The extenders were prepared by using 300 mM glucose, 10% DMSO, supplemented with different ratios of lecithin (5%, 10%, 15%, and 20%) and 10% egg yolk (control I). Negative control was made without egg yolk and soybean lecithin (control II). The pooled semen was diluted separately at ratio of 1:3 (v/v) by using egg yolk and soybean-based extenders. Diluted semen placed into 0.25 ml straws were equilibrated at 4 °C for 15 min and frozen in liquid nitrogen vapor. Fertilization was conducted using a ratio of 1 × 10(5)spermatozoa/egg. Supplementation of 10% lecithin to extender showed the best cryoprotective effect for sperm motility and duration of motility against freezing damage compared to 15%, 20% and control II groups (psperm with extender containing 10% lecithin provided a greater result in terms of fertilization success when compared to extenders containing 20% lecithin or control II (pegg yolk-based extender against freezing damages and fertilization. Therefore, soybean lecithin is a suitable alternative to avian egg yolk for the cryopreservation of fish sperm.

  5. Sperm evaluation and biochemical characterization of cat seminal plasma collected by electroejaculation and urethral catheterization.

    Science.gov (United States)

    Zambelli, Daniele; Raccagni, Ramona; Cunto, Marco; Andreani, Giulia; Isani, Gloria

    2010-11-01

    This paper aimed to evaluate cat seminal plasma protein profile (with SDS-page) and determine differences in seminal plasma composition from ejaculates obtained using urethral catheterization after pharmacological induction (UrCaPI) and electroejaculation (EE). In addition, this study evaluates whether the recovery method affected seminal plasma protein and zinc concentrations. A single ejaculation was collected from 17 mixed-breed cats by EE (5/21) or UrCaPI (12/21), while 4/21 cats underwent four sperm collections once every four days using EE and UrCaPI techniques alternately. The semen parameters evaluated were: volume, percentage of motility and progressive motility, morphology, and sperm concentration. After centrifugation, the seminal plasma obtained was stored at -80 °C and later used to measure protein and zinc concentrations, and to determine protein profile by SDS-polyacrylamide gel electrophoresis (PAGE). The results obtained indicate that cat seminal plasma protein profile is characterized by many protein bands (>30) with a molecular weight ranging from 3.5 to 200 kDa, and that the recovery method influences the seminal plasma protein profile: EE is related to the absence of two proteins (P55 and P14), and alters three protein bands (P200, P80, P28). The collection technique also affected zinc concentration (mg/dL) and protein concentration (g/dL) which were significantly higher (P < 0.01) in samples collected by UrCaPI; on the contrary the total Zn and protein amount/ejaculate were not significantly different in samples collected by both technique (P < 0.05).

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

    OpenAIRE

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

    2015-01-01

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

  7. AMP-activated kinase in human spermatozoa: identification, intracellular localization, and key function in the regulation of sperm motility.

    Science.gov (United States)

    Calle-Guisado, Violeta; de Llera, Ana Hurtado; Martin-Hidalgo, David; Mijares, Jose; Gil, Maria C; Alvarez, Ignacio S; Bragado, Maria J; Garcia-Marin, Luis J

    2016-09-27

    AMP-activated kinase (AMPK), a protein that regulates energy balance and metabolism, has recently been identified in boar spermatozoa where regulates key functional sperm processes essential for fertilization. This work's aims are AMPK identification, intracellular localization, and their role in human spermatozoa function. Semen was obtained from healthy human donors. Sperm AMPK and phospho-Thr172-AMPK were analyzed by Western blotting and indirect immunofluorescence. High- and low-quality sperm populations were separated by a 40%-80% density gradient. Human spermatozoa motility was evaluated by an Integrated Semen Analysis System (ISAS) in the presence or absence of the AMPK inhibitor compound C (CC). AMPK is localized along the human spermatozoa, at the entire acrosome, midpiece and tail with variable intensity, whereas its active form, phospho-Thr172-AMPK, shows a prominent staining at the acrosome and sperm tail with a weaker staining in the midpiece and the postacrosomal region. Interestingly, spermatozoa bearing an excess residual cytoplasm show strong AMPK staining in this subcellular compartment. Both AMPK and phospho-Thr172-AMPK human spermatozoa contents exhibit important individual variations. Moreover, active AMPK is predominant in the high motility sperm population, where shows a stronger intensity compared with the low motility sperm population. Inhibition of AMPK activity in human spermatozoa by CC treatment leads to a significant reduction in any sperm motility parameter analyzed: percent of motile sperm, sperm velocities, progressivity, and other motility coefficients. This work identifies and points out AMPK as a new molecular mechanism involved in human spermatozoa motility. Further AMPK implications in the clinical efficiency of assisted reproduction and in other reproductive areas need to be studied.

  8. Role of human- and animal-sperm studies in the evaluation of male reproductive hazards

    Energy Technology Data Exchange (ETDEWEB)

    Wyrobek, A.J.; Gordon, L.; Watchmaker, G.

    1982-04-07

    Human sperm tests provide a direct means of assessing chemically induced spermatogenic dysfunction in man. Available tests include sperm count, motility, morphology (seminal cytology), and Y-body analyses. Over 70 different human exposures have been monitored in various groups of exposed men. The majority of exposures studied showed a significant change from control in one or more sperm tests. When carefully controlled, the sperm morphology test is statistically the most sensitive of these human sperm tests. Several sperm tests have been developed in nonhuman mammals for the study of chemical spermatotoxins. The sperm morphology test in mice has been the most widely used. Results with this test seem to be related to germ-cell mutagenicity. In general, animal sperm tests should play an important role in the identification and assessment of potential human reproductive hazards. Exposure to spermatotoxins may lead to infertility, and more importantly, to heritable genetic damage. While there are considerable animal and human data suggesting that sperm tests may be used to detect agents causing infertility, the extent to which these tests detect heritable genetic damage remains unclear. (ERB)

  9. Quality and fertilizing capacity of boar spermatozoa during liquid storage in extender supplemented with different antibiotics.

    Science.gov (United States)

    Bryła, Magdalena; Trzcińska, Monika

    2015-12-01

    The aim of the study was to determine the effect of antibiotics on quality parameters and fertilizing capacity of boar sperm during liquid preservation. In the first experiment, semen was diluted in an extender containing 200 μg/mL of gentamicin as a control and diluted in a modified extenders: Ext I (contained 200 μg/mL florfenicol), Ext II (contained 200 μg/mL polymyxin B), Ext III (contained 100 μg/mL gentamicin and 100 μg/mL florfenicol) and Ext IV (contained 100 μg/mL gentamicin and 100 μg/mL polymyxin B). The semen was stored for ten days. Sperm quality was evaluated based on the motility (CASA; TM: total motility; PM: progressive motility), membrane integrity (YO-PRO-1/PI assay), mitochondrial activity (JC-1) and DNA integrity (TUNEL). The highest PM% (62.5 ± 9.6) was observed in Ext III at Day 6 of storage. The highest sperm viability and mitochondrial transmembrane potential was noticed at the end of the storage period in Ext III. Long-term storage did not induce DNA fragmentation in the extenders analyzed. In the second experiment, semen diluted in the control extender and in the extender providing the highest quality spermatozoa on Day 10 (Ext III) was used for artificial insemination (AI) of synchronized gilts. Our studies showed that the highest reproductive performance of inseminated gilts (pregnant gilts: 97.0%, litter size: 11.4 ± 1.2) occurred with Ext III semen dilution. The combination of 100 μg/mL gentamicin and 100 μg/mL florfenicol in the extender maintained sperm motility, membrane integrity and mitochondrial activity and enhanced the higher reproduction success.

  10. Evaluation of Sperm Quality, Maturation and DNA Integrity in Adult Mice Treated with Sulpiride

    Directory of Open Access Journals (Sweden)

    S Salami

    2012-07-01

    Full Text Available Background: Use of certain antipsychotic drugs has severe effects on fertility in males. Hypothalamus and hypophysial impressions and changes in plasma hormones concentration like prolactin, LH and FSH can affect sperm production. In this study, we investigated the effects of sulpiride on sperm quality, maturation and DNA damage. Methods: Twenty for adult male mice (age: 6-8 weeks were divided into three groups. The treatment group received 40 mg/kg sulpiride solution and the control sham group was given carrier of the drug intraperitoneally (IP daily for 45 days but the control group received nothing. Finally, all the mice were sacrificed by cervical dislocation and their cauda epididymis were removed surgically. The excised specimens were placed in 1 ml HTF medium and incubated for 30 min in CO2 incubator to allow the spermatozoa to swim out. Later, sperm count, motility and viability were analyzed. Additionally, sperm chromatin quality and DNA integrity were assessed by aniline blue and acridine orange staining. Results: Significant decrease in sperm motility and count were observed in the treatment group while the number of abnormal sperm increased as compared with the other two groups. Sperm viability and DNA maturation showed significant reduction and the rate of DNA damage increased in comparison with the control sham and the control groups (P<0.05. Conclusion: The study showed that sulpiride has negative effects on sperm parameters in treated animals and in some cases it could cause secondary infertility.

  11. Comparison of four methods to evaluate sperm DNA integrity between mouse caput and cauda epididymidis

    Institute of Scientific and Technical Information of China (English)

    Serafín Pérez-Cerezales; Alberto Miranda; Alfonso Gutiérrez-Adán

    2012-01-01

    It is well known that transit through the epididymis involves an increase in the compaction of sperm chromatin,which acquires fully condensed status at the caput epididymidis.The purpose of this study was to compare the terminal deoxyribonucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) assay,the comet assay,the sperm chromatin structure assay (SCSA) and the sperm chromatin dispersion (SCD) test by analysing spermatozoa from the caput and cauda epididymidis in order to demonstrate the ability of each technique to discriminate between different degrees of sperm maturity related to chromatin compaction and DNA fragmentation.Our results suggest that some populations of DNA-fragmented spermatozoa associated with immature sperm can only be identified using the comet assay and the SCSA but not with the SCD test or the TUNEL assay.

  12. 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.

  13. 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.

  14. Sperm quality evaluation in Solea senegalensis during the reproductive season at cellular level.

    Science.gov (United States)

    Beirão, J; Soares, F; Herráez, M P; Dinis, M T; Cabrita, E

    2009-12-01

    Sperm quality seems to be one of the reasons for the reproduction constraints faced by Senegalese sole (Solea senegalensis) aquaculturists. Previous studies in this species indicated that the sperm quality of individuals kept in culture varies throughout the year and that different sperm subpopulations can be identified in ejaculates according to the motility pattern of spermatozoa. Aiming to better understand factors affecting sole sperm quality in captivity, sperm of 11 males was assessed during the reproductive season using different parameters: motility characteristics using CASA analysis; cell plasma membrane resistance to seawater hyperosmolarity; DNA fragmentation with single-cell gel electrophoresis; and early apoptosis, labeled with Annexin-V FITC. Computer-assisted sperm analyses motility data were treated using multivariate analysis to identify the presence of different spermatozoa subpopulations according to their motility pattern. Four distinct sperm subpopulations were obtained: Subpop1, which includes fast linear spermatozoa; Subpop2, made up of fast nonlinear spermatozoa; Subpop3, which includes slow linear spermatozoa; and Subpop4, which contains slow nonlinear spermatozoa. The sperm subpopulation structure varied with time after activation and with male. Low cell resistance to the seawater hyperosmotic conditions was noticed. The Annexin-V assay allowed the identification of an apoptotic population ranging from 6% to 20%. A high percentage of cells (64.1%) showed a DNA fragmentation level below 30%, but these values varied significantly between males. DNA fragmentation appears to be related to cell membrane resistance to hyperosmotic conditions faced by the cells when in contact with seawater. This condition seems to modulate the composition of the motile sperm population and performance after activation. This phenomenon could be related to the spermatozoa maturation process.

  15. Glutathione Peroxidase 5 Is Expressed by the Entire Pig Male Genital Tract and Once in the Seminal Plasma Contributes to Sperm Survival and In Vivo Fertility

    Science.gov (United States)

    Barranco, Isabel; Tvarijonaviciute, Asta; Perez-Patiño, Cristina; Vicente-Carrillo, Alejandro; Parrilla, Inmaculada; Ceron, Jose J.; Martinez, Emilio A.; Rodriguez-Martinez, Heriberto; Roca, Jordi

    2016-01-01

    Glutathione peroxidase-5 (GPX5) is an H2O2-scavenging enzyme identified in boar seminal plasma (SP). This study attempted to clarify its origin and role on sperm survival and fertility after artificial insemination (AI). GPX5 was expressed (Western blot and immunocytochemistry using a rabbit primary polyclonal antibody) in testes, epididymis and accessory sex glands (6 boars). SP-GPX5 concentration differed among boars (11 boars, P boar (44 ejaculates, P sperm rich fraction (SRF, sperm-peak portion) had a significantly lower concentration (8.87 ± 0.78 ng/mL) than the rest of the SRF and the post-SRF (11.66 ± 0.79 and 12.37 ± 0.79 ng/mL, respectively, P Sperm motility of liquid-stored semen AI-doses (n = 44, at 15–17°C during 72h) declined faster in AI-doses with low concentrations of SP-GPX5 compared to those with high-levels. Boars (n = 11) with high SP-GPX5 showed higher farrowing rates and litter sizes than those with low SP-GPX5 (a total of 5,275 inseminated sows). In sum, GPX5 is widely expressed in the boar genital tract and its variable presence in SP shows a positive relationship with sperm quality and fertility outcomes of liquid-stored semen AI-doses. PMID:27627110

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

    Directory of Open Access Journals (Sweden)

    Roshanak Aboutorabi

    2014-01-01

    Conclusion: Our results showed that the correlation between CD52 labeling and sperm motility was negatively significant, but we did not observe any relation with other semen parameters, such as sperm normal morphology, sperm concentration, and semen viscosity.

  17. Development of a novel flow cytometric approach to evaluate fish sperm chromatin using fixed samples

    Science.gov (United States)

    Jenkins, Jill A.

    2013-01-01

    The integrity of the paternal DNA is essential for the accurate transmission of genetic information, yet fertilization is not inhibited by chromatin breakage. Some methods are available for the sensitive detection of DNA damage and can be applied in studies of environmental toxicology, carcinogenesis, aging, and assisted reproduction techniques in both clinical and experimental settings. Because semen samples obtained from remote locations undergo chromatin damage prior to laboratory assessment, the present study was undertaken to evaluate treatments for effective chromatin staining in the development of a DNA fragmentation assay using fixed milt from yellow perch (Perca flavescens). Similar to the sperm chromatin structure assay (SCSA), susceptibility of nuclear DNA to acid-induced denaturation was measured by flow cytometry (FCM). Use of 10% buffered formalin for milt fixation allowed easier peak discrimination than 4% paraformaldehyde. The effects of time and temperature of incubation in 0.08 N HCl were evaluated in order to determine the ideal conditions for promoting DNA decondensation and making strand breaks more available for staining and detection by FCM. The best results were obtained with incubation at 37°C for 1 minute, followed by cold propidium iodide staining for 30 minutes.

  18. Production of transgenic pigs mediated by pseudotyped lentivirus and sperm.

    Directory of Open Access Journals (Sweden)

    Yongliang Zhang

    Full Text Available Sperm-mediated gene transfer can be a very efficient method to produce transgenic pigs, however, the results from different laboratories had not been widely repeated. Genomic integration of transgene by injection of pseudotyped lentivirus to the perivitelline space has been proved to be a reliable route to generate transgenic animals. To test whether transgene in the lentivirus can be delivered by sperm, we studied incubation of pseudotyped lentiviruses and sperm before insemination. After incubation with pig spermatozoa, 62±3 lentiviral particles were detected per 100 sperm cells using quantitative real-time RT-PCR. The association of lentivirus with sperm was further confirmed by electron microscopy. The sperm incubated with lentiviral particles were artificially inseminated into pigs. Of the 59 piglets born from inseminated 5 sows, 6 piglets (10.17% carried the transgene based on the PCR identification. Foreign gene and EGFP was successfully detected in ear tissue biopsies from two PCR-positive pigs, revealed via in situ hybridization and immunohistochemistry. Offspring of one PCR-positive boar with normal sows showed PCR-positive. Two PCR-positive founders and offsprings of PCR-positive boar were further identified by Southern-blot analysis, out of which the two founders and two offsprings were positive in Southern blotting, strongly indicating integration of foreign gene into genome. The results indicate that incubation of sperm with pseudotyped lentiviruses can incorporated with sperm-mediated gene transfer to produce transgenic pigs with improved efficiency.

  19. 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.

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

    Science.gov (United States)

    Yi, Young-Joo; Lee, In-Kyoung; Lee, Sang-Myeong; Yun, Bong-Sik

    2016-03-01

    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.

  1. 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...

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

    Science.gov (United States)

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

    2011-12-01

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

  3. Evaluation on Sensitivity of the Human Sperm Motility Assay for Detecting Endotoxin in Culture Medium

    Institute of Scientific and Technical Information of China (English)

    Wei-jie ZHU; Jing LI; Wen-hong ZHANG; Kang-shou YAO

    2003-01-01

    Objective To investigate the sensitivity of the human sperm motility assay for detecting endotoxin in culture medium Materials & Methods Motile sperm were separated and exposed to different concentrations of endotoxin (0.5 ng/mL, 1 ng/mL, 10 ng/mL, 1 000 ng/mL, 10 000 ng/mL, and 50 000 ng/mL), and sperm motility was determined after incubation. Effects of endotoxin on sperm motility in media without albumin were also examined. In addition, at the same concentrations of endotoxin (0.5 ng/mL, 1 ng/mL, and 10 ng/mL), the sensitivity of the human sperm motility assay was compared to those of 1-cell and 2-cell mouse embryo bioassays.Results At levels of 0.5 ng/mL~1 000 ng/mL endotoxin in media with 2 mg/mL albumin, sperm did not show significant change in motility during 24 h of incubation when compared with the control (P>0.05). However, the sperm motility was significantly inhibited at endotoxin dosages of 10 000 and 50 000 ng/mL. In the absence of albumin supplementation, at endotoxin levels of 50 000 ng/mL, and 1 000 ng/mL, there was a marked decrease in sperm motility compared with the control after 2 h or 8 h of incubation, respectively (P<0.01). In media containing 0.5 ng/mL and 1 ng/mL endotoxin, 1-cell and 2-cell mouse embryos had significantly reduced developmental rates in all developmental stages, and at the level of 10 ng/mL, the development of the embryos was arrested.Conclusion The human sperm motility assay could detect high levels of endotoxin in culture medium but its sensitivity to endotoxin would be inferior to that of the 1-cell or 2-cell mouse embryo bioassay. In the absence of albumin supplementation, the sensitivity of the sperm motility assay could be improved.

  4. X/XY/XYY mosaicism as a cause of subfertility in boars: a single case study.

    Science.gov (United States)

    Quilter, C R; Wood, D; Southwood, O I; Griffin, D K

    2003-02-01

    Sex chromosome abnormalities are common in mammals and humans and are often associated with subfertility. In this study a boar with normal sperm parameters was indicated to have reduced prolificacy from figures obtained for return rate, farrowing rate and total number of piglets born. G-banded cytogenetic analysis of peripheral blood identified an abnormal mosaic sex chromosome constitution 39,XYY[74]/38,XY[23]/37,X[3]. Cytogenetic analysis of fibroblasts confirmed this mosaic karyotype with similar percentages of cell lines observed 39,XYY[76]/38,XY[19]/37,X[5]. External genitalia revealed a poorly developed scrotum with the right testicle being smaller than the left. To the best of our knowledge this is the first time that this chromosome constitution has been reported in the pig. It is of particular interest that this karyotype is associated with reduced boar fertility, which could lead to potential economic losses if such a boar were selected for breeding purposes.

  5. Assessing the Risk of African Swine Fever Introduction into the European Union by Wild Boar.

    Science.gov (United States)

    De la Torre, A; Bosch, J; Iglesias, I; Muñoz, M J; Mur, L; Martínez-López, B; Martínez, M; Sánchez-Vizcaíno, J M

    2015-06-01

    The presence of African swine fever (ASF) in the Caucasus region and Russian Federation has increased concerns that wild boars may introduce the ASF virus into the European Union (EU). This study describes a semi-quantitative approach for evaluating the risk of ASF introduction into the EU by wild boar movements based on the following risk estimators: the susceptible population of (1) wild boars and (2) domestic pigs in the country of origin; the outbreak density in (3) wild boars and (4) domestic pigs in the countries of origin, the (5) suitable habitat for wild boars along the EU border; and the distance between the EU border and the nearest ASF outbreak in (6) wild boars or (7) domestic pigs. Sensitivity analysis was performed to identify the most influential risk estimators. The highest risk was found to be concentrated in Finland, Romania, Latvia and Poland, and wild boar habitat and outbreak density were the two most important risk estimators. Animal health authorities in at-risk countries should be aware of these risk estimators and should communicate closely with wild boar hunters and pig farmers to rapidly detect and control ASF.

  6. A pre-breeding screening program for transgenic boars based on fluorescence in situ hybridization assay.

    Science.gov (United States)

    Bou, Gerelchimeg; Sun, Mingju; Lv, Ming; Zhu, Jiang; Li, Hui; Wang, Juan; Li, Lu; Liu, Zhongfeng; Zheng, Zhong; He, Wenteng; Kong, Qingran; Liu, Zhonghua

    2014-08-01

    For efficient transgenic herd expansion, only the transgenic animals that possess the ability to transmit transgene into next generation are considered for breeding. However, for transgenic pig, practically lacking a pre-breeding screening program, time, labor and money is always wasted to maintain non-transgenic pigs, low or null transgenic transmission pigs and the related fruitless gestations. Developing a pre-breeding screening program would make the transgenic herd expansion more economical and efficient. In this technical report, we proposed a three-step pre-breeding screening program for transgenic boars simply through combining the fluorescence in situ hybridization (FISH) assay with the common pre-breeding screening workflow. In the first step of screening, combined with general transgenic phenotype analysis, FISH is used to identify transgenic boars. In the second step of screening, combined with conventional semen test, FISH is used to detect transgenic sperm, thus to identify the individuals producing high quality semen and transgenic sperm. In the third step of screening, FISH is used to assess the in vitro fertilization embryos, thus finally to identify the individuals with the ability to produce transgenic embryos. By this three-step screening, the non-transgenic boars and boars with no ability to produce transgenic sperm or transgenic embryos would be eliminated; therefore only those boars could produce transgenic offspring are maintained and used for breeding and herd expansion. It is the first time a systematic pre-breeding screening program is proposed for transgenic pigs. This program might also be applied in other transgenic large animals, and provide an economical and efficient strategy for herd expansion.

  7. 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.

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

    Science.gov (United States)

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

    2012-04-01

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

  9. 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; Lyoo, Young S

    2016-06-01

    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.

