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Sample records for boar sperm evaluated

  1. Evaluation of sperm chromatin structure in boar semen

    Directory of Open Access Journals (Sweden)

    Banaszewska Dorota

    2015-06-01

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

  2. Factors influencing boar sperm cryosurvival.

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

    2006-10-01

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

  3. Evaluation of zearalenone and alpha-zearalenol toxicity on boar sperm DNA integrity.

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    Tsakmakidis, Ioannis A; Lymberopoulos, Aristoteles G; Khalifa, Tarek A A; Boscos, Constanten M; Saratsi, Aikaterini; Alexopoulos, Costas

    2008-07-01

    This study aimed at investigating the in vitro effects of zearalenone (zen) and alpha-zearalenol (alpha-zen) on motility and nuclear chromatin integrity (NCI) of boar spermatozoa. Mycotoxins were tested, at levels ranging from 10 to 30 microg ml(-1) of diluted semen. Four boars were used for semen collection (eight replicates per boar, four per mycotoxin). After the addition of zen or alpha-zen, semen samples were incubated for 4 h at 38.5 degrees C, 5% CO(2) and 96% humidified air. Motility and NCI were assessed at 0 and 4 h of incubation. No significant differences were noticed in motility among the experimental groups (P > 0.05) for all tested boars. Chromatin instability was significantly higher (P < 0.05) in spermatozoa of only one boar treated with zen and alpha-zen independently of the dose. In conclusion, under our experimental conditions, zen and alpha-zen did not affect the motility of boar sperm, whereas the effects of these toxins on sperm NCI were individual-dependent. 2008 John Wiley & Sons, Ltd

  4. The use of HOS test to evaluate membrane functionality of boar sperm capacitated in vitro.

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    Lechniak, D; Kedzierski, A; Stanislawski, D

    2002-12-01

    The functional and structural integrity of sperm membrane are crucial for the viability of spermatozoa. The commonly used staining test (eosin + nigrosin) for assessing sperm membrane measures only its structural integrity. The hypoosmotic swelling test (HOS) originally developed for human sperm (Jeyendran et al. 1984) has been also applied to several species of domestic animals (bull, pig, horse, dog). The test enables to evaluate the functional status of the sperm membrane. The principle of HOS is based on water transport across the sperm tail membrane under hypoosmotic conditions. It has previously been used to assess the semen quality (Revell and Mrode 1994), to analyse fertilizing capacity (Rota et al. 2000; Perez-Llano et al. 2001) and also to detect viable, immotile cells for ICSI (Intra-cytoplasmic sperm injection) in human (Zeyneloglu et al. 2000). There are two procedures commonly used for sperm capacitation in the pig-sperm washing and incubation before insemination (Nagai 1994). Capacitation involves several changes like removing molecules coating the sperm head membrane, changes in membrane fluidity and intracellular ion concentration (Green and Watson 2001). Thus the membrane integrity as well as functionality may be affected as shown by Harrison (1996). The aim of the present study was to analyse changes in sperm membrane integrity after in vitro capacitation by use of the HOS test.

  5. Alkaline phosphatase in boar sperm function.

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    Bucci, D; Isani, G; Giaretta, E; Spinaci, M; Tamanini, C; Ferlizza, E; Galeati, G

    2014-01-01

    Alkaline phosphatase (AP) catalyses the detachment of phosphate residues from different substrates. Its activity has been demonstrated in seminal plasma and spermatozoa from porcine and other mammalian species; anyway, the role of AP in male reproduction has not been clarified yet and the aim of this study was to determine AP function in boar sperm capacitation and in vitro fertilization (IVF). AP activity was assayed in seminal plasma and in uncapacitated and in vitro capacitated (IVC) spermatozoa; in addition, capacitation was studied in presence of different doses of AP (1.2 and 2.5 IU/mL). The effect of different doses of AP (1.2 and 2.5 IU/mL) on several sperm parameters after IVC (viability, acrosome integrity with FITC-PSA, capacitation status with CTC staining, tyrosine phosphorylation) and on fertilizing ability during IVF were also evaluated. High AP activity was detected in seminal plasma, in particular in sperm-rich fraction; a lower activity was detected in uncapacitated spermatozoa while a significant decrease was evidenced after IVC. Viability was not changed by AP supplementation of the capacitating medium, whereas acrosome integrity and capacitation status were significantly affected by 1.2 and 2.5 doses, with a dose-dependent decrease in acrosome-reacted cells as well as in CTC B pattern displaying cells. As for sperm head protein phosphorylation, a decrease in relative fluorescence was detected in AP 2.5 group, if compared with capacitated one. After IVF, a dose-dependent decrease in penetrated oocytes was recorded, with an increase in monospermic zygote rate. In conclusion, we demonstrated that AP activity decreases under capacitating condition and that addition of AP to spermatozoa during capacitation results in a depression of the capacitating process and IVF. We can infer that AP plays a role in keeping spermatozoa quiescent until they are ejaculated and in modulating the acquisition of the fertilizing ability. © 2013 American Society of

  6. Spermometer: electrical characterization of single boar sperm motility

    NARCIS (Netherlands)

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

    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 microfluidic

  7. 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. © 2015 Blackwell Verlag GmbH.

  8. Microscopic analysis of MTT stained boar sperm cells

    African Journals Online (AJOL)

    tulyasys

    2015-06-08

    Jun 8, 2015 ... Microscopic analysis of MTT stained boar sperm cells. B.M. van den Berg*. Barex Biochemical Products, Seb. Centenweg 45, 1602ML Enkhuizen, The Netherlands. Abstract. The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric.

  9. 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 (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. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Reducing endogenous estrogen during prepuberal life does not affect boar libido or sperm fertilizing potential.

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    Berger, Trish; Conley, Alan J

    2014-09-01

    Increasing sperm production per breeding male has economic significance with increasing use of artificial insemination. Manipulations to increase sperm production in livestock will only be useful if libido and sperm fertilizing capacity are not adversely affected. Reducing endogenous estrogens in the postnatal interval increases the number of Sertoli cells and hence testicular sperm production capacity. These experiments were designed to evaluate the effects of reducing endogenous estrogens on libido and sperm fertilizing capacity. Boars were treated with an aromatase inhibitor, letrozole, to reduce testicular estrogen production between 1 and 6 weeks of age or between 11 and 16 weeks of age, and the littermates to these boars were treated with the canola oil vehicle. Letrozole treatment did not affect time to first mount at 22 weeks of age, regardless of whether the treatment occurred from 1 to 6 weeks of age (118 seconds vs. 233 seconds, SEM = 161 for letrozole-treated and vehicle-treated boars, respectively) or from 11 to 16 weeks of age (107 seconds vs. 67 seconds, SEM = 63 for letrozole-treated and vehicle-treated boars, respectively). Similarly, sperm fertilizing ability and in vivo fertility were equivalent in letrozole-treated boars and their vehicle-treated littermates. Surprisingly, the increase in Sertoli cell numbers observed in the letrozole-treated boars at 20 weeks of age (5.8 vs. 4.3 billion, SEM = 0.5; P libido or sperm fertilizing capacity. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. 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.

  12. Immunological castration temporarily reduces testis size and function without long-term effects on libido and sperm quality in boars.

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    Lugar, D W; Rhoads, M L; Clark-Deener, S G; Callahan, S R; Revercomb, A K; Prusa, K J; Estienne, M J

    2017-04-01

    The objective was to determine the effects of immunization against gonadotropin-releasing hormone on reproductive characteristics in boars. A total of 72 boars were used in a randomized design with three treatments: single immunization (SI) (10 weeks of age) or double immunization (DI) (10 and 15 weeks of age) with Improvest® and intact controls (no Improvest®; CNT) (n=24/group). At 10, 15, 20, 25 and 40 weeks of age, blood was collected and serum harvested to evaluate testosterone concentrations. Testosterone concentrations were less for DI boars compared with CNT boars and SI boars at 20 and 25 weeks (PLibido was assessed at 32, 36, 47, 60 and 63 weeks of age and semen collected at 60 weeks of age was analyzed for indicators of quality. There were no effects of treatment (P=0.41) or treatment by week (P=0.71) on libido. Semen volume, gel weight and total number of sperm cells, determined in a subset of boars (n=3/treatment), were not different among treatments. Sperm concentration was greater for DI than SI (P=0.01), and tended to be greater for DI compared with CNT (P=0.10). Sperm motility tended to be greater for DI boars compared with CNT boars (P=0.066). In conclusion, our results show that there are no long-term effects of immunocastration on reproductive characteristics in boars.

  13. 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

  14. In vitro effect of zearalenone and alpha-zearalenol on boar sperm characteristics and acrosome reaction.

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    Tsakmakidis, I A; Lymberopoulos, A G; Alexopoulos, C; Boscos, C M; Kyriakis, S C

    2006-10-01

    This study was conducted to determine the in vitro effects of three different concentrations (125, 187.5 and 250 microM in diluted semen) of zearalenone (zen) and alpha-zearalenol (alpha-zen) on boar sperm. Semen parameters such as motility, viability and spontaneous acrosome reaction were evaluated. From the results it was shown that both zen and alpha-zen affected the sperm characteristics significantly (p < 0.05), except for alpha-zen at the low concentration which did not decrease the percentage of live reacted spermatozoa significantly. In conclusion, zen and alpha-zen are directly toxic when they affect boar semen in vitro and consequently decrease the fertilization ability of the sperm. The higher the concentration of mycotoxin tested, the greater the decline of sperm parameters noticed. The influence of mycotoxins was found to be time- and dose-dependent.

  15. Cryopreservation of boar sperm induces differential microRNAs expression.

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    Zhang, Yan; Dai, Dinghui; Chang, Yu; Li, Yuan; Zhang, Ming; Zhou, Guangbin; Peng, Zhanghua; Zeng, Changjun

    2017-06-01

    Lower conception rates and litter sizes limit the wide use of artificial insemination with frozen-thawed boar sperm, due to a lack of understanding of the mechanisms that cause cryodamage and cryoinjury to sperm during cryopreservation. CryoMiRs, a family of freeze-related microRNAs (miRNAs), are associated with freeze tolerance, and regulate metabolism in mammalian hibernators and insects. Thus, we speculate that miRNAs maybe involved in the regulation of the freeze-thaw process and may affect boar sperm function. In this study, we studied the differential expression of 46 miRNAs that have roles in spermatogenesis, sperm maturation, and sperm quality in response to cryopreservation (with or without 3% glycerol). The results indicated that, in response to cryopreservation with 3% glycerol, 14 miRNAs were significantly up-regulated, but only two miRNAs (miR-22 and miR-450b-5p) were significantly down-regulated, relative to fresh sperm. Preservation with 3% glycerol caused up-regulation of 17 miRNAs, but only caused down-regulation of one miRNA (miR-24), relative to sperm cryopreserved without glycerol. Functional annotations of these differentially expressed miRNAs indicated that these miRNAs and their targets are mainly associated with metabolic and cellular processes. Therefore, our findings show that cryopreservation results in changes in miRNA expression, and suggest that the anti-freeze mechanisms of boar sperm need to be studied further. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. 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 cytogenetic quality of boars. By analyzing a series of 25 animals judged normal by their fertility statistics and a series of seven animals with known reciprocal translocations, a model...... for identifying sperm cells from cytogenetically aberrant animals was proposed. This model was applied to a series of 50 uncharacterized animals. The model successfully identified a mosaic or chimaeric carrier of an aberrant X chromosome. However, implementation of this technique for screening purposes would...

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

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    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. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Artificial insemination with frozen-thawed boar sperm.

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    Yeste, Marc; Rodríguez-Gil, Joan E; Bonet, Sergi

    2017-09-01

    Artificial insemination with frozen-thawed semen in pigs is not a routine technique; its use is restricted to specific cases, such as preservation of valuable genetic material (germplasm banks), safety strategies in case of natural disasters, long-distance transport of sperm, and in combination with sex-sorting. Cryoinjuries resulting from freeze-thawing protocols are a major concern with regard to the fertilization capacity of the treated sperm, which is lower than that of liquid-stored semen. Here, we provide an overview of artificial insemination using cryopreserved sperm, and summarize the factors that influence cryopreservation success before, during, and after freeze-thaw (i.e., sperm selection before starting the cryopreservation process, holding time, use of cryoprotectants, and rates of freezing and thawing) and that are driving the identification of biomarkers to predict sensitivity to cryodamage. Three different artificial insemination techniques (conventional or intracervical; intrauterine; and deep intrauterine) are also discussed with regards to their relevance when using frozen-thawed semen. Finally, we review the use of additives to freezing and thawing media, given reports that they may maintain and improve the quality and fertilizing capacity of frozen-thawed sperm. In sum, artificial insemination with frozen-thawed boar sperm can provide reasonable fertility outcomes, if freezable ejaculates, specific additives, and appropriate insemination techniques are used. © 2017 Wiley Periodicals, Inc.

  19. 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 functions of spermatozoa. © 2015 American Society of Andrology and European Academy of Andrology.

  20. 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.

  1. Oxidoreductive capability of boar sperm mitochondria in fresh semen and during their preservation in BTS extender.

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    Gaczarzewicz, Dariusz; Piasecka, Małgorzata; Udała, Jan; Błaszczyk, Barbara; Laszczyńska, Maria; Kram, Andrzej

    2003-07-01

    The purpose of our study was to determine the effect of dilution and liquid-preservation of boar sperm on oxidoreductive capability of their mitochondria. The semen was diluted with BTS extender produced from water purified by destillation or by reverse osmosis. The spermatozoa were stored over a four-day period at 16-18 degrees C. The function of sperm mitochondria was assessed using the screening cytochemical test for NADH-dependent oxidoreductases (diaphorase/NADH, related to flavoprotein). Morphological assessment of cytochemical reaction was carried out using a light microscope. The intensity of the reaction was evaluated by means of a computer image analysing system (Quantimet 600S), measuring the integrated optical density (IOD) and mean optical density (MOD) of the reaction product (formazans) occurring in the sperm midpieces. In the non-diluted semen, intensive cytochemical reaction throughout the length of the sperm midpiece was observed. Furthermore, spermatozoa with the intensive reaction displayed the high optical density values. After dilution the semen with two variants of experimental extender, and as the conservation time expired, the cytochemical reaction was less intensive. Moreover, the absence of formazan deposits in various parts of the sperm midpiece was also noted. These morphological features corresponded to low values of optical density. These findings suggest that the dilution of semen and the time of sperm preservation may be critical factors that handicap energy metabolism of sperm mitochondria. The type of water used in preparing BTS extender does not have any significant effect on the oxidoreductive capability of sperm boar mitochondria.

  2. 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.

  3. 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 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. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. 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

  5. Resveratrol and Epigallocatechin-3-gallate addition to thawed boar sperm improves in vitro fertilization.

    Science.gov (United States)

    Gadani, B; Bucci, D; Spinaci, M; Tamanini, C; Galeati, G

    2017-03-01

    Thawing is one of the most delicate process after semen cryopreservation as spermatozoa pass from a dormant metabolic stage to a sudden awakening in cellular metabolism. The rapid oxygen utilization leads to an overproduction of reactive oxygen species that can damage sperm cells, thus causing a significant decrease of fertilizing potential of frozen-thawed spermatozoa. Resveratrol (Res) is a natural grape-derived phytoalexin and Epigallocatechin-3-gallate (EGCG) is the major polyphenol in green tea (Camellia sinensis); both molecules are known to possess high levels of antioxidant activity. The objective of the present study was to assess the effect of different concentrations of Res (0.5, 1 or 2 mM; Experiment 1) or EGCG (25, 50 or 100 μM; Experiment 2) supplementation to thawing boar semen extender on sperm quality parameters (viability and acrosome integrity) and in vitro fertilization (IVF). Semen after thawing and dilution with three volumes of Beltsville Thawing Solution (BTS), was immediately divided in control group without antioxidants addition (CTR) and either Res or EGCG groups. Sperm viability and acrosome integrity were evaluated in CTR, Res or EGCG groups after 1 h of incubation at 37 °C. The addition of different doses of Res or EGCG to thawing extender for 1 h did not induce any effect on boar sperm viability and acrosome integrity. However, both Res and EGCG treated samples exhibited a significantly higher penetration rate compared with CTR when used for IVF. In particular the treatment with all the EGCG concentrations increased the penetration rate (P fertilization efficiency as compared to control (EGCG 25 μM: 40.3 ± 8.2 vs 26.8 ± 9.5, P fertilization. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. 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. Copyright © 2010

  7. Boar sperm storage capacity of BTS and Androhep Plus: viability, motility, capacitation, and tyrosine phosphorylation.

    Science.gov (United States)

    Dubé, Charlotte; Beaulieu, Martin; Reyes-Moreno, Carlos; Guillemette, Christine; Bailey, Janice L

    2004-09-01

    Androhep Plus, a long-term extender (up to 7 days) and Beltsville Thawing Solution (BTS), a short-term extender (up to 3 days), are commonly used for liquid storage of porcine semen. To test the hypothesis that modifications in sperm viability, motility, chlortetracycline (CTC) fluorescence patterns, and protein tyrosine phosphorylation occur during semen storage in extenders, we compared these end points at different periods of storage in either Androhep Plus or BTS. Sperm from five boars were assessed daily over 12 days of storage (n = 5 ejaculates from different boars). Viability was not different (P BTS (P BTS. A tyrosine-phosphorylated protein of Mr 21,000 appeared after 10 days in sperm incubated in BTS, and was identified as a phospholipid hydroperoxide glutathione peroxidase. Therefore, modifications in viability, motility, CTC fluorescence patterns, and sperm protein tyrosine phosphorylation were apparent during sperm storage in extenders; these may affect the fertilizing capacity of the semen.

  8. Hyaluronic acid delays boar sperm capacitation after 3 days of storage at 15 degrees C.

    Science.gov (United States)

    Yeste, M; Briz, M; Pinart, E; Sancho, S; Garcia-Gil, N; Badia, E; Bassols, J; Pruneda, A; Bussalleu, E; Casas, I; Bonet, S

    2008-12-01

    The present study was undertaken to determine the effects of the addition of hyaluronic acid (HA), ranged from 12.5 to 200 microg/ml, on boar sperm capacitation status during a storage time (up to 3 days) at 15 degrees C in Beltsville thawing solution (BTS). The raw extender was the negative control whereas different concentrations of caffeine (CAF), ranged from 0.25 to 8mM, served as positive controls. Sperm viability, motility, morphology, and osmotic resistance were also determined before and after assessing the treatments. Samples were obtained from 28 healthy and post-pubertal Piétrain boars and sperm parameters were tested immediately after the addition of treatments and after 1, 2 and 3 days of refrigeration at 15 degrees C. Sperm capacitation status was determined by chlortetracycline (CTC) staining and sperm viability by means of a multiple fluorochrome-staining test. Sperm motility and morphology were assessed using phase-contrast microscopy accompanied by a computer assisted sperm analysis system (CASA). Whereas HA delayed sperm capacitation, CAF increased the frequency of capacitated spermatozoa after 2 days of cooling. Moreover, HA did not modify other sperm parameters, such as sperm velocity, whereas CAF increased progressive motility during the first 2 days of cooling and then decreased. It can be concluded that the addition of HA at 50 and 100 microg/ml to the BTS extender may delay sperm capacitation after 3 days of cooling.

  9. Evaluations of boar gonad development, spermatogenesis with regard to semen characteristics, libido and serum testosterone levels based on large White Duroc x Chinese Erhualian crossbred boars.

    Science.gov (United States)

    Ren, D; Xing, Y; Lin, M; Wu, Y; Li, K; Li, W; Yang, S; Guo, T; Ren, J; Ma, J; Lan, L; Huang, L

    2009-12-01

    Chinese Erhualian pigs are known for prolificacy with distinct reproductive traits compared with Western commercial breeds. In this study, a four-generation intercross resource population was constructed using White Duroc boars and Chinese Erhualian sows as founder animals, and a total of 14 male reproductive traits were recorded in 411 F2/F3 boars including the testis and epididymis weights, the seminiferous tubular diameter and spermatogenesis at 60, 90 and 300 days of age, semen characteristics, serum testosterone concentration and libido level at 300 days of age. The White Duroc-Erhualian boars showed remarkable segregations in the traits measured except for the seminiferous tubular diameter and had high ratio (13.9%) of the abnormality of spermatogenesis, providing a good experimental population for detecting quantitative trait loci affecting these male reproductive traits. Furthermore, the correlations among nine male reproductive traits at 300 days of age indicated that the testis weight and the body weight were strongly correlated with the sperm production, supporting the two traits as important parameters for boar selection to increase sperm production and ultimately improve boar fertility. The libido level in the White Duroc-Erhualian boars that was evaluated by a new and easily recorded scoring system showed a significant correlation with serum testosterone concentration. Yet, both libido and serum testosterone concentration were not correlated with the sperm production. Results of this study provided new information on the male reproductive physiology and genetics in Chinese Erhualian and White Duroc boars.

  10. Treating boar sperm with cholesterol-loaded cyclodextrins or cyclodextrins prior to cryopreservation: effects on post-thaw in vitro sperm quality of sperm cryopreserved in different freezing extenders.

    OpenAIRE

    BLANCH TORRES, EVA

    2016-01-01

    [EN] Cryopreserved boar sperm is not used extensively for artificial insemination due to poor fertility rates of the sperm after freezing and thawing. The sperm membrane is damaged when cooled from body temperature to 5 ºC (cold shock), as well as during the freeze-thaw process. Increasing the cholesterol content of boar sperm membranes could increase their post-thaw survival, similarly to other species that are cold shock sensitive. Cholesterol can be easily added to sperm membranes using ch...

  11. Crossreactivity of boar sperm monoclonal antibodies with human ...

    African Journals Online (AJOL)

    Monoclonal antibodies against the head (H mabs) and tail (Tmabs) of boar spermatozoa were produced. Spermatozoa from boar, stallion, bull, human, ram, goat and rabbit were independently incubated with the monoclonal antibodies and later stained by immunofluorescence method. There were positive reactions of the ...

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

    International Nuclear Information System (INIS)

    Berger, T.

    1990-01-01

    Exposed plasma membrane proteins were labeled with 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

  13. 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.

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

    Science.gov (United States)

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

  15. 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.

  16. 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 (Pspermatozoa express AQP7 and AQP11, and the amounts of AQP11 but not those of AQP7 are correlated with sperm motility and membrane integrity.

  17. The effects of three extenders on refrigerated boar semen

    African Journals Online (AJOL)

    sofiateixeira

    2015-03-19

    Mar 19, 2015 ... This study aims to evaluate the quality of boar sperm that was refrigerated for 14 days at 17 ºC with three extenders. ... preferred method of storage for boar semen that is to be used for artificial insemination (AI) (Johnson et al.,. 2000). Pursel et ..... Boar sperm viability after freezing and thawing. Proc. 1st Int.

  18. 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...

  19. Reproductive tract modifications of the boar sperm surface

    NARCIS (Netherlands)

    Gadella, Bart M|info:eu-repo/dai/nl/115389873

    2017-01-01

    The sperm cell has a unique, polarized and segregated surface that is modified extensively by the changing environments in both the male and the female reproductive tracts. The sperm cannot refresh its surface, as protein translation and membrane recycling by intracellular vesicular transport have

  20. Saccharide-mediated interactions of boar sperm surface proteins with components of the porcine oviduct

    Czech Academy of Sciences Publication Activity Database

    Liberda, J.; Maňásková, Pavla; Prelovská, L.; Tichá, M.; Jonáková, Věra

    2006-01-01

    Roč. 71, č. 2 (2006), s. 112-125 ISSN 0165-0378 R&D Projects: GA ČR(CZ) GP303/04/P070; GA ČR(CZ) GA303/06/0895; GA MZd(CZ) NJ7463 Institutional research plan: CEZ:AV0Z50520514 Keywords : boar seminal plasma proteins * saccharide-mediated events * sperm oviductal reservoir Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.536, year: 2006

  1. 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 (Pfunction in both media, bicarbonate induced an increase in a 'fast linear' cohort of spermatozoa in TyrBic regardless of storage (66.4% at 12h and 63.9% at 72h). These results imply a binary pattern in response of sperm 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

  2. Boar seminal plasma inhibits cryo-capacitation of frozen-thawed ram sperm and improves fertility following intracervical insemination.

    Science.gov (United States)

    Fang, Yi; Zhong, Rongzhen; Zhang, Xiaosheng; Zhang, Jinlong; Zhou, Daowei

    2018-01-01

    Cryopreservation has numerous deleterious effects on sperm structure and function, which can be reduced by adding seminal plasma (SP), either autologous or heterologous. The objective was to determine effects of adding boar SP to the freezing extender on sperm quality, tyrosine phosphorylation and fertilizing ability of frozen-thawed ram sperm. Semen was collected from eight Small-tail Han rams and extended in a glucose-egg yolk buffer supplemented with 0, 20, 40, or 60% porcine SP (from Large white boars). Compared to all other groups, 40% boar SP increased sperm viability and motility (P intracervical AI (47.5 vs 33.3%; P < 0.05), apparently due to inhibition of cryo-capacitation, although pregnancy rate for intravaginal AI was not affected (31.6 vs 30.0%). Copyright © 2017. Published by Elsevier Inc.

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

    DEFF Research Database (Denmark)

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

    2002-01-01

    Biosciences) was compared with microscopic counting using a Burker-Turk haemocytometer. In addition, sperm concentration was determined using the Corning 254 spectrophotometer which is used routinely by Danish artificial insemination stations for boars. The results show that the agreement between flow...... cytometry and microscopic counting is very high. The slope for the regression line was 1.12 (SE = 0.03) with an estimated intercept with the Y-axis of 22 x 10(6) sperm/ml (SE = 10 x 10(6) sperm/ml) and an estimated error of the model of 10 x 10(6) sperm/ml. For the spectrophotometer, the slope...... of the regression line was 1.09 (SE = 0.07) with an estimated intercept of 137 x 10(6) sperm/ml (SE = 25 x 10(6) sperm/ml). The average error made by the spectrophotometer was 55 x 10(6) sperm/ml. In addition, the results obtained using flow cytometry was highly repeatable ( CV = 2.7%) in comparison...

  4. Cryopreservation of Piau-Breed Wild Boar Sperm: Assessment of Cooling Curves and Centrifugation Regimes.

    Science.gov (United States)

    Shiomi, H H; Pinho, R O; Lima, Dma; Siqueira, J B; Santos, McR; Costa, E V; Lopes, P S; Guimarães, Sef; Guimarães, J D

    2015-08-01

    This study aimed to assess the effects of different cooling curves and centrifugation regimes used in cryopreservation protocols on the post-thaw viability of Piau-breed wild boar (Sus scrofa) sperm using in vitro assessment tests. Two centrifugations (800 g for 10 min and 2400 g for 3 min) and two cooling curves (conventional cooling using nitrogen vapour - freezing 1 and automated cooling using a programmed freezing machine - freezing 2) were tested. Therefore, the treatments were divided into M3 - centrifugation at 2400 g for 3 min and freezing 2; M10 - centrifugation at 800 g for 10 min and freezing 2; R3 - centrifugation at 2400 g for 3 min and freezing 1; and R10 - centrifugation at 800 g for 10 min and freezing 1. No significant differences (p > 0.05) between treatments occurred post-thawing regarding the total sperm motility means recorded. The mean values of the different treatments were not different from each other regarding the supravital staining (SV), hypo-osmotic test (HO), sperm-egg binding assay or sperm morphology. This study showed that both the cooling curve and the centrifugation regime affected the quality of post-thaw sperm, and centrifugation for shorter times and cooling curves using automated cooling are the most suitable for minimizing sperm injury. © 2015 Blackwell Verlag GmbH.

  5. 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

    2009-05-01

    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.

  6. Effect of L-Carnitine Supplementation on Boar Semen Quality

    Directory of Open Access Journals (Sweden)

    E. Jacyno

    2007-01-01

    Full Text Available The effect of addition of L-carnitine on boar semen quality was studied in 5 Pietrain boars at age 1.5 - 2.0 years. The boars received 500 mg of L-carnitine per day for 5 weeks. During this period, their ejaculates were collected once a week and evaluated for quality. The control ejaculates had been collected before the application of L-carnitine. It was found that the addition of L-carnitine to the boars' feed had a positive effect on the quality of boar semen. The total ejaculate volume and sperm-rich fraction volume increased by 11% and 10%, respectively; the total ejaculate sperm count increased by 11.5% (P P < 0.01. Sperm concentration and motility, as well as normal acrosome sperm percentage, did not increase considerably. The positive effect of L-carnitine on boar semen quality was observable as early as after one week of its application.

  7. 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. Copyright © 2014 Elsevier Inc. All rights reserved.

  8. Sperm-surface ATP in boar spermatozoa is required for fertilization: relevance to sperm proteasomal function.

    Science.gov (United States)

    Yi, Young-Joo; Park, Chang-Sik; Kim, Eui-Sook; Song, Eun-Sook; Jeong, Ji-Hyeon; Sutovsky, Peter

    2009-01-01

    Extracellular ATP has been implicated in a number of cellular events, including mammalian sperm function. The complement of ATP-dependent sperm proteins includes six subunits of the 26S proteasome, a multi-subunit protease specific to ubiquitinated substrate-proteins. Proteolysis of ubiquitinated proteins by the 26S proteasome is necessary for the success of mammalian fertilization, including but not limited to acrosomal exocytosis (AE) and sperm-zona pellucida (ZP) penetration. The 26S proteasome is uniquely present on the sperm acrosomal surface during mammalian, ascidian, and invertebrate fertilization. The proteasome is a multi-subunit protease complex of approximately 2 MDa composed of the 19S regulatory complex and a 20S proteolytic core. Integrity of the 19S complex is maintained by six 19S ATPase subunits (PSMC1 through PSMC6). Consequently, we hypothesized that fertilization will be blocked by the depletion of sperm-surface associated ATP (ssATP). Depletion of ssATP by the Solanum tuberosum apyrase, a 49 kDa, non-cell permeant enzyme, significantly reduced the ATP content measured by an adapted luminescence-ATP assay from which all permeabilizing agents were excluded. Addition of active apyrase to porcine in vitro fertilization (IVF) medium caused a concentration dependent reduction in the overall fertilization rate. No such outcomes were observed in control groups using heat-inactivated apyrase. Apyrase treatment altered the band pattern of 19S ATPase subunits PSMC1 (Rpt2) and PSMC4 (Rpt3) in Western blotting, suggesting that it had an effect on the integrity of the sperm proteasomal 19S complex. Apyrase only altered the proteasomal core activities slightly, since these activities are not directly dependent on external ATP. In contrast, sperm treatment with MG132, a specific inhibitor of the proteasomal core chymotrypsin-like activity, inhibited the target proteolytic activity, but also induced a compensatory elevation in proteasomal peptidyl

  9. 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

  10. Oviductal epithelial cells selected boar sperm according to their functional characteristics

    Science.gov (United States)

    López-Úbeda, Rebeca; García-Vázquez, Francisco A; Gadea, Joaquín; Matás, Carmen

    2017-01-01

    The interaction of oviductal epithelial cells (OECs) with the spermatozoa has beneficial effects on the sperm functions. The aim of this study is to evaluate the in vitro fertilizing capacity of incubating spermatozoa previously selected by density gradient in OEC and determinate some sperm characteristics that could explain the results obtained. In this study, we assessed in vitro fertilization (IVF), tyrosine phosphorylation, phosphatidylserine translocation, nuclear DNA fragmentation, and chromatin decondensation. Three experimental sperm groups, previously selected by Percoll gradient, were established according to the origin of the sperm used for IVF: (i) W30 group: spermatozoa were incubated with oocytes in the absence of OEC; (ii) NB group: after sperm incubation in OEC, the unbound spermatozoa were incubated with oocytes, in the absence of OEC; and (iii) B group: after sperm incubation with OEC, the bound spermatozoa were incubated with oocytes in the OEC plates. The results showed that sperm from the NB group led to a lower IVF yield, accompanied by low penetration rates (NB: 19.6%, B: 94.9%, and W30: 62.9%; P sperm selection, even within an already highly selected population of spermatozoa, and can capture the best functional spermatozoa for fertilization. PMID:27232850

  11. Effect of active immunization against GnRH on testosterone concentration, libido and sperm quality in mature AI boars

    Science.gov (United States)

    2012-01-01

    Background The aim of the present study was to investigate the efficacy of the Improvac on testosterone concentration in blood serum, sexual behavior and sperm quality in matured AI boars. A total of nine Danish Landrace AI boars were included in the analysis. Methods The trial period lasted for 15 weeks and was divided into four periods: Control period: three weeks before vaccination; Period I – four weeks after first vaccination; Period II – four weeks after second vaccination, Period III – four weeks after third vaccination. Blood and sperm samples were collected at weekly intervals. Freshly collected sperm samples were analyzed. Results Testosterone concentration correlated with libido (r = 0.531; p < 0.001), volume of ejaculate (r = 0.324; p < 0.001) and the percentage of morphologically normal spermatozoa (r = 0.207; p < 0.05). Testosterone concentration rised significantly (p < 0.05) in 5–6 week of trial, e. i. after the first dose of Improvac and after this peak the level of testosterone further progressively decreased (p < 0.05). Conclusions Results from this study indicate that active immunization of sexually matured boars against GnRH has negative impact on testosterone concentration, sexual behavior, volume of ejaculate and total number of normal spermatozoa in ejaculate. PMID:22640725

  12. Olfactory evaluation of boar taint: effect of factors measured at slaughter and link with boar taint compounds.

    Science.gov (United States)

    Heyrman, E; Millet, S; Tuyttens, F A M; Ampe, B; Janssens, S; Buys, N; Wauters, J; Vanhaecke, L; Aluwé, M

    2017-11-01

    There is a commitment by the European pig sector to ban surgical castration of male piglets in the European Union in 2018. One alternative to castration is to raise entire male pigs, with an increased risk of boar taint. A field study was performed to: (1) evaluate inter- and intra-farm variation in boar taint prevalence, (2) investigate factors measured at slaughter influencing boar taint and (3) evaluate the relationship between sensorial scoring by a trained panel and the concentration of boar taint components. From 34 farms, neck fat samples were collected from all entire male pigs in at least two slaughter batches per farm (78 batches; 9167 animals). In addition to olfactory boar taint analysis, data were also collected on fresh skin lesions (score 0 to 3) at the slaughter line, slaughter weight, lean meat percentage, duration of transport, time spent in lairage, total delivery duration, day length, shortening of days and outdoor mean temperature. Using the hot iron method, neck fat samples were scored (eight-point scale) for boar taint. Average boar taint prevalence (score ≥3) was 5.6±2.5% and the mean difference between the maximum and minimum prevalence per farm was 4.3±3.2%. Androstenone (AND), skatole (SKA) and indole concentrations were measured for a subset (n=254) of the samples. According to binomial univariate mixed models, entire male pigs with a higher skin lesion score had higher odds of having boar taint (P=0.031), as did fatter entire male pigs (Pmeat percentage (P<0.001) and outdoor mean temperature (P=0.005) remained as only significant factors. Based on our results, we can conclude that these statistically significant at least partially influence the prevalence of boar taint. According to the binomial univariate mixed models SKA concentration in liquid fat seems a better predictor for boar taint than AND. There were no significant synergetic effects between boar taint compounds.

  13. The effects of three extenders on refrigerated boar semen | Teixeira ...

    African Journals Online (AJOL)

    This study aims to evaluate the quality of boar sperm that was refrigerated for 14 days at 17 ºC with three extenders. For this study, samples from four boars were collected twice a week using the gloved-hand technique. After collection, only ejaculates showing total motility of 75% or greater were submitted to the refrigeration ...

  14. 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.

  15. Differences in the expression of microRNAs and their predicted gene targets between cauda epididymal and ejaculated boar sperm.

    Science.gov (United States)

    Chang, Yu; Dai, Ding-Hui; Li, Yuan; Zhang, Yan; Zhang, Ming; Zhou, Guang-Bin; Zeng, Chang-Jun

    2016-12-01

    Mammalian spermatozoa gradually mature and acquire fertility during the transition from the testis to the caput and cauda epididymis, after which they are stored at the tail of the epididymis and the ampulla of vas deferens. During ejaculation, mixing of spermatozoa with the secretions of accessory sex glands leads to their dilution and changes in their function. Although remarkable progress has been made toward the understanding of changes in spermatozoa biochemistry and function before and after ejaculation, it is unknown whether microRNAs (miRNAs) are involved in regulating the function of spermatozoa during the transition between the cauda epididymis and ejaculation. In this study, 48 miRNAs were selected for analysis on the basis of their potential involvement in spermatogenesis, sperm maturation, and quality parameters markers. The differential expression levels of these 48 miRNAs between the caudal epididymis and fresh ejaculates of boar spermatozoa were determined. We found that 15 miRNAs were significantly differentially expressed (eight downregulated and seven upregulated) between boar cauda epididymal and fresh spermatozoa. Five miRNAs hypothesized to be involved in sperm apoptosis were further tested to demonstrate their influence over the expression of their target mRNAs using quantitative reverse-transcription polymerase chain reaction. Together, our findings suggest that these differentially expressed miRNAs are associated with the functional regulation of spermatozoa between cauda epididymis and ejaculation. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. 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 (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.

  17. 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.

  18. 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. Copyright © 2015 Elsevier Inc. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Alba G C Pech-Sansores

    2011-09-01

    Full Text Available 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 + 200μg/ml VE; T3, semen + BTS + 10% SP + 200μg/ml VE; T4, semen + BTS (control. Motility (MOT, intact acrosomes (IA, membrane integrity (MI and mitochondrial activity (MA were evaluated, at 0 and 30 min after thawing. A split plot design was used and the data analyzed using a mixed model analysis of variance. There was a significant effect of SP and VE on IA and MI (P0.05. There was significant effect of incubation time on MOT (21.3 and 27.9% and IA (46.0 and 36.0%, at 0 and 30 min post-thawing (P <0.05. It is concluded that the vitamin E, seminal plasma or the combination of both, affected the thawed sperm traits here studied. Incubation time increased motility and intact acrosome percentage but did not affect the membrane integrity and mitochondrial activity.

  20. 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.

  1. Study on the in vitro effect of zearalenone and alpha-zearalenol on boar sperm-zona pellucida interaction by hemizona assay application.

    Science.gov (United States)

    Tsakmakidis, I A; Lymberopoulos, A G; Vainas, E; Boscos, C M; Kyriakis, S C; Alexopoulos, C

    2007-01-01

    The mycotoxin zearalenone (zen) impairs fertility in farm animals. The aim of the present study was to investigate the effect of zearalenone and its major metabolite (alpha-zearalenol) on boar semen binding capacity, under in vitro conditions. Extended boar semen was exposed to three different concentrations of zen and alpha-zen (40, 60 and 80 microg ml(-1) of semen) for 1 h. Afterwards, the semen was washed and incubated with homologous oocyte hemizona for 4 h. A significant decrease (P < 0.001) in the number of tightly attached spermatozoa on the hemizona was obtained at concentrations of 60 microg ml(-1) and 80 microg ml(-1) of zen and alpha-zen. In conclusion, zen and alpha-zen affected the sperm-zona interaction by reducing the ability of boar spermatozoa to bind to the zona pellucida. (c) 2007 John Wiley & Sons, Ltd.