  10. Sperm bioassay for rapid detection of cereulide-producing Bacillus cereus in food and related environments.

    Science.gov (United States)

    Andersson, Maria A; Jääskeläinen, Elina L; Shaheen, Ranad; Pirhonen, Tuula; Wijnands, Luc M; Salkinoja-Salonen, Mirja S

    2004-07-15

    A novel in vitro method, sperm micro assay for rapidly distinguishing cereulide, the emetic toxin producing Bacillus cereus from non-producers is described and its use for quantitating cereulide and screening large numbers of B. cereus strains/colonies evaluated. The assay is non-laborious and can be executed with equipment present in most laboratories. Boar spermatozoa, purchased as standard semen from artificial insemination suppliers, are used to detect toxicity. Boar sperms respond within 5 min by cessation of motility when exposed at 37 degrees C to heat-treated (100 degrees C) extract prepared from a cereulide containing B. cereus. The assay can be done on individual colonies on the primary plate, with no need for pure culture and the qualitative result is obtained within 30 min. The assay is robust, not sensitive to age or storage of the culture plates. The use of the sperm micro assay for semiquantitative estimation of cereulide in B. cereus was validated with 14 different B. cereus strains using as reference the specific chemical assay for cereulide, based on liquid chromatography-ion trap mass spectrometry (LC-ion trap MS). The cereulide contents calculated from endpoint dilutions of the sperm micro assay matched the result of the chemical analysis closely. The detection threshold of the sperm micro assay was measured as 0.3 +/- 0.1 ng of cereulide per 5.4 x 10(6) sperm cells in 0.2 ml or 0.9 ng of cereulide per mg of B. cereus biomass (wet wt.). Food-related B. cereus strains contained 4-400 ng of cereulide per mg (wet wt.). When a large number of B. cereus of food, non-food, clinical and environmental origins were screened and 107 independent strains/isolates were identified as cereulide producers, it was observed that all of these had low or no haemolytic activity when cultivated on bovine blood agar. None of the strains/isolates with wide, clear zones of haemolysis, considered typical of B. cereus, produced cereulide.

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

    Science.gov (United States)

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

    2012-02-01

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

  12. 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.

  13. The effect of the MC4R gene on boar taint compounds, sexual maturity and behaviour in growing-finishing boars and gilts.

    Science.gov (United States)

    Van den Broeke, A; Aluwé, M; Janssens, S; Wauters, J; Vanhaecke, L; Buys, N; Millet, S; Tuyttens, F A M

    2015-10-01

    Societal pressure to ban surgical castration of male piglets is rising due to animal welfare concerns, thus other methods to prevent boar taint need to be explored. Genetic selection against boar taint appears to be a long-term sustainable alternative. However, as boar taint is linked to reproductive hormones, it is important to consider possible negative side effects such as delayed sexual maturity or changes in behaviour. We reported earlier that the melanocortin-4 receptor (MC4R) marker can be used to reduce boar taint levels in fat of boars. The objective of this study was to evaluate whether MC4R marker-assisted selection for lower boar taint prevalence affects plasma levels of boar taint compounds and testosterone; sexual maturity; behaviour; skin lesions; and lameness in boars and gilts. Using an intervention study with a 2×2 design, 264 boars and gilts differing on position 893 of the MC4R gene (AA v. GG) were compared. The MC4R polymorphism did not affect the plasma concentration of either androstenone or testosterone at different time points, whereas the concentration of skatole was significantly lower (P=0.003) and the concentration of indole tended to be lower (P=0.074) in GG compared with AA boars. A higher percentage of gilts of the GG genotype were in puberty at slaughter age compared with AA gilts (Pbehaviour (P=0.015) and less passive and feeding behaviour (P=0.003). They showed more skin lesions on their back and caudal area (P=0.022), and tended to show more skin lesions on their head and anterior area (P=0.093) compared with AA animals. In conclusion, the polymorphism in the MC4R gene can be used as a marker without negative effects on reproduction characteristics in boars and gilts. Genetic selection towards a lower prevalence of boar taint will lead to more active pigs with more skin lesions. Management strategies may therefore be necessary to reduce skin lesions in the selected animals.

  14. Comportamento de monta e características seminais de suínos jovens landrace e large white Mating behavior and seminal characteristics in young landrace and large white boars

    Directory of Open Access Journals (Sweden)

    Fabiane Mendonça Ferreira

    2005-02-01

    Full Text Available O objetivo deste trabalho foi avaliar o comportamento de monta e as características seminais de machos Landrace (LD e Large White (LW dos 143 aos 232 dias de idade. A partir dos 143 dias de idade, 62 machos suínos foram avaliados quanto ao comportamento de monta. A partir dos 150 dias de idade, o sêmen foi coletado de 33 desses machos durante onze semanas para avaliar a produção e qualidade espermáticas. Não houve diferenças entre as raças em relação ao comportamento de monta (p=0,105. Durante o período de coletas de sêmen, houve aumento do volume de sêmen, da concentração espermática, do número de espermatozóides (SPTZ ejaculados e da liberação diária de SPTZ (LDE. O percentual de células espermáticas anormais diminuiu durante o período e não houve diferença entre as raças (P=0,523. Os machos LD ejacularam maior volume de sêmen (P=0,016, de espermatozóides totais por ejaculado (P=0,070 e apresentaram maior LDE (P=0,042. Esses dados indicam que machos LD apresentam maior produção espermática e maior potencial para produzir doses de sêmen no período pós-púbere.The aim of this work was to evaluate mating behavior and seminal characteristics of Landrace (LD and Large White (LW boars from 143 to 232 days of age. With 143 days of age, LD (n=28 and LW (n=34 boars were evaluated in relation to mating behavior. Being 150 days of age, semen was collected from 33 of these males during eleven weeks to evaluate sperm production and quality. No difference was observed between breeds in relation to mating behavior. During the period of semen collection, there was an increase in semen volume, sperm concentration and motility, total sperm in ejaculate, and in daily sperm output (DSO. The percent of abnormal sperm cells decreased during the period and there was no difference between breeds (P=0.523. The LD boars ejaculated higher semen volume (P=0,016, total sperm in ejaculate (P=0.070, and showed higher DSO (P=0.042. These

  15. Adaptation of ubiquitin-PNA based sperm quality assay for semen evaluation by a conventional flow cytometer and a dedicated platform for flow cytometric semen analysis.

    Science.gov (United States)

    Odhiambo, J F; Sutovsky, M; DeJarnette, J M; Marshall, C; Sutovsky, P

    2011-10-01

    The purpose of semen quality evaluation is to predict the fertility potential of the sample in an objective, rapid and inexpensive manner. However, utilization of sperm quality biomarkers such as ubiquitin and lectin Arachis hypogaea agglutinin (PNA) for flow cytometric semen evaluation might eliminate the need for visual assessment by microscopy. Herein, we demonstrate a robust ubiquitin and PNA-based semen evaluation conducted on a simple, easy to operate, dedicated sperm flow cytometer, EasyCyte Plus (IMV Technologies, L'Aigle, France). Semen samples were collected periodically from two dairy bulls, which were subjected to temporary scrotal insults to induce variable semen quality. Samples were labeled with fluorescently-conjugated anti-ubiquitin antibodies (bind exclusively to the surface of defective sperm) and lectin PNA (binds to acrosomal surface in prematurely capacitated and acrosome-damaged sperm). Fluorescent properties of the samples were measured with a conventional flow cytometer (Becton Dickinson FACScan; Becton Dickinson Corp., Franklin Lakes, NJ, USA) and by the EasyCyte (IMV Technologies) instrument. Data from the two flow cytometers were positively correlated for the percentage of PNA-positive sperm with a damaged acrosome (r = 0.47; P flow cytometric semen evaluation.

  16. Current status and potential of morphometric sperm analysis

    OpenAIRE

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

    2016-01-01

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

  17. 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...

  18. Cysteine addition on short-term cooled boar semen preservation and its relationship with swine field fertility

    Directory of Open Access Journals (Sweden)

    Carolina K Severo

    2011-12-01

    Full Text Available Artificial insemination is routinely used in the swine industry to reduce the costs of production through to increase the efficiency of the refrigerated boar semen process. The objective of this study was to evaluate the effect of different levels of cysteine (CYS added to the Beltsville Thawing Solution (BTS extender semen during cooling for up to 72 hours. Ejaculated from three boars were collected with the gloved-hand technique and semen aliquots were diluted in BTS as follow: BTS only (BTS, BTS + 0.1mM cysteine (CYS0.1, BTS + 0.5mM cysteine (CYS0.5, BTS + 1.0mM cysteine (CYS1.0, BTS + 2.5mM cysteine (CYS2.5, BTS + 5.0mM cysteine (CYS5.0, BTS + 10.0mM cysteine (CYS10.0, and BTS + 20.0mM cysteine (CYS20.0. Evaluation of sperm integrity were analyzed using 0.5mg/ml propidium iodide (plasma membrane, 100µg/ml isothiocynate-conjugated Pisum sativun agglutinin (acrosomal membrane and 153µM 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide (mitochondria potential after semen dilution at specific times (0, 24, 48 and 72 hours. Additionally, we also evaluated the effects of 5.0 mM CYS addition in the BTS extender on the maintenance of sperm quality and their influence on fertility in the swine production. After artificial insemination, animals were evaluated based on the estrous return and the number of piglet's born. Cysteine at concentrations of 10.0 and 20.0mM resulted in more pronounced reductions even at the time zero. Semen viability decreased to levels below 10% at these high levels of CYS in the first 24 hour of storage at 17ºC. At the end of the storage time, less than 65% of sperm cells had intact plasma membrane in all groups. The sperm viability decreased significantly when the semen was added at high concentrations of CYS (time "0"; CYS10.0 and CYS20.0; p<0.05, when compared to the other CYS concentrations. The BTS (10.20±0.39 treated group showed a lower rate of estrus return when compared to other (BTSCYS

  19. Comparison of Semen Quality and Spermadhesins’ mRNA Expression between Luchuan and Exotic Boars%陆川猪与外来猪精液质量及精子黏附蛋白基因家族 mRNA表达的比较

    Institute of Scientific and Technical Information of China (English)

    黄明光; 黄光云; 胡安琪; 李珣; 潘天彪; 汪燕玲; 杨尚雪; 胡传活

    2016-01-01

    Spermadhesins, as the most important components of boar semen seminoproteins, play a regulatory role in sperm motility, sperm capacitation and sperm-ovum fusion. In order to study the fertility differences among different boar breeds, the sperm motility, sperm viability and sperm deformity, as wel as the spermadhesins’ mRNA expression in fresh semen of Luchuan, Landrace and Duroc boars were compared. The results showed that the sperm motility and viability of Luchuan boar showed no significant differences with those of Landrace and Duroc boars (P>0.05), but its sperm deformity was significantly lower than those of the other two exotic breeds (P0.05), excepting that the mRNA expression level of AQN1 in Luchuan boar se-men was significantly lower than that in Duroc boar semen (P0.05),但是精子畸形率极显著低于2种外来猪种(P0.05);长白猪精液中精子黏附蛋白基因家族的 AQN3、AWN和PSP-II基因 mRNA表达水平显著(P<0.05)或极显著(P<0.01)高于杜洛克猪。总之,不同猪种精子黏附蛋白基因家族 mRNA表达水平存在明显差异,但与精子活力、活率及畸形率等精液质量指标的高低没有规律性的关联。

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

    Institute of Scientific and Technical Information of China (English)

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

    2006-01-01

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

  1. 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.

  2. 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.

  3. Plenary contribution to International Conference on Boar Semen Preservation 2011. Genetic selection for freezability and its controversy with selection for performance

    Science.gov (United States)

    Little data are available in the literature regarding freezability of boar sperm or its relationship with other traits. Existing data suggest the trait would respond favorably to selection, and information is available from other species suggesting components which might have changed. Genetic parame...

  4. Plenary contribution to International Conference on Boar Semen Preservation 2011: Genetic selection for freezability and its controversy with selection for performance

    Science.gov (United States)

    Little data are available in the literature regarding freezability of boar sperm or its relationship with other traits. Existing data suggest the trait would respond favorably to selection, and information is available from other species suggesting components which might have changed. Genetic parame...

  5. Mecobalamin promotes mouse sperm maturation.

    Science.gov (United States)

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

    1989-01-01

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

  6. 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

  7. FATAL CASE OF STREPTOCOCCUS SUIS INFECTION IN A YOUNG WILD BOAR (SUS SCROFA) FROM SOUTHWESTERN SPAIN.

    Science.gov (United States)

    Risco, David; Fernández-Llario, Pedro; Cuesta, Jesús M; García-Jiménez, Waldo L; Gonçalves, Pilar; Martínez, Remigio; García, Alfredo; Rosales, Rubén; Gómez, Luis; de Mendoza, Javier Hermoso

    2015-06-01

    Streptococcus suis is a recognized pathogen that may cause important diseases in pigs and humans. This microorganism has been repeatedly isolated from wild boar (Sus scrofa). However, its health implications for this wild species are still unknown. This article reports a detailed description of a fatal case of septicemia by S. suis affecting a young wild boar. The affected animal, about 15 days old, was found near death and exhibiting neurologic signs at a wild boar estate in southwestern Spain. Postmortem examination showed generalized congestion, brain hemorrhages and lobular pneumonia. Histopathological evaluation demonstrated the presence of meningitis and encephalitis with marked congestion and suppurative bronchopneumonia. Streptococcus suis serotype 2 isolates exhibiting important virulence factors (extracellular factor, muramidase-released protein, and suylisin) were isolated from the affected animal. This study confirms the presence of potentially virulent and zoonotic strains of S. suis in wild boar from Spain.

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

    Science.gov (United States)

    Evans, Jonathan P.

    2009-07-01

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

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

    Science.gov (United States)

    Schulze, Martin; Junkes, Christof; Mueller, Peter; Speck, Stephanie; Ruediger, Karin; Dathe, Margitta; Mueller, Karin

    2014-01-01

    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.

  10. 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.

  11. Helminth Parasites of Wild Boars, Sus scrofa, in Bushehr Prov-ince, Southwestern Iran

    Directory of Open Access Journals (Sweden)

    Majid MANSOURI

    2016-10-01

    Full Text Available Background: Wild boars, Sus scrofa, of wide distribution considered as a potential source of zoonotic parasites. The current study aimed to assess the prevalence of helminth infections in wild boars in the Persian Gulf coastal area (Bushehr Province, Southwestern Iran.Methods: Twenty-five wild boars, including 11 males and 14 females, were collected during a course of vertebrate pest control in the Bushehr Province, southwestern Iran in 2013. The specimen were immediately dissected and carefully searched for the parasites. During necropsy, each organ was examined macroscopically for presence of any helminthic agents. Tissue samples were taken from each organ. Moreover, samples were taken from the content of digestive system. Blood samples were also collected from each boar. All the samples were evaluated for helminth infections by parasitological methods.Results: Twenty-two (88% of the wild boars were infected with at least one helminth. Out of 25 wild boars, 1 (4% were infected with Cysticercus tenuicollis, the larval stage of Taenia hydatigena, 13 (52% with Macracanthorhynchus hirudinaceus, 17 (68% with Metastrongylus spp, and 20 (80% with Ascarops spp. Hydatid cyst was detected in the lung of one of the wild boars. No Trichinella spp. larvae were detected in any of the tissues of the animals when evaluated by artificial digestion method. In addition, no contamination with microfilaria was detected in any of animals when the blood samples were tested with Knott’s method.Conclusion: Wild boars are contaminated by some helminthes including zoonotic ones. These animals could be involved in the epidemiology of zoonotic helminth by acting as reservoir hosts. This in turn may bring potential risk for locals and residents of the Bushehr Province, Southwestern Iran.

  12. DNA fragmentation dynamics allows the assessment of cryptic sperm damage in human: Evaluation of exposure to ionizing radiation, hyperthermia, acidic pH and nitric oxide

    Energy Technology Data Exchange (ETDEWEB)

    Santiso, Rebeca; Tamayo, Maria [Laboratorio de Genetica Molecular y Radiobiologia, Centro Oncologico de Galicia, Doctor Camilo Veiras 1, 15009-A Coruna (Spain); Genetics Unit, INIBIC-Complejo Hospitalario Universitario A Coruna (CHUAC), As Xubias, 84, 15006-A Coruna (Spain); Gosalvez, Jaime [Genetics Unit, Facultad de Biologia, Universidad Autonoma de Madrid, Ciudad Universitaria de Cantoblanco, 28049 Madrid (Spain); Johnston, Steve [School of Agriculture and Food Science, University of Queensland, Gatton 4343 (Australia); Marino, Alfonso [Servicio de Oncologia Radioterapica, Centro Oncologico de Galicia, Doctor Camilo Veiras 1, 15009-A Coruna (Spain); Fernandez, Carlos; Losada, Carlos [Servicio de Radiofisica, Centro Oncologico de Galicia, Doctor Camilo Veiras 1, 15009-A Coruna (Spain); Fernandez, Jose Luis, E-mail: Jose.Luis.Fernandez.Garcia@sergas.es [Laboratorio de Genetica Molecular y Radiobiologia, Centro Oncologico de Galicia, Doctor Camilo Veiras 1, 15009-A Coruna (Spain); Genetics Unit, INIBIC-Complejo Hospitalario Universitario A Coruna (CHUAC), As Xubias, 84, 15006-A Coruna (Spain)

    2012-06-01

    Sperm DNA fragmentation (SDF) is not a static seminal parameter, since the longevity of sperm DNA decreases progressively with time following ejaculation or thawing. While the dynamics of SDF is a species-specific characteristic, in the case of humans, there is still significant variation within patients. To evaluate the suitability of the dynamic SDF assay to assess the adverse effects of agents that cause genetic damage, fresh semen samples from different donors were exposed in vitro to (1) increasing acute doses of ionizing radiation, (2) elevated temperature (41 Degree-Sign C and 45 Degree-Sign C), (3) acidic pH (pH 4) and (4) the nitric oxide (NO) donor sodium nitroprusside (SNP). Sperm DNA fragmentation was analyzed after an incubation period of chronic (24 h), or acute (1 h) exposure to each treatment followed by incubation at 37 Degree-Sign C over a period of 24 h. SDF was assessed using the sperm chromatin dispersion (SCD) test. Dynamic SDF for each treatment was analyzed using Kaplan-Meier survival curves. All agents, except for ionizing radiation, accelerated SDF kinetics following chronic exposure over a 24 h period. Transient exposure to NO and heat but not acidic pH increased the basal (T0) level of SDF. Despite the removal of the three toxicants, the remaining sperm following acute exposure showed a decrease in their expected DNA longevity. It is concluded that the assessment of sperm DNA fragmentation dynamics is an effective methodological approach for revealing latent damage associated with toxicants that is not initially expressed following a single initial observation of SDF.

  13. Stem-spermatogonial survival and incidence of reciprocal translocations in the. gamma. -irradiated boar

    Energy Technology Data Exchange (ETDEWEB)

    Erickson, B.H.; Martin, P.G.

    1984-01-01

    To assess the effects of ..gamma..-radiation on stem-cell survival and incidence of reciprocal translocations, boar testes were irradiated with 100, 200, or 400 rad. Stem-cell survival was markedly affected by 100 rad (51% of control) and reduced to 34% of control by 400 rad. Production of differentiating spermatogonia was all but completely interrupted by 200 rad and spermatogonial renewal was incomplete at 12 weeks. From the state of the seminiferous epithelium at 12 weeks, estimates of the percentage of permanent impairment of sperm-producing capacity ranged from 20 +/- 6 (100 rad) to 67 +/- 10 (400 rad). Incidence of translocations peaked at 200 rad and the number occurring at 100 and 400 rad was similar. Kinetics of porcine spermatogonial renewal differs considerably from those of the rodent and, relative to the rodent, this may account for the boar's higher sensitivity to stem-cell killing and lower sensitivity to translocations.

  14. Sperm Oxidative Stress Is Detrimental to Embryo Development: A Dose-Dependent Study Model and a New and More Sensitive Oxidative Status Evaluation

    Directory of Open Access Journals (Sweden)

    Letícia S. de Castro

    2016-01-01

    Full Text Available Our study aimed to assess the impact of sperm oxidative stress on embryo development by means of a dose-dependent model. In experiment 1, straws from five bulls were subjected to incubation with increasing H2O2 doses (0, 12.5, 25, and 50 μM. Motility parameters were evaluated by Computed Assisted System Analysis (CASA. Experiment 2 was designed to study a high (50 μM and low dose (12.5 μM of H2O2 compared to a control (0 μM. Samples were incubated and further used for in vitro fertilization. Analyses of motility (CASA, oxidative status (CellROX green and 2’-7’ dichlorofluorescein diacetate, mitochondrial potential (JC-1, chromatin integrity (AO, and sperm capacitation status (chlortetracycline were performed. Embryos were evaluated based on fast cleavage (30 h.p.i., cleavage (D=3, development (D=5, and blastocyst rates (D=8. We observed a dose-dependent deleterious effect of H2O2 on motility and increase on the percentages of positive cells for CellROX green, capacitated sperm, and AO. A decrease on cleavage and blastocyst rates was observed as H2O2 increased. Also, we detected a blockage on embryo development. We concluded that sperm when exposed to oxidative environment presents impaired motility traits, prooxidative status, and premature capacitation; such alterations resulting in embryo development fail.

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

    Directory of Open Access Journals (Sweden)

    Laura Gómez Montoto

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

  16. Pronuclear morphology evaluation in in vitro fertilization (IVF / intracytoplasmic sperm injection (ICSI cycles: a retrospective clinical review

    Directory of Open Access Journals (Sweden)

    Nicoli Alessia

    2013-01-01

    Full Text Available Abstract Background The assessment of the embryo quality is crucial to maintain an high pregnancy rate and to reduce the risk of multiple pregnancy. The evaluation of the pronuclear and nucleolar characteristics of human zygote have been proposed as an indicator of embryo development and chromosomal complement. The aim of the current study was to assess the role of pronuclear morphology evaluation in vitro fertilization (IVF / intracytoplasmic sperm injection (ICSI cycles. Methods Retrospective clinical analysis on 755 non-elective transfers of only one embryo (ET. Embryo assessment was performed in days 1 and 2. Clinical and biological data were recorded and analyzed according to embryo and/or pronuclear morphology. Results Both pronuclear and embryo morphology were significantly related to clinical pregnancy and live-birth rates. No significant difference in clinical pregnancy and live-birth rates was detected when the pronuclear and embryo morphology assessments were combined. Embryo morphology and maternal age were the only independent predictors of favorable outcome by logistic regression analysis. Conclusions Pronuclear evaluation is effective to select the best zygotes if ET is performed at day 1, whereas it did not improve the clinical outcomes when combined with embryo morphology evaluation in day 2.