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

    Czech Academy of Sciences Publication Activity Database

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

    2013-01-01

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

  3. 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.

  4. Sex-Sorted Boar Sperm - An Update on Related Production Methods.

    Science.gov (United States)

    Rath, D; Tiedemann, D; Gamrad, L; Johnson, L A; Klein, S; Kues, W; Mancini, R; Rehbock, C; Taylor, U; Barcikowski, S

    2015-07-01

    As in other mammals, sex sorting of pig sperm is based on quantitative flow cytometry. A major disadvantage of the technique is the relatively low efficiency to produce enough sorted sperm for artificial insemination. However, several approaches are on the way to make sexed pig sperm available for commercial application. In this context, for example, the growing field of nanotechnology may significantly contribute to these developments, as it provides highly efficient bio-nanoprobes, for example, based on plasmonic nanoparticles. Independent of the method, further development requires enormous investments and set-up of logistics to get the technology into the practical pig market. Only global players will be able to establish the necessary research projects, but in the end, a significant shift of sex ratios will be available for pig producers as it is already the case for the dairy industry. © 2015 Blackwell Verlag GmbH.

  5. Effects of long-term in vitro exposure of ejaculated boar sperm to zearalenone and α-zearalenol in sperm liquid storage medium.

    Science.gov (United States)

    Sambuu, Rentsenkhand; Takagi, Mitsuhiro; Namula, Zhao; Nii, Masahiro; Taniguchi, Masayasu; Uno, Seiich; Kokushi, Emiko; Tshering, Chenga; dos Santos, Regiane Rodrigues; Fink-Gremmels, Johanna; Otoi, Takeshige

    2013-01-01

    The effects of in vitro exposure of porcine spermatozoa to zearalenone (ZEN) and α-zearalenol (α-ZOL) were studied by evaluating several parameters of an in vitro fertilization (IVF) system. For this purpose, boar spermatozoa cultured with semen storage medium containing 0 (control), 10 and 1000 µg/L of ZEN and α-ZOL for 1 week at 5°C were used for IVF of in vitro matured oocytes. Overall, there were no significant differences in the rates of total penetration, monospermic fertilization, and polyspermic fertilization of oocytes inseminated with spermatozoa from the different groups. Similarly, ZEN and α-ZOL at 10 and 1000 µg/L did not have detrimental effects on the cleavage and development to blastocysts of oocytes after in vitro fertilization. Although the motility, viability, and plasma membrane integrity of spermatozoa significantly decreased after 3 weeks of storage compared to non-stored spermatozoa (P < 0.05), ZEN and α-ZOL at the evaluated concentrations did not exert detrimental effects on the above parameters, even after 3 weeks of storage. These results indicate that prolonged exposure of boar spermatozoa to ZEN and α-ZOL up to 1000 µg/L under reduced metabolic conditions does not affect their in vitro function. © 2012 Japanese Society of Animal Science.

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

    OpenAIRE

    Angelica Garcia-Olivares; Cesar Garzon-Perez; Oscar Gutierrez-Perez; Alfredo Medrano

    2016-01-01

    Objective: To compare different cooling temperatures before ice formation on pig sperm quality, before and after cryopreservation. Methods: Semen diluted in BF5 was cooled from 23 °C to 5 °C (1% glycerol, 200 × 106 cells/mL). Sperm were packaged in plastic straws, and maintained at +5 °C per 16 h. 1. Freezing point of diluted spermatozoa was determined by exposing straws to nitrogen vapors. 2. Straws (at +5 °C) were further cooled to −3 °C, −5 °C, and −7 °C, and rewarmed. 3. Straws (at +5 ...

  7. Cryopreservation of boar sperm comparing different cryoprotectants associated in media based on powdered coconut water, lactose and trehalose.

    Science.gov (United States)

    Silva, C G; Cunha, E R; Blume, G R; Malaquias, J V; Báo, S N; Martins, C F

    2015-04-01

    In swine spermatozoa, the damage caused by cryopreservation is more severe than other species, provoking reduced potential for fertilization. Adjustments in the freezing extender composition may be an important alternative to increase its efficiency. The objective of this study was to test the efficiency of different cryoprotectant solutions during cryopreservation of swine semen with a controlled cooling curve. Three cryoprotectant solutions (5% dimethylformamide, 3% glycerol and the combination of these two cryoprotectants) were used in association with three base media (powdered coconut water, lactose and trehalose), constituting nine different treatments. The semen was frozen using a controlled-rate freezer (TK-3000). After thawing, semen was evaluated for total sperm motility, vigor, morphology, plasma membrane integrity and acrosome integrity. Cryopreservation with the controlled curve using an automated system showed satisfactory results, guaranteeing practicality and repeatability for the process of freezing swine sperm. With this curve, the solutions of lactose, trehalose and powdered coconut water associated with glycerol, as well as the solution of coconut water containing dimethylformamide, presented higher quality of sperm compared to the other solutions. Powdered coconut water associated with dimethylformamide appears as a new solution for swine sperm cryopreservation. The freezing controlled curve used in this study allowed standardization of the cryopreservation technique. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. 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. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. The influence of hemi orchidectomy on reproductive efficiency of boars

    African Journals Online (AJOL)

    APR

    Abstract. Large White boars (n = 120) were assigned to hemi-orchidectomy at 60, 100, 140 or 300 days of age or as intact controls throughout the 450 day observation period to evaluate the effects of unilateral testicular hypertrophy on libido (reaction time - RT), ejaculation rate (EJACS) and sperm quality in hemi-.

  10. Comparative evaluation of the sperm characteristics and ...

    African Journals Online (AJOL)

    Comparative evaluation of the sperm characteristics and morphology of adult Wistar rats fed either low or normal protein-energy diets and orally dosed with ... of adult Wistar rats orally dosed aqueous extracts of C. australis seed (LPSE and NPSE) and stem (LPST and NPST), 300mg of extract/kg body weight of rat/day, ...

  11. Comparative evaluation of the sperm characteristics and ...

    African Journals Online (AJOL)

    olayemitoyin

    Comparative evaluation of the sperm characteristics and morphology of adult Wistar rats fed either low or normal protein-energy diets and orally dosed with aqueous Cuscuta australis extracts. 1. Omirinde, J.O., *. 1. Ozegbe, P.C. and. 2. Oyeyemi, M.O.. Departments of 1Veterinary Anatomy and 2Veterinary Surgery and ...

  12. Identification by affinity chromatography of boar sperm membrane-associated proteins bound to immobilized porcine zona pellucida. Mapping of the phosphorylethanolamine-binding region of spermadhesin AWN.

    Science.gov (United States)

    Ensslin, M; Calvete, J J; Thole, H H; Sierralta, W D; Adermann, K; Sanz, L; Töpfer-Petersen, E

    1995-12-01

    We have identified boar sperm membrane components recovered by affinity chromatography on a porcine zona pellucida affinity column. The major zona pellucida-bound proteins were spermadhesins AWN and AQN-3, the heparin-binding protein pAIF, and a homolog of the mouse milk fat globule membrane protein. All these proteins are phospholipid-binding proteins peripherally associated with the plasma membrane. Our data suggest that coating proteins tightly bound to the external lipid bilayer may act as major zona pellucida-binding molecules. Using a synthetic peptide approach we show that the regions of spermadhesin AWN comprising residues 6-12 and 104-108 possess affinity for phosphorylethanolamine. These two amino acid sequences are in close proximity in the predicted structural model for AWN, and in opposite location to its carbohydrate-recognition domain. Taken together, our data provide further evidence for the possible involvement of members of the porcine spermadhesin protein family in gamete interaction and suggest a model for the ultrastructural disposition of functional domains of spermadhesin AWN bound to the sperm surface.

  13. 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.

  14. 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.

  15. A comparison of mycotoxin adsorbents and their effects on some selected parameters of boar semen

    Directory of Open Access Journals (Sweden)

    Zdeněk Tvrdoň

    2006-01-01

    Full Text Available Effect of two mycotoxin adsorbents with different mechanisms of action were followed in 180 purebred and terminal combination boars reared in artificial insemination stations. The following parameters of boar sperm quality were investigated: volume in millilitres, concentration and motility, and numbers of pathological spermatozoa. When processing the boar semen, the dilution ration and numbers of AI doses were recorded. Compared were two preparations: in case of Preparation 1 the number of evaluated ejaculates was 1,037 while in case of Preparation 2 altogether 1,109 ejaculates were evaluated. Boars receiving diets with Preparation 1 produced more voluminous ejaculates (by 4.1 ml and their concentration of spermatozoa was also higher (by 39 thous./ml; P ≤ 0.001. Dilution parameters were better as well and numbers produced of AI doses were also higher. In case of Preparation 2 the motility of spermatozoa was a higher while the numbers of sperms were lower. The obtained results demonstrated that a suitable adsorbent can show a positive effect on both quantitative and qualitative parameters of boar sperm.

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

    Czech Academy of Sciences Publication Activity Database

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

    2011-01-01

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

  17. Colloid centrifugation of boar semen.

    Science.gov (United States)

    Morrell, J M; Wallgren, M

    2011-09-01

    Colloid centrifugation of boar semen has been reported sporadically for at least the last two decades, beginning with density gradient centrifugation (DGC) and progressing more recently to single layer centrifugation (SLC). Single layer centrifugation through a species-specific colloid has been shown to be effective in selecting the best spermatozoa (spermatozoa with good motility and normal morphology) from boar sperm samples. The method is easier to use and less time-consuming than DGC and has been scaled-up to allow whole ejaculates from other species, e.g. stallions, to be processed in a practical manner. The SLC technique is described, and various scale-up versions are presented. The potential applications for SLC in boar semen preservation are as follows: to improve sperm quality in artificial insemination (AI) doses for 'problem' boars; to increase the shelf-life of normal stored sperm samples, either by processing the fresh semen before preparing AI doses or by processing the stored semen dose to extract the best spermatozoa; to remove pathogens (viruses, bacteria), thus improving biosecurity of semen doses and potentially reducing the use of antibiotics; to improve cryosurvival by removing dead and dying spermatozoa prior to cryopreservation; to select spermatozoa for in vitro fertilization. These applications are discussed and practical examples are provided. Finally, a few thoughts about the economic value of the technique to the boar semen industry are presented. © 2011 Blackwell Verlag GmbH.

  18. Effects of herbal preparation on libido and semen quality in boars.

    Science.gov (United States)

    Frydrychová, S; Opletal, L; Macáková, K; Lustyková, A; Rozkot, M; Lipenský, J

    2011-08-01

    The objective of this study was to investigate the effects of a preparation from herbal extracts (PHE) on libido and semen quality in breeding artificial insemination boars. Ten fertile boars were divided into control and experimental groups according to significant difference of libido. There were no differences in semen quality between groups. Animals were fed a commercial feeding mixture for boars. The feeding mixture for the experimental group was enriched with PHE, which was prepared from Eurycoma longifolia, Tribulus terrestris and Leuzea carthamoides. Duration of the experiment was 10 weeks. Samples of ejaculate were collected weekly. Libido was evaluated according to a scale of 0-5 points. Semen volume, sperm motility, percentage of viable spermatozoa, sperm concentration, morphologically abnormal spermatozoa, daily sperm production and sperm survival were assessed. Amounts of mineral components and free amino acids were analysed in seminal plasma. Significant differences were found in these parameters: libido (4.05 ± 0.22 vs 3.48 ± 0.78; p libido was increased by 20% in comparison with the beginning of the experiment. Results of this study showed positive effect of PHE on libido and some parameters of boar semen quality. © 2010 Blackwell Verlag GmbH.

  19. Protein disulfide isomerase-P5, down-regulated in the final stage of boar epididymal sperm maturation, catalyzes disulfide formation to inhibit protein function in oxidative refolding of reduced denatured lysozyme.

    Science.gov (United States)

    Akama, Kuniko; Horikoshi, Tomoe; Sugiyama, Atsushi; Nakahata, Satoko; Akitsu, Aoi; Niwa, Nobuyoshi; Intoh, Atsushi; Kakui, Yasutaka; Sugaya, Michiko; Takei, Kazuo; Imaizumi, Noriaki; Sato, Takaya; Matsumoto, Rena; Iwahashi, Hitoshi; Kashiwabara, Shin-ichi; Baba, Tadashi; Nakamura, Megumi; Toda, Tosifusa

    2010-06-01

    In mammalian spermiogenesis, sperm mature during epididymal transit to get fertility. The pig sharing many physiological similarities with humans is considered a promising animal model in medicine. We examined the expression profiles of proteins from boar epididymal caput, corpus, and cauda sperm by two-dimensional gel electrophoresis and peptide mass fingerprinting. Our results indicated that protein disulfide isomerase-P5 (PDI-P5) human homolog was down-regulated from the epididymal corpus to cauda sperm, in contrast to the constant expression of protein disulfide isomerase A3 (PDIA3) human homolog. To examine the functions of PDIA3 and PDI-P5, we cloned and sequenced cDNAs of pig PDIA3 and PDI-P5 protein precursors. Each recombinant pig mature PDIA3 and PDI-P5 expressed in Escherichia coli showed thiol-dependent disulfide reductase activities in insulin turbidity assay. Although PDIA3 showed chaperone activity to promote oxidative refolding of reduced denatured lysozyme, PDI-P5 exhibited anti-chaperone activity to inhibit oxidative refolding of lysozyme at an equimolar ratio. SDS-PAGE and Western blotting analysis suggested that disulfide cross-linked and non-productively folded lysozyme was responsible for the anti-chaperone activity of PDI-P5. These results provide a molecular basis and insights into the physiological roles of PDIA3 and PDI-P5 in sperm maturation and fertilization. Copyright 2010 Elsevier B.V. All rights reserved.

  20. 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

  1. Effect of Selenium, Zinc, Vitamin C and E on Boar Ejaculate Quality at Heat Stress

    Directory of Open Access Journals (Sweden)

    Pavel Horký

    2016-01-01

    Full Text Available The aim of experiment was to test effect of selected antioxidants (selenium, zinc, vitamin C and E to reduce the impact of heat stress at boars. In the experiment, boars of Duroc breed were tested. The first control group (n = 10 was not supplemented with antioxidants. The second experimental group (n = 10 was supplemented with antioxidants in the following quantities of 0.5 mg of selenium (seleno-methionine, 100 mg of zinc (zinc-methionine, 70 mg of vitamin E (alpha‑tocopherol and 350 mg of vitamin C (ascorbic acid per kilogram of their feed. The experiment was carried out for 120 days and took place in summer (June to September. During the experiment, average and maximum daily temperatures, where boars were stabled, were monitored. Average daily temperature ranged from 12 to 28 °C. Maximum temperature during the day was from 13 to 32 °C. The evaluation of the semen quality has revealed increased number of abnormal spermatozoa in the control group of boars by 39 % (P < 0.05. There were observed no significant changes at other monitored parameters (ejaculate volume, total count of produced sperm, motility and sperm concentration. The results show that the addition of selenium, zinc, vitamin C and E may reduce the effect of heat stress to some extent at breeding boars.

  2. 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.

  3. Head (H) And Tail (T) Monoclonal Antibodies of Boar Spermatozoa ...

    African Journals Online (AJOL)

    Semen collected from three boars were pooled and the sperm cells separated from the semen, washed, sonicated and mixed with equal volume of Freund's adjuvant. The mixture of sperm with complete Freund's adjuvant was used to immunize five mice of the BALB/C strain intraperitoneally on Day O. The sonicated sperm ...

  4. Egg yolk and glycerol requirements for freezing boar spermatozoa treated with methyl β-cyclodextrin or cholesterol-loaded cyclodextrin.

    Science.gov (United States)

    Blanch, Eva; Tomás, Cristina; Hernández, Marta; Roca, Jordi; Martínez, Emilio A; Vázquez, Juan M; Mocé, Eva

    2014-04-24

    Egg yolk (EY) and glycerol are common constituents of extenders used for sperm cryopreservation. It has been demonstrated that using cholesterol-loaded cyclodextrins (CLC) improves sperm cryosurvival in several species. However, standard freezing extenders might not be the most appropriate for CLC-treated sperm. This study evaluated the EY and glycerol requirements for freezing CLC-treated boar spermatozoa. Semen samples from 34 ejaculates coming from 4 boars were used. Each ejaculate was split into three aliquots: one was used untreated (control), and the other two were treated with 1 mg of CLC or methyl-β-cyclodextrin/120 × 10(6) sperm for 15 min at 22 C prior to cryopreservation. Our results indicated that reducing the concentration of EY was detrimental for sperm viability after thawing (31.57 ± 2 vs. 19.89% ± 2 for 20 and 10% EY, respectively; P <0.05), even in semen treated with CLC. On the other hand, it was observed that the traditional concentration of glycerol (3%) was not the appropriate for freezing CLC-treated sperm (61.10 ± 3 vs. 47.87% ± 3 viable sperm for control and CLC-treated sperm, respectively; P <0.05). Thus, CLC-treated sperm showed a higher tolerance to high glycerol concentrations (5%) in terms of sperm viability (59.19% ± 3) than non-treated sperm (45.58% ± 3; P<0.05). Therefore, it could be necessary to modify the freezing extenders for CLC-treated sperm. Nevertheless, additional studies will be needed to evaluate alternative cryoprotectants and to determine the effect of high glycerol concentrations on sperm functionality.

  5. Methods useful for evaluation of human sperm

    Czech Academy of Sciences Publication Activity Database

    Kubátová, Alena; Čapková, Jana; Pěknicová, Jana

    2012-01-01

    Roč. 67, Issue Supplement s1 (2012), s. 27-27 ISSN 1046-7408. [13th International Symposium for Immunology of reproduction "From the roots to the tops of Reproductive Immunology". 22.06.2012-24.06.2012, Varna] R&D Projects: GA ČR(CZ) GA523/09/1793; GA ČR(CZ) GAP503/12/1834 Institutional research plan: CEZ:AV0Z50520701 Keywords : human sperm * immunofluorescence test * human seminal plasma proteins * flow cytometry Subject RIV: EC - Immunology

  6. Variability in ejaculation rate and libido of boars during reproductive

    African Journals Online (AJOL)

    You

    2015-08-29

    Aug 29, 2015 ... Time spent in preparing or collecting, constituted the period from the entry of boars into the room for collecting semen to onset of ejaculation. Ejaculation rate was defined as the volume of sperm extracted (mL) per unit of time (min). The index of boar libido was defined as the relationship between productive ...

  7. A transgenic boar model to elucidate the role of gonadotropin-releasing hormone 2 and its receptor in regulating testes and sperm function

    Science.gov (United States)

    Boar subfertility represents a major limitation to swine production, reducing conception rate and litter size. Critical to reproductive function, the classical form of GnRH (GnRH1) promotes secretion of the gonadotropins; however, the second mammalian isoform (GnRH2) is a poor stimulator of gonadotr...

  8. Quality evaluation of sperm from livebearing fishes: Standardized assessment of sperm bundles (spermatozeugmata) from Xenotoca eiseni (Goodeidae).

    Science.gov (United States)

    Liu, Yue; Torres, Leticia; Tiersch, Terrence R

    2018-02-01

    Standardized evaluation of sperm quality is essential for research, commercial-scale cryopreservation, and induced spawning. However, standardized methods for evaluation of sperm bundles (spermatozeugmata or spermatophores) have not been established. The purpose of the present study was to use Redtail Splitfin (Xenotoca eiseni) as a model for freshwater livebearing fishes to establish initial standardized methods to collect sperm bundles, and quantitatively and qualitatively evaluate quality-related attributes. No sperm or sperm bundles were able to be collected by stripping. Testes were removed, rinsed, weighed, placed in 50 μL of buffer solution on a glass slide, and crushed gently 3-5 times with angled spade-tip forceps. Sperm bundles were released into the buffer solution and collected with a pipette into 1.5-mL centrifuge tubes. To quantify size and shape, images of bundles were captured with a CCD camera connected to a microscope, and measured with computer software. There was no significant correlation between body wet weight and major bundle axis length (P = 0.6759), minor axis length (P = 0.5658), average axis length (P = 0.5869), aspect ratio (P = 0.7839), and observed area (P = 0.5727). The concentrations of sperm bundles, estimated with the three methods (Makler ® counting chamber, a hemocytometer, and direct counting) were significantly different (P < 0.0001). Hemocytometers were suitable for estimation of bundles from X. eiseni. To evaluate activation of sperm, bundles were viewed with a microscope, and classified into one of five phases by evaluating morphology of the bundles and motion of sperm within the bundles as Phase 0 through Phase 4 that represented early through late activation stages. The frequencies and duration of each activation phase were used to evaluate dissociation of sperm bundles and motility capability of sperm within the bundles. Within 180 min of activation, all five phases were observed. Overall, this study for

  9. 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.

  10. Sperm aggregation by water extracts from two Bursera species.

    Science.gov (United States)

    Serrano, H; Garcia-Suarez, M D

    2001-01-01

    Some plants have more than the common utility value, as is the case of some members of the Bursera species such as the Mexican copal, a plant used for worship. Water extracts of several plants have vaginal contraceptive properties. The authors evaluated the sperm agglutinating activity of two Bursera species on human and boar sperm. Extracts from stems and leaves were obtained. Capacitated sperm samples were used in all cases. There were different agglutinating capacities, which were not observed in the vehicle-only samples. The most frequent sperm agglutination response was that involving the heads. Agglutinating activity was higher from stem- than leaf-derived extracts. The results indicate that proteins present in the extracts are responsible for the aggregation of sperm heads.

  11. Short communication Sperm DNA damage in relation to lipid ...

    African Journals Online (AJOL)

    Leyland Fraser

    2017-03-08

    LPO) and sperm DNA damage following freezing-thawing of boar semen in different extenders. The comet assay was used to measure the extent of sperm DNA damage in a cryoprotectant-free extender or in cryoprotectant-based ...

  12. 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. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Deep-freezing of boar semen in plastic film 'cochettes'.

    Science.gov (United States)

    Eriksson, B M; Rodriguez-Martinez, H

    2000-03-01

    The motility and membrane integrity of spermatozoa from nine boars frozen with a programmable freezing machine in plastic bags, 'cochettes', and in 'maxi-straws', in total doses of 5 x 10(9) spermatozoa/5 ml with glycerol (3%) used as cryoprotectant, were assessed after thawing. A computer-based cell motion analyser was used to evaluate sperm motility, while the integrity of the plasmalemma was assessed with fluorescent supravital dyes (C-FDA/PI). The fertilizing capacity of the semen frozen in the two containers was investigated by inseminating (AI) gilts. Pregnancy was monitored by Doppler-ultrasound, and the numbers of corpora lutea and viable embryos counted at slaughter, between days 30 and 38 after AI. The cochettes sustained the overall procedure of freezing/thawing (FT), with 30 min post-thaw (PT) sperm motility being significantly higher than for straws, 46.9 vs. 39.5%. The only significant difference in motility patterns detected when comparing the packages was a higher sperm velocity (VCL) in cochettes at 30 min PT. However, percentages of FT-spermatozoa with intact membranes, detected with the supravital probes, were higher in maxi-straws than in cochettes, 46.8 vs. 43.0% (P straws and those frozen in cochettes. The results indicate that although the deep-freezing of AI-doses of boar semen in large plastic bags is feasible, problems such as their inconvenient size for storage and inconsistent thawing must be solved before this type of container can be used for the commercial cryopreservation of boar semen.

  14. 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 spermatozoa with high MMP declined drastically after exposures to ≥250 μg ml(-1) NP (P 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. © 2014 Blackwell Verlag GmbH.

  15. 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.

  16. Sensory evaluation of boar-taint-containing minced meat, dry-cured ham and dry fermented sausage by a trained expert panel and consumers.

    Science.gov (United States)

    Verplanken, Kaat; Wauters, Jella; Vercruysse, Vicky; Aluwé, Marijke; Vanhaecke, Lynn

    2017-10-15

    One of the main issues related to entire male pigs is the occurrence of boar taint, an off-odour, which compromises meat consumability. In this study, odour thresholds (indole: 24-65µgkg -1 , skatole: 44-89µgkg -1 , androstenone: 121-342µgkg -1 ) for the boar taint compounds were estimated in minced meat, dry fermented sausage and dry-cured ham. Afterwards, sensory evaluation of these products containing 10% tainted meat (minced meat and dry fermented sausage) or moderate boar taint compound levels (dry-cured ham) occurred. The beneficial effect of diluting tainted meat was demonstrated, as no significant difference in consumability was observed between gilts and 10% tainted meat by experts as well as consumers. Also dry-curing proved a promising technique for masking boar taint and preventing consumer dissatisfaction. The obtained results demonstrate the applicability of the estimated thresholds in meat as a tool for identifying masking and reducing strategies on the perception of boar taint. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Sperm cryopreservation update: Cryodamage, markers, and factors affecting the sperm freezability in pigs.

    Science.gov (United States)

    Yeste, Marc

    2016-01-01

    Cryopreservation is the most efficient method for long-term preservation of mammalian sperm. However, freeze-thawing procedures may strongly impair the sperm function and survival and thus decrease the reproductive performance. In addition, the sperm resilience to withstand cryopreservation, also known as freezability, presents a high individual variability. The present work summarizes the principles of cryoinjury and the relevance of permeating and nonpermeating cryoprotective agents. Descriptions about sperm cryodamage are mainly focused on boar sperm, but reference to other mammalian species is also made when relevant. Main cryoinjuries not only regard to sperm motility and membrane integrity, but also to the degradation effect exerted by freeze-thawing on other important components for sperm fertilizing ability, such as mRNAs. After delving into the main differences between good and poor freezability boar ejaculates, those protein markers predicting the sperm ability to sustain cryopreservation are also mentioned. Moreover, factors that may influence sperm freezability, such as season, diet, breed, or ejaculate fractions are discussed, together with the effects of different additives, like seminal plasma and antioxidants. After briefly referring to the effects of long-term sperm preservation in frozen state and the reproductive performance of frozen-thawed boar sperm, this work speculates with new research horizons on the preservation of boar sperm, such as vitrification and freeze-drying. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. 31P MR Spectroscopy of the Testes and Immunohistochemical Analysis of Sperm 
of Transgenic Boars Carried N terminal Part of Human Mutated Huntingtin

    Czech Academy of Sciences Publication Activity Database

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

    2015-01-01

    Roč. 78, Suppl 2 (2015), s. 28-33 ISSN 1210-7859. [Conference on Animal Models for neurodegenerative Diseases /3./. Liblice, 08.11.2015-10.11.2015] R&D Projects: GA MŠk ED2.1.00/03.0124; GA MŠk(CZ) 7F14308 Institutional support: RVO:67985904 Keywords : Huntington´s disease * testes * sperm Subject RIV: FH - Neurology Impact factor: 0.209, year: 2015

  19. 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.

  20. 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 (Pspermatozoa 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. PMID:24603527

  1. 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.

  2. Evaluation of epididymis storage temperature and cryopreservation conditions for improved mitochondrial membrane potential, membrane integrity, sperm motility and in vitro fertilization in bovine epididymal sperm.

    Science.gov (United States)

    Nichi, M; Rijsselaere, T; Losano, Jda; Angrimani, Dsr; Kawai, Gkv; Goovaerts, Igf; Van Soom, A; Barnabe, V H; De Clercq, Jbp; Bols, Pej

    2017-04-01

    The maintaining of the epididymis at lower temperatures during storage and transport improves sperm quality. Our study aimed to test whether epididymis storage temperature (post-mortem) and sperm cryopreservation affect sperm kinetics, membrane integrity, mitochondrial potential and fertility capacity. Thirty-six epididymides were collected from 18 bulls after slaughter and divided into two groups: at 4 or 34°C for 2-3 hr. The sperm was collected from the epididymis cauda. The evaluation consisted of computer-assisted sperm analysis (CASA), SYBR14/PI/JC1 to evaluate membrane integrity, mitochondrial membrane potential (MMP) and measurement of lipid peroxidation (TBARS). The sperm was then frozen using an automatic device. After thawing, sperm samples were evaluated by the same variables and further in vitro fertilization rates. Cryopreservation negatively affected sperm motility in samples stored at 4 and 34°C. Nevertheless, the 4°C samples yielded higher rates of blastocyst formation. Pre-freeze sperm motility, progressive motility and velocity were higher in sperm from epididymis stored at 4°C while post-thaw sperm motility, progressive motility and velocity remained the same among samples from epididymis stored at 4 or 34°C. However, with regard to the kinetic patterns, samples collected from epididymis stored at 34°C had lower values when compared to those stored at 4°C prior the cryopreservation process. Our results indicate that epididymis handling conditions after cryopreservation may affect sperm quality after thawing, especially due to compromised MMP in sperm collected from epididymis stored at higher temperatures. © 2016 Blackwell Verlag GmbH.

  3. Evaluation of sperm nuclear DNA normality by acridine orange staining technique.

    Science.gov (United States)

    Sukcharoen, N

    1995-02-01

    To determine the relationship between sperm deoxyribonucleic acid (DNA) normality and fertilizing potential, 93 semen samples from 48 fertile donors and 45 male partners of infertile couples whose major abnormalities in the female partner had been ruled out were studied. Semen samples were assessed for conventional parameters (volume, percentage of normal morphology, percentage of progressive motility, sperm concentration, and round cell concentration) according to World Health Organization (WHO) guideline and acridine orange staining. The mean sperm concentration, percentages of progressive motility, and percentage of green fluorescing sperm in semen were significantly higher in samples from fertile donors (p green-fluorescing sperm were significantly higher in fertile donors than infertile patients (65.6% vs 53.3%; p < 0.05). Therefore, the acridine orange staining technique for evaluation of the sperm DNA normality appears to give more information in infertile patients with normal semen analysis. It may be a useful addition to the conventional semen analysis.

  4. Single-cell-based evaluation of sperm progressive motility via fluorescent assessment of mitochondria membrane potential.

    Science.gov (United States)

    Moscatelli, Natalina; Spagnolo, Barbara; Pisanello, Marco; Lemma, Enrico Domenico; De Vittorio, Massimo; Zara, Vincenzo; Pisanello, Ferruccio; Ferramosca, Alessandra

    2017-12-20

    Sperm cells progressive motility is the most important parameter involved in the fertilization process. Sperm middle piece contains mitochondria, which play a critical role in energy production and whose proper operation ensures the reproductive success. Notably, sperm progressive motility is strictly related to mitochondrial membrane potential (MMP) and consequently to mitochondrial functionality. Although previous studies presented an evaluation of mitochondrial function through MMP assessment in entire sperm cells samples, a quantitative approach at single-cell level could provide more insights in the analysis of semen quality. Here we combine laser scanning confocal microscopy and functional fluorescent staining of mitochondrial membrane to assess MMP distribution among isolated spermatozoa. We found that the sperm fluorescence value increases as a function of growing progressive motility and that such fluorescence is influenced by MMP disruptors, potentially allowing for the discrimination of different quality classes of sperm cells in heterogeneous populations.

  5. Incubation of boar spermatozoa in viscous media by addition of methylcellulose improves sperm quality and penetration rates during in vitro fertilization.

    Science.gov (United States)

    González-Abreu, David; García-Martínez, Soledad; Fernández-Espín, Vanesa; Romar, Raquel; Gadea, Joaquín

    2017-04-01

    This work was designed to study whether viscous media can improve the in vitro sperm functionality in pigs by using methylcellulose as a thickener. Viscosity of porcine oviductal fluid (POF) was compared with culture medium (Tyrode's) supplemented with methylcellulose (MET 0, 0.5 and 1% w/v). Spermatozoa were incubated in the different media (0, 1 and 2 h) and sperm motion parameters, lipid membrane disorder, plasma membrane integrity and reactive oxygen species (ROS) formation were assessed. Fertilization results were assessed i) preincubating spermatozoa in the viscous media followed by gamete coculture in a non-viscous medium; and ii) gamete coculture in the viscous media. Viscosity of POF from early luteal phase was higher than late follicular phase. Medium without methylcellulose presented constant viscosity with increased shear rate, while viscosity of the POF and media with methylcellulose was reduced by increased shear rates. Methylcellulose improved sperm linearity, straightness and the proportion of fast-linear spermatozoa. Moreover, methylcellulose increased the rate of viable spermatozoa with intact acrosome and low lipid disorder, reducing the ROS generation. Preincubation in viscous media increased the penetration rate and the mean number of spermatozoa bound to the zona pellucida (both with 0.5 and 1% MET) and reduced monospermy with 1% MET. On the other hand fertilization in the viscous media reduced penetration rate and increased monospermy. The efficiency of the IVF system was not improved with the use of viscous media. The results show the relevance of increasing viscosity thus making the in vitro media more comparable to physiological conditions. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. 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. Copyright © 2012 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    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.

  8. ROLE OF APOPTOSIS IN THE EVALUATION OF SPERM QUALITY ...

    African Journals Online (AJOL)

    Results In the fertile men (control group), none of the sperms was positive for DNA strand breaks, while in the oligozoospermic patients 0.9 0.7% of the sperms were positive for DNA strand breaks. This result was statistically highly ... dans les traitements avancés de fertilisation. African Journal of Urology Vol.8(1) 2002: 6-12 ...

  9. Acrosin inhibitor detecting along the boar epididymis

    Czech Academy of Sciences Publication Activity Database

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

    2016-01-01

    Roč. 82, Jan 2016 (2016), s. 733-739 ISSN 0141-8130 R&D Projects: GA ČR(CZ) GAP503/12/1834; GA MŠk(CZ) ED1.1.00/02.0109; GA ČR GA14-05547S Institutional support: RVO:86652036 ; RVO:61388971 Keywords : Acrosin inhibitor * Boar epididymis * Sperm atozoa Subject RIV: CE - Biochemistry Impact factor: 3.671, year: 2016

  10. 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 (Psperm-oocyte interaction in 0 CLC aliquots but remained unaltered in CLC+ aliquots demonstrating a protective effect of cholesterol treatment. There was no difference between treatments in the in vitro fertilizing ability of frozen-thawed sperm or in the numbers of embryos retrieved following AI with fresh 0 CLC or CLC+ sperm. We conclude that although CLC treatment of dromedary camel sperm improves sperm motility it fails to confer an advantage to them in terms of improved in vitro sperm-oocyte interaction or in vivo fertilization under the conditions tested. Copyright © 2016 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2012-01-01

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

  12. An in vivo evaluation of induction of abnormal sperm morphology by ivermectin MSD (Mectizan).

    Science.gov (United States)

    Otubanjo, O A; Mosuro, A A; Ladipo, T F

    2007-01-01

    The in vivo effects of orally administered ivermectin (Mectizan) on sperm head morphology of albino mice were evaluated. Four different dose levels of 0.25, 0.5, 1.0 and 1.5 x the human therapeutic dose of 150 g kg(-1) body weight, were administered to the animals. The animals were exposed to a single oral treatment. The sperm of the mice from the cauda epididymes were examined 5 and 7 weeks after treatment. Ivermectin (Mectizan) induced sperm head abnormalities; however, the induction was not significantly elevated above the negative control value. Furthermore, the induction of the sperm head abnormalities was not strictly dose-dependent and there was also no correlation between dose level of administered drug and incidence of abnormal sperms. This indicates that the drug might not be mutagenic.

  13. Short communication Relationship between sperm plasma ...

    African Journals Online (AJOL)

    Matshidiso MB. Masenya

    2017-01-04

    Jan 4, 2017 ... Abstract. Sperm quality plays an important role in determining fertility. The aim of the study was to examine the relationship between sperm plasma membrane integrity and morphology, and fertility following artificial insemination (AI). A total of 16 ejaculates were collected from three Large White boars using ...

  14. Relationship between sperm plasma membrane integrity and ...

    African Journals Online (AJOL)

    Sperm quality plays an important role in determining fertility. The aim of the study was to examine the relationship between sperm plasma membrane integrity and morphology, and fertility following artificial insemination (AI). A total of 16 ejaculates were collected from three Large White boars using the gloved hand ...

  15. Semen characteristics and libido in boars treated repeatedly with PGF2alpha.

    Science.gov (United States)

    Estienne, M J; Harper, A F

    2004-05-01

    The objective was to determine the effects of repeated injections of PGF2alpha (Lutalyse; Pfizer) on semen and libido characteristics in terminal-line boars. Semen was collected once weekly from wk 0 to 15 and on four consecutive days during wk 16. Boars received an i.m. injection of 10 mg of PGF2alpha (n = 11) or 2 mL of vehicle (n = 11) immediately before entering the collection room. For the weekly collections, semen volume (220.3 +/- 3.2 mL; mean +/- SE), gel weight (38.7 +/- 0.7 g), total sperm cells (65.4 +/- 1.2 billion), motile sperm cells (67.4 +/- 0.6 %), and sperm velocity (125.9 +/- 1.2 microm/s), were affected by time (P 0.10). Sperm concentration (0.31 +/- 0.01 billion/mL) was not affected (P > 0.10) by time or treatment. The percentage of morphologically normal sperm cells, assessed at wk 16, did not differ (P = 0.39) between groups (80.8 +/- 1.0). Libido was evaluated from wk 0 to 16. There were no effects of treatment or time (P > 0.10) on the period from injection to the start of ejaculation (225.6 +/- 9.1 s). Duration of ejaculation was affected by treatment (P 0.10). The period from injection to the start of ejaculation tended to decrease (by 44%) during the intensive collection period in PGF2alpha-treated boars, but not in controls (treatment x time, P = 0.07). Regardless, the period from injection to the start of ejaculation did not differ (P = 0.63) between groups on d 4 of the intensive collection period. Duration of ejaculation was affected by treatment (P 0.10). Overall, there were no exceptional positive or negative effects of long-term treatment with PGF2alpha on semen characteristics and libido in boars.

  16. Sperm count profile of men presenting for infertility evaluation in ...

    African Journals Online (AJOL)

    Objectives: To determine the prevalence of sperm count abnormalities in men who presented at Nnamdi Azikiwe University Teaching Hospital (NAUTH) Nnewi for infertility management. Materials: Patients' seminal fluid analysis records were accessed by permission from Microbiology Laboratory Department of NAUTH.

  17. 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 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.

  18. In vivo evaluation of ovine sperm/embryo ability in mediating transgenic lamb

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    Shahin Eghbalsaied

    2015-12-01

    Full Text Available Here, we evaluated the ovine embryo ability in DNA uptake as an alternate pathway for sperm-mediated transgenesis. EGFP carrying plasmid was intra-uterus injected through laparoscopy either five days following natural insemination or mixed with sperm cells at the time of fertilization. Excluding trivial false-positive PCR results, following stringent negative control confirmation, PCR analysis of various tissues from 7 generated lambs did not reveal the transgene transfer. Therefore, non-viral vector incubation with sperm or embryo was inefficient for ewe transgenesis.

  19. 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.

  20. Supplemental effect of different levels of taurine in Modena on boar semen quality during liquid preservation at 17°C.