  17. Is there a future for the boar?

    NARCIS (Netherlands)

    Langendijk, P.

    2001-01-01

    This thesis describes several boar stimuli in their potency to elicit estrous behavior and their potency to affect uterine contractility. With different levels of boar stimuli, onset of estrus can be recorded at different time points relative to ovulation, depending on the change in responsiveness o

  18. 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.

  19. Male attitude towards masturbating: an impediment to infertility evaluation and sperm parameters.

    Science.gov (United States)

    Pottinger, A M; Carroll, K; Mason, G

    2016-09-01

    Male attitude about masturbation may influence early diagnosis and treatment of infertility and may be of particular burden in developing countries. We sought to explore attitude about masturbating and examine comfort/discomfort with masturbating and sexual history, pregnancy history and sperm quality in men investigating fertility potential. The study consisted of 83 male volunteers, 23-61 years, attending a fertility management unit in Kingston, Jamaica. Comfort with masturbation was assessed by a self-administered questionnaire. Participants also completed the unit's standard intake form for infertility investigations and produced a semen sample. T-tests, Mann-Whitney U-test and chi-square were used to compare differences in comfort level with outcome variables. We found 59% were comfortable masturbating although requiring external stimulation to produce a sample (48%); 6% (n = 5) failed to produce a sample after repeated attempts. A higher percentage of men uncomfortable with masturbating reported sexual problems (P < 0.05) and spending a longer time trying to have a baby (P < 0.05). Semen quality was not associated with masturbating comfort. Producing a sample by masturbation is standard for many assisted conception treatments. As comfort with masturbating may influence delay in infertility investigations and fertility outcome, efforts to improve men's comfort level with semen production should be considered in pre-treatment fertility counselling.

  20. Coprological tests underestimate Macracanthorhynchus hirudinaceus burden in wild boar.

    Science.gov (United States)

    Gassó, Diana; Serrano, Emmanuel; Castillo-Contreras, Raquel; Aguilar, Xavier Fernández; Cadena, Andreu Colom; Velarde, Roser; Mentaberre, Gregorio; López-Olvera, Jorge Ramón; Risco, David; Gonçalves, Pilar; Lavín, Santiago; Fernandez-Llário, Pedro; Segalés, Joaquim; Ferrer, David

    2016-05-01

    The present study evaluated the limitations of the coprological sedimentation test to assess Macracanthorhynchus hirudinaceus infestation in 59 wild boars (Sus scrofa) from central Spain. The coprological sedimentation test appeared to be a poor predictor of both prevalence of infestation and the real parasite burden due to the high number of false negative results (prevalence was reduced from 61 to 16 %). Because of the potential increased risk of this zoonosis, it is suggested that alternative techniques be used in wildlife surveillance programmes.

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

    Science.gov (United States)

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

    2015-08-01

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

  2. Human Sperm Immotility Caused by Degeneration in the Epididymis

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective To investigate whether sperm immotility was caused by degeneration in the epididymisMethods Five patients with totally immotile sperm were selected in this study. Testic ular biopsy was used to obtain testicular sperm to evaluate sperm motility. The com bined hypoosmotic swelling-eosin Y exclusion test was carried out to determine the sperm head and tail membrane integrity for the ejaculated and the testicular sperm.The ultrastructure of ejaculated sperm was examined by transmission electron microscope.Results No motile sperm were found in the ejaculated semen samples from 5 patients,whereas 2% to 11% motile testicular sperm extracted from the testicular biopsy tissues were observed. The percentage of testicular sperm with intact head and tail membranes was higher than that of the ejaculated sperm (P< 0. 01). Ultrastructure of the ejacu lated sperm showed marked degenerative features. Seminal plasma from patients did not influence the motility of normal donor sperm.Conclusion Sperm could undergo degenerative changes during transit through and /or storage in the epididymis, which led to lose sperm motility in these patients. Using motile testicular sperm would benefit the treatment for such cases.

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

    Science.gov (United States)

    Manásková, P; Jonáková, V

    2008-06-01

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

  4. Boar spermatozoa successfully predict mitochondrial modes of toxicity: implications for drug toxicity testing and the 3R principles.

    Science.gov (United States)

    Vicente-Carrillo, A; Edebert, I; Garside, H; Cotgreave, I; Rigler, R; Loitto, V; Magnusson, K E; Rodríguez-Martínez, H

    2015-04-01

    Replacement of animal testing by in vitro methods (3-R principles) requires validation of suitable cell models, preferably obtained non-invasively, defying traditional use of explants. Ejaculated spermatozoa are highly dependent on mitochondrial production and consumption of ATP for their metabolism, including motility display, thus becoming a suitable model for capturing multiple modes of action of drugs and other chemicals acting via mitochondrial disturbance. In this study, a hypothesis was tested that the boar spermatozoon is a suitable cell type for toxicity assessment, providing a protocol for 3R-replacement of animals for research and drug-testing. Boar sperm kinetics was challenged with a wide variety of known frank mito-toxic chemicals with previously shown mitochondrial effects, using a semi-automated motility analyser allied with real-time fluorescent probing of mitochondrial potential (MitoTracker & JC-1). Output of this sperm assay (obtained after 30 min) was compared to cell viability (ATP-content, data obtained after 24-48 h) of a hepatome-cell line (HepG2). Results of compound effects significantly correlated (Psperm variables and for most variables in (HepG2). Dose-dependent decreases of relative ATP content in HepG2 cells correlated to sperm speed (r=0.559) and proportions of motile (r=0.55) or progressively motile (r=0.53) spermatozoa. The significance of the study relies on the objectivity of computerized testing of sperm motility inhibition which is comparable albeit of faster output than somatic cell culture models. Sperm suspensions, easily and painlessly obtained from breeding boars, are confirmed as suitable biosensors for preclinical toxicology screening and ranking of lead compounds in the drug development processes.

  5. Pig, F1 (wild boar x pig) and wild boar meat quality

    OpenAIRE

    Ragni, M.; S. Tarricone; Pinto, F.; Dimatteo, S; G. Marsico; Rasulo, A

    2010-01-01

    Sixteen carcasses of wild boars, pigs, hybrids F1 (wild boar x pig) and reared wild boar have been examined to study the meat quality and the fatty acid composition. Four carcasses came from hunted wild boars and twelve from animals reared in outdoor pens till nine months of age. The meat produced by the hunted wild animals, although not marketable, offers the best quality and nutritional characteristics. The use of hybrids reared in outdoor pens can approximate or equalize the hunted wild bo...

  6. Motilidade espermática e integridade acrossomal em doses de sêmen suíno refrigeradas e inoculadas com Esecherichia coli e Staphylococcus aureus Sperm motility and acrossomal integrity in liquid boar semen dosis inoculated with Esecherichia coli and Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Paulo Eduardo Bennemann

    2000-04-01

    Full Text Available Foram utilizados 12 ejaculados, coletados de maneira asséptica, distribuídos em sete tratamentos, sendo um grupo controle e os demais inoculados com três diferentes concentrações de S. aureus ou E. coli (5 x 10(5, 5 x 10(6 e 5 x 10(7 UFC/ml. Durante 96 horas, foram avaliados a motilidade espermática, o percentual de acrossomas intactos (NAR, o pH e o número de unidades formadoras de colônia (UFC/ml das bactérias inoculadas. O desenvolvimento bacteriano foi decrescente ao longo das 96 horas de armazenamento. Com exceção do tratamento com a inoculação de 5 x 10(7 UFC de E. coli/ml, não foi observado efeito significativo das bactérias sobre a motilidade espermática (p>0,05. Da mesma forma, não houve efeito significativo (p>0,05 do S. aureus ou da E. coli sobre o percentual de NAR e pH nas 96 horas. Quando comparado ao controle, somente a inoculação de 5 x 10(7 UFC/ml de E. coli diferiu, às 96 horas, em relação ao percentual de NAR (pTwelve ejaculates were collected in the most aseptic manner and distributed in seven treatments (control group T1. Semen were inoculated with S. aureus (T2, T3 e T4 or E. coli (T5, T6 e T7 in three concentrations (5 x 10(5, 5 x 10(6 and 5 x 10(7 CFU/ml. The sperm motility, the percentage of normal apical range (NAR, the pH and the number of colony unit former (CFU/ml of bacteria, for 96 hours, were evaluated. The bacterial development was decreasing during the first 96 hours. Except for the treatment with the inoculation of 5 x 10(7 CFU of E. coli/ml, there was no significant effect of the bacteria on the sperm motility (p>0.05. Also, there was no significant effect (p>0.05 of the S. aureus or E. coli on the percent of NAR and the pH during the 96 hours. When compared to the control group, only the inoculation of 5 x 10(7 CFU/ml of E. coli differed at the 96 hours in relation to the percentage of NAR (p<0.05. There was no correlation among the sperm motility, NAR, pH and CFU.

  7. Sperm proteasomes degrade sperm receptor on the egg zona pellucida during mammalian fertilization.

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    Shawn W Zimmerman

    Full Text Available Despite decades of research, the mechanism by which the fertilizing spermatozoon penetrates the mammalian vitelline membrane, the zona pellucida (ZP remains one of the unexplained fundamental events of human/mammalian development. Evidence has been accumulating in support of the 26S proteasome as a candidate for echinoderm, ascidian and mammalian egg coat lysin. Monitoring ZP protein degradation by sperm during fertilization is nearly impossible because those few spermatozoa that penetrate the ZP leave behind a virtually untraceable residue of degraded proteins. We have overcome this hurdle by designing an experimentally consistent in vitro system in which live boar spermatozoa are co-incubated with ZP-proteins (ZPP solubilized from porcine oocytes. Using this assay, mimicking sperm-egg interactions, we demonstrate that the sperm-borne proteasomes can degrade the sperm receptor protein ZPC. Upon coincubation with motile spermatozoa, the solubilized ZPP, which appear to be ubiquitinated, adhered to sperm acrosomal caps and induced acrosomal exocytosis/formation of the acrosomal shroud. The degradation of the sperm receptor protein ZPC was assessed by Western blotting band-densitometry and proteomics. A nearly identical pattern of sperm receptor degradation, evident already within the first 5 min of coincubation, was observed when the spermatozoa were replaced with the isolated, enzymatically active, sperm-derived proteasomes. ZPC degradation was blocked by proteasomal inhibitors and accelerated by ubiquitin-aldehyde(UBAL, a modified ubiquitin protein that stimulates proteasomal proteolysis. Such a degradation pattern of ZPC is consistent with in vitro fertilization studies, in which proteasomal inhibitors completely blocked fertilization, and UBAL increased fertilization and polyspermy rates. Preincubation of intact zona-enclosed ova with isolated active sperm proteasomes caused digestion, abrasions and loosening of the exposed zonae, and

  8. Sperm proteasomes degrade sperm receptor on the egg zona pellucida during mammalian fertilization.

    Science.gov (United States)

    Zimmerman, Shawn W; Manandhar, Gaurishankar; Yi, Young-Joo; Gupta, Satish K; Sutovsky, Miriam; Odhiambo, John F; Powell, Michael D; Miller, David J; Sutovsky, Peter

    2011-02-23

    Despite decades of research, the mechanism by which the fertilizing spermatozoon penetrates the mammalian vitelline membrane, the zona pellucida (ZP) remains one of the unexplained fundamental events of human/mammalian development. Evidence has been accumulating in support of the 26S proteasome as a candidate for echinoderm, ascidian and mammalian egg coat lysin. Monitoring ZP protein degradation by sperm during fertilization is nearly impossible because those few spermatozoa that penetrate the ZP leave behind a virtually untraceable residue of degraded proteins. We have overcome this hurdle by designing an experimentally consistent in vitro system in which live boar spermatozoa are co-incubated with ZP-proteins (ZPP) solubilized from porcine oocytes. Using this assay, mimicking sperm-egg interactions, we demonstrate that the sperm-borne proteasomes can degrade the sperm receptor protein ZPC. Upon coincubation with motile spermatozoa, the solubilized ZPP, which appear to be ubiquitinated, adhered to sperm acrosomal caps and induced acrosomal exocytosis/formation of the acrosomal shroud. The degradation of the sperm receptor protein ZPC was assessed by Western blotting band-densitometry and proteomics. A nearly identical pattern of sperm receptor degradation, evident already within the first 5 min of coincubation, was observed when the spermatozoa were replaced with the isolated, enzymatically active, sperm-derived proteasomes. ZPC degradation was blocked by proteasomal inhibitors and accelerated by ubiquitin-aldehyde(UBAL), a modified ubiquitin protein that stimulates proteasomal proteolysis. Such a degradation pattern of ZPC is consistent with in vitro fertilization studies, in which proteasomal inhibitors completely blocked fertilization, and UBAL increased fertilization and polyspermy rates. Preincubation of intact zona-enclosed ova with isolated active sperm proteasomes caused digestion, abrasions and loosening of the exposed zonae, and significantly reduced

  9. Molecular characterization and seroprevalence of Echinococcus granulosus in wild boars (Sus scrofa) in south-western Iran.

    Science.gov (United States)

    Sarkari, Bahador; Mansouri, Majid; Khabisi, Samaneh Abdolahi; Mowlavi, Gholamreza

    2015-01-01

    This study presents the first molecular and serological evaluation of Echinococcus granulosus infections in wild boars in Iran. Twenty five wild boars were collected in south-western Iran, during authorized hunting program, from March to October 2013, necropsied and examined for E. granulosus infection. Furthermore, seroprevalence of cystic echinococcosis in hunted boars was evaluated by an ELISA system. A fertile hydatid cyst due to E. granulosus was detected in the lung of one of the animals. Genotype analysis of the isolate was determined by analyzing a mitochondrial gene, cytochrome C oxidase subunit 1 (co1). DNA was extracted from the cyst sample and polymerase chain reaction amplification and DNA sequencing of the specific region of the co1 gene was performed. Molecular evaluation confirmed the presence of a sheep strain, the G1 genotype, in the wild boar in south-western Iran. This is the first report of the presence of G1 genotype of E. granulosus in wild boar in Iran. Serological evaluation of hydatid cyst by antigen-B ELISA revealed E. granulosus antibodies in 5 (20%) of 25 wild boars. A statistically significant difference was observed between the prevalence of E. granulosus antibodies and gender while the difference between the seroprevalence of E. granulosus and age was insignificant. Findings of this study might have important implications for the prevention and control of cystic echinococcosis.

  10. Children conceived after intracytoplasmic sperm injection (ICSI)

    DEFF Research Database (Denmark)

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

    2004-01-01

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

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

    Science.gov (United States)

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

    2007-02-01

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

  12. Physical characteristics of ejaculates produced by insemination boars depending on the interval between successive ejaculate collections

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    Magdalena BAJENA

    2016-06-01

    Full Text Available The ejaculate characteristics of Polish Landrace boars showed a significant correlation with the intervals between the successive ejaculate collections. The effect of insemination use intensity was however varied. Rising frequency of ejaculate collection led to a systematic and fairly even fall in ejaculate volume. Ejaculate sperm concentration remained at a relatively high level when ejaculates were collected with a frequency of 7 to 3 days but further shortening of the interval between the successive collections led to a drastic decrease in sperm concentration. An increase in ejaculate collection frequency to every four and fewer days resulted in a significant decrease in the number of spermatozoa present in the produced ejaculates and a concomitant decrease in the number of insemination doses prepared from these ejaculates, with an escalation of such changes.

  13. Accuracy Evaluation of The Depth of Six Kinds of Sperm Counting Chambers for both Manual and Computer-Aided Semen Analyses

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    Jin-Chun Lu

    2016-12-01

    Full Text Available Background: Although the depth of the counting chamber is an important factor influencing sperm counting, no research has yet been reported on the measurement and comparison of the depth of the chamber. We measured the exact depths of six kinds of sperm counting chambers and evaluated their accuracy. Materials and Methods: In this prospective study, the depths of six kinds of sperm counting chambers for both manual and computer-aided semen analyses, including Makler (n=24, Macro (n=32, Geoffrey (n=34, GoldCyto (n=20, Leja (n=20 and Cell-VU (n=20, were measured with the Filmetrics F20 Spectral Reflectance Thin-Film Measurement System, then the mean depth, the range and the coefficient of variation (CV of each chamber, and the mean depth, relative deviation and acceptability of each kind of chamber were calculated by the closeness to the nominal value. Among the 24 Makler chambers, 5 were new and 19 were used, and the other five kinds were all new chambers. Results: The depths (mean ± SD, μm of Makler (new, Macro and Geoffrey chambers were 11.07 ± 0.41, 10.19 ± 0.48 and 10.00 ± 0.28, respectively, while those of GoldCyto, Leja and Cell-VU chambers were 23.76 ± 2.15, 20.49 ± 0.22 and 24.22 ± 2.58, respectively. The acceptability of Geoffrey chambers was the highest (94.12%, followed by Macro (65.63%, Leja (35% and Makler (20%, while that of the other two kinds and the used Makler chamber was zero. Conclusion: There existed some difference between the actual depth and the corresponding nominal value for sperm counting chambers, and the overall acceptability was very low. Moreover, the abrasion caused by the long use, as of Makler chamber, for example, may result in unacceptability of the chamber. In order to ensure the accuracy and repeatability of sperm concentration results, the depth of the sperm counting chamber must be checked regularly.

  14. Seroprevalence of Toxoplasma gondii in hunted wild boars (Sus scrofa) from southeastern France.

    Science.gov (United States)

    Roqueplo, Cedric; Blaga, Radu; Jean-Lou, Marie; Vallee, Isabelle; Davoust, Bernard

    2017-01-25

    Toxoplasma gondii (Nicolle et Manceaux, 1908) is an obligate intracellular, parasitic protozoan within the phylum Apicomplexa that causes toxoplasmosis in mammalian hosts (including humans) and birds. Since meat of wild boar, Sus scrofa (Linnaeus), has been demonstrated to be a potential source of human infection, a careful evaluation of the prevalence of infection with T. gondii in hunted animals is needed to protect public health. In the Var area in southeastern France, we performed a spatio-temporal survey in order to investigate the prevalence of IgG antibodies in wild boars shot by hunters in the Canjuers military camp during two subsequent hunting seasons. Of 841 wild boars screened, antibodies (IgG) to T. gondii (modified agglutination test, cut-off 1 : 6) were found in 141 (16.8%) muscle extract samples. A significant association (p < 0.001) was found between positivity and age, but not gender, and hunting districts. The results obtained indicate that consumption of raw or undercooked meat from wild boars carries an important risk of infection with T. gondii. Wild boars may be considered as a bioindicator of parasite circulation in this ecosystem.

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

    Science.gov (United States)

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

    2012-12-01

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

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

    Science.gov (United States)

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

    2014-06-30

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

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

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    Suherni Susilowati

    2008-12-01

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

  18. 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.

  19. EVALUATION OF A COMPUTER-AIDED SEMEN ANALYSIS SYSTEM WITH SPERM TAIL DETECTION

    NARCIS (Netherlands)

    WIJCHMAN, JG; DEWOLF, BTHM; JAGER, S

    1995-01-01

    The aim of this study was to evaluate the Stroemberg-Mika cell motion analyser (SM-CMA) which uses tail detection in order to discriminate between immotile spermatozoa and other particles. Analysis of the spermatozoa by the SM-CMA can easily be checked on a video monitor. The semen samples were from

  20. Genetic relationships between measures of sexual development, boar taint, health, and aggressiveness in pigs.

    Science.gov (United States)

    Parois, S P; Prunier, A; Mercat, M J; Merlot, E; Larzul, C

    2015-08-01

    genetic correlations were different for some pairs of traits. It would be possible to select boars based on their plasma concentration of sex hormones to decrease boar taint and aggressiveness without important consequences on the immune response. However, because of the strong links between boar taint and reproductive function, the possible consequences on the reproductive performance should be evaluated.

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

    Science.gov (United States)

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

    2014-10-15

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

  2. Dynamic quantification of intracellular calcium and protein tyrosine phosphorylation in cryopreserved boar spermatozoa during short-time incubation with oviductal fluid.

    Science.gov (United States)

    Kumaresan, A; González, R; Johannisson, A; Berqvist, A-S

    2014-11-01

    Freshly ejaculated boar spermatozoa require several hours of exposure to capacitating conditions to undergo capacitation. We hypothesized that cryopreserved boar spermatozoa might elicit a capacitation response after a relatively shorter time of exposure to capacitating conditions. Washed, frozen-thawed boar spermatozoa were incubated separately with pre-ovulatory isthmic oviductal fluid (EODF), post-ovulatory ODF (MODF), capacitation medium (CM), and noncapacitating medium (NCM) for 60 minutes. Aliquots of spermatozoa were taken at 0, 5, 15, 30, and 60 minutes during incubation and sperm kinematics, intracellular calcium [Ca2(+)]i content, and protein tyrosine phosphorylation (PTP) were studied. The proportion of motile spermatozoa increased significantly after 5 minutes of incubation with EODF. A similar increase was not observed in the other groups. During the initial 5 minutes of incubation, the proportion of spermatozoa with high [Ca(2+)]i decreased significantly in all four groups. The proportion of tyrosine phosphorylated spermatozoa increased from 6.49 ± 1.93% to 15.42 ± 3.58% and 18.41 ± 1.57% in EODF and MODF groups, respectively, at 5 minutes of incubation. Neither CM nor NCM elicited any immediate effect on PTP in spermatozoa. There was a positive and significant correlation between [Ca(2+)]i and sperm motility (P = 0.009). It may be concluded that frozen-thawed boar spermatozoa undergo capacitation-associated changes after a relatively short exposure to EODF, and there are some subpopulations of spermatozoa that undergo PTP despite possessing low [Ca(2+)]i.

  3. 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.

  4. Evaluation of Ethylene Glycol as a Cryoprotectant in the Sperm Cryopreservation of Trans-andean Shovelnose Catfish (Sorubim Cuspicaudus, Pimelodidae

    Directory of Open Access Journals (Sweden)

    Victor Julio Atencio García

    2014-02-01

    Full Text Available The catfish Sorubim cuspicaudus cryopreservation semen was evaluated using three levels (5, 10, 15% of ethylene glycol (ETG. Males (n = 13 undergoing spermiation and in final maturation females (n = 6 were induced with 0.4 ml Ovaprim®/Kg, after 12 and 14 post-induction the semen was collected in 2 ml Eppendorf vials. The different cryoprotectants solutions were prepared with glucose 6% (w/v skimmed milk powder 5% (w/v and distilled water. The semen was diluted in ratio 1:3 (semen:extender, packed in macrotubes of 2.5 ml and frozen in liquid nitrogen (NL vapor for 30 minutes, then the macrotubes were stored in cryogenic tanks submerged directly in NL. The sperm were thawed in serological bath to 35 °C for 90 seconds. The total motility, total progressivity and velocities in fresh and thawed semen were analyzed with the Sperm Class Analyzer software (SCA Microptic SL, Spain. Fertility and hatching rates were assessed with 1.0-1.5 g of oocytes in experimental up flow incubators 2 L, a completely randomized design was used. The hatching rate of fresh semen was 51.8 ± 21.0%, with no significant differences with semen cryopreserved with ETG 5% (38.6±13.9% (p>0.05, while ETG 15% (9.6±2.9%, recorded the lower hatching rate (p<0.05. The results suggest that the cryoprotectant solution composed of ETG 5%, glucose 6% and powdered milk 5% is a viable alternative for semen cryopreservation of the catfish Sorubim cuspicaudus. EVALUACIÓN DE ETILENGLICOL COMO CRIOPROTECTOR EN LA CRIOCONSERVACIÓN DE SEMEN DE BAGRE BLANCO (Sorubim cuspicaudus, PimelodidaeSe evaluó el semen crioconservado de Sorubim cuspicaudus utilizando etilenglicol (ETG a tres niveles de inclusión (5, 10, 15%. Machos (n=13 en fase de espermiación y hembras (n=6 en maduración final se indujeron con 0.4 ml de Ovaprim®/Kg, después de 12 a 14 horas post-inducción se colectó el semen en viales Eppendorf de 2 ml de capacidad. Las diferentes soluciones crioprotectoras se prepararon con

  5. 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.

  6. Effect of Using Dimethylacetamide in Combination with Trehalose on Cryopreservation of Boar Spermatozoa%海藻糖、二甲乙酰胺对公猪精液冷冻保存效果的研究

    Institute of Scientific and Technical Information of China (English)

    张伟; 易康乐; 燕海峰; 周虚

    2011-01-01

    采用5%二甲乙酰胺(DMA) (V/V)完全替代甘油,比较乳糖、海藻糖对精液冷冻保存效果的影响.结果表明,海藻糖显著提高了冷冻——解冻后精子成活力(49.32%±1.52%)与顶体完整性(47.33%±1.16%)(P<0.05).然后利用海藻糖替代乳糖,评价不同浓度的DMA对公猪精液冷冻保存的影响.结果表明,当DMA添加量为4%时,解冻后精子活率、成活力、顶体完整率分别为(45.17±0.56)%、(50.33±0.67)%、(48.30±1.44)%,均显著高于3% DMA、6% DMA添加组(P<0.05),精子活率显著高于5% DMA添加组(P<0.05),但精子成活力、顶体完整性与其差异不显著(P>0.05).因此,当利用海藻糖作为冷冻保存基础稀释液,DMA最适添加量为4%.%Two experiments were conducted to evaluate the dimethylacetylamide(DMF) and trehalose in boar semen cryopr-eservation protocols. In experiment 11DMF was added at 5 % to the diluents instead of glycerol used to freeze boar semen completely, evaluated lactose and trehalose on the effect of sperm cryopreservation. The results showed that compared with the lactose, trehalose increased significantly add frozen-thawed sperm viability(49. 32±1. 52)% and acrosome integrity(47. 33% ± 1. 16%)(P0. 05). In conclusion,these results showed that when the use of trehalose as the basis of dilution of cryopreservation, the volume fraction of 4% DMA showed a better effect on the quality of boar cryopreservation semen.

  7. 红景天多糖对藏猪精液4℃保存效果的研究%Effect of Rhodiola Polysaccharides on Tibetan Boar Semen Preserved at 4 ℃and Antioxidative

    Institute of Scientific and Technical Information of China (English)

    陈晓英; 马金英; 宋天增; 唐建华; 赵彦玲; 任子利; 朱彦宾; 王良润

    2016-01-01

    the top Tibetan pig sperm activity, sperm abnormal rate and semen quality evaluation index, to investigate the effect of Rhodiola Polysaccharides on the quality of semen quality at 4 ℃, the activity of Superoxidas dismutase(SOD) and Maleic dialdehyde(MDA) were the indexes to evaluate the protective effect of antioxidation. The results show that, to the fifth days,Tibetan boar semen preserved at 4℃, sperm motility, sperm abnormality rate, sperm integrity rate and plasma membrane integrity rate with add group of Polysaccharide 9.0 mg/L from root of the root, these 4 indicators were significantly better than other groups (P<0.01). The sperm motility was significantly (P<0.05) different between the 7.0 mg/L and 11.0 mg/L text groups, other indicators were not significant. Also, at the same time, we found that, addition of Polysaccharides from semen of low temperature dilution liquid of Tibetan boar semen, 9.0 mg/L Rhodiola Polysaccharides supplementation group sperm superox-ide dismutase (SOD) activity is the highest, but there was no difference between 5 mg/L, 7 mg/L and 11 mg/L groups. 9.0 mg/L Rhodiola Polysaccharides supplementation group sperm MDA activity is the highest, compared with 7 mg/L and 11 mg/L, 5 mg/L, 3 mg/L and control group, the difference was very significant(P<0.01), 7 mg/L and 11 mg/L add group no difference. In short adding 9.0 mg/L Rhodiola Polysaccharides to the improved pig semen diluent can significantly improve the semen quality and antioxidant protection at 4 ℃. The experimental study laid a foundation for the production and application of cryopreservation of boar semen in the future.

  8. Role of platelet-activating factor in reproduction:sperm function

    Institute of Scientific and Technical Information of China (English)

    William E. Roudebush

    2001-01-01

    Since its discovery nearly thirty years ago, platelet-activating factor has emerged as one of the more important lipid mediators known. Platelet-activating factor (PAF; 1- O-alkyl-2- O-acetyl-sn-glycero-3-phosphorylcholine) exists en dogenously as a mixture of molecular species with structural variants of the alkyl moiety. PAF is a novel potent signal ing phospholipid that has unique pleiotropic biological properties in addition to platelet activation. PAF also plays a sig nificant role in reproduction. PAF content in squirrel monkey sperm is significantly higher during the breeding season than the non-breeding season. PAF content in human sperm has a positive correlation with seminal parameters and preg nancy outcomes. High-fertility boars have significantly more PAF in their sperm than low-fertility boars. The enzymes (lyso-PAF-acetyltransferase and PAF-acetylhydrolase) necessary for PAF activation and deactivation are present in sperm. PAF-acetylhydrolase may act as a "decapacitation factor". Removal of this enzyme during capacitation may promote PAF synthesis increasing motility and fertilization. PAF also plays a significant role in the fertilization process,enhancing the fertilization rates of oocytes. Enhanced embryo development has also been reported in oocytes fertilized with PAF-treated sperm. PAF antagonists inhibit sperm motility, acrosome reaction, and fertilization, thus suggesting the presence of receptors for PAF. The PAF-receptor is present on sperm, with altered transcript levels and distribution patterns on abnormal cells. Whereas the exact mechanism of PAF in sperm function and reproduction is uncertain, its importance in normal fertility is substantial. The reproductive significance of PAF activity in sperm and fertility plus the role of PAF in the establishment of pregnancy requires further study.

  9. 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.

  10. Freeze-dried stallion spermatozoa: evaluation of two chelating agents and comparative analysis of three sperm DNA damage assays.

    Science.gov (United States)

    Olaciregui, M; Luño, V; Martí, J I; Aramayona, J; Gil, L

    2016-11-01

    During the freeze-drying procedure, sperm DNA might become damaged by both freezing and drying stresses. Sperm DNA status can be detected using well-established assays; however, most techniques are expensive and involve elaborate protocols and equipment. Indirect assessments can provide alternative strategies. The objective of this study was to compare a simple test of DNA status using Diff-Quik (DQ) with two established procedures: acridine orange test (AOT) and sperm chromatin dispersion (SCD) on freeze-dried (FD) stallion spermatozoa. Ejaculated spermatozoa from three stallions were freeze-dried in basic medium supplemented with two different chelating agents: EGTA or EDTA. After rehydration, the spermatozoa were subjected to DNA damage detection using a SCDt, AOT and DQ stain simultaneously. The results showed that the DNA damage levels in the EGTA group were significantly lower than those in the EDTA group. AOT detected a significantly higher proportion of spermatozoa with fragmented DNA than DQ and SCD. The results of the SCD test and DQ stain exhibited a significant positive correlation for DNA fragmentation (r = 0.528), whereas a negative correlation was observed between SCD, DQ and AOT (r = -0.134 and r = -0.332 respectively). The present study shows that both the SCD test and DQ assay are effective methods for detecting FD stallion sperm DNA fragmentation, whereas using of AOT is questionable.

  11. Cytological evaluation of spermatogenesis: a novel and simple diagnostic method to assess spermatogenesis in non-obstructive azoospermia using testicular sperm extraction specimens.

    Science.gov (United States)

    Hessel, M; de Vries, M; D'Hauwers, K W M; Fleischer, K; Hulsbergen-van de Kaa, C A; Braat, D D M; Ramos, L

    2015-05-01

    Most of the non-obstructive azoospermia (NOA)-patients have only focal spermatogenesis which results in insufficient numbers of spermatozoa to reach the ejaculate. In ≈50% of these NOA-patients testicular sperm extraction (TESE) is successful and intracytoplasmic sperm injection (ICSI) is pursued. We studied whether (i) spermatogenesis can be evaluated by defining the ratios between Sertoli cells, pachytene spermatocytes and spermatozoa in a testicular cell suspension, and (ii) these ratios are associated with the outcome of fertility treatment. A retrospective cohort study was conducted between June 2007 and August 2012. In this period, 441 consecutive ICSI-TESE cycles were performed in 212 couples. For each TESE biopsy, the ratios between Sertoli cells, pachytene spermatocytes and spermatozoa were calculated. A control population of 32 vasectomized men was used to define cut-off values for complete spermatogenesis. Based on the pachytene to sperm ratio (P/Sp) and number of spermatozoa per 100 Sertoli cells (#Sp/100SC) groups were defined as complete spermatogenesis, hypospermatogenesis and partial maturation arrest (MA). Validation of the cytological diagnoses was performed by comparing the results of cytology to the histological evaluation of spermatogenesis in 40 cases. In 92.5%, a perfect match was observed and in the three remaining cases cytology corresponded well with the results of TESE. Couples with complete spermatogenesis have a higher ongoing pregnancy rate after the first treatment cycle compared to couples with hypospermatogenesis (34 vs. 16%; p = 0.02) and partial MA (34 vs. 19%; p = 0.11). In conclusion, pachytene spermatocytes, spermatozoa and Sertoli cells can be easily identified and counted in a cell suspension and their ratios can be successfully used to diagnose the level of spermatogenic impairment. This pilot study indicates that once successful spermatozoa retrieval is achieved, treatment outcome declines when spermatogenesis is

  12. Use of indoor boars as models for understanding seasonal infertility: Preliminary data

    Science.gov (United States)

    This study was conducted to evaluate the potential impacts of external temperature and relative humidity (RH) variations on semen production of boars maintained in thermo-regulated barns (indoor housing). Data were collected from a local commercial hog operation. Temperature and relative humidity (R...

  13. 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

    2016-03-01

    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.

  14. Freeze-tolerance of Trichinella muscle larvae in experimentally infected wild boars.

    Science.gov (United States)

    Lacour, Sandrine A; Heckmann, Aurélie; Macé, Pauline; Grasset-Chevillot, Aurélie; Zanella, Gina; Vallée, Isabelle; Kapel, Christian M O; Boireau, Pascal

    2013-05-20

    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 to be correlated to the development of thick capsule walls which increases with age. An extended infection period and the muscle composition in some hosts (e.g. herbivores) may provide freeze-avoiding matrices due to high carbohydrate contents. The present experiment compares freeze-tolerance of 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 animals served as negative controls. All wild boars were sacrificed 24 wpi. Muscle samples of 70 g were stored at -21°C for 19, 30 and 56 h, and for 1-8 weeks. Larvae were recovered by artificial digestion. Their mobilities were recorded using Saisam(®) image analysis software and their infectivities were evaluated using mouse bioassays. Samples frozen for 19, 30 and 56 h allowed recovery of mobile ML, but samples frozen for 1 or 2 weeks did not. Correspondingly, only T. spiralis and T. britovi larvae isolated from wild boar meat frozen for 19, 30 and 56 h established in mice. This study showed that freezing at -21°C for 1 week inactivated T. spiralis and T. britovi ML encapsulated in wild boar meat for 24 weeks.

  15. Extracellular superoxide dismutase of boar seminal plasma.

    Science.gov (United States)

    Kowalowka, M; Wysocki, P; Fraser, L; Strzezek, J

    2008-08-01

    Superoxide dismutase (SOD) is an enzymatic component of the antioxidant defense system that protects spermatozoa by catalysing the dismutation of superoxide anions to hydrogen peroxide and oxygen. Age and season effects on SOD activity in the seminal plasma were measured in boars at the onset of 8 months through a 35-month period. It was found that age-related changes in SOD activity in the seminal plasma were markedly higher in boars less than 2 years of age. However, it appeared that SOD activity was established at the early sexual maturity age (8-12 months). There were variations in SOD activity throughout the season, being significantly higher in spring and autumn than in summer. A secretory extracellular form of SOD (EC-SOD) was purified to homogeneity (350-fold) from boar seminal plasma, using a three-step purification protocol (affinity chromatography followed by ion exchange and ceramic hydroxyapatite chromatography). The molecular properties and specificity of SOD (molecular mass, isoelectric point, optimum pH, thermostability and susceptibility to inhibitors) confirmed that the purified enzyme is an extracellular form of Cu/Zn-superoxide dismutase occurring in boar seminal plasma. The results of this study indicate that EC-SOD is an important antioxidant enzyme of boar seminal plasma, which plays an important physiological role in counteracting oxidative stress in spermatozoa.

  16. The use of comet assay to assess DNA integrity of boar spermatozoa following liquid preservation at 5 degrees C and 16 degrees C.

    Directory of Open Access Journals (Sweden)

    J Strzezek

    2004-03-01

    Full Text Available The comet assay, under neutral conditions, allows the assessment of DNA integrity influenced by sperm ageing, which is manifested in DNA double-strand breaks. Here, we attempted to use a modified neutral comet assay test (single-cell gel electrophoresis, to our knowledge for the first time, to assess DNA integrity of boar spermatozoa during liquid storage for 96 h at 5 degrees C and 16 degrees C. In this comet assay protocol we used 2% beta-mercaptoethanol prior to the lysis procedure, to aid in removing nuclear proteins. Ejaculates from 3 boars (designated A, C and G were diluted with a standard semen extender, Kortowo-3 (K-3, which was supplemented with lipoprotein fractions extracted from hen egg yolk (LPFh or ostrich egg yolk (LPFo. Irrespective of the extender type, the percentage of comet-detected spermatozoa with damaged DNA increased gradually during prolonged storage at 5 degrees C and 16 degrees C. Spermatozoa stored in K-3 extender exhibited elevated levels of DNA damage at both storage temperatures. Significant differences in DNA damage among the boars were more pronounced during storage in LPF-based extenders at 5 degrees C: spermatozoa of boars A and G were less susceptible to DNA damage. The percent of tail DNA in comets was lower in LPF-based extenders, and there were individual variations among the boars. We observed that changes in DNA integrity were dependent on the extender type and storage temperature. A higher level of DNA instability was observed in K-3 extended semen compared with K-3/LPFh or K-3/LPFo extended semen during storage at 5 degrees C. No significant difference in the level of DNA damage between K-3/LPFh and K-3/LPFo was observed. It seems that a long-term storage can affect genomic integrity of boar spermatozoa. The modified neutral comet assay can be used to detect low levels of DNA damage in boar spermatozoa during liquid preservation. Therefore, screening for sperm DNA damage may be used as an additional

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

    Science.gov (United States)

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

    2005-12-01

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

  18. Inhibition of sperm motility does not affect live-dead separation of bull sperm by glass beads

    Institute of Scientific and Technical Information of China (English)

    Robert H. Foote

    2001-01-01

    Aim: This study was designed to explore factors which influence binding of dead versus live sperm to glass filters.Methods: Multiple semen collections from bulls were used to explore selective filtration of bull sperm as influenced by nonlethal inhibition of sperm motility with fluoride, killing of sperm by quick-freezing, alteration of the glass surface with silicone, and different intervals of sexual rest between semen collections. Results: A comparison of glass spheres 100, 200 and 390 μm in diameter indicated that 200μm spheres were optimal for selective filtration. Quantitative separation of live from dead sperm was demonstrated with a correlation between the percentage of motile sperm and retention of sperm by the filter of r =-0.87 (P < 0.05). Up to 0.02 mol/L NaFl did not alter the proportion of sperm retained by the filter despite inhibiting sperm motility during filtration, an inhibition which was reversible. Proportions of live-dead sperm, based upon eosin staining, were unaffected by fluoride. Coating the glass spheres with silicone greatly reduced selective filtration. Dead sperm adherence to glass wasreduced and resistance to NaFl inhibition was increased by daily ejaculation versus one-week intervals of sexual rest. Conclusion: These studies indicate that the adherence of sperm to glass is primarily due to some form of physico-chemical change accompanying death of the sperm cell independent of active sperm motility. This attraction between the sperm plasma membrane and glass is modified by the age of the ejaculated sperm. This information is useful in evaluating different clinical procedures used for sperm separation.

  19. Sperm release pathway

    Science.gov (United States)

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

  20. Intracytoplasmic sperm injection

    Science.gov (United States)

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

  1. Sperm nuclear histone H2B: correlation with sperm DNA denaturation and DNA stainability

    Institute of Scientific and Technical Information of China (English)

    Armand Zini; Xiaoyang Zhang; Maria San Gabriel

    2008-01-01

    Aim: To examine the relationship between sperm DNA damage and sperm nuclear histone (H2B) staining. Methods:We evaluated sperm samples from 14 consecutive asthenoteratozoospermic infertile men and six consecutive fertile controls. Sperm nuclear histone (H2B) staining and sperm chromatin integrity (assessed by sperm chromatin structure assay and expressed using the percentage of (I) DNA fragmentation index [%DFI] and (ii) high DNA stainability [%HDS)]) were evaluated. Results: Histone H2B immunocytochemistry demonstrated two nuclear staining patterns: (I) focal punctate staining; and (ii) diffuse staining. Infertile men had a higher mean percentage of spermatozoa exhibiting diffuse H2B staining than did fertile men (7.7% ± 4.6% vs. 1.6% ± 1.2%, respectively, P < 0.01). We observed significant relationships between the proportion of spermatozoa with diffuse nuclear histone staining and both sperm %DFI (r= 0.63, P < 0.01) and sperm %HDS (r= 0.63, P < 0.01). Conclusion: The data demonstrate that infertile men have a higher proportion of spermatozoa with diffuse histone H2B than do fertile men and suggest that sperm DNA damage might, at least in part, be due to abnormally high histone H2B levels.

  2. 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

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

    Science.gov (United States)

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

    2012-01-01

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

  4. Measurement and significance of sperm morphology

    Institute of Scientific and Technical Information of China (English)

    Roelof Menkveld; Cas AG Holleboom; Johann PT Rhemrev

    2011-01-01

    The measurement or evaluation and clinical significance of human sperm morphology has always been and still is a controversial aspect of the semen analysis for the determination of a male's fertility potential.In this review the background of the development of the evaluation criteria for sperm morphology will be discussed.Aspects of criticism on the strict criteria definition and use of the criteria for sperm morphology evaluation will be discussed as well as possible reasons for the decline in normal sperm morphology values and how we can compromise for this phenomenon resulting in the very low normal reference value as published in the 2010 WHO manual for the Examination and Processing of Human Semen.One of the possible solutions may be to give more attention to a limited number of abnormal sperm morphology categories and the inclusion of sperm morphology patterns.It is concluded in this review that if done correctly and with care and with strict application of existing guidelines as outlined in the 2010 WHO manual,sperm morphology measurement still has a very important role to play in the clinical evaluation of male fertility potential.

  5. Sperm preparation for fertilization

    NARCIS (Netherlands)

    Gadella, B.M.

    2014-01-01

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

  6. Intracytoplasmic Sperm Injection (ICSI)

    Science.gov (United States)

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

  7. Effects of Cryopreservation on the Ultrastructure of Human Testicular Sperm

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Objective To investigate effects of cryopreservation on changes of the ultrastructure of human testicular sperm and evaluate the efficacy of cryopreserving testicular tissue as a source of sperm for assisted reproduction.Methods Testicular biopsy tissues were obtained from infertile patients (n=12) with obstructive azoospermia and cryopreserved. Testicular sperm motility was observed after in vitro culture procedure. The ultrastructure of testicular sperm (n=6) was examined by transmission electron microscope.Results After cryopreservation, 10 biopsy tissues frozen revealed motile sperm, and 2samples showed non-motile sperm. Some testicular sperm in frozen-thawed group had normal morphology in fine structures. Sperm head in frozen-thawed tissue showed a proportion of nuclei with more electron-dense granules of chromatin. In a few frozen-thawed sperm heads, formation of vesicles and degeneration were observed.The frozen-thawed testicular sperm frequently showed swollen or/and ruptured of the plasma membrane and acrosome membranes.Conclusion Cryopreservation of testicular tissue is simple and efficacious for testicular sperm extraction. And the freezing-thawing procedure of testicular tissue causes damage to ultrastructural morphology of human testicular sperm.

  8. Semen analysis and sperm function testing

    Institute of Scientific and Technical Information of China (English)

    Daniel R Franken; Sergio Oehninger

    2012-01-01

    Despite controversy regarding the clinical value of semen analysis,male fertility investigation still relies on a standardized analysis of the semen parameters.This is especially true for infertility clinics in both developing and developed countries.Other optional tests or sophisticated technologies have not been widely applied.The current review addresses important changes in the analysis of semen as described in the new World Health Organization (WHO) manual for semen analysis.The most important change in the manual is the use of evidence-based publications as references to determine cutoff values for normality.Apart from the above mentioned changes,the initial evaluation and handling methods remain,in most instances,the same as in previous editions.Furthermore,the review evaluates the importance of quality control in andrology with emphasis on the evaluation of sperm morphology.WHO sperm morphology training programmes for Sub-Saharan countries were initiated at Tygerberg Hospital in 1995.1he external quality control programme has ensured that the majority of participants have maintained their morphological reading skills acquired during initial training.This review reports on current sperm functional tests,such as the induced acrosome reaction,and sperm-zona pellucida binding assays,as well as the impact of sperm qualitv in terms of DNA integritv,and the relationship of sperm function tests to sperm morphologv.

  9. Seasonal-dependent variations in metabolic status of spermatozoa and antioxidant enzyme activity in the reproductive tract fluids of wild boar/domestic pig hybrids.