    Science.gov (United States)

    Li, Hao; Zhang, Xiao-Gang; Fang, Qian; Liu, Qi; Du, Ren-Rang; Yang, Gong-She; Wang, Li-Qiang; Hu, Jian-Hong

    2017-11-01

    Peroxidation damage induces sublethal injury to boar sperm during the storage process. Taurine has already been demonstrated to protect cells effectively from oxidant-induced injury. This study was aimed to evaluate the effect of different concentrations of taurine (0.5, 1, 5 and 10 mmol/L) in Modena diluent on boar sperm quality during liquid storage at 17°C. Ejaculates from sexually mature Duroc pigs were collected, pooled and preserved in the Modena containing different concentrations of taurine. Sperm motility, plasma membrane integrity, acrosome integrity, total antioxidative capacity (T-AOC) activity and malondialdehyde content (MDA) were examined every 24 h. Modena diluent containing taurine suppressed the reduction in sperm qualities during the process of liquid preservation compared with those of the control group. After 5 days of liquid preservation, the addition of taurine at 5 mmol/L had the optimal effect on survival time as well as maintenance of motility, plasma membrane integrity, acrosomal integrity, T-AOC activity and MDA content. These results may suggest the possibility that the proper addition of taurine to the semen extender improves the swine production system using artificial insemination by the suppressing of sperm damage and subsequent dysfunction during liquid preservation. © 2017 Japanese Society of Animal Science.

  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. Evaluation of genetic introgression from domesticated pigs into the Ryukyu wild boar population on Iriomote Island in Japan.

    Science.gov (United States)

    Murakami, K; Yoshikawa, S; Konishi, S; Ueno, Y; Watanabe, S; Mizoguchi, Y

    2014-08-01

    We evaluated genetic introgression from domesticated pigs into the Ryukyu wild boar (RWB) population on Iriomote Island based on their genetic structure and diversity. We used a combination of mitochondrial DNA D-loop region (596 bp) polymorphisms and 23 microsatellite markers. RWBs (n = 130) were collected from 18 locations on Iriomote Island and compared with 66 reference samples of European and Asian domestic pigs. We identified six distinct haplotypes, involving 22 single nucleotide polymorphisms (including one insertion) in the RWB population. The phylogenetic tree had two branches: the RWB group and domestic lineage. Fourteen of 130 RWBs (10.8%) belonged to the European domestic lineage, including 11 RWBs from the Panari Islands, northwest of Iriomote Main Island (IMI). The heterozygosity values, total number of alleles, number of effective alleles and polymorphism information content of the RWB groups were lower than those of the European domestic groups. The RWB population on IMI had a lower heterozygous deficiency index (FIS = 0.059) than did the other populations, which indicates that this population was more inbred. There was a large genetic distance (FST = 0.560) between RWBs on IMI and the Meishan populations. Structure analysis using the 23 microsatellite markers revealed that 16 RWBs had an admixture pattern between RWB and domesticated pig breeds. These results suggest that gene flow may have occurred from domestic pigs to RWBs and demonstrate that there was low genetic variation in the IMI population. © 2014 Stichting International Foundation for Animal Genetics.

  3. 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. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. [Effect of experimental classical swine fever (CSF) infection on libido and ejaculate parameters in boars].

    Science.gov (United States)

    Wehrend, A; Fritzemeier, J; Flögel-Niesmann, G; Mönnig, V; Hollen-Horst, M; Meinecke, B

    2006-07-01

    The objective of this study was to characterize the effect of an experimental infection with the classical swine fever (CSF) virus on libido and ejaculate parameters of adult boars. Four boars 10 month old were infected with a CSF field isolate (Visbek/Han95). Semen was collected every second day after infection and daily during the pyrexic phase. The only clinical signs in the boars were an increase in body temperature, but never above 39.9 degrees C and a temporally reduction of food intake. The libido was always good, so semen collection was performed in three boars without difficulty and the semen quality was always in the range of the minimum requirements for sperm that is used for artificial insemination. Although one boar had a good libido only a sperm free ejaculate could be collected on one day. The results show that a CSF virus infection of adult boars hardly causes any clinical symptoms and that sperm can be obtained despite fever. Insemination boars may thus be of special epidemiological relevance for the dissemination of the CSF virus.

  5. Study of sperm proteins in different mammalian species

    OpenAIRE

    Pohlová, Alžběta

    2016-01-01

    Reproduction is an essential feature of all animals and a fundamental step to produce new generations. Study of sperm proteins is crucial for understanding of the sperm-egg recognition. We searched out sperm surface proteins involving in the zona pellucida (ZP) binding and studied whether these proteins are preserved throughout mammalian species. Indirect immunofluorescent technique was used to test a panel of monoclonal antibodies prepared against boar sperm surface proteins on spermatozoa o...

  6. 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.

  7. Evaluation of sperm quality of 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

    2017-01-01

    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.

  8. Studies on varicocele III: ultrastructural sperm evaluation and 18, X and Y aneuploidies.

    Science.gov (United States)

    Baccetti, Baccio M; Bruni, Emanuele; Capitani, Serena; Collodel, Giulia; Mancini, Stefano; Piomboni, Paola; Moretti, Elena

    2006-01-01

    The idea that varicocele plays a detrimental role in fertility is supported by the presence of a higher frequency of affected men among the infertile population than among men with normal semen parameters. In this research we examined ejaculates from a large group of selected men affected by varicocele by light and electron microscopy. The effect of varicocele on chromosome meiotic segregation was investigated by fluorescence in situ hybridization (FISH). The potential benefits of varicocelectomy on sperm quality were evaluated by analyzing sperm characteristics before and after surgical correction of varicocele. Transmission electron microscopy (TEM) analysis, elaborated previously, showed that the incidence of immaturity, apoptosis, and necrosis was higher in the varicocele group than in controls. FISH analysis performed on sperm nuclei from selected patients with varicocele showed that the mean frequencies of disomies and diploidies were generally out of the normal range, indicating a severe disturbance in meiotic segregation. Sperm characteristics evaluated before and after varicocele repair showed a general improvement. As a consequence, the varicocele seem to affect sperm morphology and function concomitantly with meiotic segregation derangement. In consideration of these data, we suggest that TEM and FISH analyses should be performed for all varicocele patients.

  9. Variability in ejaculation rate and libido of boars during reproductive ...

    African Journals Online (AJOL)

    The main objective of the study was to evaluate variability in the ejaculation rate and libido of boars under various genetic and non-genetic influences. A total of 7171 semen samples were collected from Swedish Landrace, Large White and Duroc boars reared under commercial production conditions. Time spent in ...

  10. Seasonal and cryopreservation impacts on semen quality in boars

    Science.gov (United States)

    Seasonal boar infertility occurs worldwide and contributes to economic loss to the pork industry. The current study evaluated cooled vs cryopreserved semen quality of 11 Duroc boars collected in June (cool season) and August 2014 (warm season). Semen was cooled to 16°C (cooled) or frozen over liquid...

  11. 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.

  12. Evaluation of reproductive parameters of vas deferens sperms in Caucasian snake (Gloydius halys caucasicus).

    Science.gov (United States)

    Mozafari, Sayedeh Zahra; Shiravi, Abdolhossein; Todehdehghan, Fatemeh

    2012-01-01

    Reproductive parameters evaluation is considered as helpful tool for gene bank formation in ecological and economically important animals species. Gloydius halys caucasicus is venomous, viviparous pit viper of northwest of Iran. In this research, the spermatic reproductive parameters of this taxon were studied. Twenty six male snakes were collected from Takht-e-Soleiman region between September and October, 2010. Findings revealed that male snakes with body length of 45.07 ± 2.83 cm and body weight of 51.50 ± 10.42 g, and right and left gonads volume of 0.12 ± 0.03 mL and 0.06 ± 0.01 mL are mature ones and sperms concentration in first, middle and final regions of vas deferens duct were, 22.30 ± 19.34 ×10(6) mL(-1), 30.34 ± 11.55 ×10(6) mL(-1), and 37.65 ± 16.46×10(6) mL(-1), respectively. The sperms motility at three regions of duct were 60.53%, 62.07%, and 60.00% and percentage of immotile sperms in these regions were 39.46%, 37.92%, and 39.84%, respectively. Percentage of morphologically normal sperms was 69.23 ± 10.57% and abnormal sperms was 30.76 ± 10.57%; including 12.69 ± 5.25% spiral tailed, 7.33 ± 4.37% coiled tailed and 4.16 ± 2.51% folded tailed sperms. Percentage of live sperms in the first, middle and final regions of duct were 55.76 ± 10.77%, 58.84 ± 12.77%, and 57.69 ± 9.91%, respectively and percentage of dead sperm in these regions were 44.23 ± 10.77%, 41.15 ± 12.77%, and 42.30 ± 9.91%, respectively. Results suggested, mature sperms with acceptable reproductive quality could be collected from Gloydius halys caucasicus snake of Iran between September and October.

  13. Evaluation of reproductive parameters of vas deferens sperms in Caucasian snake (Gloydius halys caucasicus

    Directory of Open Access Journals (Sweden)

    Sayedeh Zahra Mozafari

    2012-06-01

    Full Text Available Reproductive parameters evaluation is considered as helpful tool for gene bank formation in ecological and economically important animals species. Gloydius halys caucasicus is venomous, viviparous pit viper of northwest of Iran. In this research, the spermatic reproductive parameters of this taxon were studied. Twenty six male snakes were collected from Takht-e-Soleiman region between September and October, 2010. Findings revealed that male snakes with body length of 45.07 ± 2.83 cm and body weight of 51.50 ± 10.42 g, and right and left gonads volume of 0.12 ± 0.03 mL and 0.06 ± 0.01 mL are mature ones and sperms concentration in first, middle and final regions of vas deferens duct were, 22.30 ± 19.34 ×106 mL-1, 30.34 ± 11.55 ×106 mL-1, and 37.65 ± 16.46×106 mL-1, respectively. The sperms motility at three regions of duct were 60.53%, 62.07%, and 60.00% and percentage of immotile sperms in these regions were 39.46%, 37.92%, and 39.84%, respectively. Percentage of morphologically normal sperms was 69.23 ± 10.57% and abnormal sperms was 30.76 ± 10.57%; including 12.69 ± 5.25% spiral tailed, 7.33 ± 4.37% coiled tailed and 4.16 ± 2.51% folded tailed sperms. Percentage of live sperms in the first, middle and final regions of duct were 55.76 ± 10.77%, 58.84 ± 12.77%, and 57.69 ± 9.91%, respectively and percentage of dead sperm in these regions were 44.23 ± 10.77%, 41.15 ± 12.77%, and 42.30 ± 9.91%, respectively. Results suggested, mature sperms with acceptable reproductive quality could be collected from Gloydius halys caucasicus snake of Iran between September and October.

  14. 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. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Sperm DNA damage in relation to lipid peroxidation following ...

    African Journals Online (AJOL)

    This study investigated the relationships between lipid peroxidation (LPO) and sperm DNA damage following freezing-thawing of boar semen in different extenders. The comet assay was used to measure the extent of sperm DNA damage in a cryoprotectant-free extender or in cryoprotectant-based extenders after single ...

  16. APPLICATION OF THE SPERM CHROMATIN STRUCTURE ASSAY TO THE TEPLICE PROGRAM SEMEN STUDIES: A NEW METHOD FOR EVALUATING SPERM NUCLEAR CHROMATIN DAMAGE

    Science.gov (United States)

    ABSTRACTA measure of sperm chromatin integrity was added to the routine semen end points evaluated in the Teplice Program male reproductive health studies. To address the hypothesis that exposure to periods of elevated air pollution may be associated with abnormalities in sp...

  17. Testicular and Related Size Evaluations in Nigerian Sahel Goats with Optimal Cauda Epididymal Sperm Reserve.

    Science.gov (United States)

    Abba, Y; Igbokwe, I O

    2015-01-01

    Testicular sizes of animals are important for identification of those with adequate sperm production. The aim of this study was to define the testicular and related size estimates that would be associated with optimal cauda epididymal sperm counts (ESC) in Sahel goats based on postmortem evaluations. A stratified quota sample population of 125 male goats inclusive of all testicular sizes was taken at a slaughterhouse in Maiduguri, Nigeria. The bucks were aged 18-30 months and weighed 17.04 ± 2.99 (12-25) kg. Body, testicular, and epididymal weights of each goat with other related size measurements were estimated. ESC was determined from homogenized tissue using a manual cytometer. At the cut-off ESC of >1.1 × 10(9) sperm heads, 66 (52.80%) of the goats had optimal ESC which was associated with testicular weight of 59.90 ± 16.10 (31.40-86.20) g, gonadosomatic index of 3.51 ± 0.69 (2.00-4.50) g/kg, and scrotal circumference of 19.07 ± 1.29 (17.00-21.80) cm. The size variables of the scrotum and testis correlated with one another and with the ESC. These findings provide data that may be used to anticipate adequate antemortem sperm reserve based on testicular size during preliminary selection of sires for breeding from a sexually mature Sahel buck population.

  18. Flow cytometric evaluation of sperm parameters in relation to fertility potential.

    Science.gov (United States)

    Gillan, Lindsay; Evans, Gareth; Maxwell, W M C

    2005-01-15

    Most laboratory methods used to evaluate semen quality have not correlated highly with fertilizing capacity. The discovery of a variety of fluorochromes and compounds conjugated to fluorescent probes has enabled a more widespread analysis of sperm attributes, and in conjunction with the flow cytometer, permit the evaluation of a large number of spermatozoa. A number of characteristics of sperm integrity, viability and function can be assessed by flow cytometry. The DNA status of spermatozoa has been determined using the metachromatic properties of acridine orange (AO). AO staining, when used in the sperm chromatin structure assay (SCSA), correlates with fertility in a number of species. DNA fragmentation can also be assessed using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, which identifies DNA strand breaks by labeling free 3'-OH termini with modified nucleotides. The status of the sperm acrosome can be determined using fluorescently labeled lectins and LysoTracker Green DND-26, a fluorescent acidotropic probe. Capacitation status has been observed through calcium-mediated changes using chlortetracycline (CTC) or by changes in membrane fluidity monitored by the binding of the fluorescent amphiphilic probe, Merocyanine 540. Fluorescently labeled annexin-V, C6NBD and Ro-09-0198 can also be used to detect changes in membrane phospholipid distribution. Cell viability can be determined using the propensity of propidium iodide (PI), ethidium homodimer-1 (EthD-1) or Yo-Pro-1 to permeate damaged membranes. These are generally more adaptable to clinical flow cytometry than the bisbenzimide membrane impermeable stain, Hoechst 33258, which excites in the ultraviolet range and requires UV laser equipment. Mitochondrial function can be determined using rhodamine 123 (R123) and MitoTracker Green FM (MITO) and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1). Flow cytometry is a tool that may be used

  19. 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

    2017-09-01

    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. © 2017 Japanese Society of Animal Science.

  20. 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...

  1. Preliminary characterization of multiple hyaluronidase forms in boar reproductive tract

    Czech Academy of Sciences Publication Activity Database

    Cibulková, Eva; Maňásková, Pavla; Jonáková, Věra; Tichá, M.

    2007-01-01

    Roč. 68, č. 7 (2007), s. 1047-1054 ISSN 0093-691X R&D Projects: GA ČR GA303/06/0895; GA MŠk 1M06011 Institutional research plan: CEZ:AV0Z50520514; CEZ:AV0Z50520701 Keywords : hyaluronidase * boar sperm * seminal plasma * epididymis * seminal vesicle Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.911, year: 2007

  2. 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.

  3. 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.

  4. 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...

  5. Strategic use of anti-GnRH vaccine allowing selection of breeding boars without adverse effects on reproductive or production performances.

    Science.gov (United States)

    Oliviero, Claudio; Ollila, Anna; Andersson, Magnus; Heinonen, Mari; Voutila, Liisa; Serenius, Timo; Peltoniemi, Olli

    2016-02-01

    Boar stations raise only entire male pigs for selection as reproductive boars, but the majority of them will fail the selection process, ending at slaughter with a high risk of boar tainted meat. The aim of this study was to investigate whether a single dose of Improvac given to 16-week old boars had a negative effect on their subsequent sperm numbers and motility in 16 artificial insemination boars. We also aimed to generate more knowledge on incidence of boar taint at slaughter among Finnish pigs, compare production performances as average daily gain, feed conversion ratio, and carcass and meat quality (lean meat percentage, back fat, pH, color, androstenone, and skatole) of immunocastrated boars (n = 248) with those of entire boars (n = 268). Moreover, we aimed also to explore whether a fat biopsy taken at 16 weeks of age could already reveal the presence of boar taint compounds and be predictive of boar taint development at slaughter age. We found that 32% of entire boars (Figen Landrace, Figen Large White, and their crossbreed) slaughtered at an age of 25 weeks presented levels of androstenone and/or skatole above the threshold for boar taint in their meat. These boars (control) had higher androstenone and skatole levels in the back fat samples at slaughter (0.77 ± 0.55 and 0.09 ± 0.06 μg/g, respectively, mean ± standard deviation) than those in the immuno group (0.20 ± 0.25 and 0.06 ± 0.03 μg/g, respectively, P artificial insemination. We found no difference in the levels of testosterone, anti-GnRH antibodies titers, testicle morphology, and sperm numbers and motility between the boars vaccinated once, at 16 weeks of age, with anti-GnRH vaccine and the control boars (no vaccination). There were no differences in average daily gain, feed conversion ratio, lean meat percentage, and back fat between the immunocastrated boars and entire boars. Meat from immunocastrated boars had a higher pH and better color than meat from entire boars (P meat quality

  6. Evaluation of cryoprotectant and cooling rate for sperm cryopreservation in the euryhaline fish medaka Oryzias latipes.

    Science.gov (United States)

    Yang, Huiping; Norris, Michelle; Winn, Richard; Tiersch, Terrence R

    2010-10-01

    Medaka Oryzias latipes is a well-recognized biomedical fish model because of advantageous features such as small body size, transparency of embryos, and established techniques for gene knockout and modification. The goal of this study was to evaluate two critical factors, cryoprotectant and cooling rate, for sperm cryopreservation in 0.25-ml French straws. The objectives were to: (1) evaluate the acute toxicity of methanol, 2-methoxyethanol (ME), dimethyl sulfoxide (Me(2)SO), N,N-dimethylacetamide (DMA), N,N-dimethyl formamide (DMF), and glycerol with concentrations of 5%, 10%, and 15% for 60min of incubation at 4°C; (2) evaluate cooling rates from 5 to 25°C/min for freezing and their interaction with cryoprotectants, and (3) test fertility of thawed sperm cryopreserved with selected cryoprotectants and associated cooling rates. Evaluation of cryoprotectant toxicity showed that methanol and ME (5% and 10%) did not change the sperm motility after 30min; Me(2)SO, DMA, and DMF (10% and 15%) and glycerol (5%, 10% and 15%) significantly decreased the motility of sperm within 1min after mixing. Based on these results, methanol and ME were selected as cryoprotectants (10%) to evaluate with different cooling rates (from 5 to 25°C/min) and were compared to Me(2)SO and DMF (10%) (based on their use as cryoprotectants in previous publications). Post-thaw motility was affected by cryoprotectant, cooling rate, and their interaction (P⩽0.000). The highest post-thaw motility (50±10%) was observed at a cooling rate of 10°C/min with methanol as cryoprotectant. Comparable post-thaw motility (37±12%) was obtained at a cooling rate of 15°C/min with ME as cryoprotectant. With DMF, post-thaw motility at all cooling rates was ⩽10% which was significantly lower than that of methanol and ME. With Me(2)SO, post-thaw motilities were less than 1% at all cooling rates, and significantly lower compared to the other three cryoprotectants (P⩽0.000). When sperm from individual males

  7. Evaluation of Cryoprotectant and Cooling Rate for Sperm Cryopreservation in the Euryhaline Fish Medaka Oryzias latipes

    Science.gov (United States)

    Yang, Huiping; Norris, Michelle; Winn, Richard; Tiersch, Terrence R.

    2017-01-01

    Medaka Oryzias latipes is a well-recognized biomedical fish model because of advantageous features such as small body size, transparency of embryos, and established techniques for gene knockout and modification. The goal of this study was to evaluate two critical factors, cryoprotectant and cooling rate, for sperm cryopreservation in 0.25-ml French straws. The objectives were to: 1) evaluate the acute toxicity of methanol, 2-methoxyethanol (ME), dimethyl sulfoxide (Me2SO), N, N- dimethylacetamide (DMA), N, N,-dimethyl formamide (DMF), and glycerol with concentrations of 5, 10, and 15% for 60 min of incubation at 4 °C; 2) evaluate cooling rates from 5 to 25 °C/min for freezing and their interaction with cryoprotectants, and 3) test fertility of thawed sperm cryopreserved with selected cryoprotectants and associated cooling rates. Evaluation of cryoprotectant toxicity showed that methanol and ME (5 and 10%) did not change the sperm motility after 30 min; Me2SO, DMA, and DMF (10 and 15%) and glycerol (5, 10 and 15%) significantly decreased the motility of sperm within 1 min after mixing. Based on these results, methanol and ME were selected as cryoprotectants (10%) to evaluate with different cooling rates (from 5 °C/min to 25 °C/min) and were compared to Me2SO and DMF (10%) (based on their use as cryoprotectants in previous publications). Post-thaw motility was affected by cryoprotectant, cooling rate, and their interaction (P ≤ 0.000). The highest post-thaw motility (50 ± 10%) was observed at a cooling rate of 10 °C/min with methanol as cryoprotectant. Comparable post-thaw motility (37 ± 12%) was obtained at a cooling rate of 15 °C/min with ME as cryoprotectant. With DMF, post-thaw motility at all cooling rates was ≤ 10% which was significantly lower than that of methanol and ME. With Me2SO, post-thaw motilities were less than 1% at all cooling rates, and significantly lower compared to the other three cryoprotectants (P ≤ 0.000). When sperm from

  8. Accurate sperm morphology assessment predicts sperm function.

    Science.gov (United States)

    Abu Hassan Abu, D; Franken, D R; Hoffman, B; Henkel, R

    2012-05-01

    Sperm morphology has been associated with in vitro as well as in vivo fertilisation. The study aimed to evaluate the possible relation between the percentage of spermatozoa with normal morphology and the following sperm functional assays: (i) zona-induced acrosome reaction (ZIAR); (ii) DNA integrity; (iii) chromatin condensation; (iv) sperm apoptosis; and (v) fertilisation rates. Regression analysis was employed to calculate the association between morphology and different functional tests. Normal sperm morphology correlated significantly with the percentages of live acrosome-reacted spermatozoa in the ZIAR (r = 0.518; P sperm apoptosis (r = -0.395; P = 0.0206; n = 34) and necrosis (r = -0.545; P = 0.0009; n = 34). Negative correlations existed between for the acrosome reaction, and DNA integrity, while negative associations were recorded with the percentages of CMA(3) -positive spermatozoa, apoptotic and necrotic spermatozoa. Sperm morphology is related to sperm dysfunction such as poor chromatin condensation, acrosome reaction and DNA integrity. Negative and significant correlations existed between normal sperm morphology and chromatin condensation, the percentage of spermatozoa with abnormal DNA and spermatozoa with apoptotic activity. The authors do not regard sperm morphology as the only test for the diagnosis of male fertility, but sperm morphology can serve as a valuable indicator of underlying dysfunction. © 2011 Blackwell Verlag GmbH.

  9. Applying the erythrocyte Pig-a assay concept to rat epididymal sperm for germ cell mutagenicity evaluation.

    Science.gov (United States)

    Ji, Zhiying; LeBaron, Matthew J

    2017-08-01

    The Pig-a assay, a recently developed in vivo somatic gene mutation assay, is based on the identification of mutant erythrocytes that have an altered repertoire of glycosylphosphatidylinositol (GPI)-anchored cell surface markers. We hypothesized that the erythrocyte Pig-a assay concept could be applied to rat cauda epididymal spermatozoa (sperm) for germ cell mutagenicity evaluation. We used GPI-anchored CD59 as the Pig-a mutation marker and examined the frequency of CD59-negative sperm using flow cytometry. A reconstruction experiment that spiked un-labeled sperm (mutant-mimic) into labeled sperm at specific ratios yielded good agreement between the detected and expected frequencies of mutant-mimic sperm, demonstrating the analytical ability for CD59-negative sperm detection. Furthermore, this methodology was assessed in F344/DuCrl rats administered N-ethyl-N-nitrosourea (ENU), a prototypical mutagen, or clofibrate, a lipid-lowering drug. Rats treated with 1, 10, or 20 mg/kg body weight/day (mkd) ENU via daily oral garage for five consecutive days showed a dose-dependent increase in the frequency of CD59-negative sperm on study day 63 (i.e., 58 days after the last ENU dose). This ENU dosing regimen also increased the frequency of CD59-negative erythrocytes. In rats treated with 300 mkd clofibrate via daily oral garage for consecutive 28 days, no treatment-related changes were detected in the frequency of CD59-negative sperm on study day 85 (i.e., 57 days after the last dose) or in the frequency of CD59-negative erythrocytes on study day 29. In conclusion, these data suggest that the epidiymal sperm Pig-a assay in rats is a promising method for evaluating germ cell mutagenicity. Environ. Mol. Mutagen. 58:485-493, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  10. Cryo-scanning electron microscopy (Cryo-SEM) of boar semen frozen in medium-straws and MiniFlatPacks.

    Science.gov (United States)

    Ekwall, Hans; Hernández, Marta; Saravia, Fernando; Rodríguez-Martínez, Heriberto

    2007-06-01

    In this study we demonstrate, in the frozen state, the architecture of frozen boar spermatozoa collected from the sperm-rich fraction of ejaculates (n=13) from four fertile boars packed and split-frozen in medium-straws (MS) and MiniFlatPacks (MFP), cross-sectioned in the frozen state and evaluated by image analysis on images obtained by use of cryo-scanning electron microscopy (Cryo-SEM). The tested hypothesis was that the degree of in situ dehydration and levels of homogeneity of boar semen either frozen in MSs or MFPs packages differ between them, with MFPs allowing for a more uniform dehydration of the spermatozoa and a higher cryosurvival, monitored by computer assisted sperm analysis (CASA) as proportion of linearly motile spermatozoa, compared to semen packaged and processed in MSs. The organization and relative surface of biological material (veins; e.g., frozen extender, bound water, solutes and spermatozoa) as well as free water (lakes) was measured as the degree of dehydration of the samples. The apparent organization of lakes and veins differed between packages, with the MFPs depicting larger lakes than the MSs. The sizes of the lakes in the latter appeared, moreover, highly asymmetrical depending on their position of the section. The relative surface of these lakes per section, respectively veins differed between packages (P<0.05), indicating a larger amount of free-water (lakes; 81.73+/-2.07% vs. 77.91+/-1.57%) in the MFPs and, consequently, thinner veins than in MSs. In conclusion, MFPs seem to allow for a more homogenous dehydration of the spermatozoa/frozen extender compared to MSs, which might account for their somewhat better sperm quality post-thaw.

  11. The Dynamic of Main Indices of Spermiogram at Boars, in Function on Some Factors of Influence

    Directory of Open Access Journals (Sweden)

    Gherasim Nacu

    2011-05-01

    Full Text Available Breeding boars used for semen production. This is conditioned by many factors. The research aim was to assess the some indices of spermogram (volume, mobility, concentration, total sperm / ejaculate at 21 boars, the breeds Large White, Landrace, Duroc and Pietrain in relation with some factors of influence: the race, age, individual and harvesting season. The age of boars was between 8 months and 30 months. The rhythm of harvesting was following: one harvesting followed by the five days break. Depending on the breed, were differences for volume (183,5± 4,88 ml at Pietrain boars and 256±5,3 ml at Landrace boars and for concentration (309,7±6,0 millions/ml at Duroc boars and 430,7±9,3 millions/ml at Pietrain boars. In function on the season, semen volume varied insignificant between summer (minimum values: 188.4 ± 5.6 ml and winter (maximum values: 222.1 ± 6.6 ml. The spermatozoon concentration was insignificantly influenced by the season (371.6 ± 11.7 million/ml - summer; 399.4 ± 12.6 million/ ml – autumn. The greatest differences for all indices of semen analysis were in function on individual. The results show that the indices analyzed are strongly influenced by internal environmental factors (breed, individual and are less conditioned by environmental factors such as the season.

  12. Isolation of total ribonucleic acid from fresh and frozen-thawed boar ...

    African Journals Online (AJOL)

    RNA) from raw fresh semen and frozen-thawed boar semen, using a protocol comprising the conventional TRIzol assay and a membrane-based technique, the PureLink RNA mini kit. Bioanalyzer profile revealed that the sperm RNA size ...

  13. 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

    , 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...

  14. 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.

  15. Sperm preparation for fertilization

    OpenAIRE

    Gadella, B.M.

    2014-01-01

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

  16. Spatiotemporal trends in Canadian domestic wild boar production and habitat predict wild pig distribution

    DEFF Research Database (Denmark)

    Michel, Nicole; Laforge, Michel; van Beest, Floris

    2017-01-01

    eradication of wild pigs is rarely feasible after establishment over large areas, effective management will depend on strengthening regulations and enforcement of containment practices for Canadian domestic wild boar farms. Initiation of coordinated provincial and federal efforts to implement population...... wild boar and test the propagule pressure hypothesis to improve predictive ability of an existing habitat-based model of wild pigs. We reviewed spatiotemporal patterns in domestic wild boar production across ten Canadian provinces during 1991–2011 and evaluated the ability of wild boar farm...... distribution to improve predictive models of wild pig occurrence using a resource selection probability function for wild pigs in Saskatchewan. Domestic wild boar production in Canada increased from 1991 to 2001 followed by sharp declines in all provinces. The distribution of domestic wild boar farms in 2006...

  17. Evaluation of Morphometrical and Histomorphometrical Changes of Testes, Fertility Potential and Sperm Quality in Mice Treated with Aflatoxin

    Directory of Open Access Journals (Sweden)

    abbas Ahamdi

    2017-03-01

    Full Text Available Introduction: Aflatoxin is the most important mycotoxin toxicity and can enter the animal or human reproductive systems and cause some problems in relation to semen quality and fertility decline. The aim of this study was to investigate the effect of aflatoxin on histological structure of the testes and sperm characteristics and cellular targets in spermatogenic compartment and blood level of testosterone and fertility potential. Methods: In this experimental study, 40 adult male mice were divided into 4 groups as the control and experimental groups. Experimental groups have received aflatoxin (100, 350, 700µg/kg by gastric intubation daily. After 45 days, the mice were sacrificed and sperm samples were collected from cauda epididyms in order to evaluate the sperm parameters and perform the in-vitro fertilization analyses. Results: Analyses of sperm parameters demonstrated that sperm motility decreased remarkably (P<0.05 in all three groups of aflatoxin in comparison with the control. Moreover, the percentage of sperms with DNA disintegrity and nuclear immaturity were significantly increased in aflatoxin groups (P<0.05. Results from IVF showed that aflatoxin have been significantly decreased the sperm fertilization potential, preimplantation embryonic development, embryonic quality and percentage of 2-cells embryos and blasocyste in comparison with the control group. Percentage of arrested embryos with high lysis and fragmantation have been increased significantly in aflatoxin-treated groups (P<0.05. Conclusion: Totally, the present results highly support the idea that aflatoxin induces testicular toxicity with adverse effect on sperm quality and fertility potential in a dose-dependent manner. 

  18. 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.

  19. Sperm evaluation of Jungle Cat (Felis chaus) obtained by urethral catheterization (CT) after medetomidine administration.

    Science.gov (United States)

    Kheirkhah, M S; Mollapour Sisakht, M; Mohammadsadegh, M; Moslemi, H R

    2017-03-15

    This study aimed to evaluate semen from Jungle Cat (Felis chaus) by urethral catheterization (CT) after medetomidine administration that offers feasible and different approaches to obtaining good quality sperm, especially in wild felids. Accordingly, this method was tested in five Jungle Cats. After general anesthesia with the α2-agonist medetomidine (which also stimulates semen release into the urethra) and ketamine, an abdomen ultrasound was performed to locate dilation of the first segment of the urethra (prostatic urethra). A commercial Tom cat urinary catheter 3-5 (depending on the size of the animal) was advanced into the urethra to reach the semen full dilated primary region of the urethra, so as to allow semen collection into the lumen of the catheter by capillary forces. After retraction, sperm volumes between 69 ± 27.92 yielded motility of 77.13 ± 14.15 (mean ± SD) with a mean sperm concentration of 75.13 ± 17.05 million/ml. The results of this study showed that semen collection in jungle cat is feasible, using this method. This study describes a simple, useful in field, inexpensive method which does not require the training of the animal and is better than other methods. Samples have normal pH, suitable color and consolidation, high concentration and lower contamination with excellent motility in Jungle Cat and potentially, other wild felid species, as an alternative to electro-ejaculation. Copyright © 2016. Published by Elsevier Inc.

  20. The application of in vitro sperm competition test to evaluate the impact of ZP-derived peptides on fertilization capacity of cat sperm.

    Science.gov (United States)

    Niu, Yuyu; Greube, Alexa; Ji, Weizhi; Jewgenow, Katarina

    2006-09-01

    The present study aimed to establish a sensitive in vitro assay to assess the binding capacity of cat spermatozoa. Cat oocytes and epididymal sperm cells were isolated from gonads and cultured for in vitro fertilization. Before fertilization, the sperm cells were incubated either in 10 microM green dye Fluo-3-AM or 10 microM orange dye CellTracker Orange CMTMR (Molecular Probes), respectively. After removing the dyes by washing, sperm cells stained with each dye were added to medium drops containing oocytes in various proportions and cultured for 16 h at 37 degrees C, 5% CO(2). The oocytes were examined using fluorescence microscopy. Sperm bound to oocytes, and stained with different colors, were counted. When fresh epididymal sperm were mixed in at a specific proportion, the number of sperm bound to the zona pellucida (ZP) of oocytes reflected the proportion of differently colored sperm in the medium. This indicated that neither dye influenced the binding capacity of cat sperm. Mixing fresh and cryopreserved sperm, however, resulted in a higher number of fresh sperm bound to the oocyte surface in comparison to frozen-thawed sperm. Also, the pre-incubation of cat sperm cells with ZP derived peptide reduced the sperm binding capacity by 40%. In conclusion, the presented sperm competition assay allows assessment of fertilizing capacity of cat spermatozoa in vitro when a mixture of two different populations is used. The applied supravital fluorescence dyes do not affect motility and binding capacity of sperm cells and were clearly distinguishable under fluorescence microscopy. We demonstrate that the assay can be used to study the impact of sperm treatment, such as cryopreservation or pre-incubation in bioactive peptides, on fertilizing capacity.

  1. 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. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. African swine fever virus transmission cycles in Central Europe: Evaluation of wild boar-soft tick contacts through detection of antibodies against Ornithodoros erraticus saliva antigen.

    Science.gov (United States)

    Pietschmann, Jana; Mur, Lina; Blome, Sandra; Beer, Martin; Pérez-Sánchez, Ricardo; Oleaga, Ana; Sánchez-Vizcaíno, José Manuel

    2016-01-04

    African swine fever (ASF) is one of the most complex viral diseases affecting both domestic and wild pigs. It is caused by ASF virus (ASFV), the only DNA virus which can be efficiently transmitted by an arthropod vector, soft ticks of the genus Ornithodoros. These ticks can be part of ASFV-transmission cycles, and in Europe, O. erraticus was shown to be responsible for long-term maintenance of ASFV in Spain and Portugal. In 2014, the disease has been reintroduced into the European Union, affecting domestic pigs and, importantly, also the Eurasian wild boar population. In a first attempt to assess the risk of a tick-wild boar transmission cycle in Central Europe that would further complicate eradication of the disease, over 700 pre-existing serum samples from wild boar hunted in four representative German Federal States were investigated for the presence of antibodies directed against salivary antigen of Ornithodoros erraticus ticks using an indirect ELISA format. Out of these samples, 16 reacted with moderate to high optical densities that could be indicative of tick bites in sampled wild boar. However, these samples did not show a spatial clustering (they were collected from distant geographical regions) and were of bad quality (hemolysis/impurities). Furthermore, all positive samples came from areas with suboptimal climate for soft ticks. For this reason, false positive reactions are likely. In conclusion, the study did not provide stringent evidence for soft tick-wild boar contact in the investigated German Federal States and thus, a relevant involvement in the epidemiology of ASF in German wild boar is unlikely. This fact would facilitate the eradication of ASF in the area, although other complex relations (wild boar biology and interactions with domestic pigs) need to be considered.

  3. The effect of breeding facility and sexual stimulation on plasma cortisol in boars.

    Science.gov (United States)

    Levis, D G; Barnett, J L; Hemsworth, P H; Jongman, E

    1995-12-01

    Nine boars were used to evaluate effects of breeding facility design and sexual activity on plasma cortisol concentrations . In one breeding facility (conventional), boars were housed individually in small pens, and female pigs were mated in those boar pens. In another breeding facility (Detection-Mating Area [DMA] system), boars were housed individually in stalls, and female pigs were mated in a specific mating pen adjacent to the front of stalls where boars were housed. After 51 d of housing treatment, a catheter was surgically implanted in the cephalic vein for collection of blood samples. Daytime profiles (hourly collections from 0900 to 1700) of cortisol did not differ among boars in the two treatment groups. Cortisol was greater (P cortisol in boars, but breeding facility did not affect the ACTH-induced changes in cortisol concentrations. There was a treatment x time interaction (P cortisol after sexual stimulation, and the magnitude and duration of increase in cortisol were greater (P Cortisol was greater (P cortisol concentration in boars seems to be a normal biological response to sexual activity. However, magnitude and duration of the increase in cortisol may be influenced by breeding facility design and mating procedure. There is no evidence, based on physiological data, that housing boars in stalls in the DMA system has any adverse effects on their welfare.

  4. 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

    2017-01-01

    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.

  5. 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

    2017-01-01

    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. PMID:27678462

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

    Directory of Open Access Journals (Sweden)

    Violeta Calle-Guisado

    2017-01-01

    Full Text Available 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.

  7. An assessment of the breeding maturity of insemination boars based on ejaculate quality changes.

    Science.gov (United States)

    Banaszewska, Dorota; Kondracki, Stanisław

    2012-01-01

    We examined the sexual development and insemination capability of boars representing five breeds used at Polish sow insemination stations. The speed of attaining full breeding maturity by the boars and their insemination usefulness was assessed based on the results of experiments analysing age-related changes in the physical characteristics of the ejaculates. Directly after sampling, the ejaculates were analysed for the following physical parameters: ejaculate volume, sperm concentration and the percentage of progressively motile spermatozoa per ejaculate. The total number of spermatozoa in the ejaculates and the number of insemination doses obtained from one ejaculate were calculated using SYSTEM SUL v. 6.1 software. The sexual development of boars continues during insemination use, leading to improved ejaculatory performance. The physical parameters of the ejaculate change along with the age of the boar. The Duroc and Hampshire boars represent a pattern of sexual development that is different from that of the PLW, PL and Pietrain males, with an evident earlier termination of growth. The American boars (Duroc and Hampshire) acquire top ejaculatory performance approximately 6 months earlier.

  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. Comparative immunolocalization of heat shock proteins (Hsp)-60, -70, -90 in boar, stallion, dog and cat spermatozoa.