    Science.gov (United States)

    Dziekońska, A; Fraser, L; Koziorowska-Gilun, M; Strzezek, J; Koziorowski, M; Kordan, W

    2014-01-01

    This study investigated seasonal changes in the metabolic performance of spermatozoa and activity of the antioxidant enzymes in the seminal plasma of three wild boar/domestic pigs (aged 1.5 to 2.5 years) and the activity of the antioxidant enzymes in fluids of the cauda epididymidis and vesicular glands from 16 wild boar/domestic pig hybrids (aged 1 to 3 years). Parameters of the sperm metabolic activity, such as total motility, mitochondrial functions, and measurements of oxygen uptake, ATP content and L-lactate production, were analyzed during the spring-summer and autumn-winter periods. Besides these sperm metabolic parameters, the sperm membrane integrity was also assessed. Total protein content and activity of the antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), were measured in the reproductive tract fluids. There were no marked significant differences (P > 0.05) between the seasonal periods in terms of sperm motility, mitochondrial function and oxygen uptake; however, spermatozoa collected during the autumn-winter period exhibited higher (P pig hybrids.

  10. Sperm retrieval outcomes with microdissection testicular sperm extraction (micro-TESE) in men with cryptozoospermia.

    Science.gov (United States)

    Alrabeeah, K; Wachter, A; Phillips, S; Cohen, B; Al-Hathal, N; Zini, A

    2015-05-01

    Several studies support of the use of testicular rather than ejaculated spermatozoa for intracytoplasmic sperm injection (ICSI) in couples with virtual azoospermia or cryptozoospermia, although this approach remains controversial. We sought to evaluate sperm retrieval outcomes with microdissection testicular sperm extraction (micro-TESE) in men with cryptozoospermia. We conducted a retrospective study of 24 consecutive micro-TESEs in men with cryptozoospermia. We also evaluated the outcomes of seven consecutive TESAs (testicular sperm aspiration) in cryptozoospermic men during the same time period (January 2007 and September 2014). Micro-TESE and TESA were performed on the day prior to ICSI. Final assessment of sperm recovery (reported on the day of ICSI) was recorded as (i) successful (available spermatozoa for ICSI) or (ii) unsuccessful (no spermatozoa for ICSI). The decision to perform a unilateral or bilateral micro-TESE was guided by the intra-operative evaluation of sperm recovery from the first testicle. A unilateral procedure was performed in 87.5% (21/24) and 57% (4/7) of the micro-TESE and TESA cohorts, respectively. Sperm recovery was successful in 96% (23/24) of the men who underwent micro-TESE and 43% (3/7) of the men who underwent TESA (p cryptozoospermia and few of these men will require a bilateral procedure. Moreover, sperm retrieval rates are higher with micro-TESE than TESA in this group of men.

  11. Comparative study on efficacy of three sperm-separation techniques

    Institute of Scientific and Technical Information of China (English)

    LanXU; Ren-KangLU; LingCHEN; Yan-LuanZHENG

    2000-01-01

    Aim: To evaluate the comparative effectiveness of real-time sperm separation technique (Wang's tube method) and other two conventional methods in isolating high-quality sperm preparation, and to compare the spouse pregnancy rate in intrauterine insemination (IUI) with sperm preparations isolated by these methods. Methods: The effectiveness of the real-time sperm separation technique, the conventional swim-up and the Percoll discontinuous density gradient methods in isolating sperm preparations from 60 infertile patients (20 with apparently normal semen and 40, abnormal semen contaminated with microorganisms and other impurities) was evaluated and compared. The microorganisms to bere moved included bacteria, virus, Chlamydia trachomaticum, Ureaplsama urealyticum, etc. The spouse pregnancy rates in IUI with sperm preparations isolated by these three techniques from 80 oligoasthenoteratospermic patients werealso compared. Results: The quality (including the percentages of normal form, normal-chromatin and motilesperm, and the grade of motility) of sperm obtained by the real-time sperm separation technique was much higher ( P<0.01 ) as compared with those by the other two methods. The Wang's tube method was also more effective in removing microorganisms and other impurities. The method provided a higher IUI pregnancy rate than the other two sperm separation techniques ( P < 0.05). Conclusion: The real-time sperm separation technique is the most effective method so far available in isolating high-quality sperm samples to be used in assisted reproduction.

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

    Science.gov (United States)

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

    2013-08-01

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

  14. First Detection of Borrelia burgdorferi sensu lato DNA in Serum of the Wild Boar (Sus scrofa) in Northern Portugal by Nested-PCR.

    Science.gov (United States)

    Faria, Ana S; Paiva-Cardoso, Maria das Neves; Nunes, Mónica; Carreira, Teresa; Vale-Gonçalves, Hélia M; Veloso, Octávia; Coelho, Catarina; Cabral, João A; Vieira-Pinto, Madalena; Vieira, Maria L

    2015-03-01

    Lyme borreliosis is the most common tick-borne zoonosis in the northern hemisphere. Several vertebrates are crucial in the epidemiological cycle of Borrelia burgdorferi sensu lato, but the role of wild boar as a reservoir is still unknown. Sera were collected from 90 wild boars shot in the Trás-os-Montes region, Northern Portugal (hunting season 2011/2012). In this study, Borrelia DNA was detected for the first time by nested-PCR in three different sera, suggesting that the wild boar may be a potential reservoir for this spirochete. Sequencing results show 100% similarity with Borrelia afzelii. Further studies are needed to evaluate the public health risks associated with boar hunting.

  15. Castration of the Vietnamese pot-bellied boar: 8 cases

    OpenAIRE

    Østevik, Liv; Elmas, Colette; Rubio-Martinez, Luis M.

    2012-01-01

    Surgical techniques for castration of the Vietnamese pot-bellied boar and outcome are described. Vietnamese pot-bellied pig (VPBP) boars (n = 8) were admitted for castration. Data retrieved from medical records (2002–2011) for these pigs included signalment, history, reason for castration, perioperative management, surgical technique, and complications. Follow-up information was obtained from owners. A scrotal approach with closed technique was used for 6 boars with normally descended testes....

  16. 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 CASA cannot accurately predict 'fertility' that will be obtained with a semen sample or subject. However, when carefully validated, current CASA systems provide information important for quality assurance of semen planned for marketing, and for the understanding of the diversity of sperm responses to changes in the microenvironment in research. The four take-home messages from this review are: (1) animal species, extender or medium, specimen chamber, intensity of illumination, imaging hardware and software, instrument settings, technician, etc., all affect accuracy and precision of output values; (2) semen production facilities probably do not need a substantially different CASA system whereas biology laboratories would benefit from systems capable of imaging and tracking sperm in deep chambers for a flexible period of time; (3) software should enable grouping of individual sperm based on one or more attributes so outputs reflect subpopulations or clusters of similar sperm with unique

  17. Effect of Different Diluents on Boar Semen Preservation at Normal Temperature%不同稀释液对猪精液常温保存效果的研究

    Institute of Scientific and Technical Information of China (English)

    朱永雄

    2011-01-01

    选取杜洛克公猪精液作为试验材料,比较5种稀释液时猪精液的保存效果,结果表明:用葡萄糖—柠檬酸稀释液,所保存精液的pH值、精子活率、精子畸形率等品质指标表现良好,且稀释液的成本低廉,自配简单易行,适于农村养猪户推行;基层常用的葡萄糖稀释液保存3d时的精子活率性状表现不佳,仅适合于现配现用.BL—液(美国)稀释液的保存效果也比较好,但该稀释液组分复杂,配制较难,成本也较高,可在规模养猪场中推广.%The Duroc boar semen was chosen as the tested material, and the preservation effect of 5 kinds of diluents on boar semen at normal temperature was compared. The results showed that the quality indexes (pH - value, sperm survival rate and sperm deformity rate, etc. ) of boar semen which was preserved in glucose -citric acid diluent expressed well; moreover, glucose -citric acid diluent was cheap, and was easily prepared, so it was suitable for being popularized in countryside. The sperm survival rate of boar semen preserved in frequently - used glucose diluent for 3 days expressed not so well, therefore the prepared glucose diluent should be quickly used for the boar semen preservation. The preservation effect of BL-liquid (made in America) was better, but this diluent had complex components and high cost, and was hardly prepared, so it could only be popularized in hoggery.

  18. Ubiquitin-activating enzyme (UBA1) is required for sperm capacitation, acrosomal exocytosis and sperm-egg coat penetration during porcine fertilization.

    Science.gov (United States)

    Yi, Y-J; Zimmerman, S W; Manandhar, G; Odhiambo, J F; Kennedy, C; Jonáková, V; Maňásková-Postlerová, P; Sutovsky, M; Park, C-S; Sutovsky, P

    2012-04-01

    Protein ubiquitination is a stable, covalent post-translational modification that alters protein activity and/or targets proteins for proteolysis by the 26S proteasome. The E1-type ubiquitin-activating enzyme (UBA1) is responsible for ubiquitin activation, the initial step of ubiquitin-protein ligation. Proteasomal proteolysis of ubiquitinated spermatozoa and oocyte proteins occurs during mammalian fertilization, particularly at the site of sperm acrosome contact with oocyte zona pellucida. However, it is not clear whether the substrates are solely proteins ubiquitinated during gametogenesis or if de novo ubiquitination also occurs during fertilization supported by ubiquitin-activating and -conjugating enzymes present in the sperm acrosome. Along this line of inquiry, UBA1 was detected in boar sperm-acrosomal extracts by Western blotting (WB). Immunofluorescence revealed accumulation of UBA1 in the nuclei of spermatogonia, spermatocytes and spermatids, and in the acrosomal caps of round and elongating spermatids. Thiol ester assays utilizing biotinylated ubiquitin and isolated sperm acrosomes confirmed the enzymatic activity of the resident UBA1. A specific UBA1 inhibitor, PYR-41, altered the remodelling of the outer acrosomal membrane (OAM) during sperm capacitation, monitored using flow cytometry of fluorescein isothiocyanate-conjugated peanut agglutinin (FITC-PNA). Although viable and motile, the spermatozoa capacitated in the presence of PYR-41, showed significantly reduced fertilization rates during in vitro fertilization (IVF; p sperm capacitation and acrosomal function during fertilization.

  19. Optimization of a sperm-oviduct binding assay mimicking in vivo conditions. Adoption of sperm separation methods and protocols for analysing sperm motility and intracellular Ca2+ level

    OpenAIRE

    Narud, Birgitte

    2014-01-01

    English: An in vitro model that mimics the interactions between spermatozoa and oviductal epithelial cells can be used to increase the knowledge about the function of the oviduct and the formation of a sperm reservoir in vivo. The aim of the present study was to optimize methods for culturing bovine epithelial cells (BOECs) bi-dimensionally on plastic and three-dimensionally on polyester membrane. These cells were used in a sperm binding assay for evaluation of sperm-BOEC binding and relea...

  20. Sperm capacitation in the porcine oviduct.

    Science.gov (United States)

    Tienthai, P; Johannisson, A; Rodriguez-Martinez, H

    2004-01-01

    In vitro studies suggests that sperm capacitation occurs in the sperm reservoir (SR) of the pig, with spermatozoa progressing towards the ampullary-isthmic junction (AIJ) around ovulation as a consequence of capacitation/hyperactivation. In contrast, in vivo studies are scarce. Consequently, we determined the degree of capacitation in boar spermatozoa that were retrieved from the SR of sows at well-defined periods of spontaneous standing oestrus, namely pre-, peri- and post-ovulation, using flow cytometry of Merocyanine-540/Yo-Pro-1-loaded spermatozoa. SR-spermatozoa retrieved and incubated in non-capacitating medium (bicarbonate-free mBO [mBO-]) were largely viable (70-85%) and uncapacitated (69-73%), irrespective of the stage of oestrus considered. Those undergoing capacitation were a minor proportion (1-5%) during pre- and peri-ovulation, but they significantly increased (14%) in post-ovulation oestrus. To clarify whether these SR-spermatozoa were able to undergo capacitation under stimuli, sperm aliquots were challenged in vitro either by incubation in a bicarbonate-rich medium (capacitation medium, mBO+), then further in mBO+ with 20% (v/v) of in vivo collected homologous pre-ovulatory isthmic oviductal fluid (IOF), or incubation with hyaluronan (HA, 500 microg/ml). Exposure to mBO+ significantly increased the sub-population of capacitated spermatozoa from the pre- and peri-ovulation SR, indicating that the uncapacitated SR-spermatozoa were responsive to the effector/inducer bicarbonate at levels recorded in peri-ovulatory AIJ/ampulla in vivo. While addition of IOF or HA to SR-spermatozoa incubated in capacitating medium (mBO+) maintained sperm viability without obviously inducing capacitation in pre- or peri-ovulatory SR-spermatozoa, they significantly increased these percentages during post-ovulation, when compared to baseline values of control incubations (mBO-). The results suggest that massive sperm capacitation does not occur in vivo in the porcine SR

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

    Science.gov (United States)

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

    2016-01-01

    ABSTRACT Mammalian spermatozoa utilize ATP as the energy source for key functions on the route to fertilization. ATP and its precursor nucleotides ADP and AMP are regularly investigated in sperm physiology studies, mostly by bioluminescence assays. Assay results vary widely, mainly due to different efficiencies in nucleotide extraction and prevention of their enzymatic degradation. Here, we describe a revised, validated protocol for efficient phosphatase inhibition and adenine nucleotide extraction resulting in consistently high ATP concentrations exceeding previously reported values for boar spermatozoa up to 20-fold. The revised assay is applicable for determining ATP concentrations and adenylate energy charge in extracts from fresh and frozen samples, thereby allowing simultaneous assessment of semen samples from long-term storage experiments. After validation, the assay was applied to liquid-preserved boar spermatozoa stored at 17°C and 5°C for 24 and 72 h. Cooling to 5°C, but not storage duration, reduced ATP concentration in spermatozoa (P<0.05), which was accompanied by the appearance of AMP and ADP in the preservation medium. ATP and energy charge were highly correlated to the proportion of membrane-intact spermatozoa, supporting the idea of nucleotides leaking through disrupted membranes in cold-shocked cells. The present assay allows highly standardized studies of energy metabolism in spermatozoa. PMID:27612509

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

    Directory of Open Access Journals (Sweden)

    Quynh Thu Nguyen

    2016-11-01

    Full Text Available Mammalian spermatozoa utilize ATP as the energy source for key functions on the route to fertilization. ATP and its precursor nucleotides ADP and AMP are regularly investigated in sperm physiology studies, mostly by bioluminescence assays. Assay results vary widely, mainly due to different efficiencies in nucleotide extraction and prevention of their enzymatic degradation. Here, we describe a revised, validated protocol for efficient phosphatase inhibition and adenine nucleotide extraction resulting in consistently high ATP concentrations exceeding previously reported values for boar spermatozoa up to 20-fold. The revised assay is applicable for determining ATP concentrations and adenylate energy charge in extracts from fresh and frozen samples, thereby allowing simultaneous assessment of semen samples from long-term storage experiments. After validation, the assay was applied to liquid-preserved boar spermatozoa stored at 17°C and 5°C for 24 and 72 h. Cooling to 5°C, but not storage duration, reduced ATP concentration in spermatozoa (P<0.05, which was accompanied by the appearance of AMP and ADP in the preservation medium. ATP and energy charge were highly correlated to the proportion of membrane-intact spermatozoa, supporting the idea of nucleotides leaking through disrupted membranes in cold-shocked cells. The present assay allows highly standardized studies of energy metabolism in spermatozoa.

  3. Effect of commercial long-term extenders on metabolic activity and membrane integrity of boar spermatozoa stored at 17 degrees C.

    Science.gov (United States)

    Dziekońska, A; Fraser, L; Majewska, A; Lecewicz, M; Zasiadczyk, Ł; Kordan, W

    2013-01-01

    This study was aimed to analyze the metabolic activity and membrane integrity of boar spermatozoa following storage in long-term semen extenders. Boar semen was diluted with Androhep EnduraGuard (AeG), DILU-Cell (DC), SafeCell Plus (SCP) and Vitasem LD (VLD) extenders and stored for 10 days at 17 degrees C. Parameters of the analyzed sperm metabolic activity included total motility (TMOT), progressive motility (PMOT), high mitochondrial membrane potential (MMP) and ATP content, whereas those of the membrane integrity included plasma membrane integrity (PMI) and normal apical ridge (NAR) acrosome. Extender type was a significant (P sperm parameters, except for ATP content. Furthermore, the storage time had a significant effect (P sperm metabolic activity and membrane integrity during semen storage. In all extenders the metabolic activity and membrane integrity of the stored spermatozoa decreased continuously over time. Among the four analyzed extenders, AeG and SCP showed the best performance in terms of TMOT and PMI on Days 5, 7 and 10 of storage. Marked differences in the proportions of spermatozoa with high MMP were observed between the extenders, particularly on Day 10 of storage. There were not any marked differences in sperm ATP content between the extenders, regardless of the storage time. Furthermore, the percentage of spermatozoa with NAR acrosomes decreased during prolonged storage, being markedly lower in DC-diluted semen compared with semen diluted with either AeG or SCP extender. The results of this study indicated that components of the long-term extenders have different effects on the sperm functionality and prolonged semen longevity by delaying the processes associated with sperm ageing during liquid storage.

  4. 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.

  5. Serologic tests for detecting antibodies against Mycobacterium bovis and Mycobacterium avium subspecies paratuberculosis in Eurasian wild boar (Sus scrofa scrofa).

    Science.gov (United States)

    Boadella, Mariana; Lyashchenko, Konstantin; Greenwald, Reena; Esfandiari, Javan; Jaroso, Raquel; Carta, Tania; Garrido, Joseba M; Vicente, Joaquín; de la Fuente, José; Gortázar, Christian

    2011-01-01

    New tools to detect exposure of free-range Eurasian wild boar (Sus scrofa scrofa) to pathogenic mycobacteria would be valuable for improved disease surveillance and wildlife management. Two hundred sera from wild boar of known Mycobacterium bovis infection status were used to evaluate test suitability for the detection of antibodies against M. bovis and Mycobacterium avium subsp. paratuberculosis (or cross-reacting members of the M. avium complex). Two traditional enzyme-linked immunosorbent assays were evaluated using M. bovis purified protein derivative (bPPD) and paratuberculosis protoplasmatic antigen 3 (PPA3) as antigens, respectively, and a new point-of-care test format for bovine tuberculosis (bTB) that uses the innovative dual-path platform (DPP TB) test. The effect of individual factors (sex, age, lesions) on the diagnostic performance of the serologic tests was also determined. Although the DPP had a sensitivity of 89.6% and a specificity of 90.4%, for bPPD, the sensitivity was 79.2% and the specificity 100%. Both tests had a kappa agreement of 0.80. Sixty-five of 68 (95.6%) wild boar sera with antibodies against the PPA3 antigen corresponded to known M. bovis-infected wild boar. Significant differences were not observed in the bPPD and DPP readings among lesion categories or between age classes. A slight sex-related difference in sensitivity toward males in the DPP was found, but it was not detected in the bPPD enzyme-linked immunosorbent assay. The results support the use of antibody-based diagnostic tests for both large-scale and individual bTB testing of Eurasian wild boar and suggest that wild boar cannot be used as sentinels for infections caused by M. avium complex members.

  6. 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.

  7. Efficacy of sperm mobility assessment in commercial flocks and the relationships of sperm mobility and insemination dose with fertility in turkeys.

    Science.gov (United States)

    King, L M; Kirby, J D; Froman, D P; Sonstegard, T S; Harry, D E; Darden, J R; Marini, P J; Walker, R M; Rhoads, M L; Donoghue, A M

    2000-12-01

    Our objectives were to evaluate: 1) the efficacy of the Sperm Mobility Test on commercial turkey farms, and 2) the influence of sperm mobility phenotype on fertility when insemination parameters are varied. In research flocks, differences in sperm mobility among toms are predictive of fertility. We wanted to test the efficacy of this sire selection test in practical, real-world situations, evaluating its usefulness in terms of assessing large numbers of toms, different strains of turkeys, and variable management practices. Utilizing field study results, controlled studies were then conducted to improve test parameters. For the field trials, semen from each of 405 breeder toms (11 strains or lines) was evaluated either in duplicate (n = 285) or in triplicate (n = 120). Sperm mobility was normally distributed among all toms tested, except for one strain. Because the sperm mobility indices for toms evaluated in these field trials were higher than those observed in research flocks, the Sperm Mobility Test was modified to increase the separation between high and low sperm mobility phenotypes by increasing the concentration of Accudenz. To determine the effects of sperm mobility and insemination dose on sustained fertility through time, hens from a research flock were inseminated twice before the onset of lay with sperm from toms classified as high-, average-, or low-mobility in concentrations of 25 to 400 million sperm per artificial insemination dose, and egg fertility was evaluated over a 5-wk period. Toms with the high-mobility sperm phenotype maintained higher fertility (P insemination doses compared with toms with low-mobility sperm. Toms with high-mobility sperm sired equal numbers of poults in a sperm competition study in which numbers favored low-mobility toms by 3:1. These results demonstrate that the Sperm Mobility Test can be used for on-farm evaluation of semen quality of toms in commercial flocks and that sperm mobility influences fertility and sire fitness.

  8. Cytological evaluation of spermatogenesis: a novel and simple diagnostic method to assess spermatogenesis in non-obstructive azoospermia using testicular sperm extraction specimens

    NARCIS (Netherlands)

    Hessel, M.L.; Vries, M. de; D'Hauwers, K.W.M.; Fleischer, K.; Hulsbergen-van de Kaa, C.A.; Braat, D.D.M.; Ramos, L.

    2015-01-01

    Most of the non-obstructive azoospermia (NOA)-patients have only focal spermatogenesis which results in insufficient numbers of spermatozoa to reach the ejaculate. In approximately 50% of these NOA-patients testicular sperm extraction (TESE) is successful and intracytoplasmic sperm injection (ICSI)

  9. 不同品种和季节对种公猪精液品质的影响%Influences of Breed and Season on Boar Semen Quality

    Institute of Scientific and Technical Information of China (English)

    朱志付; 柳云华; 周健雄; 赵国庆; 李成效; 陶劲松; 曹林; 孔凡勇

    2016-01-01

    In order to investigate the effects of breed and season on boar semen quality, the semen quality for 3 years of 3 pig breeds were detected and compared, which simpled from 28 Landrace, 87 Large Yorkshire and 98 Duroc boars. The results showed that ejaculate volume of Duroc boars was significantly lower than those of Lan-drace and Large Yorkshire (P0.05); the highest ejaculate volume of Landrace and Large Yorkshire boars was showed in winter (P>0.05), the sperm density of the 3 breeds boars in summer was lower than that in the winter (P0.05); in the same season, the e-jaculate volume of Duroc boars was lower than that of Landrace and Large Yorkshire (P>0.05), and the sperm density was higher than that of the other breeds(P>0.05), the significant difference was showed on ejaculate vol-ume between Duroc and Landrace in winter(P<0.05), and the sperm motility of Large Yorkshire boars was better than those of Landrace and Duroc. It indicated that the breed and season had some effects on boar semen quali-ty. It was advantageous to improve boar semen quality by lowering the temperature in the hot summer.%为研究不同品种和季节对公猪精液品质的影响,以28头长白公猪、87头大约克公猪、98头杜洛克公猪为研究对象,对其3年的精液品质进行了检测和分析。结果表明,杜洛克公猪的采精量显著或极显著小于长白、大约克,大约克公猪的精子活力显著优于长白公猪(P<0.05),不同品种的精子密度差异不显著(P>0.05);长白、大约克公猪的采精量以冬季最多,3个品种公猪的精子密度夏季显著或极显著小于冬季,精子活力四季差异不显著(P>0.05);在同一季节,杜洛克公猪的采精量均小于长白、大约克,且冬季显著小于长白(P<0.05),精子密度优于长白、大约克公猪(P>0.05),大约克公猪的精子活力优于长白、杜洛克。结果显示,品种和季节对公猪精液品质有一

  10. 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.