    Science.gov (United States)

    Volpe, S; Galeati, G; Bernardini, C; Tamanini, C; Mari, G; Zambelli, D; Seren, E; Spinaci, M

    2008-08-01

    Heat shock proteins (Hsp)-60, -70 and -90 are important testis chaperones that fulfil several functions during sperm cell maturation. In post-meiotic cells, their expression may change or may be undetectable and in some species it may be evident in mature spermatozoa. The aims of this study were to verify whether Hsp60, -70 and -90 are present in the sperm, and to compare their localization in boar, stallion, cat and dog spermatozoa by immunofluorescence. Hsp-60 immunoreactivity was detected in sperm midpiece in all the species examined. In stallion sperm, Hsp70 signal was localized in the sub-equatorial band, whereas immunoreactivity was evident on the neck of dog spermatozoa and on both neck and sub-equatorial region of cat spermatozoa. In agreement with our previous observations, a triangular fluorescent signal in the equatorial segment of fresh boar sperm was detected. Hsp90 immunoreactivity was present in different portions of sperm tail: in the midpiece of both boar and cat spermatozoa and in the neck and throughout the tail in dog and stallion spermatozoa, respectively. When capacitation and acrosome reaction were induced in boar, stallion and dog spermatozoa, no changes in both Hsp60 and -90 were recorded by either Western blot or immunofluorescence. After induction of acrosome reaction, a Hsp70 redistribution in boar spermatozoa and an increased percentage of stallion spermatozoa showing the post-acrosomal signal were observed although no changes were recorded by Western blot; in dog spermatozoa, no changes in Hsp70 were found by Western blot and immunofluorescence after capacitation and acrosome reaction.

  10. 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 (p0.05. The combination egg yolk-AhPlus seems to be an alternative to standard extenders, conferring stability in boar sperm cells against cold shock.

  11. Evaluation of ram semen quality using polyacrylamide gel instead of cervical mucus in the sperm penetration test.

    Science.gov (United States)

    Martínez-Rodríguez, C; Alvarez, M; Ordás, L; Chamorro, C A; Martinez-Pastor, F; Anel, L; de Paz, P

    2012-05-01

    Fertility is a very complex biological function that depends on several properties of the spermatozoa, including sperm motility. Two objectives are analyzed in this study: (1) Replace the cervical mucus by a synthetic medium in a sperm penetration test, and (2) evaluating the results of this test objectively analyzing the sperm number that migrates. In experiment 1, we have tested eight concentrations of acrylamide (1%-2%). Rheological properties of media were analyzed. The plastic straws, loaded with acrylamide, were placed vertically on the semen sample tube for 15 min at 39 °C. After, the acrylamides were placed, by segments of 5 mm, into wells of a 24-well plate, dyed with Hoechst 33342 and the number of spermatozoa were calculated by automated microscopy analysis. The 1.55% and 1.6% acrylamide gel showed a number of spermatozoa emigrating closer to that seen with natural mucus. In experiment 2, we applied the sperm penetration in acrylamide 1.6% and 1.55% using fresh semen and cooled semen at 15 °C and 5 °C. The spermatozoa counts were performed for each segment of 10 mm. Semen chilled at 15 °C presented intermediate values of sperm counts in comparison with fresh semen (higher) and 5 °C chilled semen. The sperm counts do not differ between acrylamides but the rheological properties of acrylamide 1.6% were more similar to those of the natural cervical mucus. In experiment 3, we have observed significant correlations between the number of spermatozoa and several sperm quality parameters (positive: progressive motility and velocity according to the straight path; negative: damaged acrosomes and apoptotic cells) in 1.6% acrylamide media. We conclude that the size of the cell subpopulation, objectively calculated, that migrate beyond 20 mm in 0.5-mL straws filled with acrylamide is a useful parameter in ram sperm quality assessment and further studies are needed to evaluate its relationship with field fertility. Copyright © 2012 Elsevier Inc. All rights

  12. Cryopreservation of boar semen. II: Effect of cooling rate and duration of freezing point plateau on boar semen frozen in mini- and maxi-straws and plastic bags.

    Science.gov (United States)

    Bwanga, C O; Einarsson, S; Rodriguez-Martinez, H

    1991-01-01

    The post-thaw motility and the acrosome integrity of semen from 4 boars frozen with a programmable freezing machine, in mini (0.25 ml) and maxi (5 ml) plastic straws and in 10 x 5 cm Teflon FEP-plastic bags (0.12 mm thick, 5 ml), were compared. The freezing of the semen was monitored by way of thermo-couples placed in the straws and the bags. Three freezing programmes were used, namely A: from +5 degrees C, at a rate of 3 degrees C/min, to -6 degrees C, held for 1 min at -6 degrees C, and followed by a cooling rate of 20 degrees C/min to -100 degrees C; B: a similar curve except that there was no holding time at -6 degrees C and that the cooling rate was 30 degrees C/min, and C: from +5 degrees C to -100 degrees C, with a cooling rate of 35 degrees C/min, followed by storage in liquid N2. Despite the freezing curve assayed, both the mini-straws and the bags depicted much shorter freezing point plateaus as compared to the maxi-straws. Post-thaw sperm motility as well as the amount of normal apical ridges were equally significantly higher when semen was frozen in mini-straws or in bags than in maxi-straws. Significant differences in these post-thawing parameters were obtained between the freezing curves used. The stepwise freezing procedure A appeared as the best alternative for boar semen, considering this in vitro evaluation.

  13. Experimental Evaluation of Faecal Escherichia coli and Hepatitis E Virus as Biological Indicators of Contacts Between Domestic Pigs and Eurasian Wild Boar.

    Science.gov (United States)

    Barth, S; Geue, L; Hinsching, A; Jenckel, M; Schlosser, J; Eiden, M; Pietschmann, J; Menge, C; Beer, M; Groschup, M; Jori, F; Etter, E; Blome, S

    2017-04-01

    Domestic pigs and Eurasian wild boar (Sus scrofa) share several important viral and bacterial pathogens. Therefore, direct and indirect contacts between domestic pigs and wild boar present a risk of pathogen spillover and can lead to long-term perpetuation of infection. Biological indicators could be a powerful tool to understand and characterize contacts between wild boar and domestic pigs. Here, faecal Escherichia coli and Hepatitis E virus (HEV) were explored as potential biological indicators under experimental conditions. The data gained in our pilot study suggest that faecal E. coli can be used as biological indicator of contact between wild boar and domestic pig. For HEV, faecal transmission was also confirmed. However, molecular studies on full-genome basis did not reveal markers that would allow tracing of transmission direction. Based on these promising results, future field studies will especially target the practicability of E. coli microbiome molecular typing as surrogate of contacts at the wildlife-livestock interface. © 2015 Blackwell Verlag GmbH.

  14. In vivo fertilizing capacity of deep frozen boar semen packaged in plastic bags and maxi-straws.

    Science.gov (United States)

    Bwanga, C O; Hofmo, P O; Grevle, I S; Einarsson, S; Rodriguez-Martinez

    1991-05-01

    Pooled ejaculates from six fertile boars were frozen under controlled conditions in Teflon FEP-film plastic bags (5 ml) and maxi-straws (2.5 ml) using 3% glycerol as cryoprotectant. The percentages of both post-thaw motility and normal apical ridges were significantly higher (P less than 0.001) for the bags (54.5 and 75%) than for the maxi-straws (40.1 and 59.4%) respectively. For evaluation of the in vivo fertilizing capacity of the frozen-thawed spermatozoa, 26 gilts were inseminated once 24 h after the first observation of standing reflex in their second oestrus, with 5 ml of semen (containing 5 billion spermatozoa) reconstituted in 80 ml of BTS from either bags or maxi-straws. Ova were recovered from the oviducts/uteri 2-4 days following insemination and examined for cleavage and sperm binding to the zona pellucida (ZP). Significantly higher rates (P less than 0.02) of fertilized ova were found in the bag-inseminated (75%) than in maxi-straw inseminated gilts (63%); and similarly their ova had significantly more spermatozoa in the ZP, irrespective of whether they were fertilized or nonfertilized. This study confirmed that the plastic bags are suitable and may be used for packaging single insemination doses of deep frozen boar semen for routine A.I. work.

  15. Dietary l-arginine supplementation improves semen quality and libido of boars under high ambient temperature.

    Science.gov (United States)

    Chen, J Q; Li, Y S; Li, Z J; Lu, H X; Zhu, P Q; Li, C M

    2017-12-04

    l-Arginine is a nutritionally essential amino acid for spermatogenesis and plays versatile roles in animal health and can be utilized as a potential agent to improve reproductive performance of boars under high ambient temperature. The present study aimed to determine whether dietary l-arginine could alleviate heat stress-induced infertility in boars. In all, 20 boars (PIC 1040; 248.59±3.84 kg BW and 407.65±6.40 days of age) were selected and randomly assigned to four groups (group 0.0%, basal diet; group 0.6%, 0.8% or 1.0%, basal diet added with 0.6%, 0.8% or 1.0% l-arginine (wt:wt), respectively.) The four diets were made isonitrogenous by addition of appropriate amounts of l-alanine. Boars were pre-fed the corresponding experimental diet for 42 days. Then, the semen characteristics and libido were accessed for 6 weeks during the hot summer period (25.5° to 33.0°C). Results show that dietary l-arginine remarkably improved sperm motility, normality, total sperm number and effective total sperm number. Also, dietary l-arginine improved semen antioxidant capacity, such as decrease of malondialdehyde and 8-Hydroxy-2'-deoxyguanosine content in sperm (Pl-arginine-supplemented group which also accompany with higher ATP content than the 0.0% group. The boars fed 0.8% l-arginine show increased levels of estradiol-17β and testosterone and exhibit improved libido performance than boars in the 0.0% group. Adding dietary l-arginine linearly increased (P=0.002) nitric oxide content (as l-arginine increased). The scrotal surface temperature in the 0.6%, 0.8% and 1.0% group were decreased by 0.9°C, 0.9°C and 0.4°C, respectively, compared with the 0.0% group. l-Arginine levels caused linear effect on semen quality and antioxidant capacity, also caused quadratic effect on libido performance. During the hot summer months, the predicted optimal l-arginine levels for best semen quality and antioxidant capacity was 0.8% to 1.0% and for best libido performance was 0.8%. It can

  16. Cryopreservation of boar semen in mini- and maxi-straws.

    Science.gov (United States)

    Bwanga, C O; de Braganca, M M; Einarsson, S; Rodriguez-Martinez, H

    1990-10-01

    Split ejaculates from four boars were frozen with a programmable freezing machine, in mini- (0.25 ml) and maxi- (5 ml) plastic straws with an extender at either acidic (6.3) or alkaline (7.4) pH. Glycerol (3%) was used as cryoprotectant. The freezing of the semen was monitored by way of thermocouples placed in the straws. Post-thaw motility and acrosome integrity were evaluated; the latter using phase contrast microscopy, eosin-nigrosin stain and electron microscopy. Post-thaw sperm motility was significantly higher when semen was frozen in mini-straws than in maxi-straws. For the mini-straws, the motility was better when semen was exposed to an acidic environment during freezing, but this beneficial effect of the low extracellular pH was not evident when maxi-straws were thawed. The motility of the spermatozoa diminished significantly during the thermoresistance test (0 h and 2 h time) at 37 degrees C in a similar way for both straws and extracellular pH's. The freezing procedure, no matter the extracellular pH, did not cause major acrosomal damages, but significantly more normal apical ridges were present in the mini-straws than in the maxi-straws. This in vitro evaluation indicated that the freezing method employed was better for mini- than for maxi-straws since the freezing of the 5 ml volumes was not homogeneous, due to the large section area between the surface and the core of the straw.

  17. 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 (Pslaughter, boars and gilts of the GG genotype showed more playing behaviour (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

  18. Evaluation of sperm DNA quality in men presenting with testicular cancer and lymphoma using alkaline and neutral Comet assays.

    Science.gov (United States)

    Kumar, K; Lewis, S; Vinci, S; Riera-Escamilla, A; Fino, M-G; Tamburrino, L; Muratori, M; Larsen, P; Krausz, C

    2018-01-01

    Despite more cancers in young men over the past two decades, improvements in therapies give a greater chance to live full lives following treatment. Sperm genomic quality is variable following cancer diagnosis, so its assessment is important if sperm cryopreservation is being considered. Here, we evaluated DNA damage using two DNA damage assays: an alkaline and for the first time, a neutral Comet assays in men presenting with testicular cancer (n = 19 for alkaline and 13 for neutral group) and lymphoma (n = 13 for alkaline and 09 for neutral group) compared with fertile donors (n = 20 for alkaline and 14 for neutral group). No significant differences were observed in any semen analysis parameters. In contrast, sperm DNA damage was higher in men with testicular cancer than in donors as assessed by both the alkaline (12.4% vs. 37.4%, p alkaline (35.0% vs. 12.4%) and neutral (10.7% against 7.5% (p alkaline and neutral Comet assays is more sensitive than semen analysis in detecting impaired sperm quality in men presenting with cancer. It may provide a useful adjunct when considering storage prior to cancer investigations and assisted reproductive techniques (ART)-based treatment. © 2017 American Society of Andrology and European Academy of Andrology.

  19. Human sperm bioassay has potential in evaluating the quality of cumulus-oocyte complexes.

    Science.gov (United States)

    Hossain, A M; Rizk, B; Huff, C; Helvacioglu, A; Thorneycroft, I H

    1996-01-01

    Human sperm bioassay is routinely used as a quality control check for the culture media. This is one of the three bioassays chosen by the College of American Pathologists (CAP) for interlaboratory proficiency testing to assess the standards of in vitro fertilization (IVF) and andrology laboratories. This study utilized sperm bioassay to assess the quality of cumulus-oocyte complexes (COCs) retrieved in IVF procedures COCs, harvested from the female partner of IVF couples, undergoing identical ovarian stimulation protocols, were individually inseminated with the sperm of the corresponding male partner. Sperm motility in sperm-COC cocultures were compared. Cocultures were established by inseminating the 103 COCs, retrieved from 18 IVF couples with 1 x 10(5) to 2 x 10(5) sperm of the corresponding male partners of the couples. In all 18 cases, the sperm were prepared identically using the Percoll wash method. The cocultures were maintained for 48 h but the oocytes were removed immediately after the fertilization check (approximately 16 h). The motility of sperm in the cocultures and in the insemination stocks were noted and 17 of 18 sperm stocks used for insemination had similar high preinsemination motility (90.2 +/- 5.0%). At 48 h the sperm motility had significantly decreased in the cocultures compared to the insemination stocks; 52.7 +/- 19.9% versus 67.2 +/- 10.4%. There was no difference in the motility among the small, medium, and large COCs (56.4 +/- 24.6%, 52.5 +/- 17.9%, and 50.8 +/- 20.9%, respectively). In 45% of IVF cases, the motility in cocultures varied widely, falling below as well as above that of their corresponding insemination stocks. Furthermore, the sperm motility varied among the cocultures in both pregnant and nonpregnant patients but the extent of variation appears to be greater in the latter. The inter-COC coculture sperm motility variation most likely is due to the differences in the quality of cumulus-oocyte complexes.

  20. Effect of liquid storage on sorted boar spermatozoa.

    Science.gov (United States)

    Spinaci, M; Vallorani, C; Bucci, D; Bernardini, C; Tamanini, C; Seren, E; Galeati, G

    2010-09-15

    A routine use of boar sexed semen is far from being a reality due to many limiting factors among which is the long sorting time necessary to obtain the adequate number of sexed spermatozoa for artificial insemination and the high susceptibility to damages induced by cryopreservation. The aim of this study was to evaluate the modification induced by 24-26 h storage on sorted boar spermatozoa on the basis of their viability, acrosome status, Hsp70 presence, and in vitro fertilizing ability. The percentage of viable cells, according to SYBR green/PI staining, was negatively affected (P fertilization, these parameters were negatively (P fertility and function of sex-sorted boar spermatozoa, the mechanisms by which sorting and liquid storage cause these injures require further study. Copyright 2010 Elsevier Inc. All rights reserved.

  1. Involvement of glucocorticoids in testicular involution after active immunization of boars against GnRH.

    Science.gov (United States)

    Wagner, A; Claus, R

    2004-02-01

    Active GnRH immunization of boars inhibits LH and testicular steroids but the consequences for spermatogenesis are unknown. Six boars were immunized three times against GnRH at 20, 24 and 28 weeks. Another six boars served as controls. Plasma LH and FSH were determined at 28 and 31 weeks. Testosterone and cortisol were determined before killing the pigs at 32 weeks. Tissue samples were taken for histology and fluid from the seminiferous tubuli for steroid determination. Individual germ cells were counted in histological sections. The glucocorticoid receptor (GCR), mitosis of spermatogonia and apoptosis were characterized by immunocytochemistry. Immunization reduced LH and testosterone to base levels whereas FSH was not changed. Testis weight was reduced by 64% due to a loss of Leydig cell cytoplasm (90.3%) and a decrease of tubule diameters (60.6%). Except for A-spermatogonia, all other spermatogenic cells were reduced by about 60%. Mitosis was reduced in immunized boars. Expression of GCRs was limited to spermatogonia and differed between immunized boars (8% of spermatogonia) and controls (2%). In the controls, androgen concentrations in tubular fluid were tenfold higher compared with immunized boars. Cortisol concentrations were of the order of 40 nmol/l both in the tubular fluid and blood plasma. These concentrations did not differ between groups. Apoptosis occurred only in spermatogonia and pachytene spermatocytes and was twofold higher in immunized boars compared with controls. Thus the availability of glucocorticoids in the tubuli and the expression of GCRs initiate apoptosis, which in turn reduces sperm yield. Testosterone is known to be an inhibitor of GCR expression, thus increasing the efficiency of spermatogenesis.

  2. 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.

  3. 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 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 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. © 2015 American Society of Andrology and European Academy of Andrology.

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

    Science.gov (United States)

    CHANAPIWAT, Panida; OLANRATMANEE, Em-On; KAEOKET, Kampon; TUMMARUK, Padet

    2014-01-01

    ABSTRACT The aim of the present study was to determine the conception rate and litter size in sows after fixed time intra-uterine insemination using frozen-thawed boar semen in a commercial swine herd in Thailand. Sixty-nine Landrace multiparous sows were randomly allocated into two groups, including control (n=36) and treatment (n=33). The control sows were inseminated with extended fresh semen (3 × 109 motile sperm/dose, 100 ml) at 24, 36 and 48 hr after the onset of estrus. The treatment sows were inseminated with frozen-thawed semen (2 × 109 motile sperm/dose, 20 ml) at 24 and 36 hr after induction of ovulation by human chorionic gonadotropin. All inseminations were carried out by using an intra-uterine insemination technique. The time of ovulation was determined by using transrectal real-time B-mode ultrasonography. The conception rate, farrowing rate, total number of piglets born/litter (TB) and number of piglets born alive/litter (BA) were evaluated. The sows inseminated with extended fresh semen yield a higher TB (10.8 versus 9.0 piglets/l, P=0.015) and tended to have a higher conception rate (88.9% versus 75.8%, P=0.150) than sows inseminated with frozen-thawed semen. In conclusion, insemination using frozen-thawed boar semen can be practiced with convinced fertility under field conditions by fixed-time intrauterine insemination with 2 × 109 sperm/ dose of 20 ml at 24 and 36 hr after the onset of estrus. PMID:24954517

  5. Assessment of sperm DNA fragmentation in stallion (Equus caballus) and donkey (Equus asinus) using the sperm chromatin dispersion test.

    Science.gov (United States)

    Cortés-Gutiérrez, E I; Crespo, F; Serres-Dalmau, C; Gutiérrez de las Rozas, A L; Dávila-Rodríguez, M I; López-Fernández, C; Gósalvez, J

    2009-10-01

    Sperm DNA fragmentation (sDF) is an important parameter to assessing sperm quality. Information about sperm quality is not available for donkeys, especially in some breeds at risk of extinction. The objectives of this research were to test the four commercial variants of sperm chromatin dispersion test (SCD; sperm Halomax test), originally developed to assess sDF in boars, bulls, rams and stallions, in order to scrutinize their applicability in the study of sDF in a donkey breed at risk of extinction (Zamorano-Leonesa), for which there is no specific test available to analyze sperm at present. Only the SCD test, originally developed for stallions, produced stable and consistent results, and was deemed suitable to assess DNA fragmentation in sperm samples from donkeys. Image analysis was used to compare differences between the SCD methodology applied to stallion and donkey semen samples processed under the same experimental conditions. The extent of SCD in the SCD test was approximately 20% lower in donkey sperm than in stallion sperm. Yet, the ratio of chromatin sperm dispersion achieved in fragmented and unfragmented nuclei did not differ significantly between species. These data suggest that a similar protein depletion treatment can cause differences in protein removal in equivalent cells from different species and that sperm chromatin may be organized differently in stallions and donkeys.

  6. 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.

  7. Is there a relationship between the chromatin status and DNA fragmentation of boar spermatozoa following freezing-thawing?

    Science.gov (United States)

    Fraser, L; Strzezek, J

    2007-07-15

    In this study a radioisotope method, which is based on the quantitative measurements of tritiated-labeled actinomycin D ((3)H-AMD) incorporation into the sperm nuclei ((3)H-AMD incorporation assay), was used to assess the chromatin status of frozen-thawed boar spermatozoa. This study also tested the hypothesis that frozen-thawed spermatozoa with altered chromatin were susceptible to DNA fragmentation measured with the neutral comet assay (NCA). Boar semen was diluted in lactose-hen egg yolk-glycerol extender (L-HEY) or lactose ostrich egg yolk lipoprotein fractions-glycerol extender (L-LPFo), packaged into aluminum tubes or plastic straws and frozen in a controlled programmable freezer. In Experiment 1, the chromatin status and DNA fragmentation were measured in fresh and frozen-thawed spermatozoa from the same ejaculates. There was a significant increase in sperm chromatin destabilization and DNA fragmentation in frozen-thawed semen as compared with fresh semen. The proportions of spermatozoa labeled with (3)H-AMD were concurrent with elevated levels of sperm DNA fragmentation in K-3 extender, without cryoprotective substances, compared with L-HEY or L-LPFo extender. Regression analysis revealed that the results of the (3)H-AMD incorporation assay and NCA for frozen-thawed spermatozoa were correlated. Boars differed significantly in terms of post-thaw sperm DNA damage. In Experiment 2, the susceptibility of sperm chromatin to decondensation was assessed using a low concentration of heparin. Treatment of frozen-thawed spermatozoa with heparin revealed enhanced (3)H-AMD binding, suggesting nuclear chromatin decondensation. The deterioration in post-thaw sperm viability, such as motility, mitochondrial function and plasma membrane integrity, was concurrent with increased chromatin instability and DNA fragmentation. This is the first report to show that freezing-thawing procedure facilitated destabilization in the chromatin structure of boar spermatozoa, resulting in

  8. Niche conservatism and the invasive potential of the wild boar.

    Science.gov (United States)

    Sales, Lilian Patrícia; Ribeiro, Bruno R; Hayward, Matt Warrington; Paglia, Adriano; Passamani, Marcelo; Loyola, Rafael

    2017-09-01

    Niche conservatism, i.e. the retention of a species' fundamental niche through evolutionary time, is cornerstone for biological invasion assessments. The fact that species tend to maintain their original climate niche allows predictive maps of invasion risk to anticipate potential invadable areas. Unravelling the mechanisms driving niche shifts can shed light on the management of invasive species. Here, we assessed niche shifts in one of the world's worst invasive species: the wild boar Sus scrofa. We also predicted potential invadable areas based on an ensemble of three ecological niche modelling methods, and evaluated the performance of models calibrated with native vs. pooled (native plus invaded) species records. By disentangling the drivers of change on the exotic wild boar population's niches, we found strong evidence for niche conservatism during biological invasion. Ecological niche models calibrated with both native and pooled range records predicted convergent areas. Also, observed niche shifts are mostly explained by niche unfilling, i.e. there are unoccupied areas in the exotic range where climate is analogous to the native range. Niche unfilling is expected as result of recent colonization and ongoing dispersal, and was potentially stronger for the Neotropics, where a recent wave of introductions for pig-farming and game-hunting has led to high wild boar population growth rates. The invasive potential of wild boar in the Neotropics is probably higher than in other regions, which has profound management implications if we are to prevent their invasion into species-rich areas, such as Amazonia, coupled with expansion of African swine fever and possibly great economic losses. Although the originally Eurasian-wide distribution suggests a pre-adaptation to a wide array of climates, the wild boar world-wide invasion does not exhibit evidence of niche evolution. The invasive potential of the wild boar therefore probably lies on the reproductive, dietary and

  9. A new laboratory model using bull and boar spermatozoa and fluorescent beads to assess a membrane's occlusive potential.

    Science.gov (United States)

    Szellö, M; Janett, F; Ewald, C; Music, M; Sener, B; Attin, T; Schmidlin, P R

    2016-11-01

    The objective of the present study is to assess the potential of bull and boar spermatozoa and fluorescent beads to be used as a surrogate cell model to determine the cell occlusive potential in vitro using membranes of standardized porosities. A two-chamber model system consisting of upper and lower chambers, which could be separated by membranes, was constructed. Isopore polycarbonate membranes with different standardized pore diameters were used to assess the mobile cellular penetration behavior of spermatozoa or the more passive non-cellular permeability of fluorescent particles (beads) of different diameter and color. In a first experiment, spermatozoa were placed in the lower chamber, whereas semen extender only was placed in the upper chamber. After 10 min of incubation at 37 °C, the sperm number was assessed in the latter. In a second experiment, a bead solution was drawn through resorbable collagen membranes from the upper into the lower chamber by vacuum using a syringe and bead number and size was analyzed by flow cytometry. All experiments were carried out in triplicates. A non-porous polyester membrane was used as negative control to assess the overall tightness of the setup. Boar and bull spermatozoa had average cell body lengths and widths of 9 × 5 μm and were unable to pass through pores ≤2 μm, whereas they were detectable at pore sizes ≥3 μm. Their number increased with increasing pore diameters, i.e., from minimal concentrations of 0.1 × 10 6 /ml for boar and 0.5 × 10 6 /ml for bull spermatozoa at 3 μm to maximal concentrations of 2.1 × 10 6 /ml for boar and 13.1 × 10 6 /ml for bull spermatozoa at 8 μm. The fluorescent beads followed the expected pattern of permeability reliably correlating bead and pore diameter. Within the limitations of this laboratory study and the xenogeneic cell surrogate material, the model allows to easily assess cell and particle penetration through porous structures like

  10. Effects of feeding regimen on weight gain, semen characteristics, libido, and lameness in 170- to 250-kilogram Duroc boars.

    Science.gov (United States)

    Wang, C; Li, J L; Wei, H K; Zhou, Y F; Tan, J J; Sun, H Q; Jiang, S W; Peng, J

    2016-11-01

    A 2-period field trial was conducted to determine the effects of feeding regimen on weight gain, semen characteristics, libido, and lameness in 170- to 200-kg (period 1) and 200- to 250-kg (period 2) boars. Sixty-one Duroc boars were allotted to 1 of 3 dietary treatments and 15 g/d fish oil was also provided for each experimental boar, to maintain the n-6:n-3 intake ratio at approximately 6.1:1. The energy intakes of the 3 treatment groups were controlled by basing the feed intake on a corn-soybean meal-based diet (3.11 Mcal/kg of ME) to create: 1) low energy intake group (L, = 20, 7.3 Mcal/d of ME), 2) medium energy intake group (M, = 20, 7.7 Mcal/d of ME), and 3) high energy intake group (H, = 21, 8.3 Mcal/d of ME) in period 1. Feed intake was then increased to 7.6, 8.2, and 8.6 Mcal/d of ME for the 3 groups, respectively, in period 2. During the 28-wk experimental period, boar weight gain, testis volume, semen characteristics, libido, toe measurements, claw lesions, and lameness were examined, and the number of boars culled in each group was recorded to calculate the culling rate. Although there were no differences in testis volume, sperm concentration, and motility, the percentage of abnormal sperm, the percentage of claw lesions, and claw lesion scores differed among treatments ( > 0.05), and ADG was significantly increased with the increase of energy intake between the 2 periods ( 0.05). Nonetheless, the H boars took more time to mount the collection dummy and produce an ejaculate and, in particular, showed a greater percentage of lameness than the L and M boars ( < 0.05). Therefore, the medium energy intake regimen (energy intakes of 7.7 and 8.2 Mcal/d of ME with ADG of 454.5 and 375.3 g/d in 2 periods, respectively) improved the total sperm number and functional sperm number, and meanwhile decreased the occurrence of lameness in 170- to 250-kg Duroc boars.

  11. Sperm head's birefringence: a new criterion for sperm selection.

    Science.gov (United States)

    Gianaroli, Luca; Magli, M Cristina; Collodel, Giulia; Moretti, Elena; Ferraretti, Anna P; Baccetti, Baccio

    2008-07-01

    To investigate the characteristics of birefringence in human sperm heads and apply polarization microscopy for sperm selection at intracytoplasmic sperm injection (ICSI). Prospective randomized study. Reproductive Medicine Unit, Società Italiana Studi Medicina della Riproduzione, Bologna, Italy. A total of 112 male patients had birefringent sperm selected for ICSI (study group). The clinical outcome was compared with that obtained in 119 couples who underwent a conventional ICSI cycle (control group). The proportion of birefringent spermatozoa was evaluated before and after treatment in relation to the sperm sample quality. Embryo development and clinical outcome in the study group were compared with those in the controls. Proportion of birefringent sperm heads, rates of fertilization, cleavage, pregnancy, implantation, and ongoing implantation. The proportion of birefringent spermatozoa was significantly higher in normospermic samples when compared with oligoasthenoteratospermic samples with no progressive motility and testicular sperm extraction samples. Although fertilization and cleavage rates did not differ between the study and control groups, in the most severe male factor condition (oligoasthenoteratospermic with no progressive motility and testicular sperm extraction), the rates of clinical pregnancy, ongoing pregnancy, and implantation were significantly higher in the study group versus the controls. The analysis of birefringence in the sperm head could represent both a diagnostic tool and a novel method for sperm selection.

  12. First report of Mycoplasma hyopneumoniae seroprevalence in farmed wild boars in China.

    Science.gov (United States)

    Liang, Qin-Li; Zou, Yang; Gao, Yun-Hang; Nie, Lan-Bi; Zhang, Xiao-Xuan; Hu, Gui-Xue; Du, Rui; Zhu, Xing-Quan

    2018-06-01

    Porcine enzootic pneumonia caused by Mycoplasma hyopneumoniae affects the global pig industry with significant economic losses. It is yet to know whether wild boars in China were infected with M. hyopneumoniae. The present study was conducted to examine the seroprevalence and to evaluate risk factors of M. hyopneumoniae infection in farmed wild boars in China. A total of 882 serum samples were collected from farmed wild boars in Jilin City, Siping City and Baishan City in Jilin Province, northeastern China from April 2015 to February 2016, and were examined by the double sandwich enzyme-linked immunosorbent assay (ELISA). Seventy-eight out of 882 (8.8%) serum samples were M. hyopneumoniae-seropositive. Among region groups, wild boars from Jilin city (11.7%, 33/281) had the highest seropositivity, followed by Siping city (11%, 29/263) and Baishan city (4.7%, 16/338), and the difference was statistically significant (P = 0.0031). The M. hyopneumoniae seroprevalence in the female wild boars (9.0%, 75/831) was higher than that in the male wild boars (5.9%, 3/51) (P = 0.4429). The results of this investigation showed that farmed wild boars were susceptible to M. hyopneumoniae. Logistic regression analysis showed that there is a significant correlation between the geographical area and M. hyopneumoniae infection, which may be related to the regional environment. This is the first report of M. hyopneumoniae seroprevalence in farmed wild boars in China, which provided baseline information for further studies and control of M. hyopneumoniae infection in wild boars in China. Copyright © 2018 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2016-07-01

    The main objective of this study was to investigate the effects of cultured wild ginseng root extracts (cWGRE) on the sperm of boars and the reproductive system of guinea pigs. Firstly, semen collected from boars (n=10) were incubated in 38°C for 1h with xanthine and xanthine oxidase to generate ROS. The cWGRE was added to the sperm culture system to test its antioxidant effect on the boar sperm. The amount of Reactive Oxygen Species (ROS) was measured by a chemiluminescence assay using luminol. The results indicated that the addition of cWGRE to boar sperm culture inhibited xanthine and xanthine oxidase-induced ROS concentrations. Treatment with cWGRE also had a positive effect on maintaining sperm motility. Effects of cWGRE administration on vitamin C-deficient guinea pigs were further investigated. Hartley guinea pigs (n=25) at 8 weeks of age were randomly divided into five groups. With the exception of the positive control group, each group was fed vitamin C-deficient feed for 21days (d). Respective groups were also orally administered cWGRE, ginseng extract, or mixed ginsenosides for 21 days. In comparison to the control group, oral administration of cWGRE reduced (P<0.05) amount of lipid peroxidation and increased (P<0.05) both glutathione peroxidase concentrations and the trolox equivalent antioxidant capacity. In addition, administration of cWGRE induced increases (P<0.05) in body weight, testosterone concentrations, and spermatid populations. The results of the present study support our hypothesis that cWGRE has positive effects on male reproductive functions via suppression of ROS production. Copyright © 2016 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    B.M. van den Berg

    2014-02-01

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

  15. The adult boar testicular and epididymal transcriptomes

    Directory of Open Access Journals (Sweden)

    Guyonnet Benoît

    2009-08-01

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

  16. Current status and potential of morphometric sperm analysis

    OpenAIRE

    Maroto-Morales, Alejandro; García-Álvarez, Olga; Ramón, Manuel; Martínez-Pastor, Felipe; Fernández-Santos, M. Rocío; Soler, Ana J.; Garde, José Julián

    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. Effect of estrogens on boar sperm capacitation in vitro

    Czech Academy of Sciences Publication Activity Database

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

    2010-01-01

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

  18. Effect of estrogens on boar sperm capacitation in vitro

    Czech Academy of Sciences Publication Activity Database

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

    2010-01-01

    Roč. 33, Supplement 1 (2010), s. 93-93 ISSN 0105-6263. [6th European Congress of And rology. 29.09.2010-01.10.2010, Athens] R&D Projects: GA MŠk(CZ) 1M06011; GA ČR(CZ) GA523/08/H064; GA ČR(CZ) GA523/09/1793 Institutional research plan: CEZ:AV0Z50520701 Keywords : spermatozoa * capacitation * monoclonal antibody * estrone * 17beta-estradiol * estriol * 17alpha-ethynylestradiol Subject RIV: EI - Biotechnology ; Bionics

  19. Effect of Monoclonal Antibodies to Boar Sperm on Conception Rate ...

    African Journals Online (AJOL)

    The Conception Rates (CR) were 100%, 30% and 50% for control, Hmab and Tmab respectively. The litter sizes were 8.4±1.71, 5.0+1.0 and 5.4 ±1.14for the control, Hmab and Tmab group respectively. Immunization with both Hmab and Tmab significantly reduced CR (P<0.001), the reduction in CR was significantly lower ...

  20. Cryopreservation of microencapsulated canine sperm.

    Science.gov (United States)

    Shah, Shambhu; Otsuki, Tsubasa; Fujimura, Chika; Yamamoto, Naoki; Yamashita, Yasuhisa; Higaki, Shogo; Hishinuma, Mitsugu

    2011-03-01

    The objective was to develop a method for cryopreserving microencapsulated canine sperm. Pooled ejaculates from three beagle dogs were extended in egg yolk tris extender and encapsulated using alginate and poly-L-lysine at room temperature. The microcapsules were cooled at 4 °C, immersed in pre-cooled extender (equivalent in volume to the microcapsules) to reach final concentration of 7% (v/v) glycerol and 0.75% (v/v) Equex STM paste, and equilibrated for 5, 30 and 60 min at 4 °C. Thereafter, microcapsules were loaded into 0.5 mL plastic straws and frozen in liquid nitrogen. In Experiment 1, characteristics of microencapsulated canine sperm were evaluated after glycerol addition at 4 °C. Glycerol exposure for 5, 30 and 60 min did not significantly affect progressive motility, viability, or acrosomal integrity of microencapsulated sperm compared with pre-cooled unencapsulated sperm (control). In Experiment 2, characteristics of frozen-thawed canine microencapsulated sperm were evaluated at 0, 3, 6, and 9 h of culture at 38.5 °C. Pre-freeze glycerol exposure for 5, 30, and 60 min at 4 °C did not influence post-thaw quality in unencapsulated sperm. Post-thaw motility and acrosomal integrity of microencapsulated sperm decreased more than those of unencapsulated sperm (P < 0.05) following glycerol exposure for 5 min. However, motility, viability and acrosomal integrity of microencapsulated sperm after 30 and 60 min glycerol exposure were higher than unencapsulated sperm cultured for 6 or 9 h (P < 0.05). In conclusion, since microencapsulated canine sperm were successfully cryopreserved, this could be a viable alternative to convention sperm cryopreservation in this species. Copyright © 2011 Elsevier Inc. All rights reserved.

  1. Boar genotype as a factor shaping age-related changes in semen parameters and reproduction longevity simulations.

    Science.gov (United States)

    Knecht, Damian; Jankowska-Mąkosa, Anna; Duziński, Kamil

    2017-08-01

    The aim of this study was a detailed analysis of the boar genotypes used in AI stations with an indication of their production capacity, including age and a precise analysis of their culling time and reason. The study included 334 boars: 81 Polish Large White (PLW), 108 Polish Landrace (PL), 49 Pietrain (P), 56 Duroc × Pietrain (D × P) and 40 Hampshire × Pietrain (H × P). Semen volume, spermatozoa concentration, total number of spermatozoa, number of motile spermatozoa, and number of insemination doses were analyzed. Quadratic regression was used to illustrate the selected sperm parameters at specific ages. Among all the studied boars the lowest motilities of spermatozoa were identified in white breeds PLW and PL, and the difference between motility extremes was 3.53% (P ≤ 0.01). The highest number of insemination doses were produced from D × P crossbreed boars: about 0.7 portions more compared to PL, 1.13 to PLW, 1.18 to H × P and 1.8 to P (all differences P ≤ 0.01). It has been shown in the case of ejaculate volume that for PLW and H × P boars the culling moment was far too early in terms of production capacity and differences were, respectively, 16.35 ml for PLW and 12.61 ml for H × P. Based on the developed regression equations, the earliest maximum number of motile sperm (73.82 × 10 9 ) was obtained by H × P crossbreed boars as early as at age 24 months. The highest values for this parameter were achieved, however, by other D × P crossbreed boars: 74.30 × 10 9  at the later age of 32 months. A consequence of the high number of motile sperm in young H × P boars was that the theoretical maximum value of the number of AI doses was produced as early as the 14th month (25.59 portions). Curves of similar shape were obtained for PL and D × P boars; the difference in maximal values was 0.54 portions in favor of crossbreeds, at a later age of 7 months. It was noted that for PLW and D × P boars the highest number

  2. 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.