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

    Institute of Scientific and Technical Information of China (English)

    张洁; 陈敏恒; 李宝红; 王均亮; 苏文昌; 彭国良; 李剑豪

    2015-01-01

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

  12. Bovine tuberculosis in a wild boar (Sus scrofa) in Poland.

    Science.gov (United States)

    Krajewska, Monika; Lipiec, Marek; Zabost, Anna; Augustynowicz-Kopeć, Ewa; Szulowski, Krzysztof

    2014-10-01

    Poland is officially tuberculosis free and bovine tuberculosis (BTB) cases are rarely found except in bovids. We found BTB in a wild boar (Sus scrofa) in the Bieszczady Mountains, southeastern Poland. Studies suggest possible transmission of infection between free-living European bison (Bison bonasus caucasicus) and wild boar in this area.

  13. Perceptual masking of boar taint in Swedish fermented sausages.

    Science.gov (United States)

    Stolzenbach, Sandra; Lindahl, Gunilla; Lundström, Kerstin; Chen, Gang; Byrne, Derek V

    2009-04-01

    Surgical castration of male piglets has traditionally been practiced to avoid development of boar taint in pork meat which can occur if entire male pigs are raised. Boar taint is commonly characterised as exhibiting the odour and flavour of urine and manure. This study involves sensory characterisation of the possibilities to mask boar taint in meat from entire male pigs by fermentation and smoking to maintain high sensory quality in meat products if castration is prohibited. Model and commercial type Swedish fermented sausage products based on low or high levels of boar tainted fat, three different starter cultures and two different levels of smoking were studied. In the model sausages, liquid smoke masked the perception of boar taint. In contrast, the smoking procedure of the commercial sausages was insufficient to totally mask the perception of boar taint. In both the model and commercial sausages, the aroma development from the starter cultures lowered the perception of boar taint but was insufficient for total perceptual masking. Due to the total masking effect of smoking in the model sausages, it was clear that smoke may present a potential solution to remove the perception of boar taint in fermented sausages if the smoking procedure is optimised.

  14. First production of larvae using cryopreserved sperm: Effects of preservation temperature and cryopreservation on European eel sperm fertilization capacity

    DEFF Research Database (Denmark)

    Asturiano, J.F.; Sørensen, Sune Riis; Perez, L.

    2016-01-01

    Sperm cryopreservation is a useful tool in captive fish reproduction management, that is to synchronize gamete production, especially in the case of species as the European eel, where the time of female spawning readiness is unpredictable. Several protocols to cryopreserve sperm of this species...... have been described, but until recently fertilization trials were not feasible. This study evaluated the effect of cold storage of diluted sperm prior to fertilizations and tested whether a previously defined protocol for European eel sperm cryopreservation can be successfully applied in fertilization...... trials to produce viable offspring. In our experiment, the sperm motility was evaluated after the extraction and the best samples were selected and pooled. Until stripping of eggs and fertilization, diluted sperm samples were maintained at either 4 or 20°C, or cryopreserved, following existing protocols...

  15. I know what you did last summer - High resolution mapping of wild boar damages with drones

    OpenAIRE

    2014-01-01

    Wild boar ability to cause important damages to agricultural lands is not anymore to be demonstrated. These damages often raise conflicts between farmers and hunters due to the associated economic losses. Objective and accurate method for real impact assessment of losses suffered by farmers is thus needed. Currently,in Wallonia damage assessment is performed from the ground by experts, asked to evaluate precisely the area impacted. This task is time and man power consuming. Moreover, damaged ...

  16. Different computer-assisted sperm analysis (CASA) systems highly influence sperm motility parameters.

    Science.gov (United States)

    Boryshpolets, S; Kowalski, R K; Dietrich, G J; Dzyuba, B; Ciereszko, A

    2013-10-15

    In this study, we examined different computer-assisted sperm analysis (CASA) systems (CRISMAS, Hobson Sperm Tracker, and Image J CASA) on the exact same video recordings to evaluate the differences in sperm motility parameters related to the specific CASA used. To cover a wide range of sperm motility parameters, we chose 12-second video recordings at 25 and 50 Hz frame rates after sperm motility activation using three taxonomically distinct fish species (sterlet: Acipenser ruthenus L.; common carp: Cyprinus carpio L.; and rainbow trout: Oncorhynchus mykiss Walbaum) that are characterized by essential differences in sperm behavior during motility. Systematically higher values of velocity and beat cross frequency (BCF) were observed in video recordings obtained at 50 Hz frame frequency compared with 25 Hz for all three systems. Motility parameters were affected by the CASA and species used for analyses. Image J and CRISMAS calculated higher curvilinear velocity (VCL) values for rainbow trout and common carp at 25 Hz frequency compared with the Hobson Sperm Tracker, whereas at 50 Hz, a significant difference was observed only for rainbow trout sperm recordings. No significant difference was observed between the CASA systems for sterlet sperm motility at 25 and 50 Hz. Additional analysis of 1-second segments taken at three time points (1, 6, and 12 seconds of the recording) revealed a dramatic decrease in common carp and rainbow trout sperm speed. The motility parameters of sterlet spermatozoa did not change significantly during the 12-second motility period and should be considered as a suitable model for longer motility analyses. Our results indicated that the CASA used can affect motility results even when the same motility recordings are used. These results could be critically altered by the recording quality, time of analysis, and frame rate of camera, and could result in erroneous conclusions.

  17. Serological anthrax surveillance in wild boar (Sus scrofa) in Ukraine.

    Science.gov (United States)

    Bagamian, Karoun H; Skrypnyk, Artem; Rodina, Yana; Bezymennyi, Maksym; Nevolko, Oleg; Skrypnyk, Valeriy; Blackburn, Jason K

    2014-08-01

    Anthrax, caused by Bacillus anthracis, is an acute disease affecting wildlife, livestock, and humans worldwide, although its impact on these populations is underappreciated. In Ukraine, surveillance is passive, and anthrax is often detected in livestock. However, wildlife is not subject to surveillance, although anthrax deaths (such as in wild boar, Sus scrofa) have been documented. The wild boar is a plentiful and widespread species in Ukraine and is frequently hunted. We initiated a screening study testing Ukrainian wild boar blood samples for antibodies to B. anthracis. We mapped results relative to known livestock anthrax hotspots. We discovered evidence of exposure in wild boar up to 35 km from livestock anthrax hotspots and over 400 km from previous anthrax reports in boars. We make recommendations about using wildlife species as biosentinels for anthrax in Ukraine.

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

    DEFF Research Database (Denmark)

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

    2003-01-01

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

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

    Science.gov (United States)

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

    2016-01-15

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

  20. 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.

  1. Effects of hepatitis B virus infection on human sperm chromosomes

    Institute of Scientific and Technical Information of China (English)

    Jian-Min Huang; Tian-Hua Huang; Huan-Ying Qiu; Xiao-Wu Fang; Tian-Gang Zhuang; Hong-Xi Liu; Yong-Hua Wang; Li-Zhi Deng; Jie-Wen Qiu

    2003-01-01

    AIM: To evaluate the level of sperm chromosome aberrations in male patients with hepatitis B, and to directly detect whether there are HBV DNA integrations in sperm chromosomes of hepatitis B patients.METHODS: Sperm chromosomes of 14 tested subjects (5healthy controls, 9 patients with HBV infection, including 1with acute hepatitis B, 2 with chronic active hepatitis B, 4with chronic persistent hepatitis B, 2 chronic HBsAg carriers with no clinical symptoms) were prepared using interspecific in vitro fertilization between zona-free golden hamster ova and human spermatozoa, and the frequencies of aberration spermatozoa were compared between subjects of HBV infection and controls. Fluorescence in situ hybridization (FISH) to sperm chromosome spreads was carried out with biotin-labeled full length HBV DNA probe to detect the specific HBV DNA sequences in the sperm chromosomes.RESULTS: The total frequency of sperm chromosome aberrations in HBV infection group (14.8%, 33/223) was significantly higher than that in the control group (4.3%,5/116). Moreover, the sperm chromosomes in HBV infection patients commonly presented stickiness, clumping, failure to staining, etc, which would affect the analysis of sperm chromosomes. Specific fluorescent signal spots for HBV DNA were seen in sperm chromosomes of one patient with chronic persistent hepatitis. In 9 (9/42) sperm chromosome complements containing fluorescent signal spots, one presented 5 obvious FISH spots, others presented 2 to 4signals. There was significant difference of fluorescence intensity among the signal spots. The distribution of signal sites among chromosomes was random.CONCLUSION: HBV infection can bring about mutagenic effects on sperm chromosomes. Integrations of viral DNA into sperm chromosomes which are multisites and nonspecific, can further increase the instability of sperm chromosomes. This study suggested that HBV infection can create extensively hereditary effects by alteration genetic constituent and

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

    Directory of Open Access Journals (Sweden)

    D.J. Ackerman

    1996-02-01

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

  3. Measurement and Analysis on the Semen Quality of Dahe Black Pigs with Boar%大河乌猪种公猪精液品质测定与分析

    Institute of Scientific and Technical Information of China (English)

    李祥; 李娇艳; 许泽现; 龙罡; 李俊

    2016-01-01

    The Dahe black boars were divided into three group according to the kind of the age in this paper, and the semen volume,semen specific gravity,sperm activity,sperm density,sperm abnormality rate and sperm morphology were systematically analyzed.The result of the indicators of the Dahe black boars' semen showed that semen volumes were (233.23±78.93) mL,(228.66±80.06) g,the proportion of sperm was (1.02±0.01) g/mL,sperm activity was(71.26±8.48)%,sperm density was(3.27±1.17)×108 per mL,sperm abnormality rate was (7.89±7.51)%, respectively.The semen volume,sperm activity and sperm density were significantly effected by the age.Sperm motility and sperm concentration was moderate positive correlation with the coefficient of 0.465 (P<0.01).The indexes of sperm morphology among different age groups were not significantly different.%试验按大河乌猪种公猪年龄分3组,系统测定采精量、精液比重、精子活力、精子密度、精子畸形率和精子形态.结果显示,大河乌猪采精量按体积测定为(233.23±78.93) mL、按质量测定为(228.66±80.06) g,精液比重为(1.02±0.01) g/mL,精子活力为(71.26±8.48)%,精子密度为(3.27±1.17)亿个/mL,畸形率为(7.89±7.51)%. 采精量、精子活力、精子密度受年龄的影响明显,精子活力与精子密度呈中等正相关,相关系数为0.465(P<0.01);不同年龄间精子形态各项指标差异不显著.

  4. Sperm dilution ratio affects post-thaw motility rate and velocity of Prochilodus lineatus (Characiformes) sperm.

    Science.gov (United States)

    Viveiros, Ana T M; Leal, Marcelo C

    2016-10-01

    There is a lack of standardization in sperm cryopreservation of aquatic organisms and, thus, a necessity of more accurate investigations in all steps of this process. In this study, the effects of sperm dilution ratio on post-thaw sperm quality of Prochilodus lineatus were evaluated. Sperm was diluted in a standard freezing medium (glucose and methyl glycol) at four different ratios (sperm to final volume = 1:5, 1:10, 1:50 or 1:100), frozen in a nitrogen vapour vessel at -170°C and then stored in liquid nitrogen vessel at -196°C. Post-thaw motility rate and velocities (curvilinear = VCL; average path = VAP; straight line = VSL) were determined using a Computer-Assisted Sperm Analyzer (CASA) at 10 and 40 s post-activation. The highest motility rates were observed when sperm was frozen at a ratio of 1:5 (76%) and 1:10 (75%). The highest VCL (225 μm/s) and VAP (203 μm/s) were observed at a ratio of 1:10, while VSL was similar among samples frozen at 1:5, 1:10 and 1:50 (97-124 μm/s). When those parameters were evaluated again 30 s later, motility decreased significantly in samples frozen at a ratio of 1:5 (57%) and 1:10 (61%), while velocities decreased significantly in all samples regardless of dilution ratio (75-85 μm/s of VCL, 38-53 μm/s of VAP and 25-39 μm/s of VSL). P. lineatus sperm should be frozen at a ratio of 1:10, where both the number of loaded sperm per straw and the post-thaw quality are maximized.

  5. Effects of Aging on Spermatogenesis, Sperm Maturation and Fertility in Mice

    Institute of Scientific and Technical Information of China (English)

    Qiu-ju CHEN; Wei-jie ZHU; Jing LI

    2006-01-01

    Objective To investigate effects of aging on spermatogenesis in testis, sperm maturation in epididymis, and fertility in mice.Methods Testicular specimens, caput epididymal sperm and cauda epididymal sperm were obtained from Kuming mice (18-month aged group, n=15; 6-month young group as control, n=15). The testicular histological examinations and quantitative evaluations on spermatogenesis were performed. Sperm parameters including sperm density, sperm viability, sperm motility, and normal morphological rate were assessed. The fertilization rate and embryo development were measured by in vitro fertilization and embryo culture.Results The histological changes of testes in aged mice were mainly seminiferous tubule atrophy and hypospermatogenesis. In aged testes, a significant decline was found in the numbers of round spermatids and elongated spermatids per Sertoli cell (P<0.01). Sperm density, sperm motility and normal morphological rate in caput epididymis and cauda epididymis in aged mice significantly decreased (P<0. 05). The fertilization rate and embryo development of aged group were lower than those in the control(P< 0.01).Conclusions Spermatogenesis and sperm functions could be maintained in the aging male. However, aging affects spermatogenesis and sperm maturation, which leads to lower the quality of sperm, including sperm fertilizing capacity. The development of embryo from aging sperm would have more abnormalities.

  6. 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

  7. The Expression and Tyrosine Phosphorylation of Sperm Protein 32 Regulate the Activation of the Boar Proacrosin/Acrosin System%精子蛋白32的表达及酪氨酸磷酸化调控猪精子顶体蛋白的活化

    Institute of Scientific and Technical Information of China (English)

    孙培亮; 崔明勋; 姜园园; 曹立朋; 金一

    2013-01-01

    为了探究猪精子蛋白32(Sperm protein,sp32)表达及酪氨酸磷酸化调控猪精子顶体蛋白活化的关系,本研究对不同处理(鲜精、冷冻-解冻、获能、顶体反应)的猪精子顶体膜蛋白进行分离,通过考马斯亮蓝染色、SDS-PAGE电泳和Western blot检测和分析.结果表明,猪精子经过获能处理、冷冻-解冻处理以及顶体反应处理后,前顶体蛋白(Proacrosin)和顶体蛋白(Acrosin)在转化过程中,sp32表达有所差异,获能和顶体反应处理的sp32的表达量略高于冷冻-解冻处理组,而显著高于鲜精处理组.与其他处理组相比,冷冻-解冻精子处理组sp32酪氨酸磷酸化水平产生显著的差异.但在SDS-PAGE电泳中,猪鲜精处理组在蛋白质分子质量38~170 ku区间的蛋白表达条带较其他对比组明显,这表明猪精子在受精前所必需经历的获能和顶体反应过程中,顶体膜蛋白伴随着大分子的蛋白质修饰和降解.结论:sp32作为一种前顶体蛋白结合蛋白,在前顶体蛋白活化过程中它的表达水平及酪氨酸磷酸化水平上调.

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

    Directory of Open Access Journals (Sweden)

    Wiebke Garrels

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

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

    Directory of Open Access Journals (Sweden)

    Jesús L Yániz

    2016-01-01

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

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

    Science.gov (United States)

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

    2016-01-01

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

  11. Effect of semen preparation technique and its incubation on sperm quality in the Moroccan population.

    Science.gov (United States)

    Aboulmaouahib, S; Madkour, A; Kaarouch, I; Saadani, B; Sefrioui, O; Louanjli, N; Copin, H; Cadi, R; Benkhalifa, M

    2016-09-05

    In in vitro fertilisation (IVF), sperm preparation as critical part and influencing the sperm quality is especially dependent on the chosen technique itself and incubation parameters including temperature and CO2. In this study, we compared firstly density-gradient centrifugation technique (DGC) to the adapted DGC using the sperm pellet of 80% fraction (DGC/80P) in order to improve the sperm yield. Secondly, this study led to evaluate different sperm incubation conditions based on temperature effect (room temperature (RT = 23°C) versus 35°C) and in the other hand, with or without 5% CO2 during 24 hrs. Based on evaluating sperm conventional parameters and the DNA damage using TUNEL assay, our result showed that DGC/80P increased sperm quality compared to DGC with 25% of improvement. For temperature incubation effect after 24 hrs, 35°C increased the DNA damage and decreased the sperm quality while RT could improve sperm motility by 38%. Moreover, the sperm incubation with 5% CO2 after 24 hrs realised a negative impact on sperm parameters and its DNA damage. Indeed, for current IVF practice, a good sperm quality can be maintained for several hours at room temperature, while the sperm preparation is processed using the DGC/80P without CO2.

  12. A new method for sperm characterization for infertility treatment: hypothesis testing by using combination of watershed segmentation and graph theory.

    Science.gov (United States)

    Shojaedini, Seyed Vahab; Heydari, Masoud

    2014-10-01

    Shape and movement features of sperms are important parameters for infertility study and treatment. In this article, a new method is introduced for characterization sperms in microscopic videos. In this method, first a hypothesis framework is defined to distinguish sperms from other particles in captured video. Then decision about each hypothesis is done in following steps: Selecting some primary regions as candidates for sperms by watershed-based segmentation, pruning of some false candidates during successive frames using graph theory concept and finally confirming correct sperms by using their movement trajectories. Performance of the proposed method is evaluated on real captured images belongs to semen with high density of sperms. The obtained results show the proposed method may detect 97% of sperms in presence of 5% false detections and track 91% of moving sperms. Furthermore, it can be shown that better characterization of sperms in proposed algorithm doesn't lead to extracting more false sperms compared to some present approaches.

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

    Science.gov (United States)

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

    2014-09-01

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

  14. Stable Variants of Sperm Aneuploidy among Healthy Men Show Associations between Germinal and Somatic Aneuploidy

    OpenAIRE

    Rubes, Jiri; Vozdova, Miluse; Robbins, Wendie A.; Rezacova, Olga; Perreault, Sally D; Andrew J. Wyrobek

    2002-01-01

    Repeated semen specimens from healthy men were analyzed by sperm fluorescence in situ hybridization (FISH), to identify men who consistently produced elevated frequencies of aneuploid sperm and to determine whether men who were identified as stable variants of sperm aneuploidy also exhibited higher frequencies of aneuploidy in their peripheral blood lymphocytes. Seven semen specimens were provided by each of 15 men over a 2-year period and were evaluated by the X-Y-8 multicolor sperm FISH met...

  15. Effects of wild boar grazing on the yield of summer truffle (Tuscany, Italy

    Directory of Open Access Journals (Sweden)

    Elena Salerni

    2013-12-01

    Full Text Available The research presented here seeks to describe the impact of wild boar to a natural truffle ground of Tuber aestivum Vittad. on Monte Amiata (Tuscany – Italy. Pedoclimatic analyses indicated that the selected area could be considered suitable for the truffle production. Then classification of the vegetation of a Quercus cerris forest was carried out exploring the possibility of the BACI (Before-After-Control-Impact sampling design. Finally 10 plots were selected, half of which have been fenced. For the first time the impact of wild boar was evaluated by estimating the surface area turned over by its activity. Moreover in each plot the number and weight of summer truffles was performed every 10 days during the fruiting period (June-November 2006-2008. The hypothesis that the presence of Sus scrofa has a strong negative influence on truffle harvesting has been amply confirmed by the data presented here, given the large increase of fruiting bodies of the summer truffle collected in the fenced plots. Consequently the destructive behaviour of the wild boar imply not only an ecological but also an economic damage in areas in which non-wood forest products are an important source of income.

  16. Chemotactic Maneuverability of Sperm

    CERN Document Server

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

    2011-01-01

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

  17. Effect of Cysteamine on Tibetan Boar Semen Preserved at 4℃and Antioxidative%半胱胺对4℃保存藏猪精液指标的影响

    Institute of Scientific and Technical Information of China (English)

    陈晓英; 马金英; 宋天增; 唐建华; 赵彦玲; 任子利; 朱彦宾; 王良润

    2016-01-01

    This study was conducted to investigate the effects of Cysteamine on Tibetan boar semen preserved at 4℃and antioxidative. The semen was collected from Tibetan boar by electrical stimulation. Add Cysteamine 0 g, 0.02 g, 0.04 g, 0.06 g, 0.08 g per 1 000 mL, in the improvement of pig semen low temperature dilution liquid. Save to fifth days at 4℃after dilution of fresh semen. Body integrity and membrane integrity rate of the top Tibetan pig sperm activity, sperm abnormal rate and semen quality evaluation index, to investigate the effect of Cysteamine on the quality of se-men quality at 4℃, the activity of Superoxidas dismutase (SOD) and Maleic dialdehyde (MDA) were the indexes to e-valuate the protective effect of antioxidation. The results show that, The sperm motility, sperm deformity rate, sperm motility rate and plasma membrane integrity rate of the Cysteamine 0.06 g/L addition group were significantly better than other test groups(P<0.05), Seminal superoxide dismutase(SOD) activity and malondialdehyde(MDA) content were significantly better than other experimental groups (P<0.05), added Cysteamine was in pig semen at low temperature. Also, at the same time, we found that, 0.08 g/L Cysteamine supplementation group sperm superoxide dismutase (SOD) activity is the highest, But the difference was not significant between the groups. And The content of MDA in sperm was highest in 0.06 g/L group. In short, adding 0.06 g/L Cysteamine to the improved pig semen diluent and semen to do 4 times dilution, it can significantly improve the semen quality and antioxidant protection at 4℃. The experimental study laid a foundation for the production and application of cryopreservation of boar semen in the future.%试验旨在研究半胱氨酸对4℃藏猪精液品质和抗氧化保护作用影响,采用电刺激法采集健康藏猪精液,在改进的猪精液低温稀释液为基础液中,每1000 mL添加半胱氨酸0、0.02、0.04、0.06、0.08 g,将

  18. Improved risk assessment by screening sperm parameters.

    Science.gov (United States)

    Plassmann, S; Urwyler, H

    2001-02-28

    treatment, but mating trials were necessary for confirmation of the toxic effect. Without sperm motility examination, this effect would have been missed in early drug development causing problems in clinical studies. Further comparisons of the validity of 4 or 9 weeks treatment require the testing of other compounds with different targets/mechanism of actions, as well as evaluation of dose-response relationships.