  3. Cryopreservation of epididymal stallion sperm.

    Science.gov (United States)

    Olaciregui, M; Gil, L; Montón, A; Luño, V; Jerez, R A; Martí, J I

    2014-02-01

    Any event that makes semen collection or mating impossible, such as death, castration, or injury, may terminate a stallion's breeding career. Fortunately, stallion sperm which are capable of fertilization can be harvested from the epididymis, and frozen for future use. However, the fertility of frozen-thawed epididymal sperm has been found to be lower than that of ejaculated sperm. Therefore, this study aimed to optimize the fertility of frozen epididymal stallion sperm by investigating the effects of different cryoprotectants and freezing protocols on sperm quality. Dimethylformamide was tested alone or combination with pasteurized egg yolk as substitute of fresh egg yolk. In addition, the effect of the pre-freeze stabilization on sperm quality was analyzed. Heterospermic samples obtained from stallion epididymis were collected and cryopreserved in lactose-egg-yolk extender or in the same extender with varying content of cryoprotectant and content of egg yolk, stabilized and no-stabilized. Sperm motility, viability, hypoosmotic swelling test (HOST) and acrosome integrity were evaluated post-thawing. No improvement was observed on the replacement of fresh yolk by pasteurized egg yolk, whereas the results suggest that dimethylformamide is a cryoprotectant suitable for cryopreservation of equine epididymal semen, even better than glycerol. In addition, we found that the stabilization before freezing on epididymal stallion sperm, can improve sperm quality parameters. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. Current status and potential of morphometric sperm analysis

    Directory of Open Access Journals (Sweden)

    Alejandro Maroto-Morales

    2016-01-01

    Full Text Available 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 cryoresistance and male fertility is also discussed. For this, the evaluation of patterns of change of sperm head morphometry throughout a process, instead of the study of the morphometric characteristics of the sperm head at different stages, allows a better identification of the males with different sperm cryoconservation ability. These new approaches, together with more studies employing a greater number of individuals, are needed to obtain novel results concerning the role of sperm morphometry on sperm function. Future studies should aim at understanding the causes of sperm design diversity and the mechanisms that generate them, giving increased attention to other sperm structures besides the sperm head. The implementation of scientific and technological advances could benefit the simultaneous examination of sperm phenotype and sperm function, demonstrating that sperm morphometry could be a useful tool for sperm assessment.

  5. Current status and potential of morphometric sperm analysis.

    Science.gov (United States)

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

    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 cryoresistance and male fertility is also discussed. For this, the evaluation of patterns of change of sperm head morphometry throughout a process, instead of the study of the morphometric characteristics of the sperm head at different stages, allows a better identification of the males with different sperm cryoconservation ability. These new approaches, together with more studies employing a greater number of individuals, are needed to obtain novel results concerning the role of sperm morphometry on sperm function. Future studies should aim at understanding the causes of sperm design diversity and the mechanisms that generate them, giving increased attention to other sperm structures besides the sperm head. The implementation of scientific and technological advances could benefit the simultaneous examination of sperm phenotype and sperm function, demonstrating that sperm morphometry could be a useful tool for sperm assessment.

  6. 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.

  7. 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.

  8. 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...... containing saturated (myristoyl, palmitoyl and stearoyl) fatty acyl residues. Glycerophosphatidylcholine and glycerophosphatidylethanolamine with alk(en)yl ether residues in the sn-1 position and unsaturated long chained fatty acyl residues in sn-2 position were identified as the most prominent polar lipids....... The only glycoglycerolipid was sulfogalactosylglycerolipid carrying 16:0-alkyl- and 16:0-acyl chains. Using stable isotope-labelling, the metabolic incorporation of exogenously supplied fatty acids was analysed. Boar spermatozoa incorporated hexadecenoic (16:1), octadecenoic (18:1), octadecadienoic (18...

  9. Requirement for an intact cytoskeleton for volume regulation in boar spermatozoa.

    Science.gov (United States)

    Petrunkina, A M; Hebel, M; Waberski, D; Weitze, K F; Töpfer-Petersen, E

    2004-01-01

    Osmotically induced cell swelling triggers a chain of events leading to a net loss of major cell ions and water, resulting in cell volume recovery, a process known as regulatory volume decrease (RVD). In many cell types, there is an evidence that the cytoskeleton may play a role in the initial sensing and transduction of the signal of volume change. In this study, we tested the hypothesis that an intact microfilament and microtubule network is required for volume response and RVD in boar sperm before and after capacitation treatment and whether addition of cytochalasin D and colchicine to the capacitation medium would affect volumetric behaviour. Capacitation is a series of cellular and molecular alterations that enable the spermatozoon to fertilize an oocyte. Cell volume measurements of washed sperm suspensions were performed electronically in Hepes-buffered saline solutions of 300 and 180 mosmol/kg. After exposure to hypoosmotic conditions, boar sperm showed initial swelling (up to 150% of initial volume within 5 min), which was subsequently partially reversed (to about 120-130% after 20 min). Treatment with cytochalasin D led to reduced initial swelling (1 micromol/l) and loss of RVD in washed sperm (1-10 micromol/l) and at the beginning of incubation under capacitating conditions (5 micromol/l). Short treatment with 500 micromol/l colchicine affected the volume regulatory ability in sperm under capacitating conditions but not in washed sperm. No significant differences in cell volume response were observed after subsequent addition of cytochalasin D and colchicine to the suspensions of sperm incubated for 3 h under capacitating conditions. However, the incubation under capacitating conditions in the presence of cytochalasin D led to improved volume regulation at the end of the incubation period (23%). The microfilament network appears to be important for volume regulation in washed boar spermatozoa while intact microtubules do not seem to be necessary for

  10. Nuclear chromatin variations in human spermatozoa undergoing swim-up and cryopreservation evaluated by the flow cytometric sperm chromatin structure assay.

    Science.gov (United States)

    Spanò, M; Cordelli, E; Leter, G; Lombardo, F; Lenzi, A; Gandini, L

    1999-01-01

    The sperm chromatin structure assay (SCSA) is a flow cytometric (FCM) technique which exploits the metachromatic properties of Acridine Orange to monitor the susceptibility of sperm chromatin DNA to in-situ acid denaturation. SCSA was used to study the chromatin structure variations of human spermatozoa in semen, both before and after swim-up and after cryopreservation. Semen samples were provided by 19 healthy normozoospermic subjects attending pre-marriage checks. Each sample was divided into three aliquots: the first aliquot was evaluated without further treatment, the second underwent swim-up, and the third was stored according to standard cryopreservation techniques in liquid nitrogen at -196 degrees C. Samples were also analysed by light and fluorescence microscopy (after Acridine Orange staining to evaluate the number of green fluorescent sperm heads), and by computer-assisted semen analysis. The results showed that post-rise spermatozoa represent a subpopulation characterized by a general improvement of the morphological (reduction of the percentage of abnormal forms and heads, increase of the green head sperm percentage) and kinetic parameters. This subpopulation also exhibited improved chromatin structure properties, confirming that these cells have the best structural and functional characteristics, indicative of optimal fertilizing ability. On the other hand, overall sperm quality deteriorates after cryopreservation. When thawed spermatozoa underwent an additional swim-up round, a general improvement of nuclear maturity was seen in the post-rise spermatozoa.

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

    Czech Academy of Sciences Publication Activity Database

    Čapková, Jana; Kubátová, Alena; Děd, Lukáš; Teplá, O.; Pěknicová, Jana

    2016-01-01

    Roč. 18, č. 1 (2016), s. 108-113 ISSN 1008-682X R&D Projects: GA MŠk(CZ) ED1.1.00/02.0109; GA ČR(CZ) GAP503/12/1834 Institutional support: RVO:86652036 Keywords : asthenozoospermia * flow cytometry * fluorescence microscopy * monoclonal antibodies * sperm proteins Subject RIV: CE - Biochemistry Impact factor: 2.996, year: 2016 http://www.ajandrology.com/article.asp?issn=1008-682X;year=2016;volume=18;issue=1;spage=108;epage=113;aulast=Capkova

  12. Mitochondrial respiratory efficiency is positively correlated with human sperm motility.

    Science.gov (United States)

    Ferramosca, Alessandra; Provenzano, Sara Pinto; Coppola, Lamberto; Zara, Vincenzo

    2012-04-01

    To correlate sperm mitochondrial respiratory efficiency with variations in sperm motility and with sperm morphologic anomalies. Sperm mitochondrial respiratory activity was evaluated with a polarographic assay of oxygen consumption carried out in hypotonically-treated sperm cells. A possible relationship among sperm mitochondrial respiratory efficiency, sperm motility, and morphologic anomalies was investigated. Mitochondrial respiratory efficiency was positively correlated with sperm motility and negatively correlated with the percentage of immotile spermatozoa. Moreover, midpiece defects impaired mitochondrial functionality. Our data indicate that an increase in sperm motility requires a parallel increase in mitochondrial respiratory capacity, thereby supporting the fundamental role played by mitochondrial oxidative phosphorylation in sperm motility of normozoospermic subjects. These results are of physiopathological relevance because they suggest that disturbances of sperm mitochondrial function and of energy production could be responsible for asthenozoospermia. Copyright © 2012 Elsevier Inc. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Ivanović Snežana D.

    2013-01-01

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

  14. Cryo-scanning electron microscopy (Cryo-SEM) of semen frozen in medium-straws from good and sub-standard freezer AI-boars.

    Science.gov (United States)

    Hernández, Marta; Ekwall, Hans; Roca, Jordi; Vazquez, Juan Maria; Martinez, Emilio; Rodríguez-Martínez, Heriberto

    2007-02-01

    A major limiting factor for commercial cryopreservation of boar semen for artificial insemination (AI) is the large individual variation to cooling, where the degree of cell dehydration during ice (re)shaping seems to play a major role. This study investigated, in the frozen state, the degree of dehydration and ice crystal distribution in boar semen doses whose spermatozoa displayed different viability after thawing. Cross-sectioned medium-straws (0.5 mL, n=10) from a total of 10 stud boars classified as "good"(n=5) or sub-standard (e.g., "bad" freezers, n=5) by conventional analyses (computer assisted motility and sperm viability) were examined by Cryo-scanning electron microscopy (Cryo-SEM) to determine whether differences between groups could be already distinguishable prior to thawing. The degree of hydration was monitored in relation to the areas of ice crystal formed extracellularly (lakes), the areas of frozen, concentrated extender (veins) where spermatozoa were located and the degree of compartmentalization (number of lakes) present. Irrespectively of the region studied, the gradient of main dehydration (as lakes) observed along the cross-section area of the straws was very irregular. Most spermatozoa were enclosed in the freezing extender matrix and no obvious signs of external membrane damage were observed. None of the Cryo-SEM variables significantly correlated with post-thaw sperm parameters (p>0.05). However, we identified significant differences (p<0.0001) among boars for all ultrastructure variables studied, including the size of the veins, where differences in solute concentration is expected. We concluded that despite the large variability in ice crystal formation during the conventional freezing process among boars, this is unrelated to inter-boar post-thaw sperm differences.

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

    Directory of Open Access Journals (Sweden)

    Laura Gómez Montoto

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

  16. Sperm Production Rate, Gonadal and Extragonadal Sperm ...

    African Journals Online (AJOL)

    Five healthy West African Dwarf (WAD) rams, 1.5 to 2.5 years of age and weighing between 15 kg to 20 kg were used to determine daily sperm production, gonadal and exragonadal sperm reserves. Gonadal and extragonadal sperm reserves were estimated by the haemocytometric method, while the daily sperm production ...

  17. 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.

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

    International Nuclear Information System (INIS)

    Santiso, Rebeca; Tamayo, María; Gosálvez, Jaime; Johnston, Steve; Mariño, Alfonso; Fernández, Carlos; Losada, Carlos; Fernández, José Luis

    2012-01-01

    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 °C and 45 °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 °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.

  19. Relationship between sperm parameters and intracytoplasmic sperm injection outcome

    Directory of Open Access Journals (Sweden)

    Shahla Chaichian

    2015-12-01

    Full Text Available Objectives: With the adventure of intracytoplasmic sperm injection (ICSI technique, great progresses have developed in the treatment of infertility. Concentration on the properties of male’s gamete has been encouraged by the increasing concerns about the causes of ICSI failure. We hence conducted this study to investigate the probable association of sperm parameters with ISCI outcome. Methods: A total of 523 couples referred to Isfahan Fertility and Sterility Center from January 2007 to June 2008 for ICSI. Semen analysis was performed before ICSI procedure according to the WHO criteria. Patients were assigned into successful ICSI (case and failed ICSI (control groups. Sperm parameters were then compared between the 2 groups. Results: One hundred and six patients (20% had successful ICSI results (case group compared with 417 couples (80% with undesirable ICSI outcomes (control group. Among evaluated factors, sperm agglutination (p = 0.007, sperm concentration (p = 0.043, leukocytospermia (p = 0.026 and head abnormality of sperm (p = 0.019 showed statistically significant differences between two groups with differing ICSI results. None of the other semen parameters revealed significant differences between these two groups. Conclusion: Our study showed that some sperm parameters are associated with desirable ICSI outcome. However, it is unclear whether these associations are causal.

  20. 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

    2014-05-01

    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 (p0.05; mientras que ETG 15% (9.6±2.9% reportó la menor eclosión (p<0.05. Los resultados sugieren que la solución crioprotectora compuesta por ETG 5%, glucosa 6% y leche en polvo 5% es una alternativa viable para la crioconservación de semen de Sorubim cuspicaudus con fecundaciones similares al usar semen fresco.

  1. Effect of production management on semen quality during long-term storage in different European boar studs.

    Science.gov (United States)

    Schulze, M; Kuster, C; Schäfer, J; Jung, M; Grossfeld, R

    2018-03-01

    The processing of ejaculates is a fundamental step for the fertilizing capacity of boar spermatozoa. The aim of the present study was to identify factors that affect quality of boar semen doses. The production process during 1 day of semen processing in 26 European boar studs was monitored. In each boar stud, nine to 19 randomly selected ejaculates from 372 Pietrain boars were analyzed for sperm motility, acrosome and plasma membrane integrity, mitochondrial activity and thermo-resistance (TRT). Each ejaculate was monitored for production time and temperature for each step in semen processing using the special programmed software SEQU (version 1.7, Minitüb, Tiefenbach, Germany). The dilution of ejaculates with a short-term extender was completed in one step in 10 AI centers (n = 135 ejaculates), in two steps in 11 AI centers (n = 158 ejaculates) and in three steps in five AI centers (n = 79 ejaculates). Results indicated there was a greater semen quality with one-step isothermal dilution compared with the multi-step dilution of AI semen doses (total motility TRT d7: 71.1 ± 19.2%, 64.6 ± 20.0%, 47.1 ± 27.1%; one-step compared with two-step compared with the three-step dilution; P < .05). There was a marked advantage when using the one-step isothermal dilution regarding time management, preservation suitability, stability and stress resistance. One-step dilution caused significant lower holding times of raw ejaculates and reduced the possible risk of making mistakes due to a lower number of processing steps. These results lead to refined recommendations for boar semen processing. Copyright © 2018 Elsevier B.V. All rights reserved.

  2. Myoinositol: does it improve sperm mitochondrial function and sperm motility?

    Science.gov (United States)

    Condorelli, Rosita A; La Vignera, Sandro; Bellanca, Salvatore; Vicari, Enzo; Calogero, Aldo E

    2012-06-01

    To evaluate whether an improvement in mitochondrial membrane potential was associated with sperm motility amelioration and greater sperm recovery after the swim-up procedure. A second purpose was to evaluate the effects of myoinositol (MYO) on sperm apoptosis, quality of chromatin compaction, and DNA integrity. Spermatozoa from 20 normozoospermic men and 20 patients with oligo-astheno-teratozoospermia were incubated in vitro with 2 mg/mL of MYO or phosphate-buffered saline as a control for 2 hours. After this incubation period, sperm motility was evaluated. Flow cytometry was used to analyze the mitochondrial membrane potential, phosphatidylserine externalization, chromatin compactness, and DNA fragmentation. We also evaluated the total number of motile spermatozoa recovered after swim-up after incubation with MYO or phosphate-buffered saline. MYO significantly increased the percentage of spermatozoa with progressive motility in both normozoospermic men and patients with oligo-astheno-teratozoospermia. Motility improvement in the latter group was associated with a significant increase in the percentage of spermatozoa with high mitochondrial membrane potential. MYO had no effects on mitochondrial function in spermatozoa from normozoospermic men. Sperm phosphatidylserine externalization, chromatin compactness, and DNA fragmentation were unaffected by MYO in both groups. After incubation with MYO, the total number of spermatozoa recovered after swim-up had improved significantly in both groups. These data show that MYO increases sperm motility and the number of spermatozoa retrieved after swim-up in both normozoospermic men and patients with abnormal sperm parameters. In patients with oligo-astheno-teratozoospermia, the improvement in these parameters was associated with improved sperm mitochondrial function. These findings support the use of MYO in both in vivo- and in vitro-assisted reproductive techniques. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. 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

  4. Decline of semen quality among Chinese sperm bank donors within 7 years (2008-2014

    Directory of Open Access Journals (Sweden)

    Li Wang

    2017-01-01

    Full Text Available Semen from 5210 sperm bank donors was analyzed and trends in semen quality were evaluated at Shandong Human Sperm Bank between 2008 and 2014. After 2-7 days of abstinence, semen samples were collected. Measurements of semen volume, sperm concentration, sperm forward motility, and total sperm count were performed. There were significant declining trends in semen volume, sperm concentration, sperm forward motility, and total sperm count. Our results indicate that the quality of semen in this cohort of sperm donors had decreased during the study period.

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

    Directory of Open Access Journals (Sweden)

    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

  6. Ractopamine hydrochloride improves growth performance and carcass composition in immunocastrated boars, intact boars, and gilts.

    Science.gov (United States)

    Rikard-Bell, C; Curtis, M A; van Barneveld, R J; Mullan, B P; Edwards, A C; Gannon, N J; Henman, D J; Hughes, P E; Dunshea, F R

    2009-11-01

    The beta-agonist ractopamine is a dietary ingredient that improves growth and increases the lean mass with little change in fat mass in gilts and barrows. Limited data in boars indicate that dietary ractopamine may increase lean tissue and decrease fat deposition, whereas there are no data for immunocastrated boars. The aims of this investigation were 1) to assess whether the growth performance of all sexes could be maintained over 31 d by using a step-up dietary ractopamine feeding program of 5 mg/kg of ractopamine for the first 14 d, then increasing the dose to 10 mg/kg for a further 17 d, and 2) to determine if dietary ractopamine would increase lean mass in all sexes and decrease fat mass in boars and immunocastrated boars. The study involved 286 pigs randomized and proportionally allocated by breed into 24 groups of 11 or 12 pigs at 17 wk of age, with equal groups of boars, immunocastrated boars, and gilts. Dietary ractopamine decreased (P = 0.005) ADFI during the first 2 wk, particularly in the intact and immunocastrated boars, with the reduction in ADFI being maintained in the immunocastrated boars after the increment in dietary ractopamine. Daily BW gain was not altered by dietary ractopamine during the first 2 wk, but was increased (P ractopamine. Dietary ractopamine decreased (P ractopamine. Carcass weight was increased (P ractopamine in all sexes, whereas back fat tended (P = 0.076) to be reduced in the immunocastrated boars. Dietary ractopamine increased (P = 0.018) lean tissue mass by 4.0, 4.8, and 6.5 kg in the intact boars, gilts, and immunocastrated boars, respectively. In the entire and immunocastrated boars, the increase in lean tissue was accompanied with a decrease (P = 0.004) in fat mass. There was little effect of dietary ractopamine on fat mass in gilts. However, carcass percent fat was decreased (P = 0.004) and percent lean increased (P = 0.006) in all sexes. Immunocastration caused a decrease in lean tissue mass and an increase in fat mass

  7. Coupling sperm mediated gene transfer and sperm sorting techniques: a new perspective for swine transgenesis.

    Science.gov (United States)

    De Cecco, Marco; Spinaci, Marcella; Zannoni, Augusta; Bernardini, Chiara; Seren, Eraldo; Forni, Monica; Bacci, Maria Laura

    2010-09-15

    Flow cytometric separation of X and Y chromosome-bearing spermatozoa has been demonstrated to be effective in pigs, allowing the use of boar sexed semen in in vitro trials. Sperm Mediated Gene Transfer (SMGT) is a widely used and efficient technique for the creation of transgenic animals. The present research intended to prove that it is possible to associate sperm sexing with the SMGT technique in order to speed up the assessment of homozygous lines of transgenic pigs. In the first experiment, the sorting protocol was modified in order to obtain the highest DNA uptake by sorted spermatozoa. In the second experiment, spermatozoa that had undergone only sperm sorting, only SMGT, or both procedures (Sorted-SMGT) were used for in in vitro fertilization of in vitro matured oocytes. In the third experiment, transformed blastocysts of the desired gender (male) were obtained with Sorted-SMGT in an in vitro fertilization trial. The method we developed here allowed us to produce transgenic swine blastocysts of pre-determined gender, giving a positive answer at the aim to couple SMGT and sperm sorting in swine, obtaining fertile spermatozoa able to produce transgenic embryos of pre-determined gender. Copyright 2010 Elsevier Inc. All rights reserved.

  8. Prevalence of Toxoplasma gondii and potentially zoonotic helminths in wild boars (Sus scrofa hunted in central Italy

    Directory of Open Access Journals (Sweden)

    Roberto Amerigo Papini

    2018-03-01

    Full Text Available Our aim was to evaluate the risk of human toxoplasmosis via meat consumption from wild boars by estimating the seroprevalence of Toxoplasma gondii in animals hunted in central Italy. Using a modified agglutination test, 213 sera from wild boars were examined for anti-Toxoplasma IgG antibodies. Diaphragm samples (n=65 from seropositive and seronegative animals were tested by nested-PCR to detect T. gondii DNA. Toxoplasma DNA from diaphragms was genotyped by PCR-RFLP using 12 genetic markers. Moreover, the aim of the study was also to identify helminth infections of wild boars in the selected area and to evaluate their hazard for humans. Examination of sera revealed a seroprevalence of 12.2%. Only one T. gondii strain could be genotyped from a seropositive animal and PCR-RFLP revealed that it belonged to type II. Analysis of 50 samples of faeces and 32 small intestines revealed that 78% and 15.6% of the samples harboured parasites, respectively, with the occurrence of parasites potentially dangerous for humans. These latter included Ascaris suum, Macracanthorhynchus hirudinaceus, Trichuris suis, and Metastrongylus spp. A significant association was found between coprological positivity and male sex. These results indicate that T. gondii infection may be present in wild boar tissues and consumption of undercooked or raw wild boar meat may expose humans to risk of toxoplasmosis in the study area. Furthermore, the study highlights that wild boars are hosts of helminths of veterinary and medical importance transmissible to pigs and humans.

  9. 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....

  10. Oxidative stress negatively affects human sperm mitochondrial respiration.

    Science.gov (United States)

    Ferramosca, Alessandra; Pinto Provenzano, Sara; Montagna, Daniela Domenica; Coppola, Lamberto; Zara, Vincenzo

    2013-07-01

    To correlate the level of oxidative stress in serum and seminal fluid and the level of sperm deoxyribonucleic acid (DNA) fragmentation with sperm mitochondrial respiratory efficiency. Sperm mitochondrial respiratory activity was evaluated with a polarographic assay of oxygen consumption carried out in hypotonically treated sperm cells. A possible relationship between sperm mitochondrial respiratory efficiency, the level of oxidative stress, and the level of sperm DNA fragmentation was investigated. Sperm motility was positively correlated with mitochondrial respiration but negatively correlated with oxidative stress and DNA fragmentation. Interestingly, sperm mitochondrial respiratory activity was negatively affected by oxidative stress and DNA fragmentation. Our data indicate that sperm mitochondrial respiration is decreased in patients with high levels of reactive oxygen species by an uncoupling between electron transport and adenosine triphosphate synthesis. This reduction in mitochondrial functionality might be 1 of the reasons responsible for the decrease in spermatozoa motility. Copyright © 2013 Elsevier Inc. All rights reserved.

  11. 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.

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

    Science.gov (United States)

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

    2014-09-01

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

  13. BOAR EFFECT“ INFLUENCE ON GILTS ESTRUAL REACTION

    OpenAIRE

    Ivan Stančić; Ivan Radović; Blagoje Stančić; Ognjen Stevančević; Milenko Stevančević

    2012-01-01

    Induction of synchronised estrus in prepubertal gilts by the sexually mature boar contact is a phenomenon known as “boar effect”. This effect is used for more efficient estrus detection in sexually mature (cyclic) gilts. The best results of application of “boar effect” are achieved if the stimulation is performed by direct contact between gilts and a sexually mature boar, at least two times a day every 12 hours. This paper presents the results of foreign and domestic authors on the “boar effe...

  14. 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.

  15. Varicocele Negatively Affects Sperm Mitochondrial Respiration.

    Science.gov (United States)

    Ferramosca, Alessandra; Albani, Denise; Coppola, Lamberto; Zara, Vincenzo

    2015-10-01

    To evaluate the effect of varicocele on oxidative stress, sperm mitochondrial respiratory efficiency, sperm morphology, and semen parameters. A total of 20 patients with varicocele and 20 normozoospermic subjects without varicocele (control group) were recruited from a medical center for reproductive biology. The levels of serum reactive oxygen metabolites and seminal lipid peroxides were assessed for both control and varicocele subjects. Sperm deoxyribonucleic acid fragmentation was measured by sperm chromatin dispersion test. Mitochondrial respiratory activity was evaluated with a polarographic assay of oxygen consumption carried out in hypotonically treated sperm cells. In this study, varicocele patients were compared with men without varicoceles. Oxidative stress was observed in the serum and seminal fluid of varicocele patients. These patients showed an increase of 59% (P <.05) in serum reactive oxygen metabolites and a 3-fold increase in the level of sperm lipid peroxides. A parallel and significant increase (a 2-fold increase; P <.05) in the degree of sperm deoxyribonucleic acid fragmentation was also observed. Varicocele patients showed a 27% decrease (P <.05) in mitochondrial respiratory activity in comparison to the control group. A 32% increase (P <.05) in sperm midpiece defects and a 41% decrease (P <.05) in sperm concentration and motility were also observed. Men with varicocele have increased markers of oxidative stress and decreased mitochondrial respiratory activity. These results correlated with abnormalities in semen parameters. For morphology, these correlated with midpiece defects. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. 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. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Cytometry of mammalian sperm

    International Nuclear Information System (INIS)

    Gledhill, B.L.

    1983-01-01

    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

  18. 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.

  19. 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

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Evaluation of the qualitative and quantitative effectiveness of three media of centrifugation (Maxifreeze, Cushion Fluid Equine, and PureSperm 100) in preparation of fresh or frozen-thawed brown bear spermatozoa.

    Science.gov (United States)

    Nicolas, M; Alvarez, M; Borragán, S; Martinez-Pastor, F; Chamorro, C A; Alvarez-Rodriguez, M; de Paz, P; Anel, L

    2012-04-01

    Centrifugation is a crucial procedure in sperm cryopreservation protocols of brown bear (Ursus arctos), because the semen must be processed to increase sperm concentration and/or clean urine-contaminated samples. The efficacy of three media for centrifugation (Maxifreeze [IMV technologies, L'Aigle, France], Cushion Fluid Equine (Minitübe, Tiefenbach, Germany), and PureSperm [Nidacon, Gothenburg, Sweden]) on the quality of bear spermatozoa was evaluated. In experiment one, two cushioned media used for protecting against mechanical stress during centrifugation were analyzed. In experiment two, a density gradient based on PureSperm was assessed in relation to the maximum retrieval and the quality of fresh spermatozoa, and the freezability of the spermatozoa selected in this density gradient was studied in experiment three. Finally, the selection of frozen-thawed sperm using PureSperm was analyzed in experiment four. Our results indicate that the use of dense isotonic cushion solutions (Maxifreeze, Cushion Fluid Equine) in centrifugation did not improve the quality of recovered spermatozoa compared with standard centrifugation. However, a density gradient prepared with PureSperm improved the quality of spermatozoa in fresh semen and frozen-thawed semen, but the spermatozoa selected from the fresh sample with this density gradient did not show a better resistance to freezing with this density gradient in comparison with the control sample. Copyright © 2012 Elsevier Inc. All rights reserved.

  2. 137Cs monitoring in the meat of wild boar population in Slovakia

    Directory of Open Access Journals (Sweden)

    Katarina Beňová

    2016-06-01

    Full Text Available Currently, due to the elapsed time and the nature of the Chernobyl accident, the only artificial radionuclide present in the soil is 137Cs, with a physical half-life conversion of 30.17 years. The 137Cs is quickly integrated into a biological cycle, similar to potassium. Generally, radionuclides are characterized by their mobility in soil. Contamination of materials and food by radionuclides represent a serious problem and has a negative impact on human health. The threat of international terrorism and the inability to forestall the impact of natural disasters on nuclear energetic (Fukushima accident, are also reasons for continuous monitoring of food safety. According screening measurement performed in European countries, high radioactivity levels were reported in the wild boars muscles from Sumava (Czech Republic. Seasonal fluctuation of 137Cs activity in the wild boar meat samples was observed in the forests on the southern Rhineland. Monitoring of 137Cs activity in the wild boar meat samples in the hunting grounds in Slovakia was initiated based on the reports on exceeding limits of the content of radiocaesium in the meat of wild boar from the surrounding countries. The aim of this study was to determine the 137Cs post Chernobyl contamination of wild boars population in different hunting districts of Slovakia during 2013 - 2014. A total of 60 thigh muscle samples from wild boars of different age categories (4 months - 2 years were evaluated. 137Cs activity was measured by gamma spectrometry (Canberra. Despite the fact Slovakia is closer to Chernobyl as Czech Republic and Germany, the 137Cs activity measured was very low and far below the permitted limit. The highest radiocaesium activity level measured in muscle was 37.2 Bq.kg-1 ±4.7%. Wild boar originated from Zlate Moravce district. The measurement results show, that 137Cs contamination levels of game in Slovakia are low. Radiocaesium activity in examined samples was very low and

  3. Epizootic and epidemiological situation of Trichinella sp. infection in Poland in 2006–2015 in view of wild boar population dynamics

    Directory of Open Access Journals (Sweden)

    Flis Marian

    2017-06-01

    Full Text Available Introduction: The objective was to evaluate the epizootic and epidemiological situation of Trichinella sp. infection in Poland between 2006 and 2015 against the dynamics of the wild boar population and its primary reservoir host.

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

    NARCIS (Netherlands)

    WIJCHMAN, JG; DEWOLF, BTHM; JAGER, S

    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

  5. Influence of dietary amino acid level on chemical body composition and performance of growing-finishing boars of two sire lines.

    Science.gov (United States)

    Otten, Caroline; Berk, Andreas; Müller, Simone; Weber, Manfred; Dänicke, Sven

    2013-12-01

    There is only little information available concerning the chemical body composition of growing-finishing boars. For that reason, a total of 26 entire male pigs (boars) of two different Piétrain sire lines were fed with different levels of dietary essential amino acids (EAA) and the influence of this treatment on performance and chemical body composition was evaluated. In addition, an initial group of eight boars (n = 4 per sire line) was slaughtered at approximately 21 kg live weight (LW). The other 26 boars were fed three different diets containing 11.5, 13.2 and 14.9 g lysine/kg during the grower period and 9.0, 10.4, 11.7 g lysine/kg during the finisher period, respectively. Other EAA were added in relation to lysine (Lys: Met + Cys: Thr: Trp: Val = 1: 0.60: 0.65: 0.18: 0.75). At a LW of approximately 122 kg these 26 boars (six groups with three to seven animals each) were also slaughtered. The effects of EAA level and sire line on fattening and slaughter performance was recorded, and body and weight gain composition were analysed. There were no significant effects of EAA level on performance or on chemical body composition. Boars sired with Piétrain line 1 demonstrated increased lean meat content and protein body content (p < 0.05) as compared to Piétrain line 2-sired boars.

  6. Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices

    Science.gov (United States)

    Matsuura, Koji; Huang, Han-Wei; Chen, Ming-Cheng; Chen, Yu; Cheng, Chao-Min

    2017-04-01

    Mammalian sperm motility has traditionally been analyzed to determine fertility using computer-assisted semen analysis (CASA) systems. To develop low-cost and robust male fertility diagnostics, we created a paper-based MTT assay and used it to estimate motile sperm concentration. When porcine sperm motility was inhibited using sperm enzyme inhibitors for sperm enzymes related to mitochondrial activity and glycolysis, we simultaneously recorded sperm motility and enzymatic reactivity using a portable motility analysis system (iSperm) and a paper-based MTT assay, respectively. When using our paper-based MTT-assay, we calculated the area mean value signal intensity (AMV) to evaluate enzymatic reactivity. Both sperm motility and AMV decreased following treatment with iodoacetamide (IODO) and 3-bromopyruvic acid (3BP), both of which are inhibitors of glycolytic enzymes including glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a correlation between recorded motility using iSperm and AMV from our paper-based assay (P coupled with catalysis of GAPDH and was promoted by electron transfer from NADH. Based on this inhibitor study, sperm motility can be estimated using our paper-based MTT-assay.

  7. Testicular Damage following Testicular Sperm Retrieval

    DEFF Research Database (Denmark)

    Fedder, Jens; Marcussen, Niels; Fedder, Maja D.K.

    2017-01-01

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

  8. 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

  9. How accurate is sperm morphology as an indicator of sperm function?

    Science.gov (United States)

    Franken, D R

    2015-08-01

    Sperm morphology has been consistently correlated with fertilisation success or failure. The clinical relevance of the percentage normal spermatozoa has been a widely discussed topic amongst infertility specialists and scientists. This study aimed to evaluate the role of sperm morphology as an indicator of additional sperm functions among 114 andrology referrals. The sperm functions that were investigated included chromatin packaging quality (CMA3 test (n = 109), zona-induced acrosome reaction (ZIAR test; n = 36), hemizona assay (HZI; n = 36) and progressive motility (n = 47). Chromatin packaging quality had a negative and significant (P = 0.0001, r = -0.74) correlation with the percentage normal spermatozoa, while progressive motility had a significant and positive correlation (P = 0.0001, 0.59). Accurate sperm morphology scoring as described by the WHO 2010 manual can therefore be used as an indicator of specific sperm functions. © 2014 Blackwell Verlag GmbH.

  10. Sperm Surface Proteomics 2

    NARCIS (Netherlands)

    Brewis, I.A.; Gadella, B.M.|info:eu-repo/dai/nl/115389873

    2017-01-01

    This contribution will focus exclusively on the total (global) protein composition (the proteome) of the sperm surface. Immune responses directed towards sperm surface proteins may cause infertility since functionally intact sperm are under immune attack. The immune attack can be achieved directly

  11. Comparison of effects of cryopreservation diluents on the ability of Ram sperm to reduce resazurin dye

    Science.gov (United States)

    Resazurin dye is an effective way to test the metabolism of sperm. As sperm move, they create metabolic waste which is detected by the dye. Another way sperm are evaluated is by Computer-Assisted Sperm Analysis (CASA). CASA detects motility, progression, curvilinear velocity, lateral head amplitude,...

  12. Effect of alpha-lipoic acid on boar spermatozoa quality during freezing-thawing.

    Science.gov (United States)

    Shen, Tao; Jiang, Zhong-Liang; Li, Cong-Jun; Hu, Xiao-Chen; Li, Qing-Wang

    2016-04-01

    Alpha-lipoic acid (ALA) is known to be a natural antioxidant. The aim of the present study was to evaluate the cryoprotective effect of ALA on the motility of boar spermatozoa and its antioxidant effect on boar spermatozoa during freezing-thawing. Different concentrations (2.0, 4.0, 6.0, 8.0 or 10.0 mg/ml) of ALA were added to the extender used to freeze boar semen, and the effects on the quality and endogenous antioxidant enzyme activities of frozen-thawed spermatozoa were assessed. The results indicated that the addition of ALA to the extender resulted in a higher percentage of motile spermatozoa post-thaw (P < 0.05). The activities of superoxide dismutase, lactate dehydrogenase, glutamic-oxaloacetic transaminase and catalase improved after adding ALA to the extender (P < 0.05). Artificial insemination results showed that pregnancy rate and litter size were significantly higher at 6.0 mg/ml in the ALA group than in the control group (P < 0.05). In conclusion, ALA conferred a cryoprotective capacity to the extender used for boar semen during the process of freezing-thawing, and the optimal concentration of ALA for the frozen extender was 6.0 mg/ml.