  19. Thyroxin Is Useful to Improve Sperm Motility

    Directory of Open Access Journals (Sweden)

    Mendeluk Gabriela Ruth

    2016-07-01

    Full Text Available Background The aim of this study was to evaluate the non-genomic action of thyroxin on sperm kinetic and its probable use to improve sperm recovery after applying an en- richment method like “swim-up” in comparison with the available one, pentoxifylline. Materials and Methods This is an experimental study. A total of 50 patients were re- cruited, followed by infertility consultation. Conventional sperm assays were performed according to World Health Organization criteria-2010 (WHO-2010. A Computer Aided Semen Analysis System was employed to assess kinetic parameters and concentrations. Number of the motile sperm recovered after preparation technique was calculated. Results Addition of T4 (0.002 µg/ml to semen samples increased hypermotility at 20 minutes (control: 14.18 ± 5.1% vs. 17.66 ± 8.88%, P<0.03, data expressed as mean ± SD and remained unchanged after 40 minutes. Significant differences were found in the motile sperm recovered after swim-up (control: 8.93×106 ± 9.52× 06vs. 17.20×106 ± 21.16×106, P<0.03, achieving all of the tested samples a desirable threshold value for artificial insemination outcome, while adding pentoxifylline increased the number of recovered sperm after swim-up in 60% of the studied cases. No synergism between two treatments could be determined. Conclusion We propose a new physiological tool to artificially improve insemination. The discussion opens windows to investigate unknown pathways involved in sperm ca- pacitation and gives innovative arguments to better understand infertility mechanisms.

  20. Sperm variation within a single ejaculate affects offspring development in Atlantic salmon

    Science.gov (United States)

    Immler, Simone; Hotzy, Cosima; Alavioon, Ghazal; Petersson, Erik; Arnqvist, Göran

    2014-01-01

    It is generally believed that variation in sperm phenotype within a single ejaculate has no consequences for offspring performance, because sperm phenotypes are thought not to reflect sperm genotypes. We show that variation in individual sperm function within an ejaculate affects the performance of the resulting offspring in the Atlantic salmon Salmo salar. We experimentally manipulated the time between sperm activation and fertilization in order to select for sperm cohorts differing in longevity within single ejaculates of wild caught male salmon. We found that within-ejaculate variation in sperm longevity significantly affected offspring development and hence time until hatching. Whether these effects have a genetic or epigenetic basis needs to be further evaluated. However, our results provide experimental evidence for transgenerational effects of individual sperm function. PMID:24522632

  1. Sperm banking for male cancer patients: social and semen profiles

    Directory of Open Access Journals (Sweden)

    Tatiana C.S. Bonetti

    2009-04-01

    Full Text Available PURPOSE: Report the characteristics of cryopreserved semen from a cohort of male cancer patients, attitudes towards cryopreservation and outcomes of semen samples based on a 12-year cryopreservation program. MATERIAL AND METHODS: Data from 98 male cancer patients whose sperm samples were banked were evaluated. Demographic parameters, semen characteristics, destination of sperm banked samples and questionnaires answered by the patients regarding cryopreservation time were evaluated. RESULTS: The cancer diagnoses were testicle (56.1%, prostate (15.3%, Hodgkin’s lymphomas (9.2%, non-Hodgkin’s lymphomas (7.1%, leukemia (3.1% and other malignancies (9.2%. The patients with testicular cancer presented lower sperm concentration (p < 0.001; however, there were no differences with the percentage of normozoospermic patients among cancer type groups (p = 0.185. A shorter time between cancer diagnosis and sperm banking was observed for testicular and prostate cancer patients (p < 0.001. Most of the patients (89.5% favored sperm banking as a fertility preservation method. CONCLUSIONS: Although less than 20% of banked sperm samples were disposed of, the majority of patients related sperm banking with safe for fertility preservation. Our results show that all male cancer patients of reproductive age facing cancer treatment could be offered sperm banking.

  2. Correlation between Different Patterns of Hypo-Osmotic Swelling and Sperm Functional Tests

    Directory of Open Access Journals (Sweden)

    Farzaneh Bassiri

    2013-01-01

    Full Text Available Background: Sperm membrane integrity is not only important as a barrier between intraandextra-cellular spaces, but also it can be considered as a sign of DNA integrity. Hypoosmoticswelling test reflects membrane integrity and has been used to evaluate spermquality. Intracytoplasmic sperm injection (ICSI in adjunct with hypo-osmotic swellingtest (HOST has been used for treatment of males with asthenozoospermia. Therefore,this study aims to evaluate correlation of different pattern of HOST with sperm parameters,protamine deficiency and apoptosis.Materials and Methods: In this case-control study, sixteen semen samples were randomlycollected from infertile normozospermic men. Semen samples were divided intotwo portions as follows: one portion was assessed for sperm parameters according toWorldHealth Organization (WHO-2010, while the other portion, after applying HOSTprocedure, was used for assessment of sperm morphology, protamine deficiency and lateor early apoptosis. Statistical analysis was carried out using the Statistical Package forthe Social Studies (SPSS 11.5.Results: Our results showed that, the lowest odds ratio (OR of abnormal sperm headmorphology and abnormal acrosome was in d-sperm as compared to a-pattern or nonviablespermatozoa (p=0.00, p=0.01. In addition, a significant correlation was observedbetween d-sperm with sperm concentration and percentage of DNA damage (p=0.03and p=0.04, respectively. A significant correlation was observed between percentageof sperm motility and DNA fragmentation (r=-0.56; p=0.01. Furthermore, significantcorrelations were observed between percentages of early apoptotic sperm with protaminedeficiency and sperm concentration (p=0.009 and p=0.01, respectively.Conclusion: Significant correlations exist between d-pattern and sperm DNA integrity.Semen samples with low sperm concentration have low percentage of d-sperm which aremature and intact sperms.

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

    Science.gov (United States)

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

    2016-10-01

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

  4. 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.

  5. Test on Factors Affecting Boar and Its Semen Quality%不同因素对种公猪及其精液品质的影响试验

    Institute of Scientific and Technical Information of China (English)

    刘政; 余佳维; 刘万钧

    2012-01-01

    The effects of untritional level, preservation temperature and semen collection frequency on the semen quality of boars was studied. The results indicated that feeding boars with protein-rich feed and properly green feed could improve the semen quality. Preserving and transporting under the environmental conditions of 18~22℃ could ensure the semen quality and prolong the sperm survival time. The rational frequency of semen collection and proper exercise of boar also played important roles in enhancing the boar physical fitness and providing high quality semen.%通过营养水平、保存温度和采精频率3个因素对公猪精液品质的影响研究,结果表明,饲喂含蛋白质丰富的饲料和适量的青绿饲料可以提高其精液品质;精液在18~22℃环境条件下保存、运输,可以保证精液质量、延长精子存活时间;合理采精频率与适当运动是增强公猪体质、提供优质精液的重要环节.

  6. 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...

  7. Effect of sorafenib on sperm count and sperm motility in male Swiss albino mice

    Science.gov (United States)

    Shetty, Surekha Devadasa; Bairy, Laxminarayana Kurady

    2015-01-01

    The issue of male germ line mutagenesis and the effects on developmental defects in the next generation has become increasingly high profile over recent years. Mutagenic substance affects germinal cells in the testis. Since the cells are undergoing different phases of cell division and maturation, it is an ideal system to study the effect of chemotherapeutic agents. There are lacunae in the literature on the effect of sorafenib on gonadal function. With background, a study was planned to evaluate the effects of sorafenib on sperm count and sperm motility in male Swiss albino mice. Male Swiss albino mice were used for the study. The animals were segregated into control, positive control (PC) and three treatment groups. PC received oral imatinib (100 mg/kg body weight) and treatment groups received 25, 50, and 100 mg/kg body weight of sorafenib orally for 7 consecutive days at intervals of 24 h between two administrations. The control group remained in the home cage for an equal duration of time to match their corresponding treatment groups. The animals were sacrificed at the end of 1st, 2nd, 4th, 5th, 7th, and 10th weeks after the last exposure to drug, respectively. Sperm suspensions were prepared and introduced into a counting chamber. Total sperm count and motility were recorded. There was a significant decrease in sperm count and sperm motility by sorafenib which was comparable with the effect of PC imatinib. Sorafenib adversely affects sperm count and sperm motility which are reversible after discontinuation of treatment. PMID:26605157

  8. Sperm Motility in Flow

    Science.gov (United States)

    Guasto, Jeffrey; Juarez, Gabriel; Stocker, Roman

    2012-11-01

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

  9. Sperm-egg interaction.

    Science.gov (United States)

    Evans, Janice P

    2012-01-01

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

  10. Prognosis for sperm fertilizability: analysis of different variables in men.

    Science.gov (United States)

    Check, J H; Check, M L; Katsoff, D

    2002-01-01

    An overview of various sperm tests is presented. The standard semen analysis obtained by most clinicians evaluating infertility usually consists of sperm concentration, percent motility, quality of motility, and sperm morphology. Unfortunately, unless the motile density is extremely low, the count and motility are not good prognosticators of fertility potential. Values above the norm for normal fertile couples unfortunately cannot reliably predict normal fertility potential. It is important to find sperm tests that are easy to perform, are relatively inexpensive, and provide an accurate prognosis. Strict morphology was hoped to be such a tool with initial optimism that it was far superior to standard morphology. Unfortunately, this test also failed to be the ideal inexpensive prognostic test after further evaluation. One test that is inexpensive and highly correlates with fertilizability is the presence of antisperm antibodies since their presence frequently does not alter count, motility, or morphology. This test should be performed as part of the routine semen analysis. Other tests highly correlate with the achievement of pregnancy and are simple and inexpensive to perform, but, interestingly, do not correlate with fertilizability. These include the hypoosmotic swelling test (HOST) and the sperm stress test. Abnormalities in these tests imply a different abnormality of sperm that leads to conception failure and that is the transfer of a toxic factor from the sperm to oocyte to embryo that prevents the embryo from implanting. Certainly, the simple, inexpensive HOST should be performed routinely. Other tests of sperm function, e.g., sperm penetration assay, sperm zona pellucida binding assay, and acrosome reaction, have their definite place in the evaluation of the infertile male. However, because they are expensive and difficulty to perform they lend themselves to certain specific circumstances but not to routine testing.

  11. Effects of dilution rates, animal species and instruments on the spectrophotometric determination of sperm counts.

    Science.gov (United States)

    Rondeau, M; Rouleau, M

    1981-06-01

    Using semen from bull, boar and stallion as well as different spectrophotometers, we established the calibration curves relating the optical density of a sperm sample to the sperm count obtained on the hemacytometer. The results show that, for a given spectrophotometer, the calibration curve is not characteristic of the animal species we studied. The differences in size of the spermatozoa are probably too small to account for the anticipated specificity of the calibration curve. Furthermore, the fact that different dilution rates must be used, because of the vastly different concentrations of spermatozoa which is characteristic of those species, has no effect on the calibration curves since the dilution rate is shown to be artefactual. On the other hand, for a given semen, the calibration curve varies depending upon the spectrophotometry used. However, if two instruments have the same characteristic in terms of spectral bandwidth, the calibration curves are not statistically different.

  12. Cryopreservation of mutton snapper ( Lutjanus analis) sperm.

    Science.gov (United States)

    Sanches, Eduardo G; Oliveira, Idili R; Serralheiro, Pedro C Da Silva; Cerqueira, Vinicius R

    2013-09-01

    This study aimed to develop a protocol of semen cryopreservation of the mutton snapper Lutjanus analis. The interaction between three extenders ( pH 6.1; 7.8 and 8.2) , two concentrations of dimethyl sulfoxide ( DMSO, 5 and 10%) and three cooling rates ( -90; -60 and -30°C.min-1) on the sperm motility rate and motility time were analyzed by a factorial experiment. A sample of 30 fishes ( 1,261 ± 449 g) collected in the nature was kept in floating net cages. The semen was frozen by using cryogenic straws, in nitrogen vapour and transferred, later, to liquid nitrogen. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The highest sperm motility rate and motility time ( P snapper.

  13. Efficient Boar Semen Production and Genetic Contribution: The Impact of Low-Dose Artificial Insemination on Fertility.

    Science.gov (United States)

    Broekhuijse, M L W J; Gaustad, A H; Bolarin Guillén, A; Knol, E F

    2015-07-01

    Diluting semen from high fertile breeding boars, and by that inseminating many sows, is the core business for artificial insemination (AI) companies worldwide. Knowledge about fertility results is the reason by which an AI company can lower the concentration of a dose. Efficient use of AI boars with high genetic merit by decreasing the number of sperm cells per insemination dose is important to maximize dissemination of the genetic progress made in the breeding nucleus. However, a potential decrease in fertility performance in the field should be weighed against the added value of improved genetics and, in general, is not tolerated in commercial production. This overview provides some important aspects that influence the impact of low-dose AI on fertility: (i) the importance of monitoring field fertility, (ii) the need for accurate and precise semen assessment, (iii) the parameters that are taken into account, (iv) the application of information from genetic and genomic selection and (v) the optimization when using different AI techniques. Efficient semen production, processing and insemination in combination with increasing use of genetic and genomic applications result in maximum impact of genetic trend.

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

    Science.gov (United States)

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

    2015-01-01

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

  15. Sorting of Sperm by Morphology

    Science.gov (United States)

    Koh, James; Marcos, Marcos

    2016-11-01

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

  16. 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.

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

    Directory of Open Access Journals (Sweden)

    Necmettin TEKİN

    2016-01-01

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

  18. [Eosin Y-water test for sperm function examination].

    Science.gov (United States)

    Zha, Shu-wei; Lü, Nian-qing; Xu, Hao-qin

    2015-06-01

    Based on the principles of the in vitro staining technique, hypotonic swelling test, and water test, the Eosin Y-water test method was developed to simultaneously detect the integrity of the sperm head and tail and sperm membrane structure and function. As a widely used method in clinical laboratories in China, the Eosin Y-water test is methodologically characterized by three advantages. Firstly, both the sperm head and tail can be detected at the same time, which allows easy and comprehensive assessment of membrane damage in different parts of sperm. Secondly, distilled water is used instead of the usual formula solution to simplify and standardize the test by eliminating any potential effects on the water molecules through the sperm membrane due to different osmotic pressure or different sugar proportions and electrolyte solutions. Thirdly, the test takes less time and thus can be repeated before and after treatment. This article focuses on the fundamental principles and modification of the Eosin Y-water test and its application in sperm function examination and routine semen analysis for male infertility, assessment of the quality of sperm retrieved by testicular fine needle aspiration, semen cryopreservation program development, and evaluation of sperm membrane integrity after microwave radiation.

  19. Impact of external factors on sperm motility of Sepiella maindroni

    Institute of Scientific and Technical Information of China (English)

    WANG Jia; JIANG Xiamin; FENG Xiandong

    2011-01-01

    Sperm motility is important in reproductive biology. To understand the physiological ecology characteristics of sperm of Sepiella maindroni, we studied the activation rate, active time, and lifespan of sperm to evaluate the influence of external factors, including temperature, salinity, pH, glucose, NaC1, KC1, MgCl2, CaCl2, EDTA-2Na and artificial seawater on sperm motility. The results show that the appropriate activation and motility condition for sperm were: temperature 20-30掳C (optimum 25掳C), salinity 10.6-30.6 (optimum 15.9) and pH 8.0-8.6 (optimum 8.0-8.4). Sperm activity varied in different concentrations of glucose, NaCl, K.C1, MgCl2 and CaCl2. None of the selected concentrations of EDTA-2Na solution could activate the sperm. The activation rate in artificial seawater devoid of Ca+2, Mg+2 or HCO-3 was low. The results should help further studies on the preservation and activation of squid sperm.

  20. 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.

  1. Effect of different disaccharides on the integrity and fertilising ability of freeze-dried boar spermatozoa: a preliminary study.

    Science.gov (United States)

    Garcia, A; Gil, L; Malo, C; Martinez, F; Kershaw-Young, C; de Blas, I

    2014-01-01

    Freeze-drying spermatozoa is a developing technique that facilitates semen storage and transport. However, freeze-dried sperm exhibits impaired DNA integrity, which is associated with reduced fertilizing ability. Boar spermatozoa were freeze-dried in three different freeze-drying EDTA buffers with trehalose (75mM) and lactose (75mM) (EDTA-TL), (2) with sucrose (75mM) and lactose (75mM) (EDTA-SL) or just lactose (150mM) (EDTA-LL) using two freeze-drying protocols. In experiment 1 a one-step protocol was used and in experiment 2 a two-steps protocol was used. Spermatozoa were stored in1.5 mL cryo-tubes and 1.5 mL glass ampules at both 16 degree C and 25 degree C for 1 month. Successfully freeze-dried spermatozoa were stained with acridine-orange to assess chromatin stability. Freeze-drying was most successful when the 2-step protocol was used (experiment 2). Chromatin stability was greater in samples stored in glass ampules compared to cryo tubes. Chromatin stability was also greater in samples freeze-dried in EDTA-LL compared to EDTA-SL or EDTA-TL buffers. Spermatozoa freeze-dried in EDTA-LL and stored for 14 and 28 days at either 16 degree C or 25 degree C were utilized for ICSI. Two pronuclear formation wasgreatest using spermatozoa stored at 25 degree C (69.23%) and for 28 days (50%). Although 16 degree C spermatozoa samples had better stable chromatin, 25 degree C spermatozoa samples offered better two pronuclear formation results. In conclusion, boar spermatozoa freeze-dried using media containing disaccharides exhibit high chromatin stability and are able to fertilise oocytes following ICSI. Disaccharides may therefore advance the development of freeze-drying techniques for spermatozoa enabling ease of sperm storage and transportation.

  2. The quality of sperm preparation medium affects the motility, viability, and DNA integrity of human spermatozoa

    Directory of Open Access Journals (Sweden)

    Fatemeh Anbari

    2016-01-01

    Full Text Available Aim: The goal was to compare the effects of three different sperm preparation media on sperm motility, viability, and DNA integrity of semen samples from normozoospermic men. Methods: A total of 15 normozoospermic males were included in the study. The semen analysis (SA was performed in accordance with the WHO guidelines (2010. After SA, each sample was divided into three aliquots, and swim-up was performed with three different sperm preparation media (Sperm Preparation Media, Origio, Denmark; Ham′s F10, Biochrome, Berlin, Germany; and VitaSperm TM , Innovative Biotech, Iran. Sperm motility, viability, and DNA fragmentation were evaluated at 0, 1, 2, and 24 h after swim-up. Results: There were no significant differences, at any time intervals, in the total sperm motility between the different sperm preparation media. However, the rate of progressive motility was significantly higher in spermatozoa prepared using the media from Origio in comparison with VitaSperm TM (P = 0.03, whereas no significant difference was found against Ham′s F10 medium. No significant differences in sperm viability were seen between the media products. However, 1 h after swim-up, the extent of sperm DNA fragmentation was lower in the medium from Origio versus VitaSperm TM (P = 0.02. Conclusions: The data showed that the quality of medium for preparation of semen samples from normozoospermic men significantly affects the performance of spermatozoa in assisted conception programs.

  3. Effect of methoxychlor on antioxidant system of goat epididymal sperm in vitro

    Institute of Scientific and Technical Information of China (English)

    Bindu Gangadharan; M. Arul Murugan; P.P. Mathur

    2001-01-01

    Aim: To evaluate the effect of methoxychlor on the antioxidant system of goat epididymal sperm. Methods:Epididymis of adult goat was obtained from local slaughter houses and sperm were collected by chopping the epididymis in modified Ringer's phosphate solution (RPS). After several washings, the sperm samples were dispersed in RPS and incubated with methoxychlor (1 μnol/L, 10 μmol/L and 100 μmol/L) and methoxychlor + vitamin C (100μmol/L each) for 3 h at 32℃. After incubation, the sperm motility and viability were assessed. An aliquot of sperm sample was homogenized, centrifuged and used for the assay of superoxide dismutase, glutathione peroxidase, glutathione reductase and lipid peroxidation. Results: In methoxychlor-incubated sperm and in sperm co-incubated with methoxychlor and vitamin C, the sperm motility and viability showed no significant changes as compared to the corresponding controls. In methoxychlor-incubated sperm the activity of superoxide dismutase, glutathione reductase and glutathione peroxidase were decreased while lipid peroxidation was increased in a dose-dependent manner. Co-incubation of sperm with methoxychlor and vitamin C showed no changes in the activity of superoxide dismutase, glutathione reductase and glutathione peroxidase and in the level of lipid peroxidation. Conclusion: Methoxychlor induced oxidative stress in epididymal sperm of goats by decreasing the levels of antioxidant enzymes. Co-incubation of sperm with methoxychlor and vitamin C, a natural antioxidant, reversed the effect of methoxychlor.

  4. Evaluation of fertility outcome as a consequence of intravaginal inoculation with sperm-impairing micro-organisms in a mouse model.

    Science.gov (United States)

    Vander, Harpreet; Prabha, Vijay

    2015-04-01

    The concept of infertility as a result of asymptomatic microbial colonization of the female reproductive tract has been neglected to date. However, increasing incidence of infertility and advanced research has drawn attention towards this idea. Many of these micro-organisms have been reported to bring about adverse changes in sperm parameters in vitro, but their in vivo potential to cause infertility is still a controversy. The present study was carried out to observe what effect the intravaginal inoculation of sperm-agglutinating Serratia marcescens and sperm-immobilizing Candida albicans had in the reproductive tract and consequently in fertility outcome. When these strains were intravaginally inoculated into female BALB/c mice at 10(4), 10(6) and 10(8) c.f.u. in 20 µl PBS for 10 consecutive days, with mating of mice on day 12, the results showed 100 % decrease in fertility in all groups as compared with control mice receiving PBS alone. Furthermore, no clinical or histopathological changes were observed in the reproductive organs (i.e. ovary, uterus and vagina), suggesting that colonization of the genital tract with sperm-impairing micro-organisms could be a feasible reason for female infertility.