  13. Severity of bovine tuberculosis is associated with co-infection with common pathogens in wild boar.

    Directory of Open Access Journals (Sweden)

    David Risco

    Full Text Available Co-infections with parasites or viruses drive tuberculosis dynamics in humans, but little is known about their effects in other non-human hosts. This work aims to investigate the relationship between Mycobacterium bovis infection and other pathogens in wild boar (Sus scrofa, a recognized reservoir of bovine tuberculosis (bTB in Mediterranean ecosystems. For this purpose, it has been assessed whether contacts with common concomitant pathogens are associated with the development of severe bTB lesions in 165 wild boar from mid-western Spain. The presence of bTB lesions affecting only one anatomic location (cervical lymph nodes, or more severe patterns affecting more than one location (mainly cervical lymph nodes and lungs, was assessed in infected animals. In addition, the existence of contacts with other pathogens such as porcine circovirus type 2 (PCV2, Aujeszky's disease virus (ADV, swine influenza virus, porcine reproductive and respiratory syndrome virus, Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae, Haemophilus parasuis and Metastrongylus spp, was evaluated by means of serological, microbiological and parasitological techniques. The existence of contacts with a structured community of pathogens in wild boar infected by M. bovis was statistically investigated by null models. Association between this community of pathogens and bTB severity was examined using a Partial Least Squares regression approach. Results showed that adult wild boar infected by M. bovis had contacted with some specific, non-random pathogen combinations. Contact with PCV2, ADV and infection by Metastrongylus spp, was positively correlated to tuberculosis severity. Therefore, measures against these concomitant pathogens such as vaccination or deworming, might be useful in tuberculosis control programmes in the wild boar. However, given the unexpected consequences of altering any community of organisms, further research should evaluate the impact of such measures

  14. Aujeszky’s Disease and Hepatitis E Viruses Transmission between Domestic Pigs and Wild Boars in Corsica: Evaluating the Importance of Wild/Domestic Interactions and the Efficacy of Management Measures

    Science.gov (United States)

    Charrier, François; Rossi, Sophie; Jori, Ferran; Maestrini, Oscar; Richomme, Céline; Casabianca, François; Ducrot, Christian; Jouve, Johan; Pavio, Nicole; Le Potier, Marie-Frédérique

    2018-01-01

    Wildlife species as reservoirs of infectious pathogens represent a serious constraint in the implementation of disease management strategies. In the Mediterranean island of Corsica, the dynamics of hepatitis E virus (HEV) and Aujeszky’s disease virus (ADV) are suspected to be influenced by interactions between wild and domestic pigs. To improve our understanding of these influences, we first compared the seroprevalences of both viruses in domestic pig populations from different locations with contrasted levels of wild–domestic interactions, ADV vaccination, biosafety, and farm husbandry. Second, we performed an analysis at a more restricted geographical scale, to assess the matching of ADV or HEV prevalence between sympatric wild boar and outdoor pig farms most exposed to interactions with wildlife. Logistic models were adjusted to the observed data. A high seroprevalence of HEV (>80%) and ADV (40%) in pigs, with no significant difference according to the region, confirms that both pathogens are enzootic in Corsica. Vaccination against ADV had a strong protective effect, even when performed voluntarily by farmers. Farm biosafety had an additional effect on pigs’ exposure, suggesting that contact between wild boars and pigs were involved in disease transmission. A strong correlation in HEV seroprevalence was observed between pigs and wild boars that were in close contact, and significantly lower seroprevalence was observed in pigs when they had little contact with wild boars due to spatial segregation. These results suggest a regular HEV circulation between sympatric wild boar and domestic pigs. The high HEV seroprevalence observed in domestic pigs (>80%) suggests a spillover of the virus from domestic to wild populations through environmental contamination, but this hypothesis has to be confirmed. Conversely, even though avoiding sows’ release on pasture during estrus showed some protecting effect in the free ranging pig farms regarding ADV, ADV

  15. Aujeszky’s Disease and Hepatitis E Viruses Transmission between Domestic Pigs and Wild Boars in Corsica: Evaluating the Importance of Wild/Domestic Interactions and the Efficacy of Management Measures

    Directory of Open Access Journals (Sweden)

    François Charrier

    2018-01-01

    Full Text Available Wildlife species as reservoirs of infectious pathogens represent a serious constraint in the implementation of disease management strategies. In the Mediterranean island of Corsica, the dynamics of hepatitis E virus (HEV and Aujeszky’s disease virus (ADV are suspected to be influenced by interactions between wild and domestic pigs. To improve our understanding of these influences, we first compared the seroprevalences of both viruses in domestic pig populations from different locations with contrasted levels of wild–domestic interactions, ADV vaccination, biosafety, and farm husbandry. Second, we performed an analysis at a more restricted geographical scale, to assess the matching of ADV or HEV prevalence between sympatric wild boar and outdoor pig farms most exposed to interactions with wildlife. Logistic models were adjusted to the observed data. A high seroprevalence of HEV (>80% and ADV (40% in pigs, with no significant difference according to the region, confirms that both pathogens are enzootic in Corsica. Vaccination against ADV had a strong protective effect, even when performed voluntarily by farmers. Farm biosafety had an additional effect on pigs’ exposure, suggesting that contact between wild boars and pigs were involved in disease transmission. A strong correlation in HEV seroprevalence was observed between pigs and wild boars that were in close contact, and significantly lower seroprevalence was observed in pigs when they had little contact with wild boars due to spatial segregation. These results suggest a regular HEV circulation between sympatric wild boar and domestic pigs. The high HEV seroprevalence observed in domestic pigs (>80% suggests a spillover of the virus from domestic to wild populations through environmental contamination, but this hypothesis has to be confirmed. Conversely, even though avoiding sows’ release on pasture during estrus showed some protecting effect in the free ranging pig farms regarding ADV

  16. Daily sperm production

    DEFF Research Database (Denmark)

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

    2013-01-01

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

  17. 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

    Domestication promotes the emergence of novel phenotypic and behavioural traits in domesticated animals compared to their wild ancestors. We analysed variation at the melanocortin receptor I (MC1R) and nuclear receptor subfamily 6, group A, member 1 (NR6A1) genes in European wild boar populations......, 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...... of hybridization between wild and domestic forms. Most of the wild boars (94%) were homozygous for the European wild-type (E+) MC1R allele. We did not observe any synonymous substitution in the European E+ allele, confirming its monomorphism even in areas known to be hotspots of wild boar genetic diversity...

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

    Science.gov (United States)

    Svetlichnyy, Valentin; Müller, Peter; Pomorski, Thomas G; Schulze, Martin; Schiller, Jürgen; Müller, Karin

    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 containing saturated (myristoyl, palmitoyl and stearoyl) fatty acyl residues. Glycerophosphatidylcholine and glycerophosphatidylethanolamine with alk(en)yl ether residues in the sn-1 position and unsaturated long chained fatty acyl residues in sn-2 position were identified as the most prominent polar lipids. The only glycoglycerolipid was sulfogalactosylglycerolipid carrying 16:0-alkyl- and 16:0-acyl chains. Using stable isotope-labelling, the metabolic incorporation of exogenously supplied fatty acids was analysed. Boar spermatozoa incorporated hexadecenoic (16:1), octadecenoic (18:1), octadecadienoic (18:2) and octadecatrienoic (18:3) acids primarily in the diacylglycerols and glycerophosphatidylcholines. In contrast, incorporation of eicosapentaenoic acid (20:5) was not detected. The analysis of molecular species composition subsequent to the incorporation of exogenous [(14)C]-octadecadienoic acid suggests two pathways for incorporation of exogenous fatty acids into glycerophosphatidylcholine: (1) de novo synthesis of glycerophosphatidylcholine via the CDP-choline pathway and (2) reacylation of lysophosphatidylcholine via an acyltransferase. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  19. 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.

  20. Characterization of proteins from boar prostate

    Czech Academy of Sciences Publication Activity Database

    Maňásková, Pavla; Ryšlavá, H.; Tichá, M.; Jonáková, Věra

    2002-01-01

    Roč. 48, - (2002), s. 283-290 ISSN 8755-8920. [International Congress of Reproductive Immunology /8./. Opatija, 02.07.2001-06.07.2001] R&D Projects: GA ČR GA310/00/0205; GA ČR GP303/02/P069 Institutional research plan: CEZ:AV0Z5052915 Keywords : boar prostate * beta-microseminoprotein * serotransferrin Subject RIV: CE - Biochemistry Impact factor: 1.413, year: 2002

  1. Relationships among seminal culture, seminal white blood cells, and the percentage of primary sperm abnormalities in bulls evaluated prior to the breeding season.

    Science.gov (United States)

    Sprecher, D J; Coe, P H; Walker, R D

    1999-04-15

    Semen samples from 100 beef breed bulls were evaluated for sperm morphology (phased contrast microscopy), seminal white blood cells, and the presence of potential reproductive pathogens. Eligibility required visualization of the glans penis throughout semen collection. Based on clinical spermiograms, bulls were grouped into normal, marginal, or unsatisfactory morphology classifications. The 3 experimental groups were similar in age and scrotal circumference and differed significantly in the percentage of primary sperm abnormalities. Most semen samples (94%) contained one or more potential reproductive pathogens (Hemophilus somnus. Mycoplasma bovigenitalium, Arcanobacterium pyogenes and Ureaplasma diversum). No significant relationship could be demonstrated between primary abnormalities and the assigned culture score. Our experimental results suggest that clinicians should interpret clinical semen culture results with great care. No significant relationship could be demonstrated between primary abnormalities and assigned white blood cell (WBC) score, although, only 1% of the samples was scored >5 WBC per high power field. The use of seminal WBC score may be valid adjunct to routine semen evaluation when that threshold is the basis for clinical decisions.

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

    Science.gov (United States)

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

    2015-08-01

    In this study, the effect of conjugated linoleic acid (10 trans, 12 cis) (CLA) on refrigerated boar sperm quality parameters up to 14 days at 17°C was assessed. Semen was extended in Androhep and divided into four treatments supplemented with CLA (25, 50, 100 and 200 μm) and control group, then kept for 2 h at 22°C. Afterwards an aliquot of each treatment was removed, and mitochondrial activity, viability, lipid membrane peroxidation (LPO) and stability of the sperm plasma membrane were assessed by flow cytometry. The remaining extended semen was maintained at 17°C until 336 h, repeating the same analysis every 48 h. Regarding percentage of live spermatozoa, no statistical differences were observed among treatments up to 96 h. After this time, viability decreased significantly (p refrigerated boar spermatozoa. © 2015 Blackwell Verlag GmbH.

  3. Advanced sperm selection techniques for assisted reproduction.

    Science.gov (United States)

    McDowell, Simon; Kroon, Ben; Ford, Emily; Hook, Ysanne; Glujovsky, Demián; Yazdani, Anusch

    2014-10-28

    Assisted reproductive technologies (ART) such as in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI) bring together gametes outside of the body to enhance the probability of fertilisation and pregnancy. Advanced sperm selection techniques are increasingly being employed in ART, most commonly in cycles utilising ICSI. Advanced sperm selection techniques are thought to improve the chance that structurally intact and mature sperm with high DNA integrity are selected for fertilisation. Advanced sperm selection strategies include selection according to surface charge; sperm apoptosis; sperm birefringence; ability to bind to hyaluronic acid; and sperm morphology under ultra-high magnification. These techniques theoretically improve ART outcomes. To evaluate the impact of advanced sperm selection techniques on ART outcomes. Systematic search of electronic databases (Cochrane Menstrual Disorders and Subfertility Group Specialised Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, PsycINFO, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Latin American and Caribbean Health Science Information Database (LILACS)), trials registers (ClinicalTrials.gov, Current Controlled Trials, World Health Organization International Clinical Trials Registry Platform), conference abstracts (Web of Knowledge) and grey literature (OpenGrey) for relevant randomised controlled trials. We handsearched the reference lists of included studies and similar reviews. The search was conducted in May 2014. We included randomised controlled trials (RCTs) comparing an advanced sperm selection technique versus standard IVF or ICSI or versus another advanced sperm selection technique. We excluded studies of sperm selection using ultra-high magnification (intracytoplasmic morphologically selected sperm injection, or IMSI), as they are the subject of a separate Cochrane review. Quasi-randomised and pseudo-randomised trials were

  4. Feeding of wild boar (Sus scrofa L.) in the exclusion zone of the Chernobyl accident in the territory of Ukraine

    International Nuclear Information System (INIS)

    Petrov, M.F.

    1996-01-01

    Investigations of seasonal feed choice of a wild boar have been carried out during a 3-year period on the basis of the stomach content. Results of the investigations are given. A list of forage plants and forage animals is presented. Seasonal intensity of their consumption is evaluated. Main plant assemblages of the exclusion zone of the Chernobyl accident are analyzed for their significance in the diet of a wild boar. Special attention is paid to the role of underground forage (35% of average annual forage), and to consumption of Oenotera biennis roots comprising 26% of average annual forage. Recent state of the forage base and of the wild boar population is estimated. An attempt to predict its dynamics for the nearest 10-15 years is made

  5. The influence of boar breed and season on semen parameters

    African Journals Online (AJOL)

    Kamil

    2014-01-29

    Jan 29, 2014 ... Abstract. The aim of the present study was to demonstrate the influence of boar breed and season on semen parameters. The research material consisted of 31 boars: Polish Large White (PLW), Polish Landrace (PL), and Duroc × Pietrain (D × P), aged 8 to 24 months. The analysed material consisted of ...

  6. Effects of supplemental seminal plasma on cryopreserved boar ...

    African Journals Online (AJOL)

    To analyse the effects of supplemental autologous seminal plasma on boar semen quality before freezing and after thawing, thirty ejaculates were collected from six Pietrain boars. The main factors of a 2 x 2 x 2 factorial arrangement of treatments were Beltsville thawing solution (BTS), seminal plasma before freezing, and ...

  7. The influence of boar breed and season on semen parameters ...

    African Journals Online (AJOL)

    The aim of the present study was to demonstrate the influence of boar breed and season on semen parameters. The research material consisted of 31 boars: Polish Large White (PLW), Polish Landrace (PL), and Duroc × Pietrain (D × P), aged 8 to 24 months. The analysed material consisted of 1390 ejaculates, collected ...

  8. Effect of transfection and co-incubation of bovine sperm with exogenous DNA on sperm quality and functional parameters for its use in sperm-mediated gene transfer.

    Science.gov (United States)

    Arias, María Elena; Sánchez-Villalba, Esther; Delgado, Andrea; Felmer, Ricardo

    2017-02-01

    Sperm-mediated gene transfer (SMGT) is based on the capacity of sperm to bind exogenous DNA and transfer it into the oocyte during fertilization. In bovines, the progress of this technology has been slow due to the poor reproducibility and efficiency of the production of transgenic embryos. The aim of the present study was to evaluate the effects of different sperm transfection systems on the quality and functional parameters of sperm. Additionally, the ability of sperm to bind and incorporate exogenous DNA was assessed. These analyses were carried out by flow cytometry and confocal fluorescence microscopy, and motility parameters were also evaluated by computer-assisted sperm analysis (CASA). Transfection was carried out using complexes of plasmid DNA with Lipofectamine, SuperFect and TurboFect for 0.5, 1, 2 or 4 h. The results showed that all of the transfection treatments promoted sperm binding and incorporation of exogenous DNA, similar to sperm incorporation of DNA alone, without affecting the viability. Nevertheless, the treatments and incubation times significantly affected the motility parameters, although no effect on the integrity of DNA or the levels of reactive oxygen species (ROS) was observed. Additionally, we observed that transfection using SuperFect and TurboFect negatively affected the acrosome integrity, and TurboFect affected the mitochondrial membrane potential of sperm. In conclusion, we demonstrated binding and incorporation of exogenous DNA by sperm after transfection and confirmed the capacity of sperm to spontaneously incorporate exogenous DNA. These findings will allow the establishment of the most appropriate method [intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF)] of generating transgenic embryos via SMGT based on the fertilization capacity of transfected sperm.

  9. Single bovine sperm sex typing by amelogenin nested PCR.

    Science.gov (United States)

    Colley, A; Buhr, M; Golovan, S P

    2008-10-01

    Sex-sorted bovine semen has become a valuable tool in animal production for sex preselection. Development of novel sperm sexing technologies, or evaluation of the quality of existing methods, often requires a single-sperm, sex-typing method that is reliable and easy to perform. In the present study, we report the development, validation, and application of a simple, reliable, and cost-effective method for single-sperm sex typing using nested polymerase chain reaction (PCR), based on the amelogenin gene. Several hundred single sperm were isolated using a simple manual technique, or a high-speed flow-sorter, and were successfully sex-typed using the amelogenin nested PCR. Based on the pooled results of individual sperm, there was no significant difference in the semen sex ratio of unsorted (44.6% X-sperm and 55.4% Y-sperm) or X/Y-sorted semen (91.4% X-sperm and 94.0% Y-sperm), as compared to the expected ratio in unsorted semen or the post-sorting reanalysis data, respectively. The amelogenin single-sperm sexing method was an adaptable, accurate, and reliable tool for single-sperm sex typing.

  10. 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.

  11. Rheotaxis guides mammalian sperm

    Science.gov (United States)

    Miki, Kiyoshi; Clapham, David E

    2013-01-01

    Background In sea urchins, spermatozoan motility is altered by chemotactic peptides, giving rise to the assumption that mammalian eggs also emit chemotactic agents that guide spermatozoa through the female reproductive tract to the mature oocyte. Mammalian spermatozoa indeed undergo complex adaptations within the female (the process of capacitation) that are initiated by agents ranging from pH to progesterone, but these factors are not necessarily taxic. Currently, chemotaxis, thermotaxis, and rheotaxis have not been definitively established in mammals. Results Here, we show that positive rheotaxis, the ability of organisms to orient and swim against the flow of surrounding fluid, is a major taxic factor for mouse and human sperm. This flow is generated within 4 hours of sexual stimulation and coitus in female mice; prolactin-triggered oviductal fluid secretion clears the oviduct of debris, lowers viscosity, and generates the stream that guides sperm migration in the oviduct. Rheotaxic movement is demonstrated in capacitated and uncapacitated spermatozoa in low and high viscosity medium. Finally, we show that a unique sperm motion we quantify using the sperm head's rolling rate reflects sperm rotation that generates essential force for positioning the sperm in the stream. Rotation requires CatSper channels, presumably by enabling Ca2+ influx. Conclusions We propose that rheotaxis is a major determinant of sperm guidance over long distances in the mammalian female reproductive tract. Coitus induces fluid flow to guide sperm in the oviduct. Sperm rheotaxis requires rotational motion during CatSper channel-dependent hyperactivated motility. PMID:23453951

  12. 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.

  13. Sperm Na+, K+-ATPase and Ca2+-ATPase activity: A preliminary study of comparison of swim up and density gradient centrifugation methods for sperm preparation

    Science.gov (United States)

    Lestari, Silvia W.; Larasati, Manggiasih D.; Asmarinah, Mansur, Indra G.

    2018-02-01

    As one of the treatment for infertility, the success rate of Intrauterine Insemination (IUI) is still relatively low. Several sperm preparation methods, swim-up (SU) and the density-gradient centrifugation (DGC) are frequently used to select for better sperm quality which also contribute to IUI failure. Sperm selection methods mainly separate the motile from the immotile sperm, eliminating the seminal plasma. The sperm motility involves the structure and function of sperm membrane in maintaining the balance of ion transport system which is regulated by the Na+, K+-ATPase, and Ca2+-ATPase enzymes. This study aims to re-evaluate the efficiency of these methods in selecting for sperm before being used for IUI and based the evaluation on sperm Na+,K+-ATPase and Ca2+-ATPase activities. Fourteen infertile men from couples who underwent IUI were involved in this study. The SU and DGC methods were used for the sperm preparation. Semen analysis was performed based on the reference value of World Health Organization (WHO) 2010. After isolating the membrane fraction of sperms, the Na+, K+-ATPase activity was defined as the difference in the released inorganic phosphate (Pi) with and without the existence of 10 mM ouabain in the reaction, while the Ca2+-ATPase was determined as the difference in Pi contents with and without the existence of 55 µm CaCl2. The prepared sperm demonstrated a higher percentage of motile sperm compared to sperm from the whole semen. Additionally, the percentage of motile sperm of post-DGC showed higher result than the sperm from post-SU. The velocity of sperm showed similar pattern with the percentage of motile sperm, in which the velocity of prepared sperm was higher than the sperm from whole semen. Furthermore, the sperm velocity of post-DGC was higher compared to the sperm from post-SU. The Na+, K+-ATPase activity of prepared sperm was higher compared to whole semen, whereas Na+, K+-ATPase activity in the post DGC was higher than post SU. The Ca2

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

    Science.gov (United States)

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

    2014-10-15

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

  15. Evaluation of semen parameters of boars (Sus scrofa experimentally infected with Toxoplasma gondii/ Avaliação dos parâmetros seminais de cachaços (Sus scrofa experimentalmente infectados com Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Alvimar José da Costa

    2004-05-01

    Full Text Available Aiming to investigate the influence of T. gondii on semen parameters and spermatozoa morphology, eight boars were inoculated with T. gondii. Experimental groups consisted of: GI (n=3 1,5 x 104 oocysts of P strain; GII (n=3 1,0 x 106 tachyzoites of RH strain and GIII (n=2, control noninoculated. Evaluations of semen parameters (volume, motility, strength, concentration, study of spermatozoa morphology, serology (RIFI, parasitemia and hemograms were performed. For this purpose, blood and semen collection were carried out on days -2, -1, 1, 3, 5, 7, 9, 11, 14 and weekly until 84 days post-inoculation. Non hematimetrics alterations and clinical signs were observed on animals. Parasitemia was detected in an animal inoculated with oocysts, on the 7th day post inoculation (DPI and in a two pigs of GII (tachyzoites, on the 3rd and 49th DPI. Serology results revealed the presence of antibody anti-T. gondii on the animals inoculated with oocysts or tachyzoites since 7th DPI, with tittles of 256 and 64, reaching a maximal level of 4096 on days 11 and 9 post inoculation, respectively. The GIII (control was negative through out all experimental period. The semen parametersevaluated did not present any alteration due to toxoplasmosis. Significative differences (PCom o objetivo de investigar a influência do Toxoplasma gondii nos parâmetros seminais e na morfologia espermática de suínos, oito reprodutores foram inoculados com T. gondii, sendo constituídos os seguintes grupos experimentais: GI (n=3 1,5 x 104 oocistos da cepa P, via oral; GII (n=3 1,0 x 106 taquizoítos da cepa RH, via subcutânea e GIII (n=2, controle. Foram realizadas avaliações de parâmetros espermáticos (volume, motilidade, vigor, concentração, estudo da morfologia dos espermatozóides, exames sorológicos(RIFI, parasitemia e hemogramas. Para tanto, colheitas de sangue e sêmen foram realizadas nos dia -2, -1, 1, 3, 5, 7, 9, 11, 14 e semanalmente até 84 dias p

  16. Single layer centrifugation-selected boar spermatozoa are capable of fertilization in vitro

    Science.gov (United States)

    2013-01-01

    Background Good quality spermatozoa are important to achieve fertilization, viable embryos and offspring. Single Layer Centrifugation (SLC) through a colloid (Androcoll-P) selects good quality spermatozoa. However, it has not been established previously whether porcine spermatozoa selected by this method maintain their fertility. Methods The semen was prepared either by SLC or by standard centrifugation (control) and used for in vitro fertilization (IVF) at oocyte:spermatozoa ratios of 1:50; 1:100 and 1:300 (or 4 x 103, 8 x 103 and 24 x 103 spermatozoa/ml) to evaluate their subsequent ability to generate blastocysts. In addition, sperm motility was assessed by computer assisted sperm motility analysis. Results Total and progressive motility were significantly higher in sperm samples prepared by SLC compared to uncentrifuged samples. Sperm binding ability, polyspermy, cleavage and blastocyst rates were affected by the oocyte:sperm ratio, but not by sperm treatment. Conclusion The use of SLC does not adversely affect the in vitro fertilizing and embryo-generating ability of the selected spermatozoa compared to their unselected counterparts, but further modifications in the IVF conditions would be needed to improve the monospermy in IVF systems. Since SLC did not appear to have a negative effect on sperm fertilizing ability, and may in fact select for spermatozoa with a greater potential for fertilization, an in vivo trial to determine the usefulness of this sperm preparation technique prior to artificial insemination is warranted. PMID:23497680

  17. Sperm storage potential and daily sperm production of brown male ...

    African Journals Online (AJOL)

    Sperm storage potential and daily sperm production of brown male Japanese quails for three different physiological age groups. ... positively correlated (r=0.91 P<0.01). This study revealed that the brown male Japanese quail has its peak reproduction potential at puberty. Keywords: Quail sperm storage, sperm production ...

  18. Sperm form and function in the absence of sperm competition.

    Science.gov (United States)

    van der Horst, Gerhard; Maree, Liana

    2014-03-01

    Sperm competition is a post-copulatory, sexual selection force that, together with phylogeny and fertilization mode, has been regarded as one of the main factors explaining the diversity in sperm size across species. This universal sperm selection mechanism favors traits that enhance a male's fertilizing ability and paternity success. Surprisingly, however, sperm characteristics and semen quality in monogamous species, with low risk of sperm competition, have barely received any attention. In this review, we consider sperm competition and monogamy as two ends of the selective spectrum, and discuss its effect on sperm structure and function. We address the issue of a lack of sperm competition by comparing sperm traits of essentially monogamous species--their largely degenerative sperm features and high degree of polymorphisms could be norms for monogamous species. Further, the level of sperm competition in humans is discussed by comparing its mating strategy, relative testis size, and sperm traits to other primate species. In terms of sperm concentration, sperm swimming speed, and sperm morphology, humans seem to be closer aligned to the low-risk sperm competition situation in gorillas than to promiscuous chimpanzees. © 2013 Wiley Periodicals, Inc.

  19. Immunization against recombinant GnRH-I alters testicular structure in an experimental boar model.

    Science.gov (United States)

    Li, Yunsheng; Liu, Ya; Su, Shiping; Pu, Yong; Zhang, Xiaorong; Fang, Fugui

    2015-02-01

    The aim of this study was to evaluate and to compare testicular tissue in immunized and control boars. Eighteen male piglets, aged 12 weeks, were vaccinated twice intramuscularly with a maltose-binding protein-gonadotropin-releasing hormone I hexamer peptide (MBP-GnRH-I6). Blood samples were taken at 12, 18, 21 and 24 weeks of age. Serum concentrations of testosterone and GnRH-I antibodies were determined by radioimmunoassay. The pigs were sacrificed 6 weeks after the second immunization. Testicular weight and size were recorded and tissue samples were collected for histological examination. The results demonstrated that active immunization against MBP-GnRH-I6 increased serum GnRH-I antibody levels (P immunized pigs, and a significant reduction (P immunized boars.

  20. Familial Hepatitis E Outbreak Linked to Wild Boar Meat Consumption.

    Science.gov (United States)

    Rivero-Juarez, A; Frias, M; Martinez-Peinado, A; Risalde, M A; Rodriguez-Cano, D; Camacho, A; García-Bocanegra, I; Cuenca-Lopez, F; Gomez-Villamandos, J C; Rivero, A

    2017-11-01

    An HIV-infected patient was diagnosed with acute hepatitis E infection in our hospital. An epidemiological inquiry was performed to collect demographic, food and animal exposure variables in order to identify the potential route of transmission. The patient reported that his family traditionally hunted wild boar for food. All family members were analysed for hepatitis E virus infection. Additionally, route of transmission by wild boar meat consumption and prevalence of HEV infection among wild boar from the same hunting area were investigated. In all-family members (n = 8), HEV-RNA was amplified. Two wild boar meat slices consumed was analysed, showing the presence of HEV. The virus isolated was consistent with genotype 3, revealing 100% homology between family members and meat. Additionally, we tested nine wild boar hunted in the same hunting area. All of them were RNA-HEV positive, isolating the same HEV genotype 3 viral strain. We demonstrated by phylogenetic analysis zoonotic transmission of HEV by wild boar meat consumption. The prevalence of HEV infection among wild boar found in our study suggests that this species is an important route of transmission to human. © 2017 Blackwell Verlag GmbH.

  1. Improving sperm banking efficiency in endangered species through the use of a sperm selection method in brown bear (Ursus arctos) thawed sperm.

    Science.gov (United States)

    Anel-Lopez, L; Ortega-Ferrusola, C; Álvarez, M; Borragán, S; Chamorro, C; Peña, F J; Morrell, J; Anel, L; de Paz, P

    2017-06-26

    Sperm selection methods such as Single Layer Centrifugation (SLC) have been demonstrated to be a useful tool to improve the quality of sperm samples and therefore to increase the efficiency of other artificial reproductive techniques in several species. This procedure could help to improve the quality of genetic resource banks, which is essential for endangered species. In contrast, these sperm selection methods are optimized and focused on farm animals, where the recovery task is not as important as in endangered species because of their higher sperm availability. The aim of this study was to evaluate two centrifugation methods (300 x g/20 min and 600 x g/10 min) and three concentrations of SLC media (Androcoll-Bear -80, 65 and 50%) to optimise the procedure in order to recover as many sperm with the highest quality as possible. Sperm morphology could be important in the hydrodynamic relationship between the cell and centrifugation medium and thus the effect of sperm head morphometry on sperm yield and its hydrodynamic relationship were studied. The samples selected with Androcoll-Bear 65% showed a very good yield (53.1 ± 2.9) although the yield from Androcoll-Bear 80% was lower (19.3 ± 3.3). The latter showed higher values of motility than the control immediately after post-thawing selection. However, both concentrations of colloid (65 and 80%) showed higher values of viable sperm and viable sperm with intact acrosome than the control. After an incubation of 2 h at 37 °C, the samples from Androcoll-Bear 80% had higher kinematics and proportion of viable sperm with intact acrosome. In the morphometric analysis, the sperm selected by the Androcoll-Bear 80% showed a head with a bigger area which was more elongated than the sperm from other treatments. We conclude that sperm selection with Androcoll-Bear at either 65% or 80% is a suitable technique that allows a sperm population with better quality than the initial sample to be obtained. We recommend the

  2. Genotype-Independent Transmission of Transgenic Fluorophore Protein by Boar Spermatozoa

    Science.gov (United States)

    Garrels, Wiebke; Holler, Stephanie; Taylor, Ulrike; Herrmann, Doris; Struckmann, Christina; Klein, Sabine; Barg-Kues, Brigitte; Nowak-Imialek, Monika; Ehling, Christine; Rath, Detlef; Ivics, Zoltán; Niemann, Heiner; Kues, Wilfried A.

    2011-01-01

    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. PMID:22110672

  3. Intestinal protozoa in wild boars (Sus scrofa) in western Iran.

    Science.gov (United States)

    Solaymani-Mohammadi, S; Rezaian, M; Hooshyar, H; Mowlavi, G R; Babaei, Z; Anwar, M A

    2004-10-01

    A total of 12 gastrointestinal tracts of wild boars (Sus scrofa) from western Iran (Luristan) were examined for protozoan infection between September 2000 and November 2001. Of 12 boars examined, 67% harbored one or more species of the following protozoa: Balantidium coli (25%), Tritrichomonas suis (25%), Blastocystis sp. (25%), Entamoeba polecki (17%), Entamoeba suis (8%), Iodamoeba butschlii (17%), and Chilomastix mesnili (8%). Four of these protozoan species also are reported in humans, and persons living in rural areas where wild boars are abundant should take precaution to avoid infection.

  4. How does supplementary feeding affect endoparasite infection in wild boar?

    DEFF Research Database (Denmark)

    Oja, Ragne; Velstrom, Kaisa; Moks, Epp

    2017-01-01

    , as well as accumulation of parasites in the environment, may increase parasite prevalence. Our aim was to investigate whether host density or the number of feeding sites drives endoparasite infection in an Estonian wild boar (Sus scrofa) population. For this, we collected wild boar faeces from forests...... contamination or indicate that supplementary feeding sites are suitable habitat for soil-dwelling nematodes. These results suggest that the effects of supplementary feeding on parasite prevalence in wild boar are mediated by the characteristics of parasite life cycles....

  5. 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 TESE and TESA groups were comparable [33% (6/18) and 50% (1/2), respectively]. The data indicate that micro-TESE is a highly successful sperm retrieval technique for men with 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. © 2015 American Society of Andrology and European Academy of Andrology.

  6. A dictionary learning approach for human sperm heads classification.

    Science.gov (United States)

    Shaker, Fariba; Monadjemi, S Amirhassan; Alirezaie, Javad; Naghsh-Nilchi, Ahmad Reza

    2017-12-01

    To diagnose infertility in men, semen analysis is conducted in which sperm morphology is one of the factors that are evaluated. Since manual assessment of sperm morphology is time-consuming and subjective, automatic classification methods are being developed. Automatic classification of sperm heads is a complicated task due to the intra-class differences and inter-class similarities of class objects. In this research, a Dictionary Learning (DL) technique is utilized to construct a dictionary of sperm head shapes. This dictionary is used to classify the sperm heads into four different classes. Square patches are extracted from the sperm head images. Columnized patches from each class of sperm are used to learn class-specific dictionaries. The patches from a test image are reconstructed using each class-specific dictionary and the overall reconstruction error for each class is used to select the best matching class. Average accuracy, precision, recall, and F-score are used to evaluate the classification method. The method is evaluated using two publicly available datasets of human sperm head shapes. The proposed DL based method achieved an average accuracy of 92.2% on the HuSHeM dataset, and an average recall of 62% on the SCIAN-MorphoSpermGS dataset. The results show a significant improvement compared to a previously published shape-feature-based method. We have achieved high-performance results. In addition, our proposed approach offers a more balanced classifier in which all four classes are recognized with high precision and recall. In this paper, we use a Dictionary Learning approach in classifying human sperm heads. It is shown that the Dictionary Learning method is far more effective in classifying human sperm heads than classifiers using shape-based features. Also, a dataset of human sperm head shapes is introduced to facilitate future research. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Dilution of boar ejaculates with BTS containing HEPES in place of bicarbonate immediately after ejaculation can reduce the increased inducibility of the acrosome reaction by treatment with calcium and calcium ionophore A23187, which is potentially associated with boar subfertility.

    Science.gov (United States)

    Murase, Tetsuma; Imaeda, Noriaki; Yamada, Hiroto; Takasu, Masaki; Taguchi, Kazuo; Katoh, Tsutomu

    2010-06-01

    The present study investigated whether substitution of HEPES for bicarbonate in BTS (BTS-H) used to dilute boar ejaculates immediately after ejaculation could reduce the increased inducibility of the acrosome reaction by calcium and calcium ionophore A23187. When an ejaculate was split, diluted 5-fold with regular BTS (BTS-B) and BTS-H and stored at 17 C for 12 h or 60 h, the extender or storage time had no significant influence on sperm motility or viability measured by the eosin-nigrosin method. When spermatozoa diluted serially with BTS-B and stored (36 h) were stimulated with Ca2+ (3 mM) and A23187 (0.3 microM), the proportion of spermatozoa that underwent the acrosome reaction (% acrosome reactions) significantly increased as the magnifications of dilution increased (bicarbonate content almost unchanged by dilution). By contrast, the % acrosome reactions in spermatozoa similarly diluted and stored with BTS-H decreased with the increasing magnifications of dilution (bicarbonate decreased). Sperm motility immediately after the end of incubation without A23178 tended to be lower for BTS-H than BTS-B, and the ejaculates for BTS-H had a tendency to have a lower total protein in seminal plasma than those for BTS-B. These results implied that the samples for BTS-H could be used as a model for ejaculates possibly collected during summer and showing subfertility. When an ejaculate was split, diluted serially with BTS-B and BTS-H and stored, viability measured by staining with propidium iodide was extremely similar between the 2 extenders and among the different dilution magnifications, regardless of whether spermatozoa were washed (stored for 36-66 h) or not (stored for 66-72 h). These results suggest that boar ejaculate can be stored with BTS-H at least according to the results for sperm motility and viability and that hypersensitivity of spermatozoa to Ca2+ and A23187 potentially associated with boar subfertility could be lessened by diluting ejaculates with BTS-H.

  8. Implementing an open-access CASA software for the assessment of stallion sperm motility: Relationship with other sperm quality parameters.

    Science.gov (United States)

    Giaretta, Elisa; Munerato, Mauro; Yeste, Marc; Galeati, Giovanna; Spinaci, Marcella; Tamanini, Carlo; Mari, Gaetano; Bucci, Diego

    2017-01-01

    Setting an open-access computer assisted sperm analysis (CASA) may benefit the evaluation of motility in mammalian sperm, especially when economic constraints do not allow the use of a commercial system. There have been successful attempts to develop such a device in Zebra fish sperm and the system has been used in very few studies on mammalian spermatozoa. Against this background, the present study aimed at developing an open-access CASA system for mammalian sperm using the horse as a model and based upon the Image J software previously established for Zebra fish sperm. Along with determining the sperm progressive motility and other kinetic parameters (such as amplitude of lateral head displacement), the "results" window was adjusted to simplify subsequent statistical analyses. The path window was enriched with colored sperm trajectories on the basis of the subpopulation they belong to and a number that allowed the sperm track to be associated to the sperm motility data shown in the "results" window. Data obtained from the novel plugin (named as CASA_bgm) were compared with those of the commercial CASA Hamilton-Thorn IVOS Vers.12, through Bland Altman's plots. While the percentage of total and progressive motile sperm, VCL, VAP, VSL, LIN and STR and ALH were in agreement with those obtained with the commercial system, BCF significantly differed between the two systems probably due to their settings. Interestingly, a positive and significant correlation between the percentages of total motile sperm evaluated through CASA_bgm and those showing high mitochondrial membrane potential evaluated by JC-1 staining was found. In conclusion, CASA_bgm ImageJ plugin could be useful and reliable for stallion sperm motility analysis and it is our aim to apply this system to other mammalian species. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Bionic Design for Reducing Adhesive Resistance of the Ridger Inspired by a Boar's Head

    Science.gov (United States)

    Li, Jianqiao; Yan, Yunpeng; Chirende, Benard; Wu, Xuejiao; Wang, Zhaoliang

    2017-01-01

    The main feature of the boar's head used to root around for food is the front part, which is similar to the ridger in terms of function, load, and environment. In this paper, the boar's head was selected as the biological prototype for developing a new ridger. The point cloud of the head was captured by a 3D scanner, and then, the head surface was reconstructed using 3D coordinates. The characteristic curves of the front part of the boar's head were extracted, and then, five cross-sectional curves and one vertical section curve were fitted. Based on the fitted curves, five kinds of bionic ridgers were designed. The penetrating resistances of the bionic ridgers and traditional ridger were tested at different speeds in an indoor soil bin. The test results showed that bionic ridger B had the best penetrating resistance reduction ratio of 16.67% at 4.2 km/h velocity. In order to further evaluate the performance of the best bionic ridger (bionic ridger B), both the bionic ridger and traditional ridger were tested in a field under the same working conditions. The field results indicate that the bionic ridger reduces penetrating resistance by 6.91% compared to the traditional ridger, and the test results validate that the bionic ridger has an effect on reducing penetrating resistance. PMID:28757796

  10. Mycobacterium tuberculosis Infection in a Domesticated Korean Wild Boar ( Sus scrofa coreanus).

    Science.gov (United States)

    Seo, Min-Goo; Ouh, In-Ohk; Kim, Munki; Lee, Jienny; Kim, Young-Hoan; Do, Jae-Cheul; Kwak, Dongmi

    2017-06-01

    Tuberculosis, a chronic progressive disease, has been reported in bovine, swine, and primate species. Here, we report the first case of Mycobacterium tuberculosis infection in a Korean wild boar ( Sus scrofa coreanus). The owners this domesticated boar brought it to the Gyeongbuk Veterinary Service Laboratory in Korea after it was found dead and severely emaciated. Demarcated yellowish white nodules were found around the larynx and retropharyngeal lymph node during necropsy. The lungs had diffuse fibrinous pleuritis, severe congestion, and scattered nodules. More nodules were found in the spleen. Tuberculosis is characterized by massive macrophage infiltration and central caseous necrosis; both characteristics were found in the lungs. Histopathologic examination revealed that the alveolar lumen had marked fibrosis and exudates. Examination of the fluid revealed extensive macrophage permeation. To confirm a Mycobacterium infection, PCR was performed using two primer sets specific to the rpoB gene of Mycobacterium; Mycobacterium was detected in the lungs and spleen. To identify the species of Mycobacterium, immunohistochemical evaluation was performed using antibodies against Mycobacterium tuberculosis and Mycobacterium bovis . The results revealed immunoreactivity against M. tuberculosis but not against M. bovis . The consumption of undercooked or raw meat from game animals may expose humans and other animals to sylvatic infection. Consequently, Koreans who ingest wild boar may be at risk of a tuberculosis infection. To reduce the risk of foodborne infection and maintain public health, continuous monitoring and control strategies are required.

  11. 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.

  12. Vitrification of neat semen alters sperm parameters and DNA integrity.

    Science.gov (United States)

    Khalili, Mohammad Ali; Adib, Maryam; Halvaei, Iman; Nabi, Ali

    2014-05-06

    Our aim was to evaluate the effect of neat semen vitrification on human sperm vital parameters and DNA integrity in men with normal and abnormal sperm parameters. Semen samples were 17 normozoospermic samples and 17 specimens with abnormal sperm parameters. Semen analysis was performed according to World Health Organization (WHO) criteria. Then, the smear was provided from each sample and fixed for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Vitrification of neat semen was done by plunging cryoloops directly into liquid nitrogen and preserved for 7 days. The samples were warmed and re-evaluated for sperm parameters as well as DNA integrity. Besides, the correlation between sperm parameters and DNA fragmentation was assessed pre- and post vitrification. Cryopreserved spermatozoa showed significant decrease in sperm motility, viability and normal morphology after thawing in both normal and abnormal semen. Also, the rate of sperm DNA fragmentation was significantly higher after vitrification compared to fresh samples in normal (24.76 ± 5.03 and 16.41 ± 4.53, P = .002) and abnormal (34.29 ± 10.02 and 23.5 ± 8.31, P neat ejaculates has negative impact on sperm parameters as well as DNA integrity, particularly among abnormal semen subjects. It is, therefore, recommend to process semen samples and vitrify the sperm pellets.

  13. [Classical swine fever in wild boars in Switzerland].

    Science.gov (United States)

    Hofmann, M A; Thür, B; Vanzetti, T; Schleiss, W; Schmidt, J; Griot, C

    1999-01-01

    In May 1998, wild boars with classical swine fever (CSF) symptoms were detected in the southern part (Canton Ticino) of Switzerland. CSF virus was isolated from the submitted samples and RT-PCR followed by direct nucleotide sequencing of the 5' non-translated region showed that this virus was identical to the isolate previously recognized in wild boars from the area of Varese (Italy). In most animals, antibodies to CSF virus were detected as well. An immediate measurement was taken by limiting the movement of pigs and identifying both risk and surveillance zones. In order not to disturb potentially infected wild boars within their habitat a complete hunting prohibition for 2 months was enforced. The different possibilities of the control of CSF outbreaks in wild boars are discussed.

  14. A 38-year study on Trichinella spp. in wild boar (Sus scrofa) of Latvia shows a stable incidence with an increased parasite biomass in the last decade.

    Science.gov (United States)

    Kirjušina, Muza; Deksne, Gunita; Marucci, Gianluca; Bakasejevs, Eduards; Jahundoviča, Inese; Daukšte, Anžela; Zdankovska, Aleksandra; Bērziņa, Zanda; Esīte, Zanda; Bella, Antonino; Galati, Fabio; Krūmiņa, Angelika; Pozio, Edoardo

    2015-03-01

    Trichinella spp. are zoonotic parasites transmitted to humans by the consumption of raw or insufficiently cooked meat of different animal species. The most common source of infection for humans is meat from pigs and wild boar (Sus scrofa). The aim of the present work was to evaluate the incidence of Trichinella spp. infections in wild boar hunted in Latvia over a 38 year interval (1976 to 2013). A total 120,609 wild boars were individually tested for Trichinella spp. by trichinoscopy and, in case of negativity, by artificial digestion of 25 g muscles, in the 1976-2005 period, and by artificial digestion of 25-50 g muscles in the 2006-2013 period. Trichinella spp. larvae were identified at the species level by multiplex PCR. In the study period, the overall prevalence of infected wild boar was 2.5%. Trichinella britovi was the predominant (90%) species. The incidence of Trichinella spp. infection in wild boar exhibited two different trends. From 1976 to 1987, the incidence of infected/hunted wild boar increased from 0.23% to 2.56%, then it decreased to 0.19 in 1994. Thereafter, the incidence fluctuated between 0.05% and 0.37%. A statistically significant (P Latvia increased by 4.9 times and the hunting bag by 9.7 times, with a stable incidence of Trichinella spp. in the population. It follows that the biomass of Trichinella spp. larvae and of T. britovi, in particular, increased. The incidence trends of Trichinella spp. in wild boar could be related to the role played by the snow in reducing the thermal shock and muscle putrefaction which increases the survival of the larvae in muscle tissues of carrion in the 1976-1993 period; and, in the 1997-2013 period, to the increased biomass of Trichinella spp. due to the increased carnivore populations, which are the main reservoirs of these parasites.

  15. Helminth parasites of wild boar, Sus scrofa, in Iran.

    Science.gov (United States)

    Eslami, A; Farsad-Hamdi, S

    1992-04-01

    Fifty-seven wild boars (Sus scrofa) from protected regions of Iran were examined for helminths. Sixteen species of helminths were collected; there were ten nematodes, one acanthocephalan, two trematodes and three larval cestodes. New host and distribution records were established for all helminths except of Taenia solium cysticerci. Wild boar shared nine of these helminths with domestic pigs, six with ruminants and three with human beings in Iran. Metastrongylus pudendotectus and M. salmi are reported for the first time from Iran.

  16. Analysis of intravital assessment results in purebred and crossbred boars

    Directory of Open Access Journals (Sweden)

    Wanda Milewska

    2005-01-01

    Full Text Available 1879 purebred boars: Duroc (D, Hampshire (H and Pietrain (Pn as well as their two-crossbreds (D x H, H x D, Pn x H, H x Pn, Pn x D, D x Pn were analysed after a living assessment completed between 1995–1998 within the area of activity of the Animal Breeding Station in Olsztyn. The standardi- zed daily gains in body weight ranged between 594 g (H and 628 g (Pn x D. The thinnest backfat layer was observed in Pietrain boars and their crossbred boars. The height of loin eye was the best for boars Pn and for all crossbred animals. The lean content in the body of the assessed boars ranged from 56.2% (H to 60.0% (Pn. The highest value of selection index was found for Pn boars (124 points and crossbred boars Pn x D and Pn x H, the lowest index value was observed for purebred H boars (109 points. The correlation coefficient between the index and daily gains range from 0.69 (P ≤ 0.01 (H to 0.91 (P ≤ 0.01 (H x D, between the index and the height of loin eye range from 0.57 (P ≤ 0.01 (H to 0.75 (P ≤ 0.01 (Pn and between the index and the lean content range from 0.13 (P ≤ 0.05 (H to 0.71 (P ≤ 0.01 (Pn.

  17. Low Sperm Count

    Science.gov (United States)

    ... problem such as an inherited chromosomal abnormality, a hormonal imbalance, dilated testicular veins or a condition that blocks ... Elevated temperatures impair sperm production and function. Although studies are limited and are inconclusive, frequent use of ...

  18. Sperm quality after swim up and density gradient centrifugation sperm preparation with supplementation of alpha lipoic acid (ALA): A preliminary study

    Science.gov (United States)

    Lestari, Silvia W.; Lestari, Sarah H.; Pujianto, Dwi A.

    2018-02-01

    Intra uterine insemination (IUI) as one of the treatment for infertility, persists low success rate. A factor that contributes to the unsuccessful of IUI is sperm preparation, performed through Swim-up (SU) and Density Gradient Centrifugation (DGC) methods. Furthermore, studies have shown that Alpha Lipoic Acid (ALA) is a potent antioxidant that could enhance the sperm motility and protect the DNA integrity of the sperm [1]. This study is aimed to re-evaluate the efficiency of the DGC and SU methods in selecting sperm before being transferred for IUI by the supplementation of ALA based on the sperm DNA integrity. Semen samples were obtained from 13 men from partners of women who are infertile (normozoospermia) and underwent IUI. Semen analysis based on the guideline of World Health Organization (WHO) 2010 was performed to measure the sperm motility and velocity, before and after sperm preparation. Then, samples were incubated with Alpha Lipoic Acid (ALA) in 0.625 mg (ALA 1), 1.25 mg (ALA 2) and 2.5 mg (ALA 3). The Sperm Chromatin Dispersion (SCD) test was performed to evaluate the sperm DNA Fragmentation Index (DFI). The percentage of motile sperm was higher in prepared sperm (post-DGC and post-SU) than in whole semen. Furthermore, the percentage of motile sperm was higher in post-DGC compared to post-SU. The level of DFI after the supplementation of ALA was decreased in prepared sperm compared to the whole semen. ALA was proved capable to select the better sperm quality with decreased sperm DNA fragmentation of prepared sperm in the all of DFI category.

  19. 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.

  20. Reproductive tissue expression and sperm localization of porcine beta-microseminoprotein

    Czech Academy of Sciences Publication Activity Database

    Maňásková-Postlerová, Pavla; Davidová, Nina; Šulc, Miroslav; Philimonenko, Anatoly; Hozák, Pavel; Jonáková, Věra

    2011-01-01

    Roč. 344, č. 2 (2011), s. 341-353 ISSN 0302-766X R&D Projects: GA ČR(CZ) GA303/09/1285; GA ČR GD523/08/H064; GA MZd(CZ) NS10009; GA MŠk(CZ) 1M06011; GA MŠk(CZ) LC07017; GA MŠk(CZ) LC545 Institutional research plan: CEZ:AV0Z50520701; CEZ:AV0Z50520514; CEZ:AV0Z50200510 Keywords : porcine beta-microseminoprotein * reproductive organs * spermatozoa * immunofluorescence * sperm acrosome * boar Subject RIV: CE - Biochemistry Impact factor: 3.114, year: 2011

  1. A Study of Anti-Sperm Antibodies among Infertile Subjects ...

    African Journals Online (AJOL)

    Background: Anti-sperm antibodies (ASA) can interfere with sperm functions and fertility and may be found in the blood, lymph or local secretions such as seminal and cervico-vaginal fluids in both men and women. Objective: to evaluate the contribution of ASA to infertility in male and female subjects investigated for ...

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

    Science.gov (United States)

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

    2015-03-31

    Relaxin is detected in seminal plasma of many species and its association with sperm motility may be beneficial in some aspects of assisted reproduction. Here, we immunolocalized relaxin receptors and investigated the effects of exogenous relaxin on motility characteristics, viability, and cAMP content of boar spermatozoa after storage. Commercial doses of boar semen were obtained on the collection day (Day 0) and kept in shipping containers at room temperature for up to 4 days (Day 4). On Day 0, spermatozoa were fixed for immunofluorescence detection of relaxin receptors RXFP1 and RXFP2 (Experiment 1). Semen aliquots were taken from the same dose at Day 0, Day 1, and Day 2 (Experiment 2a), and Day 2 and Day 4 (Experiment 2b) for analyses. Alive spermatozoa were purified and incubated (1 h-37°C) with 0, 50, or 100 ng relaxin/ml (Experiment 2a) and 0, 100, or 500 ng relaxin/ml (Experiment 2b). Afterward, aliquots of each treatment group were subjected to motility (Experiments 2), viability (Experiment 3) analyses, and cAMP quantification (Experiment 4). Data (3-4 independent replicates) were statistically analyzed (ANOVA followed by pairwise comparisons) and p values less or equal to 0.05 was set for significant difference. Both RXFP1 and RXFP2 receptors were immunolocalized on the entire spermatozoon. Relaxin concentration of 100 ng/ml significantly improved the proportions of motile, progressive, and rapid spermatozoa up to Day 2. Only 500 ng relaxin/ml provided beneficial effects on Day 4. The viability of spermatozoa was not affected by relaxin (100 ng/ml) during storage, but the extent of mitochondria membrane damages was significantly decreased. Furthermore, relaxin did not affect the cAMP contents of spermatozoa during storage, in our conditions. Relaxin could be a valuable motility booster of stored- or aged-spermatozoa for assisted reproduction techniques. However, the related-intracellular signaling cascades of relaxin in boar spermatozoa remain

  3. 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. © 2015 Blackwell Verlag GmbH.

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

    DEFF Research Database (Denmark)

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

    2003-01-01

    Multiple insemination induces sperm competition and may select for longer, faster moving sperm in species where sperm is short-lived and egg fertilization takes place almost immediately after ejaculation. Here we report the first detailed analysis of sperm length in social insects with long......-term storage of sperm, using three bumblebee species with different mating systems as models. We show that individual males produce only one size-class of sperm, but that sperm length is highly variable among brothers, among unrelated conspecific males, and among males of different species. Males of Bombus...... hypnorum, a species with multiple-mating queens, have longer sperm than males of B. terrestris and B. lucorum whose queens are single mated. Although the sample size on the species level was too small to perform a phylogenetic analysis, this finding supports the hypothesis that, all other things being...

  5. 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.

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

    Directory of Open Access Journals (Sweden)

    Jens Fedder

    2017-01-01

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

  7. Use of laptop computers connected to internet through Wi-Fi decreases human sperm motility and increases sperm DNA fragmentation.

    Science.gov (United States)

    Avendaño, Conrado; Mata, Ariela; Sanchez Sarmiento, César A; Doncel, Gustavo F

    2012-01-01

    To evaluate the effects of laptop computers connected to local area networks wirelessly (Wi-Fi) on human spermatozoa. Prospective in vitro study. Center for reproductive medicine. Semen samples from 29 healthy donors. Motile sperm were selected by swim up. Each sperm suspension was divided into two aliquots. One sperm aliquot (experimental) from each patient was exposed to an internet-connected laptop by Wi-Fi for 4 hours, whereas the second aliquot (unexposed) was used as control, incubated under identical conditions without being exposed to the laptop. Evaluation of sperm motility, viability, and DNA fragmentation. Donor sperm samples, mostly normozoospermic, exposed ex vivo during 4 hours to a wireless internet-connected laptop showed a significant decrease in progressive sperm motility and an increase in sperm DNA fragmentation. Levels of dead sperm showed no significant differences between the two groups. To our knowledge, this is the first study to evaluate the direct impact of laptop use on human spermatozoa. Ex vivo exposure of human spermatozoa to a wireless internet-connected laptop decreased motility and induced DNA fragmentation by a nonthermal effect. We speculate that keeping a laptop connected wirelessly to the internet on the lap near the testes may result in decreased male fertility. Further in vitro and in vivo studies are needed to prove this contention. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  8. Sperm DNA fragmentation: An early and reliable marker of air pollution.

    Science.gov (United States)

    Bosco, Liana; Notari, Tiziana; Ruvolo, Giovanni; Roccheri, Maria C; Martino, Chiara; Chiappetta, Rosanna; Carone, Domenico; Lo Bosco, Giosuè; Carrillo, Laura; Raimondo, Salvatore; Guglielmino, Antonino; Montano, Luigi

    2018-03-01

    Environmental factors could have a key role in the continuous and remarkable decline of sperm quality observed in the last decades. This study compared the seminal parameters and sperm DFI in men living in areas with different levels of air pollution. Results demonstrate that both steel plants workers and patients living in a high polluted area show a mean percentage of sperm DNA fragmentation above 30%, highlighting a clear sperm damage. In this work, two different techniques were used to measure sperm DNA damage in patients' groups, finding in both cases a high sperm DFI in patients living in polluted areas. We candidate sperm DNA fragmentation as a valuable early marker of the presence and harmful effects of pollution. We suggest that sperm DNA evaluation could be both an indicator of individual health and reproductive capacity, and a suitable datum to connect the surrounding environment with its effects. Copyright © 2018 Elsevier B.V. All rights reserved.

  9. 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...

  10. Cryo-scanning electron microscopy discloses differences in dehydration of frozen boar semen stored in large containers.

    Science.gov (United States)

    Ekwall, H

    2009-02-01

    In general, freezing in flat plastic polyethylene terephthalate (PET) bags (FlatPacks) at 50 degrees C/min gives better post-thaw viability, in terms of sperm motility and membrane integrity, than does freezing in plastic maxi-straws, probably owing to differences in cryobiology. To test the hypothesis that this better survival post-thaw relates to the degree of sperm dehydration during freezing, the present study investigated the structure of boar semen in a frozen state using cryo-scanning electron microscopy (cryo-SEM) to compare two different packages (FlatPacks and maxi-straws) for single artificial insemination (AI) doses, and three different freezing rates. The semen was split-sample frozen in maxi-straws or FlatPacks (both holding 5 ml) using 3% glycerol as cryoprotectant. Three freezing rates were applied from -5 degrees C to -100 degrees C, namely 2 degrees C/min, 50 degrees C/min and 1200 degrees C/min, the lattermost by plunging the samples into liquid nitrogen (LN(2)). The samples were thereafter fractured into LN(2) and larger areas of extra-cellular, unbound frozen water ('ice lakes') were measured to determine the degree of dehydration of the spermatozoa. These areas decreased in size with an increase in cooling rate, the differences in size being more dramatic for maxi-straws than for FlatPacks. Size of ice lakes was also influenced by location within package in relation to cooling rate, the central values being always smaller in maxi-straws than in Flatpacks (p < 0.05 at 2 degrees C/min and 50 degrees C/min) but not at 1200 degrees C/min, which suggested the FlatPack allows for more homogenous freezing of boar semen.

  11. Healthy Sperm: Improving Your Fertility

    Science.gov (United States)

    Healthy Lifestyle Getting pregnant Healthy sperm aren't always a given. Understand how lifestyle factors can affect your ... of retrieving and storing sperm before treatment. Adopting healthy lifestyle practices to promote your fertility — and avoiding things ...

  12. 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.

  13. Serological investigation of wild boars (Sus scrofa) and red foxes (Vulpes vulpes) as indicator animals for circulation of Francisella tularensis in Germany.

    Science.gov (United States)

    Otto, Peter; Chaignat, Valerie; Klimpel, Diana; Diller, Roland; Melzer, Falk; Müller, Wolfgang; Tomaso, Herbert

    2014-01-01

    Tularemia outbreaks in humans have recently been reported in many European countries, but data on the occurrence in the animal population are scarce. In North America, seroconversion of omnivores and carnivores was used as indicator for the presence of tularemia, for the European fauna, however, data are barely available. Therefore, the suitability of wild boars (Sus scrofa) and red foxes (Vulpes vulpes) as indicators for the circulation of F. tularensis in Germany was evaluated. Serum samples from 566 wild boars and 457 red foxes were collected between 1995 and 2012 in three federal states in Central Germany (Hesse, Saxony-Anhalt, and Thuringia). The overall rate of seropositive animals was 1.1% in wild boars and 7.4% in red foxes. In conclusion, serological examination of red foxes is recommended, because they can be reliably used as indicator animals for the presence of F. tularensis in the environment.

  14. Intracellular translocation and differential accumulation of cell-penetrating peptides in bovine spermatozoa: evaluation of efficient delivery vectors that do not compromise human sperm motility

    Science.gov (United States)

    Jones, Sarah; Lukanowska, Monika; Suhorutsenko, Julia; Oxenham, Senga; Barratt, Christopher; Publicover, Steven; Copolovici, Dana Maria; Langel, Ülo; Howl, John

    2013-01-01

    STUDY QUESTION Do cell penetrating peptides (CPPs) translocate into spermatozoa and, if so, could they be utilized to deliver a much larger protein cargo? SUMMARY ANSWER Chemically diverse polycationic CPPs rapidly and efficiently translocate into spermatozoa. They exhibit differential accumulation within intracellular compartments without detrimental influences upon cellular viability or motility but they are relatively ineffective in transporting larger proteins. WHAT IS ALREADY KNOWN Endocytosis, the prevalent route of protein internalization into eukaryotic cells, is severely compromised in mature spermatozoa. Thus, the translocation of many bioactive agents into sperm is relatively inefficient. However, the delivery of bioactive moieties into mature spermatozoa could be significantly improved by the identification and utility of an efficient and inert vectorial delivery technology. STUDY DESIGN CPP translocation efficacies, their subsequent differential intracellular distribution and the influence of peptides upon viability were determined in bovine spermatozoa. Temporal analyses of sperm motility in the presence of exogenously CPPs utilized normozoospermic human donor samples. MATERIALS AND METHODS CPPs were prepared by manual, automated and microwave-enhanced solid phase synthesis. Confocal fluorescence microscopy determined the intracellular distribution of rhodamine-conjugated CPPs in spermatozoa. Quantitative uptake and kinetic analyses compared the translocation efficacies of chemically diverse CPPs and conjugates of biotinylated CPPs and avidin. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) conversion assays were employed to analyse the influence of CPPs upon sperm cell viability and sperm class assays determined the impact of CPPs on motility in capacitated and non-capacitated human samples. MAIN RESULTS Chemically heterogeneous CPPs readily translocated into sperm to accumulate within

  15. Effect of sorafenib on sperm count and sperm motility in male Swiss albino mice

    Directory of Open Access Journals (Sweden)

    Surekha Devadasa Shetty

    2015-01-01

    Full Text Available 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 1 st , 2 nd , 4 th , 5 th , 7 th , and 10 th 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.

  16. Study of Sperm Parameters and Sperm Fertility in Mice were Exposed to Tamoxifen during Embryonic Development

    Directory of Open Access Journals (Sweden)

    J Soleimanirad

    2017-05-01

    Full Text Available Introduction: Tamoxifen is steroidal drug, which mainly treats breast cancer and also used to stimulate ovulation. The purpose of the present study was the evaluation of sperm parameters and fertility of mice whose mothers had received tamoxifen during pregnancy. Methods: In this study, 30 female and 15 male mice of NMRI were selected for mating. After mating female mice were randomly divided into two groups, the first group (control and second group (experimental. All of which contained 15 mice. From the day 13th day of pregnancy, experimental group has received tamoxifen with the dosage of 5 mg/kg for 7 days. After childbirth of the mated mice, male infants were selected. After reaching the age of puberty (6-8Weeks, adult mice were sacrificed by the cervical dislocation. After take sperm, sperm parameters (count, normality and motility, and sperm fertility was performed. In this study SPSS software and statistical t-test was used (p <0.001. Results: Studies showed that sperm parameters and sperm fertilization were significantly different. The number of sperm in the control group was 83.50±28.20 million, and in the experimental group was 60±14.14 million. There was a decrease in average sperm count in the experimental group compared with the control group (p <0.001. Our findings from in vitro fertilization culture media showed that embryos formation and oocyte disruption between control and experimental groups significantly different (p <0.001. Conclusion: The results showed that tamoxifen exposure during development can cause histological changes in the seminiferous tubules, which can lead to infertility.

  17. Validation of a spectrophotometer-based method for estimating daily sperm production and deferent duct transit.

    Science.gov (United States)

    Froman, D P; Rhoads, D D

    2012-10-01

    The objectives of the present work were 3-fold. First, a new method for estimating daily sperm production was validated. This method, in turn, was used to evaluate testis output as well as deferent duct throughput. Next, this analytical approach was evaluated in 2 experiments. The first experiment compared left and right reproductive tracts within roosters. The second experiment compared reproductive tract throughput in roosters from low and high sperm mobility lines. Standard curves were constructed from which unknown concentrations of sperm cells and sperm nuclei could be predicted from observed absorbance. In each case, the independent variable was based upon hemacytometer counts, and absorbance was a linear function of concentration. Reproductive tracts were excised, semen recovered from each duct, and the extragonadal sperm reserve determined by multiplying volume by sperm cell concentration. Testicular sperm nuclei were procured by homogenization of a whole testis, overlaying a 20-mL volume of homogenate upon 15% (wt/vol) Accudenz (Accurate Chemical and Scientific Corporation, Westbury, NY), and then washing nuclei by centrifugation through the Accudenz layer. Daily sperm production was determined by dividing the predicted number of sperm nuclei within the homogenate by 4.5 d (i.e., the time sperm with elongated nuclei spend within the testis). Sperm transit through the deferent duct was estimated by dividing the extragonadal reserve by daily sperm production. Neither the efficiency of sperm production (sperm per gram of testicular parenchyma per day) nor deferent duct transit differed between left and right reproductive tracts (P > 0.05). Whereas efficiency of sperm production did not differ (P > 0.05) between low and high sperm mobility lines, deferent duct transit differed between lines (P < 0.001). On average, this process required 2.2 and 1.0 d for low and high lines, respectively. In summary, we developed and then tested a method for quantifying male

  18. Sperm whale clicks

    DEFF Research Database (Denmark)

    Møhl, Bertel; Wahlberg, Magnus; Madsen, Peter T.

    2000-01-01

    . A sound generator weighing upward of 10 tons and with a cross-section of 1 m is expected to generate high-intensity, directional sounds. This prediction from the Norris and Harvey theory is not supported by published data for sperm whale clicks ~source levels of 180 dB re 1 mPa and little, if any....... This implicates sonar as a possible function of the clicks. Thus, previously published properties of sperm whale clicks underestimate the capabilities of the sound generator and therefore cannot falsify the Norris and Harvey theory.......In sperm whales ~Physeter catodon L. 1758! the nose is vastly hypertrophied, accounting for about one-third of the length or weight of an adult male. Norris and Harvey @in Animal Orientation and Navigation, NASA SP-262 ~1972!, pp. 397–417# ascribed a sound-generating function to this organ complex...

  19. Microsurgical and Percutaneous Epididymal Sperm Aspiration for Sperm Collection from Live Mice.

    Science.gov (United States)

    Boersma, Auke; Olszanska, Olga; Walter, Ingrid; Rülicke, Thomas

    2015-09-01

    Spermatozoa for in vitro fertilization of mouse oocytes and other methods of assisted reproduction typically are collected from the cauda epididymis of euthanized male mice. As an alternative to this terminal protocol, we developed and examined 2 methods for collecting sperm from anesthetized male mice without decreasing subsequent fertility: microsurgical epididymal sperm aspiration and, as a refinement, percutaneous epididymal sperm aspiration. Collected sperm was evaluated in terms of motility, concentration and in vitro fertilization ability. After recovery, both treated and untreated control male mice underwent in vivo fertility testing and subsequent histologic analysis of the treated male reproductive tract (epididymis and testis). In vitro fertilization using sperm recovered by the 2 collection methods was successfully achieved in all cases. The in vivo fertility test and the histologic analysis revealed no impairment of fertility and no permanent histologic alteration in the treated mice. Therefore, we recommend both techniques as simple and effective methods for recovering high-quality epididymal mouse sperm without having to euthanize fertile male mice.

  20. Calibration of redox potential in sperm wash media and evaluation of oxidation-reduction potential values in various assisted reproductive technology culture media using MiOXSYS system.

    Science.gov (United States)

    Panner Selvam, M K; Henkel, R; Sharma, R; Agarwal, A

    2018-03-01

    Oxidation-reduction potential describes the balance between the oxidants and antioxidants in fluids including semen. Various artificial culture media are used in andrology and IVF laboratories for sperm preparation and to support the development of fertilized oocytes under in vitro conditions. The composition and conditions of these media are vital for optimal functioning of the gametes. Currently, there are no data on the status of redox potential of sperm processing and assisted reproduction media. The purpose of this study was to compare the oxidation-reduction potential values of the different media and to calibrate the oxidation-reduction potential values of the sperm wash medium using oxidative stress inducer cumene hydroperoxide and antioxidant ascorbic acid. Redox potential was measured in 10 different media ranging from sperm wash media, freezing media and assisted reproductive technology one-step medium to sequential media. Oxidation-reduction potential values of the sequential culture medium and one-step culture medium were lower and significantly different (p media. Calibration of the sperm wash media using the oxidant cumene hydroperoxide and antioxidant ascorbic acid demonstrated that oxidation-reduction potential and the concentration of oxidant or antioxidant are logarithmically dependent. This study highlights the importance of calibrating the oxidation-reduction potential levels of the sperm wash media in order to utilize it as a reference value to identify the physiological range of oxidation-reduction potential that does not have any adverse effect on normal physiological sperm function. © 2017 American Society of Andrology and European Academy of Andrology.

  1. Pentoxifylline effects on capacitation and fertility of stallion epididymal sperm.

    Science.gov (United States)

    Guasti, P N; Monteiro, G A; Maziero, R R D; Carmo, M T; Dell'Aqua, J A; Crespilho, A M; Rifai, E A; Papa, F O

    2017-04-01

    The aims of this study were to determinate whether pentoxifylline (PTX) increases the motion parameters of fresh and frozen-thawed equine epididymal spermatozoa, to evaluate the tyrosine phosphorylation of frozen-thawed epididymal sperm in the presence of PTX and to determine whether the PTX-treatment of stallion epididymal sperm prior to freezing improves the fertility response of mares to a reduced number of spermatozoa per insemination dose. Fifty epididymis were flushed with a skim milk based extender with or without PTX. The pre-treatment with PTX enhanced the sperm motility after being harvested (Psperm kinematics between control and treated samples (P>0.05). Plasma membrane integrity did not differ between control and PTX group after recovery and after thawing (P>0.05), as observed in tyrosine phosphorylation, which the PTX treatment did not alter the percentage of tail-associated immunofluorescence of cryopreserved epididymal sperm (P>0.05). For the fertility trial, different insemination groups were tested: 800×10 6 epididymal sperm (C800); 100×10 6 epididymal sperm (C100); 100×10 6 epididymal sperm recovered in an extender containing PTX (PTX100). The conception rates for C800; C100 and PTX100 were 68.7% (11/16); 31.5% (5/16) and 50% (8/16), respectively. The conception rate did not differ among groups (P>0.05), however, a low number of animals was used in this study. A trend toward significance (P=0.07) was observed between C800 and C100 groups. In conclusion, PTX has no deleterious effect on sperm motility, viability and capacitation of cryopreserved stallion epididymal sperm. The conventional artificial insemination with 100×10 6 sperm recovered with PTX ensures acceptable conception rates and maximize the limited number of doses of cryopreserved stallion epididymal sperm. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. The cryoprotective effects of soybean lecithin on boar spermatozoa ...

    African Journals Online (AJOL)

    Semen parameters including sperm motion characteristics (the percentage of total motile sperm and motility), plasma membrane integrity and acrosome integrity were assessed with a computer-aided semen analysis (CASA) system, hypoosmotic swelling test (HOST) and fluorescein isothiocyanateconjugated

  3. First Production of Larvae Using Cryopreserved Sperm: Effects of Preservation Temperature and Cryopreservation on European Eel Sperm Fertilization Capacity.

    Science.gov (United States)

    Asturiano, J F; Sørensen, S R; Pérez, L; Lauesen, P; Tomkiewicz, J

    2016-08-01

    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. Fertilization of two egg batches was attempted. Diluted sperm caused a similar percentage of fertilized eggs and a similar number of embryos and larvae, independently of storage temperature (4 or 20°C). The cryopreserved sperm resulted in a lower percentage of fertilized eggs, but embryos developed and a few larvae ('cryolarvae') were obtained 55 h after fertilization in one of the two egg batches. This result evidences that the tested cryopreservation protocol is applicable for eel reproduction management, although improvements will be required to enhance fertilization success. © 2016 Blackwell Verlag GmbH.

  4. Association of GH Gene Polymorphism with Semen Parameters of Boars

    Directory of Open Access Journals (Sweden)

    M. Kmieć

    2007-01-01

    Full Text Available Relations between polymorphism of the Growth Hormone gene and semen characters were analyzed. The DNA for the purpose of examination was isolated from the peripheral blood of 173 boars. In the boar herd under study the frequency of allele occurrence for the GH/MspI was as follows: allele GHA - 0.79 and allele GHB - 0.21. As far as the GH/HaeII polymorphism is concerned, the relevant frequency was as follows: allele GHA - 0.53 and allele GHB - 0.47, respectively. The relationship between the GH genotypes and semen characteristic traits were analyzed. The study showed that boars with GHBGHB genotype of both polymorphous loci of the GH gene produced ejaculates of larger volume, higher percentage, number of normozosperms in the ejaculate and number of insemination as compared to GHA GHA and GHAGHB boars. Our current findings suggested that polymorphism of the GH/MspI and GH/HaeII might have potential effect for reproductive performance traits of boars.

  5. Analysis of epididymal sperm maturation by MALDI profiling and top-down mass spectrometry.

    Science.gov (United States)

    Labas, Valérie; Spina, Lucie; Belleannee, Clémence; Teixeira-Gomes, Ana-Paula; Gargaros, Audrey; Dacheux, Françoise; Dacheux, Jean-Louis

    2015-01-15

    The fertilization ability of male gametes is achieved after their transit through the epididymis where important post-gonadal differentiation occurs in different cellular compartments. Most of these maturational modifications occur at the protein level. The epididymal sperm maturation process was investigated using the ICM-MS (Intact Cell MALDI-TOF MS) approach on boar spermatozoa isolated from four different epididymal regions (immature to mature stage). Differential and quantitative MALDI-TOF profiling for whole cells or sub-cellular fractions was combined with targeted top-down MS in order to identify endogenous biomolecules. Using this approach, 172m/z peaks ranging between 2 and 20kDa were found to be modified during maturation of sperm. Using top-down MS, 62m/z were identified corresponding to peptidoforms/proteoforms with post-translational modifications (MS data are available via ProteomeXchange with identifier PXD001303). Many of the endogenous peptides were characterized as N-, C-terminal sequences or internal fragments of proteins presenting specific cleavages, suggesting the presence of sequential protease activities in the spermatozoa. This is the first time that such proteolytic activities could be evidenced for various sperm proteins through quantification of their proteolytic products. ICM-MS/top-down MS thus proved to be a valid approach for peptidome/degradome studies and provided new contributions to understanding of the maturation process of the male gamete involved in the development of male fertility. This peptidomic study (i) characterized the peptidome of epididymal spermatozoa from boar (Sus scrofa); (ii) established characteristic molecular phenotypes distinguishing degrees of maturation of spermatozoa during epididymal transit, and (iii) revealed that protease activities were at the origin of numerous peptides from known and unknown proteins involved in sperm maturation and/or fertility processes. Copyright © 2014 Elsevier B.V. All

  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. Valosine vontaining protein is a substrate of c-AMP-activated boar sperm tyrosine kinase

    Czech Academy of Sciences Publication Activity Database

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

    2002-01-01

    Roč. 2002, č. 63 (2002), s. 366-375 ISSN 1040-452X R&D Projects: GA ČR GA204/02/1373; GA ČR GA303/00/1651; GA MZd NJ5851 Institutional research plan: CEZ:AV0Z5052915 Keywords : gamete biology * spermatozoa, capacitation * phosphorylation Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.322, year: 2002

  8. Effect of freezing and thawing rates on the post-thaw viability of boar spermatozoa frozen in FlatPacks and Maxi-straws.

    Science.gov (United States)

    Eriksson, B M; Rodriguez-Martinez, H

    2000-11-01

    The effects of different freezing and thawing rates on the post-thaw motility and membrane integrity of boar spermatozoa, processed as split samples in Maxi-straws or flat PET-plastic packages (FlatPack) were studied. A programmable freezing device was used to obtain freezing rates of either 20, 50 or 80 degrees C/min. Thawing of the samples was performed in a bath of circulating water; for 40s at 50 degrees C or 27s at 70 degrees C for Maxi-straws and 23s at 35 degrees C, 13s at 50 degrees C or 8s at 70 degrees C for the FlatPacks. Sperm motility was assessed both visually and with a computer assisted semen analysis (CASA) apparatus, while plasma membrane integrity was assessed using the fluorescent probes Calcein AM and ethidium homodimer-1. Temperature changes during freezing and thawing were monitored in both forms of packaging. Values for motile spermatozoa, sperm velocity and lateral head displacement variables were significantly (pstraws, with superior results at higher thawing rates. Freezing at 50 degrees C/min yielded better motility than 20 or 80 degrees C/min, although the effect was rather small. Neither freezing rate nor thawing rate had any effect on membrane integrity (p>0.05). A significant boar effect was seen for several parameters. The most striking difference in temperature courses between containers was a 4-5-fold lowering of the thawing rate, between -20 and 0 degrees C, in the center of the Maxi-straw, compared with the FlatPack. This is apparently due to the insulating effect of the thawed water in the periphery of the Maxi-straw. The improvement in sperm motility seen when using the FlatPack appears to be related to the rapid thawing throughout the sample, which decreases the risk of cell damage due to recrystallization during thawing. Since sperm motility patterns have been reported to be correlated with fertility both in vitro and in vivo it is speculated that the use of the FlatPack might improve the results when using frozen

  9. Effects of Panax ginseng, zearalenol, and estradiol on sperm function.

    Science.gov (United States)

    Gray, Sandra L; Lackey, Brett R; Boone, William R

    2016-07-01

    Estrogen signaling pathways are modulated by exogenous factors. Panax ginseng exerts multiple activities in biological systems and is classified as an adaptogen. Zearalenol is a potent mycoestrogen that may be present in herbs and crops arising from contamination or endophytic association. The goal of this study was to investigate the impact of P. ginseng, zearalenol and estradiol in tests on spermatozoal function. The affinity of these compounds for estrogen receptor (ER)-alpha and beta (ERα and ERβ)-was assessed in receptor binding assays. Functional tests on boar spermatozoa motility, movement and kinematic parameters were conducted using a computer-assisted sperm analyzer. Tests for capacitation, acrosome reaction (AR), and chromatin decondensation in spermatozoa were performed using microscopic analysis. Zearalenol-but not estradiol (E2)- or ginseng-treated spermatozoa-decreased the percentage of overall, progressive, and rapid motile cells. Zearalenol also decreased spontaneous AR and increased chromatin decondensation. Ginseng decreased chromatin decondensation in response to calcium ionophore and decreased AR in response to progesterone (P4) and ionophore. Zearalenol has adverse effects on sperm motility and function by targeting multiple signaling cascades, including P4, E2, and calcium pathways. Ginseng protects against chromatin damage and thus may be beneficial to reproductive fitness.

  10. Investigations on the contamination of Saxonian wild boars with radiocaesium

    International Nuclear Information System (INIS)

    Heinrich, T.; Abraham, A.; Preusse, W.; Pianski, J.; Alisch-Mark, M.; Lange, S.

    2016-01-01

    As a result of the Chernobyl fallout some parts of the free state of Saxony were contaminated with radioactive caesium. Based on published maps of the soil contamination and on additional investigations some regions of elevated contamination could be localized. Parallel to soil investigations a game monitoring to wild boars and roe deer was performed. For both types of game typical seasonal variations of contamination were found. In Saxony only the contamination of wild boars is important. In the south of the Vogtland a region was found, where in all seasons the recommended high value of 600 Bq/kg was exceeded in game. In this region the investigation on radiocaesium is now obligatory for wild boars. The hunter can combine this analysis with the analysis on trichina. After three years measurements the region for obligatory analysis was adapted and expanded to neighbouring counties.

  11. 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.

  12. First TBEV serological screening in Flemish wild boar

    Science.gov (United States)

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

    2016-01-01

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

  13. Effect of ACTH and CRH on Plasma Levels of Cortisol and Prostaglandin F2α Metabolite in Cycling Gilts and Castrated Boars

    Directory of Open Access Journals (Sweden)

    Kindahl H

    2005-12-01

    Full Text Available The present study was designed to evaluate the effects of synthetic ACTH (1–24, tetracosactid and porcine CRH on the plasma levels of cortisol and PGF2α metabolite in cycling gilts (n = 3 and castrated boars (n = 3. The experiments were designed as crossover studies for each gender separately. Each animal received, during three consecutive days; 1 ACTH (Synacthen® Depot at a dose of 10 μg/kg body weight in 5 ml physiological saline, 2 porcine CRH at a dose 0.6 μg/kg body weight in 5 ml physiological saline or 3 physiological saline (5 ml. The test substances were administered via an indwelling jugular cannula in randomized order according to a Latin square. The administration of ACTH to cycling gilts resulted in concomitant elevations of cortisol and PGF2α metabolite with peak levels reached at 70.0 ± 10.0 and 33.3 ± 6.7 min, respectively. Similarly, the administration of ACTH to castrated boars resulted in concomitant elevation of cortisol and PGF2α metabolite with peak levels reached at 60.0 ± 0.0 and 20.0 ± 0.0 min, respectively. Cortisol peaked at 20 min after administration of CRH in both cycling gilts and castrated boars with maximum levels of 149.3 ± 16.5 nmol/1 and 138.3 ± 10.1 nmol/1, respectively. It can be concluded that administration of synthetic ACTH (tetracosactid to pigs caused a concomitant elevation of cortisol and PGF2α metabolite levels in both cycling gilts as well as castrated boars. The administration of CRH to pigs resulted in an elevation of cortisol levels in both cycling gilts and castrated boars. Conversely, PGF2α metabolite levels were not influenced by the administration of CRH either in cycling gilts or in castrated boars.