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

    Directory of Open Access Journals (Sweden)

    D.J. Ackerman

    1996-02-01

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

  6. Subversive practices of sperm donation - globalizing Danish sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

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

  7. Morphology of the Testis and Epididymis of Large White Boars

    Directory of Open Access Journals (Sweden)

    Samuel Gbadebo Olukole

    2016-06-01

    Full Text Available The testis and epididymis of twenty five adult Large White boars were used to investigate the biometric and histomorphometric parameters of the testis and epididymis of the boars. The aim of the study was to provide information which could be useful in the comparative regional anatomy of the male reproductive organs of domestic animals and thus an improved assessment of breeding soundness and fertility potential in boars. The average weight of the animals was 71.3 ± 10.7 kg. The average weights of the right and left testes were 170 ± 0.7.60 g and 179±6.48g, respectively with no significant dif¬ference. The average weights of the right and left epididymis were 40.9 ± 6.81 g and 43.7 ± 8.55 g, respectively, with no significant difference. The relative testicular and epididymal weights were 0.49% and 0.12%, respectively. This study shows that the testis is about four times the size of the epididymis. The ductal diameter of the head, body and tail of the epididymis were 418 ± 22.6 µm, 432 ± 20.3 µm and 939 ± 50.6 µm, respectively. The mean relative volume of the germinal epithelium, interstitium and lumen of the seminiferous tubules of the boars rats were 68.4 ± 3.46%, 5.5 ± 0.66% and 78.0 ± 4.81%, respectively. It can be concluded that the morphology of the testis and epididymis of the Large White boar are similar to those of most mammals. This work provides information the testis and epididymis of the Large White boar which could be useful in the comparative regional anatomy of the male reproductive organs of domestic animals and thus an improved assessment of breeding soundness and fertility potential in boars.

  8. Evaluación del ADN espermático de llamas utilizando azul de toluidina Evaluation of llama sperm dna using toluidine blue

    Directory of Open Access Journals (Sweden)

    M.I Carretero

    2009-06-01

    Full Text Available El colorante azul de toluidina (AT se une al ADN permitiendo diferenciar espermatozoides de acuerdo al grado de condensación de la cromatina. Los objetivos de este trabajo fueron: poner a punto una técnica que evalúe la condensación de la cromatina espermática de llama, determinar los patrones de condensación para la especie mediante la tinción con AT y determinar si es posible utilizar ditíotreitol (DTT como control positivo de la tinción. Se ensayaron 2 tiempos de fijación de las muestras con etanol 96 º (2 y 30 minutos y 3 tiempos de incubación con DTT al 1% (30 seg, 1,5 min y 3 min. Los patrones de coloración observados fueron: coloración celeste (negativos, sin alteración en la condensación normal de la cromatina, violeta claro (intermedios, algún grado de descondensación, violeta oscuro (positivos, alto grado de descondensación. No se observaron diferencias significativas entre los tiempos de fijación tanto en las muestras con y sin DTT. En conclusión, se logró simplificar la técnica de AT y determinar los diferentes patrones en espermatozoides de llama. Se comprobó que la incubación con DTT se puede utilizar como control positivo de la técnica y para evaluar la susceptibilidad de cada individuo a la descondensación in vitro.Toluidine blue stain (TB binds to DNA, allowing differentiation of spermatozoa according to the degree of chromatin condensation. The objectives of this study were to adapt a technique for evaluating sperm chromatin in llamas, determine chromatin condensation patterns in llamas using TB and determine if it is possible to use dithiothreitol (DTT as a positive control for the stain. Two fixation times with ethanol 96° (2 and 30 minutes and 3 incubation times with 1% DTT (30 s, 1.5 min and 3 min were studied. Staining patterns observed were: light blue (negative, without alteration in the normal chromatin condensation, light violet (intermediate, some degree of decondensation, dark violet

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

    Science.gov (United States)

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

    2011-05-01

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

  10. Efeito de fontes de óleo e níveis de suplementação de vitamina E na ração sobre as características físicas e morfológicas do sêmen in natura de suínos Effect of oil sources and vitamin E supplementation in the diet on physical and morphological characteristics of fresh boar semen

    Directory of Open Access Journals (Sweden)

    E.A. Moraes

    2010-06-01

    Full Text Available Avaliou-se o efeito da adição de diferentes fontes de óleo e níveis de suplementação de vitamina E na ração sobre as características do sêmen in natura de suínos reprodutores. Foram utilizados 24 reprodutores Dalboar 85, com idades entre 12 e 18 meses, distribuídos em delineamento inteiramente ao acaso, em arranjo fatorial 2 x 3, com duas fontes de óleo, soja e salmão, e três níveis de antioxidantes, 150, 300 e 450mg de vitamina E/kg. Volume, motilidade espermática total, teste hiposmótico, porcentagem de espermatozoides vivos e morfologia espermática não diferiram (P>0,05 entre os tratamentos. Óleo de salmão (P0,05. Os animais tratados com óleo de salmão apresentaram menor (PThe addition of oil sources and dietary vitamin E supplementation was evaluated on the characteristics of fresh boar sperm. Twenty-four mature Dalboar 85 boars of proven fertility and in routine semen production for artificial insemination were randomly distributed in a factorial arrangement 2 X 3, with two oil sources, soy and salmon, and three antioxidant levels: 150, 300, and 450mg/kg of vitamin E. Volume, total motile sperm, hyposmotic swelling test, percentage of live cell, and morphology did not differ (P0.05. Salmon oil treatment showed the lowest (P<0.05 concentration of total antioxidants in the semen. Vitamin E and concentration of total antioxidant showed a linear effect (P<0.05. The semen vigor and the concentration of total antioxidants were improved by oil sources, but the sperm concentration was dependent on the levels of vitamin E. Therefore, salmon oil can be used in the diet of male pigs.

  11. Microfluidic single sperm analysis

    NARCIS (Netherlands)

    Wagenaar, de Bjorn

    2016-01-01

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

  12. In vitro human immunodeficiency virus and sperm cell interaction mediated by the mannose receptor.

    Science.gov (United States)

    Cardona-Maya, Walter; Velilla, Paula A; Montoya, Carlos Julio; Cadavid, Ángela; Rugeles, María T

    2011-12-01

    Leukocytes are considered to be the main source of HIV-1 infection in semen. However, HIV-1 interaction with spermatozoa has also been demonstrated, suggesting that both spermatozoa and leukocytes might play a role during sexual transmission of HIV-1. The purpose of the present study was to evaluate if HIV-1 particles interact with sperm cells through the mannose receptor (MR), and then to determine the ability of "infected" sperm cells to transmit the virus to susceptible targets. The expression of classical HIV-1 receptor and co-receptors and the MR by sperm cells was determined by flow cytometry; the interaction in vitro between sperm and HIV-1 was evaluated by fluorescence microscopy. Additionally, the in vitro interaction of sperm cells and HIV-1 was determined detecting viral nucleic acids by PCR. D-Mannose was used to block HIV-1-sperm cell interaction. Sperm cells preincubated with HIV-1 particles and activated mononuclear cells were co-cultured to determine viral transmission. The presence of viral RNA was detected in 28% of the samples in which sperm cells were preincubated with HIV-1 particles. Mannose was able to block interaction in 75% of the cases. Finally, we demonstrated that "infected" sperm cells were able to transmit the HIV-1 infection to susceptible targets. In conclusion, these results indicate that the MR is involved in sperm cell-HIV-1 interaction. Our results also suggest that sperm cells could be an important source of infection.

  13. Lack of polymorphism at MC1R wild-type allele and evidence of domestic allele introgression across European wild boar populations

    DEFF Research Database (Denmark)

    Canu, Antonio; Vilaça, Sibelle T.; Iacolina, Laura;

    2016-01-01

    , two loci which have been under strong artificial selection during domestication. These loci influence coat colour and number of vertebrae, respectively. A total of 145 wild boars were sampled throughout Europe, to evaluate frequency and spatial distribution of domestic alleles and patterns...

  14. Sperm cryopreservation of lane snapper Lutjanus synagris(Linnaeus, 1758

    Directory of Open Access Journals (Sweden)

    EG Sanches

    Full Text Available AbstractThis study aims developing and evaluate a protocol of semen cryopreservation of the lane snapper Lutjanus synagris. Firstly, sperm motility rate, motility time, density and spermatocrit were appraised to characterize the sperm quality of the lane snapper. The effect of three extenders with distinct ionic compositions and pH values combined with seven concentrations of cryoprotector dimethylsulfoxide (0; 2.5; 5.0; 7.5; 10.0; 12.5 e 15.0%, five cooling rates (110, 90, 60, 45 e 30°C –min, nine equilibration time (1; 2,5; 5; 10; 15; 20; 25; 30 e 60 minutes e five dilutions ratio (1:1; 1:3; 1:6; 1:10 e 1:20 on the sperm motility rate and motility time were analyzed. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The higher sperm motility rate and motility time (P60% validating the present protocol for lane snapper. The cryoconserved sperm of lane snapper is a viable alternative, being possible to maintain appropriate sperm viability.

  15. Sperm cryopreservation of lane snapper Lutjanus synagris (Linnaeus, 1758).

    Science.gov (United States)

    Sanches, E G; Oliveira, I R; Serralheiro, P C S; Cerqueira, V R

    2015-08-01

    This study aims developing and evaluate a protocol of semen cryopreservation of the lane snapper Lutjanus synagris. Firstly, sperm motility rate, motility time, density and spermatocrit were appraised to characterize the sperm quality of the lane snapper. The effect of three extenders with distinct ionic compositions and pH values combined with seven concentrations of cryoprotector dimethylsulfoxide (0; 2.5; 5.0; 7.5; 10.0; 12.5 e 15.0%), five cooling rates (110, 90, 60, 45 e 30°C -min), nine equilibration time (1; 2,5; 5; 10; 15; 20; 25; 30 e 60 minutes) e five dilutions ratio (1:1; 1:3; 1:6; 1:10 e 1:20) on the sperm motility rate and motility time were analyzed. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The higher sperm motility rate and motility time (P60% validating the present protocol for lane snapper. The cryoconserved sperm of lane snapper is a viable alternative, being possible to maintain appropriate sperm viability.

  16. Different sperm sources and parameters can influence intracytoplasmic sperm injection outcomes before embryo implantation

    Institute of Scientific and Technical Information of China (English)

    Yue-hong LU; Hui-juan GAO; Bai-jia LI; Ying-ming ZHENG; Ying-hui YE; Yu-li QIAN; Chen-ming XU; He-feng HUANG; Fan JIN

    2012-01-01

    To evaluate the effects of sperm with different parameters and sources on the outcomes of intracytoplasmic sperm injection (ICSI),1972 ICSI cycles were analyzed retrospectively.Groups 1 to 5 were composed of cycles using ejaculated sperm and were grouped according to sperm quantity,quality,and morphology into normal (288 cycles),or mild (329 cycles),moderate (522 cycles),severe (332 cycles),and extremely severe (171 cycles)oligozoospermia and/or asthenozoospermia and/or teratozoospermia (OAT) groups.Group 6 was composed of 250 cycles using testicular or epididymal sperm,and Group 7 consisted of 80 cycles using frozen-thawed sperm.We found that fertilization rates were gradually reduced from Groups 1 to 6,and reached statistical difference in Groups 5 and 6 (P<0.05).The high-quality embryo rate was higher in Group 1 than in Groups 2,3,5,6,and 7 (P<0.05).No statistical differences were observed in the rates of embryo cleavage,clinical pregnancy,miscarriage,live-birth,premature birth,low birth weight,weeks of premature birth,average birth weight,or sex ratio for all seven groups (P>0.05).A total of nine cases of malformation were observed,with a malformation rate of 1.25% (9/719).In conclusion,different sperm sources and parameters can affect ICSI outcomes before embryo implantation.A full assessment of offspring malformation will require further study using a larger sample size.

  17. 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.

  18. Linear model analysis of the influencing factors of boar longevity in Southern China.

    Science.gov (United States)

    Wang, Chao; Li, Jia-Lian; Wei, Hong-Kui; Zhou, Yuan-Fei; Jiang, Si-Wen; Peng, Jian

    2017-04-15

    This study aimed to investigate the factors influencing the boar herd life month (BHLM) in Southern China. A total of 1630 records of culling boars from nine artificial insemination centers were collected from January 2013 to May 2016. A logistic regression model and two linear models were used to analyze the effects of breed, housing type, age at herd entry, and seed stock herd on boar removal reason and BHLM, respectively. Boar breed and the age at herd entry had significant effects on the removal reasons (P linear models (with or without removal reason including) showed boars raised individually in stalls exhibited shorter BHLM than those raised in pens (P introduction.

  19. Effect of the spirulina platensis included in the main diet on the boar sperm quality

    OpenAIRE

    Kistanova E.; Marchev Y.; Nedeva R.; Kacheva D.; Shumkov K.; Georgiev B.; Shimkus A.

    2009-01-01

    Microalgae Spirulina platensis accumulates many chemical components which are suitable for all higher organisms as food and forage raw material. There are a lot of vitally important for the organisms minerals and macroelements such as iron, calcium, sodium, potassium, copper, magnesium, phosphorus, selenium, vitamins, carotin, nucleic acids, enzymes and other active substances. That should explain the value of Spirulina as a feed additive for the agricultural animals. In the present work the ...

  20. Viability and Risk Assessment in Species Restoration: Planning Reintroductions for the Wild Boar, a Potential Disease Reservoir

    Directory of Open Access Journals (Sweden)

    Hans-Hermann Thulke

    2006-06-01

    Full Text Available The reintroduction of large mammals is often considered a priority conservation action in highly industrialized countries in which many of these species have been depleted. However, species reintroduction after decades of absence may involve important risks for human activities and ecological communities, such as favoring the spread of diseases. An example of a potentially troublesome reintroduction is the wild boar, which may act as a reservoir of diseases, e.g., classical swine fever, and cause high economic losses, and has become a species of concern in several European countries for both ecological and recreational reasons. Failure to prevent the disease consequences of species restoration can negate its conservation benefits. Here we evaluated the probability of both successfully reintroducing wild boar into Denmark and limiting their contact with domestic pig farms to which they might spread disease. For this purpose, we developed a spatially explicit, individual-based population model that incorporates information on boar habitat and demography information from Central European populations. We then compared model predictions with the spatial distribution of farms to achieve a spatial assessment of the contact risk. The most restrictive model scenario predicted that nearly 6% of Denmark provides habitat conditions that would allow wild boar to reproduce. The best habitats for reintroduction were aggregated in seven different areas throughout the country in which the extinction probability was < 5%. However, the expected population expansion was very limited in most of these areas. Both the number of suitable areas and the potential for population expansion greatly increased when we relaxed our habitat assumptions about boar forest requirements; this provided a more conservative scenario for a cautious risk analysis. We additionally found that part of the risk of contact with piggeries was associated with the magnitude of the expansion

  1. First TBEV serological screening in Flemish wild boar

    Directory of Open Access Journals (Sweden)

    Sophie Roelandt

    2016-04-01

    Full Text Available 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.

  2. Zinc and magnesium in bull and boar spermatozoa.

    Science.gov (United States)

    Arver, S; Eliasson, R

    1980-11-01

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

  3. 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.

  4. 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.

  5. Syntaxin and VAMP association with lipid rafts depends on cholesterol depletion in capacitating sperm cells.

    Science.gov (United States)

    Tsai, Pei-Shiue; De Vries, Klaas J; De Boer-Brouwer, Mieke; Garcia-Gil, Nuria; Van Gestel, Renske A; Colenbrander, Ben; Gadella, Bart M; Van Haeften, Theo

    2007-01-01

    Sperm cells represent a special exocytotic system since mature sperm cells contain only one large secretory vesicle, the acrosome, which fuses with the overlying plasma membrane during the fertilization process. Acrosomal exocytosis is believed to be regulated by activation of SNARE proteins. In this paper, we identified specific members of the SNARE protein family, i.e., the t-SNAREs syntaxin1 and 2, and the v-SNARE VAMP, present in boar sperm cells. Both syntaxins were predominantly found in the plasma membrane whereas v-SNAREs are mainly located in the outer acrosomal membrane of these cells. Under non-capacitating conditions both syntaxins and VAMP are scattered in well-defined punctate structures over the entire sperm head. Bicarbonate-induced in vitro activation in the presence of BSA causes a relocalization of these SNAREs to a more homogeneous distribution restricted to the apical ridge area of the sperm head, exactly matching the site of sperm zona binding and subsequent induced acrosomal exocytosis. This redistribution of syntaxin and VAMP depends on cholesterol depletion and closely resembles the previously reported redistribution of lipid raft marker proteins. Detergent-resistant membrane isolation and subsequent analysis shows that a significant proportion of syntaxin emerges in the detergent-resistant membrane (raft) fraction under such conditions, which is not the case under those conditions where cholesterol depletion is blocked. The v-SNARE VAMP displays a similar cholesterol depletion-dependent lateral and raft redistribution. Taken together, our results indicate that redistribution of syntaxin and VAMP during capacitation depends on association of these SNAREs with lipid rafts and that such a SNARE-raft association may be essential for spatial control of exocytosis and/or regulation of SNARE functioning.

  6. Cytometric analysis of shape and DNA content in mammalian sperm

    Energy Technology Data Exchange (ETDEWEB)

    Gledhill, B.L.

    1983-10-10

    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. Sperm were analyzed by flow cytometry and slit-scan flow analysis for injury following the exposure of testes to mutagens. The utility of flow cytometry in genotoxin screening and monitoring of occupational exposure was evaluated. The technique proved valuable in separation of X- and Y-chromosome bearing sperm and the potential applicability of this technique in artificial insemination and a solution, of accurately assessing the DNA content of sperm were evaluated-with reference to determination of X- and Y-chromosome bearing sperm.

  7. Cryopreservation of mutton snapper ( Lutjanus analis sperm

    Directory of Open Access Journals (Sweden)

    EDUARDO G. SANCHES

    2013-09-01

    Full Text Available This study aimed to develop a protocol of semen cryopreservation of the mutton snapper Lutjanus analis. The interaction between three extenders ( pH 6.1; 7.8 and 8.2 , two concentrations of dimethyl sulfoxide ( DMSO, 5 and 10% and three cooling rates ( -90; -60 and -30°C.min−1 on the sperm motility rate and motility time were analyzed by a factorial experiment. A sample of 30 fishes ( 1,261 ± 449 g collected in the nature was kept in floating net cages. The semen was frozen by using cryogenic straws, in nitrogen vapour and transferred, later, to liquid nitrogen. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The highest sperm motility rate and motility time ( P < 0.05 was achieved by combining extender C ( pH 8.2 with DMSO ( 10% and cooling rate of -60°C.min−1 ( P < 0.05 . The use of cryopreserved sperm presented fertilization rates higher than 59% validating the present protocol for mutton snapper.

  8. 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.

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

    DEFF Research Database (Denmark)

    Lützen, Jørgen; Jespersen, Åse; Takahashi, Tohru

    2004-01-01

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

  10. Magneto-optical characteristics of human sperms: normal and deformed.

    Science.gov (United States)

    Sakhnini, Lama; Dairi, Maheen; Manaa, Hacene

    2008-08-01

    In this study we report on magnetic orientation of human sperms. Samples were taken from 17 donors. Normal human sperms became oriented with their long axis perpendicular to the magnetic field (1 T maximum). Total orientation was achieved with magnetic field of about 1 T, while for abnormal sperms the magnetic behavior was different. The dependence of the measured degree of orientation on the intensity of the magnetic field was in good agreement with the theoretical equation for the magnetic orientation of diamagnetic substances. As a result of a numerical analysis based on the equation, the anisotropic diamagnetic susceptibility of normal sperm was found to be Delta(chi) = 8 x 10(-20) J/T(2). The degree of orientation was influenced by the alterations in the shape of the head, body or the tail. It has been suggested that the DNA in the sperm head retain the strong magnetic anisotropy to counterbalance the magnetic anisotropy retained by flagellum microtubules. Recent studies demonstrated a well-defined nuclear architecture in human sperm nucleus, where the head morphology has significant correlation with sperm chromatin structure assay SCSA. Then, as the methods to evaluate SCSA can be difficult and expensive our simple magnetic orientation technique can be an alternative to diagnose alteration in DNA.

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

    Science.gov (United States)

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

    2015-08-01

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

  12. Predictive factors of successful microdissection testicular sperm extraction.

    Science.gov (United States)

    Bernie, Aaron M; Ramasamy, Ranjith; Schlegel, Peter N

    2013-01-01

    Azoospermia in men requires microsurgical reconstruction or a procedure for sperm retrieval with assisted reproduction to allow fertility. While the chance of successful retrieval of sperm in men with obstructive azoospermia approaches >90%, the chances of sperm retrieval in men with non-obstructive azoospermia (NOA) are not as high. Conventional procedures such as fine needle aspiration of the testis, testicular biopsy and testicular sperm extraction are successful in 20-45% of men with NOA. With microdissection testicular sperm extraction (micro-TESE), the chance of successful retrieval can be up to 60%. Despite this increased success, the ability to counsel patients preoperatively on their probability of successful sperm retrieval has remained challenging. A combination of variables such as age, serum FSH and inhibin B levels, testicular size, genetic analysis, history of Klinefelter syndrome, history of cryptorchidism or varicocele and histopathology on diagnostic biopsy have provided some insight into the chance of successful sperm retrieval in men with NOA. The goal of this review was to evaluate the preoperative factors that are currently available to predict the outcome for success with micro-TESE.

  13. Study on Effect of Storage in Ambient Temperature of Boar Semen Using Different Diluent%不同稀释液对公猪精液常温保存效果的研究

    Institute of Scientific and Technical Information of China (English)

    陈玉霞; 孙克宁; 林峰; 杨婷; 高汉婷; 高腾云

    2012-01-01

    In order to improve the storage effect of boar semen, three diluent formulas in ambient temperature was designed to preserve the boar semen, and the acrosome of the sperms was stained after they were preserved.The results showed that the storage time of the sperms stored by formula II was obviously different(P0.05) and all more than 95%.The storage effect of formula Ⅱ for sperms was better than that of formula I and formula Ⅲ in ambient temperature.This study provids the reference basis for the research of the storage technology in ambient temperature of boar semen and the semen diluents formula.%为提高猪精液的保存效果,设计了3种常温保存稀释液配方,并对保存后的精子进行了顶体染色,为猪精液常温保存技术及稀释液配方的研究提供参考依据.结果表明:当精子活率为50%与30%时,配方Ⅱ的精子保存时间均显著高于(P<0.05)配方Ⅰ与配方Ⅲ,且配方Ⅱ所保存精子的总存活时间也显著高于(P<0.05)配方Ⅰ与配方Ⅲ;在精液保存24h后,3种配方保存的精子顶体完整率均在95%以上,且差异不显著(P>0.05),配方Ⅱ对精子的常温保存效果要优于配方Ⅰ与配方Ⅲ.

  14. 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 mo