  14. Effect of ACTH and CRH on Plasma Levels of Cortisol and Prostaglandin F2α Metabolite in Cycling Gilts and Castrated Boars

    Directory of Open Access Journals (Sweden)

    Madej A

    2006-12-01

    Full Text Available The present study was designed to evaluate the effects of synthetic ACTH (1–24, tetracosactid and porcine CRH on the plasma levels of cortisol and PGF2α metabolite in cycling gilts (n = 3 and castrated boars (n = 3. The experiments were designed as crossover studies for each gender separately. Each animal received, during three consecutive days; 1 ACTH (Synacthen® Depot at a dose of 10 μg/kg body weight in 5 ml physiological saline, 2 porcine CRH at a dose 0.6 μg/kg body weight in 5 ml physiological saline or 3 physiological saline (5 ml. The test substances were administered via an indwelling jugular cannula in randomized order according to a Latin square. The administration of ACTH to cycling gilts resulted in concomitant elevations of cortisol and PGF2α metabolite with peak levels reached at 70.0 ± 10.0 and 33.3 ± 6.7 min, respectively. Similarly, the administration of ACTH to castrated boars resulted in concomitant elevation of cortisol and PGF2α metabolite with peak levels reached at 60.0 ± 0.0 and 20.0 ± 0.0 min, respectively. Cortisol peaked at 20 min after administration of CRH in both cycling gilts and castrated boars with maximum levels of 149.3 ± 16.5 nmol/1 and 138.3 ± 10.1 nmol/1, respectively. It can be concluded that administration of synthetic ACTH (tetracosactid to pigs caused a concomitant elevation of cortisol and PGF2α metabolite levels in both cycling gilts as well as castrated boars. The administration of CRH to pigs resulted in an elevation of cortisol levels in both cycling gilts and castrated boars. Conversely, PGF2α metabolite levels were not influenced by the administration of CRH either in cycling gilts or in castrated boars.

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

    OpenAIRE

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

    2015-01-01

    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 ca...

  16. Threshold detection of boar taint chemicals using parasitic wasps.

    Science.gov (United States)

    Olson, Dawn; Wäckers, Felix; Haugen, John-Erik

    2012-10-01

    Surgical castration has been long used to prevent consumers from experiencing taint in meat from male pigs, which is a large problem in the pig husbandry industry. Due to obvious animal welfare issues, the EU now wants an alternative for castration, suggesting an urgent need for novel methods of boar taint detection. As boar taint is only a problem when taint chemicals exceed a well-defined threshold, detection methods should be concentration-specific. The wasp, Microplitis croceipes' ability to learn and respond to particular concentrations of the boar taint compounds, skatole, androstenone, and indole was tested. Also tested was the wasps' ability to discriminate between known concentrations of indole, skatole, and androstenone in real boar fat samples at room temperature. Wasps were trained using associative learning by providing food-deprived wasps with sucrose-water in the presence of specific odor concentrations. Trained wasps' responses were tested to a range of concentrations of 3 compounds. Wasps showed unidirectional generalization of learned concentration responses, whereby the direction of concentration generalization was shown to be chemical-dependent. Through both positive (sucrose) and negative feeding experiences (water only) with varying compound concentrations, the wasps can also be conditioned to respond to concentrations exceeding a defined threshold, and they were successful in reporting low, medium, and high concentrations of indole, skatole, and androstenone in boar fat at room temperature. The need for threshold detection rather than simple detection of absence/presence applies to many food quality issues, including the detection of spoilage or pest damage in crops or stored foods. An inexpensive and reliable means of detecting boar tainted pork at slaughter to avoid tainted meat on the market and dissatisfied consumers. Journal of Food Science © 2012 Institute of Food Technologists® No claim to original US government works.

  17. Antibody Prevalence to Influenza Type A in Wild Boar of Northern Ukraine.

    Science.gov (United States)

    Kovalenko, Ganna; Molozhanova, Alona; Halka, Ihor; Nychyk, Serhiy

    2017-12-01

    A preliminary serological survey was carried out to assess the likelihood of influenza A (IA) infection in wild boar and begin to characterize the role of wild boar in the epidemiology of the IA virus (IAV). Sera collected from 120 wild boar that were hunted in 2014 were tested. To detect antibodies to IA, a blocking the enzyme-linked immunosorbent assay (ELISA) was used. Thirty boar were collected from each of four oblasts in the north central and northwestern regions of Ukraine. Antibodies against IAV were detected in 27 samples (22.5%; 95% confidence interval 16.0-30.8) and in at least some of the wild boar from all of the four oblasts. This preliminary survey of IA antibodies in wild boar populations of northern Ukraine indicates a substantial frequency of exposure to IAV throughout the region. Infection of wild boar populations could provide an alternative or additional route for spillover from wild populations to domestic animals and humans.

  18. Genetic variability among the wild boars (Sus scrofa scrofa), crossbred animals and pigs using microsatellite markers (STRs)

    OpenAIRE

    Silva, Paula Viana Correa da; Lui, Jeffrey Frederico; Band, Guilherme de Oliveira; Regitano, Luciana Correia de Almeida; Grossi, Selma de Fátima; Sollero, Bruna Pena; Nunes, Cleujosí da Silva

    2011-01-01

    The aim of this work was to study the genetic variability among the wild boars, crossbred animals and pigs using microsatellite markers. Five genetic groups were studied. The fragments of three microsatellites developed for Sus scrofa domestica - IGF1, ACTG2 and TNFB - were amplified through PCR technique to evaluate the expected intra populacion variability (He) and observed (Ho) heterozygosity, and endogamy coefficient (F IS ) within each population and inter population variability F IS , t...

  19. 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.

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

    Directory of Open Access Journals (Sweden)

    D.J. Ackerman

    1996-08-01

    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.

  1. Clinical and structural features of sperm head vacuoles in men included in the in vitro fertilization programme.

    Science.gov (United States)

    Fekonja, Nina; Štrus, Jasna; Tušek Žnidarič, Magda; Knez, Katja; Vrtacnik Bokal, Eda; Verdenik, Ivan; Virant-Klun, Irma

    2014-01-01

    The human sperm head vacuoles and their role in male infertility are still poorly understood. The aim of this study was to identify the clinical and ultrastructural features of human sperm head vacuoles in men included in the in vitro fertilization programme: men with normal (normozoospermia) and impaired sperm morphology (teratozoospermia). The sperm samples were observed under 6000-time magnification using motile sperm organelle morphology examination (MSOME). The proportion of sperm with head vacuoles was evaluated and related to the outcome of in vitro fertilization. The sperm of men with impaired sperm morphology was characterized by a higher proportion of sperm head vacuoles. The sperm head vacuoles were related to impaired semen quality (sperm concentration, motility, and morphology) but were not influenced by male factors (semen volume, height, age, weight, or body mass index). Moreover, sperm head vacuoles were related to impaired fertilization rate merely after classical in vitro fertilization (IVF), while there was no relation to pregnancy. In a subgroup of men, the sperm was fixed and observed by transmission electron microscopy (TEM). The ultrastructural study revealed that sperm head vacuoles are large nuclear indentations of various sizes and positions, packed with membranous material organized in membrane whorls (MW).

  2. Genomic selection in a pig population including information from slaughtered full sibs of boars within a sib-testing program.

    Science.gov (United States)

    Samorè, A B; Buttazzoni, L; Gallo, M; Russo, V; Fontanesi, L

    2015-05-01

    introduction of genomic selection. None of the considered genomic models provided improvements in prediction ability of pigs with no recorded phenotype. However, a few advantages were found for traits with low heritability (h 2=0.10). These heritability levels are characteristic for meat quality traits recorded after slaughtering or for reproduction or health traits, typically recorded on field and not in performance stations. Other scenarios of data recording and genotyping should be evaluated before considering the implementation of genomic selection in a pig-selection scheme based on sib testing of boars.

  3. Axicon-based annular laser trap for studies on sperm activity

    Science.gov (United States)

    Shao, Bing; Vinson, Jaclyn M.; Botvinick, Elliot L.; Esener, Sadik C.; Berns, Michael W.

    2005-08-01

    As a powerful and noninvasive tool, laser trapping has been widely applied for the confinement and physiological study of biological cells and organelles. Researchers have used the single spot laser trap to hold individual sperm and quantitatively evaluated the motile force generated by a sperm. Early studies revealed the relationship between sperm motility and swimming behavior and helped the investigations in medical aspects of sperm activity. As sperm chemotaxis draws more and more interest in fertilization research, the studies on sperm-egg communication may help to explain male or female infertility and provide exciting new approaches to contraception. However, single spot laser trapping can only be used to investigate an individual target, which has limits in efficiency and throughput. To study the chemotactic response of sperm to eggs and to characterize sperm motility, an annular laser trap with a diameter of several hundred microns is designed, simulated with ray tracing tool, and implemented. An axicon transforms the wavefront such that the laser beam is incident on the microscope objective from all directions while filling the back aperture completely for high efficiency trapping. A trapping experiment with microspheres is carried out to evaluate the system performance. The power requirement for annular sperm trapping is determined experimentally and compared with theoretical calculations. With a chemo-attractant located in the center and sperm approaching from all directions, the annular laser trapping could serve as a speed bump for sperm so that motility characterization and fertility sorting can be performed efficiently.

  4. Effects of coffee husk as floor covering on the behavior of boars

    Directory of Open Access Journals (Sweden)

    Mariele Cristina Teles

    Full Text Available ABSTRACT The objective was to evaluate the influence of coffee husks as floor covering on the aspects of animal welfare such as behavioral characteristics, body surface temperature, and salivary cortisol levels of stabled boars. Sixteen boars were housed in individual stalls; eight were maintained in a conventional system with a concrete floor and eight were maintained on a concrete floor lined with coffee husks. The experimental period was 60 days. All animals were filmed two days prior to the start of the experiment, on both the 7th and 60th days after exposure to coffee husks, and finally two days after the removal of the material. During this period, the number of times that the animals ate, drank, stood, sat, lay down, and dug was recorded. Furthermore, both body surface temperature and salivary cortisol levels were measured at the beginning and end of the experiment. The use of coffee husks did not influence body surface temperature. Salivary cortisol levels increased during the experimental period only in the animals maintained on coffee husks. In the morning, the coffee husks decreased the number of times that the animals sat and increased the number of times that they lay down. In the afternoon, the use of coffee husks decreased the number of times that the animals stood, sat, or dug and increased the number of times that the animals lay down. Although coffee husks do not change the behavior of the animals in an expressive way, they should not be used as floor covering for boars.

  5. Roles of Intracellular Cyclic AMP Signal Transduction in the Capacitation and Subsequent Hyperactivation of Mouse and Boar Spermatozoa

    Science.gov (United States)

    HARAYAMA, Hiroshi

    2013-01-01

    It is not until accomplishment of a variety of molecular changes during the transit through the female reproductive tract that mammalian spermatozoa are capable of exhibiting highly activated motility with asymmetric whiplash beating of the flagella (hyperactivation) and undergoing acrosomal exocytosis in the head (acrosome reaction). These molecular changes of the spermatozoa are collectively termed capacitation and promoted by bicarbonate, calcium and cholesterol acceptors. Such capacitation-promoting factors can stimulate intracellular cyclic AMP (cAMP) signal transduction in the spermatozoa. Meanwhile, hyperactivation and the acrosome reaction are essential to sperm fertilization with oocytes and are apparently triggered by a sufficient increase of intracellular Ca2+ in the sperm flagellum and head, respectively. Thus, it is necessary to investigate the relationship between cAMP signal transduction and calcium signaling cascades in the spermatozoa for the purpose of understanding the molecular basis of capacitation. In this review, I cover updated insights regarding intracellular cAMP signal transduction, the acrosome reaction and flagellar motility in mammalian spermatozoa and then account for possible roles of intracellular cAMP signal transduction in the capacitation and subsequent hyperactivation of mouse and boar spermatozoa. PMID:24162806

  6. Effect of climate on the seminal characteristics of boars in a region of humid tropical forest

    International Nuclear Information System (INIS)

    Henao Restrepo, Guillermo; Trujillo Aramburo, Luis Emilio; Buritica Henao, Maria Elizabet; Sierra Perez, Carlos Ignacio; Correa Londono, Guillermo; Gonzalez Boto, Oscar Domingo

    2004-01-01

    In a region of humid tropical forest, ten boars of from 12 to 24 months of age were selected to evaluate the effect of climatic variables measured on the day of semen collection and for each of preceding 45 days. On seminal characteristics, the variability of each characteristic was separated into an intra individual component and an interindividual component, using maximum likelihood estimators (PROC VARCOMP of SAS). In order to relate the seminal characteristics with the climatic variables, morphological abnormalities were grouped according to the affected spermatic region, into head. Midsection and main section abnormalities; the other characteristics were evaluated without any modification. Possible correlations between seminal characteristics and climatic variables were evaluated. In a total of 298 ejaculates collected weekly during a period of 30 weeks, except for total volume and morphological abnormalities. The seminal characteristics presented low or moderate intra and interindividual variation and were similar to those found in other latitudes, with a tendency to present greater seminal volumes and concentrations maximum temperature minimum temperature. Range among temperatures. Relative humidity and precipitation of the day of the semen collection and on each of the preceding 45 days had low effects on the seminal characteristics. It is possible that the boars in warm humid tropical areas develop a high level of adaptation that permits an adequate testicular thermoregulation that favors the spermatogenic function of the seminiferous tubules in a way that does not perceptibly affect production the seminal quality

  7. Comparative evidence for the evolution of sperm swimming speed by sperm competition and female sperm storage duration in passerine birds.

    Science.gov (United States)

    Kleven, Oddmund; Fossøy, Frode; Laskemoen, Terje; Robertson, Raleigh J; Rudolfsen, Geir; Lifjeld, Jan T

    2009-09-01

    Sperm swimming speed is an important determinant of male fertility and sperm competitiveness. Despite its fundamental biological importance, the underlying evolutionary processes affecting this male reproductive trait are poorly understood. Using a comparative approach in a phylogenetic framework, we tested the predictions that sperm swim faster with (1) increased risk of sperm competition, (2) shorter duration of female sperm storage, and (3) increased sperm length. We recorded sperm swimming speed in 42 North American and European free-living passerine bird species, representing 35 genera and 16 families. We found that sperm swimming speed was positively related to the frequency of extrapair paternity (a proxy for the risk of sperm competition) and negatively associated with clutch size (a proxy for the duration of female sperm storage). Sperm swimming speed was unrelated to sperm length, although sperm length also increased with the frequency of extrapair paternity. These results suggest that sperm swimming speed and sperm length are not closely associated traits and evolve independently in response to sperm competition in passerine birds. Our findings emphasize the significance of both sperm competition and female sperm storage duration as evolutionary forces driving sperm swimming speed.

  8. Diagnosis of tuberculosis in the wild boar (Sus scrofa: a comparison of methods applicable to hunter-harvested animals.

    Directory of Open Access Journals (Sweden)

    Nuno Santos

    Full Text Available BACKGROUND: To obtain robust epidemiological information regarding tuberculosis (TB in wildlife species, appropriate diagnostic methods need to be used. Wild boar (Sus scrofa recently emerged as a major maintenance host for TB in some European countries. Nevertheless, no data is available to evaluate TB post-mortem diagnostic methods in hunter-harvested wild boar. METHODOLOGY/PRINCIPAL FINDINGS: Six different diagnostic methods for TB were evaluated in parallel in 167 hunter-harvested wild boar. Compared to bacteriological culture, estimates of sensitivity of histopathology was 77.8%, gross pathology 72.2%, PCR for the MPB70 gene 66.7%, detection of acid-fast bacilli (AFB in tissue contact smears 55.6% and in histopathology slides 16.7% (estimated specificity was 96.7%, 100%, 100%, 94.4% and 100%, respectively. Combining gross pathology with stained smears in parallel increased estimated sensitivity to 94.4% (94.4% specificity. Four probable bacteriological culture false-negative animals were identified by Discriminant Function Analysis. Recalculating the parameters considering these animals as infected generated estimated values for sensitivity of bacteriology and histopathology of 81.8%, gross pathology 72.7%, PCR for the MPB70 gene 63.6%, detection of AFB in tissue contact smears 54.5% and in histopathology slides 13.6% (estimated specificity was 100% for gross pathology, PCR, bacteriology and detection of AFB in histopathology slides, 96.7% for histopathology and 94.4% for stained smears. CONCLUSIONS/SIGNIFICANCE: These results show that surveys for TB in wild boar based exclusively on gross pathology considerably underestimate prevalence, while combination of tests in parallel much improves sensitivity and negative predictive values. This finding should thus be considered when planning future surveys and game meat inspection schemes. Although bacteriological culture is the reference test for TB diagnosis, it can generate false

  9. Sperm cell proteomics.

    Science.gov (United States)

    Oliva, Rafael; de Mateo, Sara; Estanyol, Josep Maria

    2009-02-01

    The spermatozoon is an accessible cell which can be easily purified and therefore it is particularly well suited for proteomic analysis. It is also an extremely differentiated cell with very marked genetic, cellular, functional and chromatin changes as compared to other cells, and has profound implications for fertility, embryo development and heredity. The recent developments in MS have boosted the potential for identification and study of the sperm proteins. Catalogues of hundreds to thousands of spermatozoan proteins in human and in model species are becoming available setting up the basis for subsequent research, diagnostic applications and the development of specific treatments. The present article reviews the available scientific publications dealing with the composition and function of the sperm cell using an MS proteomic approach.

  10. Cryopreservation of Fish Sperm

    Science.gov (United States)

    Kurokura, Hisashi

    Present status of research activities in cryopreservation of fish gamete in aquaculture field was introduced. More than 59 fish species have been reported in the research histories and nearly half of them were studied during recent 10 years. This means that the research activities are increasing, though commercial profit have not obtained yet. Fish species of which sperm can successfully cryopreserved is still limited comparing to numerous species in telost. One of the major obstacle for improvement of the technique is existence of wide specie specific variance in the freezing tolerance of fish sperm. The varianc can possibly be explaind thorugh the informations obtained by the studies in comparative spermatology, which is recently activated field in fish biology.

  11. Effectiveness of the sperm quality analyzer (SQA-Vp) for porcine semen analysis.

    Science.gov (United States)

    Rodríguez, Alfonso López; Rijsselaere, Tom; Bijttebier, Jo; Vyt, Philip; Van Soom, Ann; Maes, Dominiek

    2011-03-15

    The Sperm Quality Analyzer (SQA-Vp) was evaluated for assessing concentration and motility of porcine semen. Both fresh and diluted semen from 50 different boars from a commercial artificial insemination (AI) centre were investigated. For the fresh ejaculate, the concentration obtained with SQA-Vp was compared with a photometer and a haemocytometer. For the diluted samples, the concentration and motility were compared with computer assisted semen analysis (CASA) and visual sperm analysis. The agreement between methods was studied with Bland-Altman plots and the repeatability with coefficient of variation (CV) as well as Bland-Altman plots. The sperm concentration (x10(6)/ml) obtained with SQA-Vp (379.3 ± 134.9) for fresh ejaculates agreed well with concentration by the photometer (447.2 ± 154.2; difference= -67.9 x 10(6)/ml; difference + 2SD = 55.3 x 10(6)/ml; difference - 2SD = -191.1 x 10(6)/ml) and with the haemocytometer (332.8 ± 141.11; d = 92.8; d + 2SD = 448.6; d - 2SD = -263). For diluted semen, the agreement between the concentration (x10(6)/ml) assessed with SQA-Vp (20.4 ± 4.3) was good with CASA (23.2 ± 5.8; d = -2.8; d + 2SD = 6.2; d - 2 SD = -11.8) but poor with the haemocytometer (18.8 ± 5.0; d = 1.6; d+ 2SD = 12.2; d - 2SD = -9). The % motile spermatozoa assessed by SQA-Vp (65.8 ± 10.0) in diluted semen agreed well with CASA (72.2 ± 13.7; d = -6.4; d+ 2SD = 20; d - 2SD = -32.8) and with visual assessment (64.1 ± 11.6; d = 1.7; d+ 2SD = 30.9; d - 2SD = -27.5). The SQA-Vp showed a good repeatability (CV; repeatability coefficient) for measuring the concentration of both fresh (3.9%; d = 10.7; d + 2SD = 30.9; d - 2SD = -9.5) and diluted semen (2.6%; d = 1.0; d + 2SD = 2.38; d - 2SD = -0.42) and for motility (3.2%; d = 0.9; d + 2SD = 8.5; d - 2SD = -6.7). The mean SQA-Vp values fell between the other methods' results for both fresh and diluted semen. Moreover the repeatability was acceptable. Therefore SQA-Vp can be used as a valid device for

  12. The effects of three extenders on refrigerated boar semen

    African Journals Online (AJOL)

    sofiateixeira

    2015-03-19

    Mar 19, 2015 ... Ambrogi, M., Ballester, J., Saravia, F., Caballero, I., Johannisson, A., Wallgren, M., Andersson, M. &. Rodriguez-Martinez, H., 2006. Effect of storage in short and long term commercial semen extenders on the motility, plasma membrane and chromatin integrity of boar spermatozoa. Int. J. Androl. 29,. 543-552.

  13. Parasitic infections in wild ruminants and wild boar

    Directory of Open Access Journals (Sweden)

    Ilić Tamara

    2011-01-01

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

  14. Gene expression during testis development in Duroc boars

    DEFF Research Database (Denmark)

    Lervik, Siri; Kristoffersen, Anja Bråthen; Conley, Lene

    2015-01-01

    Androstenone is a steroid pheromone occurring in the pubertal Leydig cells. Breeding against androstenone can decrease pheromone odour in swine meat but appears to cause unwanted side effects such as delayed onset of puberty. To study causality, global gene expression in developing boar testes...

  15. 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...

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

    OpenAIRE

    Gage, Matthew J G; Freckleton, Robert P

    2003-01-01

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

  17. Effects of Synthetic Serum Supplementation in Sperm Preparation Media on Sperm Capacitation and Function Test Results

    Directory of Open Access Journals (Sweden)

    Ying-Fu Shih

    2016-01-01

    Full Text Available Albumin supplementation of culture media induces sperm capacitation in assisted reproduction technique cycles. Synthetic serum supplementation is clinically used to replace albumin for preventing transmission of infectious agents. However, the effects of synthetic serum supplementation on sperm capacitation have rarely been investigated. Spermatozoa from 30 men with normal basic semen analysis results were collected, divided into five aliquots, and cultured in capacitating conditions in four combinations of two synthetic serum supplements, serum substitute supplement (SSS and serum protein substitute (SPS, and two fertilization media, Quinns Advantage™ Fertilization (QF and human tubular fluid (HTF media. Reactive oxygen species (ROS levels in spermatozoa were measured through chemiluminescence. Furthermore, acrosome reaction and western blotting for tyrosine phosphorylation were used to evaluate sperm capacitation. HTF+SSS had significantly higher ROS levels than QF+SPS did (11,725 ± 1,172 versus 6,278 ± 864 relative light units. In addition, the spermatozoa cultured in QF+SPS had lower motility, acrosome reaction rates, and tyrosine phosphorylation levels compared with those cultured in HTF+SSS. In conclusion, the effects of synthetic serum supplementation on sperm capacitation varied according to the combination of media. These differences may lead to variations in spermatozoon ROS levels, thus affecting sperm function test results.

  18. 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.

  19. Motile sperm organelle morphology examination (MSOME: intervariation study of normal sperm and sperm with large nuclear vacuoles

    Directory of Open Access Journals (Sweden)

    Baruffi Ricardo LR

    2010-06-01

    Full Text Available Abstract Background Although the motile sperm organelle morphology examination (MSOME was developed only as a selection criterion, its application as a method for classifying sperm morphology may represent an improvement in evaluation of semen quality, with potential clinical repercussions. The present study aimed to evaluate individual variations in the motile sperm organelle morphology examination (MSOME analysis after a time interval. Methods Two semen samples were obtained from 240 men from an unselected group of couples undergoing infertility investigation and treatment. Mean time interval between the two semen evaluations was 119 +/- 102 days. No clinical or surgical treatment was realized between the two observations. Spermatozoa were analyzed at greater than or equal to 8400× magnification by inverted microscope equipped with DIC/Nomarski differential interference contrast optics. At least 200 motile spermatozoa per semen sample were evaluated and percentages of normal spermatozoa and spermatozoa with large nuclear vacuoles (LNV/one or more vacuoles occupying >50% of the sperm nuclear area were determined. A spermatozoon was classified as morphologically normal when it exhibited a normal nucleus (smooth, symmetric and oval nucleus, width 3.28 +/- 0.20 μm, length 4.75 +/- 0.20 μm/absence of vacuoles occupying >4% of nuclear area as well as acrosome, post-acrosomal lamina, neck and tail, besides not presenting cytoplasm around the head. One examiner, blinded to subject identity, performed the entire study. Results Mean percentages of morphologically normal and LNV spermatozoa were identical in the two MSOME analyses (1.6 +/- 2.2% vs. 1.6 +/- 2.1% P = 0.83 and 25.2 +/- 19.2% vs. 26.1 +/- 19.0% P = 0.31, respectively. Regression analysis between the two samples revealed significant positive correlation for morphologically normal and for LNV spermatozoa (r = 0.57 95% CI:0.47-0.65 P P Conclusions The significant positive correlation and

  20. Sexing of dog sperm by fluorescence in situ hybridization.

    Science.gov (United States)

    Oi, Maya; Yamada, Keisuke; Hayakawa, Hiroyuki; Suzuki, Hiroshi

    2013-01-01

    Effective preselection of sex has been accomplished in several species of livestock and also in humans using the flow cytometric sperm sorting method. A guaranteed high sorting accuracy is a key prerequisite for the widespread use of sperm sexing. The standard validation method is flow cytometric remeasurement of the DNA content of the sexed sperm. Since this method relies on the same instrument that produced the original sperm separation, it is not truly independent. Therefore, to be able to specifically produce either male or female offspring in the dog, we developed a method of direct visualization of sex chromosomes in a single sperm using fluorescence in situ hybridization (FISH) as a validation method. Denaturation of canine spermatozoa by immersion in 1 M NaOH for 4 min yielded consistent hybridization results with over 97% hybridization efficiency and a good preservation of sperm morphology. There was no significant difference between the theoretical ratio (50:50) and the observed ratio of X- and Y-chromosome-bearing spermatozoa in any of the three dogs. In addition, the mean purities of flow-sorted sex chromosomes in spermatozoa of the three dogs were 90.8% for the X chromosome fraction and 89.6% for the Y chromosome fraction. This sorting was evaluated by using the dual color FISH protocol. Therefore, our results demonstrated that the FISH protocol worked reliably for both unsorted and sexed sperm samples.

  1. Autocrine regulation of human sperm motility by tachykinins

    Directory of Open Access Journals (Sweden)

    Pinto Francisco M

    2010-08-01

    Full Text Available Abstract Background We examined the presence and function of tachykinins and the tachykinin-degrading enzymes neprilysin (NEP and neprilysin-2 (NEP2 in human spermatozoa. Methods Freshly ejaculated semen was collected from forty-eight normozoospermic human donors. We analyzed the expression of substance P, neurokinin A, neurokinin B, hemokinin-1, NEP and NEP2 in sperm cells by reverse-transcriptase polymerase chain reaction (RT-PCR, western blot and immunocytochemistry assays and evaluated the effects of the neprilysin and neprilysin-2 inhibitor phosphoramidon on sperm motility in the absence and presence of tachykinin receptor-selective antagonists. Sperm motility was measured using WHO procedures or computer-assisted sperm analysis (CASA. Results The mRNAs of the genes that encode substance P/neurokinin A (TAC1, neurokinin B (TAC3, hemokinin-1 (TAC4, neprilysin (MME and neprilysin-2 (MMEL1 were expressed in human sperm. Immunocytochemistry studies revealed that tachykinin and neprilysin proteins were present in spermatozoa and show specific and differential distributions. Phosphoramidon increased sperm progressive motility and its effects were reduced in the presence of the tachykinin receptor antagonists SR140333 (NK1 receptor-selective and SR48968 (NK2 receptor-selective but unmodified in the presence of SR142801 (NK3 receptor-selective. Conclusion These data show that tachykinins are present in human spermatozoa and participate in the regulation of sperm motility. Tachykinin activity is regulated, at least in part, by neprilysins.

  2. Autocrine regulation of human sperm motility by tachykinins.

    Science.gov (United States)

    Pinto, Francisco M; Ravina, Cristina G; Subiran, Nerea; Cejudo-Román, Antonio; Fernández-Sánchez, Manuel; Irazusta, Jon; Garrido, Nicolas; Candenas, Luz

    2010-08-26

    We examined the presence and function of tachykinins and the tachykinin-degrading enzymes neprilysin (NEP) and neprilysin-2 (NEP2) in human spermatozoa. Freshly ejaculated semen was collected from forty-eight normozoospermic human donors. We analyzed the expression of substance P, neurokinin A, neurokinin B, hemokinin-1, NEP and NEP2 in sperm cells by reverse-transcriptase polymerase chain reaction (RT-PCR), western blot and immunocytochemistry assays and evaluated the effects of the neprilysin and neprilysin-2 inhibitor phosphoramidon on sperm motility in the absence and presence of tachykinin receptor-selective antagonists. Sperm motility was measured using WHO procedures or computer-assisted sperm analysis (CASA). The mRNAs of the genes that encode substance P/neurokinin A (TAC1), neurokinin B (TAC3), hemokinin-1 (TAC4), neprilysin (MME) and neprilysin-2 (MMEL1) were expressed in human sperm. Immunocytochemistry studies revealed that tachykinin and neprilysin proteins were present in spermatozoa and show specific and differential distributions. Phosphoramidon increased sperm progressive motility and its effects were reduced in the presence of the tachykinin receptor antagonists SR140333 (NK1 receptor-selective) and SR48968 (NK2 receptor-selective) but unmodified in the presence of SR142801 (NK3 receptor-selective). These data show that tachykinins are present in human spermatozoa and participate in the regulation of sperm motility. Tachykinin activity is regulated, at least in part, by neprilysins.

  3. Cytogenetic Analysis of Sperm Nucleous Components of Iranian Normal and Sub-Fertile Individuals Using Zona Free Hamster Oocytes

    OpenAIRE

    Mozdarani, H.; Mohseni.Meybodi, A.

    2004-01-01

    Purpose: The purpose of this study was to investigate whether infertility is affected by sperm chromatin and cytogenetic abnormalities. To this purpose, the frequency of sperm premature chromosome condensation (PCC) induction and numerical chromosome abnormalities in the sperm of normal and sub-fertile men were analyzed. PCC rate was studied for evaluating the role of sperm chromatin abnormalities in the process of nuclear decondensation.

  4. Sperm vacuoles cannot help to differentiate fertile men from infertile men with normal sperm parameter values.

    Science.gov (United States)

    Gatimel, N; Léandri, R D; Marino, L; Esquerre-Lamare, C; Parinaud, J

    2014-11-01

    Can the assessment of sperm vacuoles at high magnification contribute to the explanation of idiopathic infertility? The characteristics of sperm head vacuoles (number, area, position) are no different between fertile controls and patients with unexplained infertility. Until now, the assessment of sperm head vacuoles has been focused on a therapeutic goal in the intracytoplasmic morphologically selected sperm injection (IMSI) procedure, but it could be pertinent as a new diagnostic tool for the evaluation of male fertility. This diagnostic test study with blind assessment included a population of 50 fertile men and 51 men with idiopathic infertility. They were selected from September 2011 to May 2013. Fertile men were within couples who had a spontaneous pregnancy in the last 2 years. Infertile men were within couples who had unexplained infertility and were consulting in our centre. After analysis of conventional sperm parameters, we investigated the number, position and area of sperm head vacuoles at high magnification (×6000) with interference contrast using an image analysis software. We also carried out a nuclear status analysis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling assay (TUNEL), sperm chromatin structure assay (SCSA) and aniline blue staining. Concerning the vacuoles data, we did not find any significant difference between the two populations. We found no significant correlation between the vacuolar parameters (mean number of vacuoles, relative vacuole area and percentage of spermatozoa with large vacuoles) and either conventional semen parameters, male age or the data from the aniline blue staining, SCSA assay and TUNEL assay. Despite the fact all of the vacuole parameters values were identical in fertile and infertile men, we cannot totally exclude that a very small cause of unexplained infertilities could be related to an excess of sperm vacuoles. In line with its widely debated use as a therapeutic tool, sperm vacuole

  5. 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.

  6. Cytometric analysis of shape and DNA content in mammalian sperm

    International Nuclear Information System (INIS)

    Gledhill, B.L.

    1983-01-01

    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. Post-Natal Persistent Infection With Classical Swine Fever Virus in Wild Boar: A Strategy for Viral Maintenance?

    Science.gov (United States)

    Cabezón, O; Colom-Cadena, A; Muñoz-González, S; Pérez-Simó, M; Bohórquez, J A; Rosell, R; Marco, I; Domingo, M; Lavín, S; Ganges, L

    2017-04-01

    In this study, fifteen wild boar piglets were intranasally inoculated classical swine fever virus (CSFV) strain Catalonia 01. At 5 days post-inoculation, seven other animals within 48 h of birth were put in contact with them. Viral replication and innate and specific immune responses were evaluated. Of the inoculated animals, 46.67% remained post-natally persistently infected and were apparently healthy with neither humoral nor cellular immunological responses specific to CSFV and with high viral loads in their blood, organs and body secretions. Moreover, the present data extend the time period to 48 h after birth when a moderately virulent CSFV strain could lead to post-natal persistent infection given the generation of persistently infected wild boars in the contact group (33.33%). The innate immune response to the virus, as measured by type I IFN-α in serum, was mostly not impaired in the persistently infected wild boars. Interestingly, a decrease and lack of IFN-γ-producing cells against CSFV and PHA was observed. In endemic countries where wild swine species are increasing and low and moderate virulence CSFV strains are prevalent, the possible generation of this form of disease cannot be ruled out. © 2015 Blackwell Verlag GmbH.

  8. Immunization against recombinant GnRH-I alters ultrastructure of gonadotropin cell in an experimental boar model.

    Science.gov (United States)

    Fang, Fugui; Su, Shiping; Liu, Ya; Zhang, Yunhai; Pu, Yong; Zhao, Xijie; Li, Yunsheng; Cao, Hongguo; Wang, Juhua; Zhou, Jie; Zhang, Xiaorong

    2013-07-15

    Gonadotropin cell is the main responsible for the secretion of follicle stimulating hormone (FSH) and luteinizing hormone (LH), and immunocastration reduces the concentrations of serum FSH and LH. A few studies have reported the histological structure of gonadotropin cells obtained from immunocastration animals at the light microscopy level. However, the ultrastructure of gonadotropin cells remains largely unexplored. The aim of this study was to evaluate and to compare ultrastructure of gonadotropin cell in gonadally intact boars and immunologically castrated male animals. In this study, serum and adenohypophysis tissue were collected from nine gonadally intact boars and nine male pigs treated with recombinant gonadotropin releasing hormone I (GnRH-I). Anti-GnRH-I antibodies in serum and the ultrastructure of gonadotropin cell in adenohypophysis were determined by enzymelinked immunosorbent assay and electron microscopy, respectively. The results demonstrated that active immunization against recombinant GnRH-I increased serum GnRH-I antibody levels (Pimmunized animals. We conclude that immunization against recombinant GnRH-I induces severe atrophy of granules in gonadotropin cell of boars, possibly reflecting GnRH-I regulation of gonadotropin cell.

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

    African Journals Online (AJOL)

    SH

    significantly (P<0.05) higher in rabbits fed prebiotic and symbiotic diets than those fed diet 3 and the control. The daily sperm production and sperm ... The amount of good quality live spermatozoa produced by the testis and the ... production and reproductive performance of rabbits. Effects of nutrition on reproduction.

  10. Sperm ultrastructure, morphometry, and abnormal morphology in American black bears (Ursus americanus).

    Science.gov (United States)

    Brito, L F C; Sertich, P L; Stull, G B; Rives, W; Knobbe, M

    2010-11-01

    The objective of this study was to describe sperm ultrastructure, morphometry, and abnormal morphology in American black bears. Electroejaculation was successful in 53.8% (7/13) of the attempts, but urine contamination was common. Epididymal sperm samples were also obtained from five bears. Sperm had a paddle-like head shape and the ultrastructure was similar to that of most other mammals. The most striking particularity of black bear sperm ultrastructure was a tightening of the nucleus in the equatorial region. Although the differences were not significant in all bears, the overall decrease in sperm nucleus dimensions during transport from the caput epididymis to the cauda suggested increasing compaction of the nucleus during maturation. For ejaculated sperm, nucleus length, width, and base width were 4.9, 3.7, and 1.8 μm, respectively, whereas sperm head length, width, and base width were 6.6, 4.8, and 2.3 μm, and midpiece, tail (including midpiece), and total sperm lengths were 9.8, 68.8, and 75.3 μm. Evaluation of sperm cytoplasmic droplets in the epididymis revealed that proximal droplets start migrating toward a distal position in the caput epididymis and that the process was mostly completed by the time sperm reached the cauda epididymis. The proportion of morphologically normal sperm in the ejaculate was 35.6%; the most prevalent sperm defects were distal cytoplasmic droplets and bent/coiled tails. The morphology of abnormal sperm and the underlying ultrastructural defects were similar to that in other large domestic animals thus suggesting similar underlying pathogenesis of specific sperm defects and similar effects on fertility. Copyright © 2010 Elsevier Inc. All rights reserved.

  11. Unraveling the Sperm Bauplan: Relationships Between Sperm Head Morphology and Sperm Function in Rodents.

    Science.gov (United States)

    Varea-Sánchez, María; Tourmente, Maximiliano; Bastir, Markus; Roldan, Eduardo R S

    2016-07-01

    Rodents have spermatozoa with features not seen in other species. Sperm heads in many rodent species bear one or more apical extensions known as "hooks." The process by which hooks have evolved, together with their adaptive significance, are still controversial issues. In order to improve our understanding of the biological meaning of these sperm head adaptations, we analyzed hook curvature angles, hook length, and overall hook shape in muroid rodents by using geometric morphometrics. We also searched for relationships between hook design and measurements of intermale competition to assess whether postcopulatory sexual selection was an important selective force driving changes in this sperm structure. Finally, we sought possible links between aspects of sperm hook design and sperm velocity as a measure of sperm performance. Results showed that one hook curvature angle is under strong selective pressure. Similarly, hook length appears to be strongly selected by sexual selection, with this selective force also exhibiting a stabilizing role reducing intermale variation in this trait. The adaptive significance of changes in hook structure was supported by the finding that there are strong and significant covariations between hook dimensions and shape and between hook design and sperm swimming velocity. Overall, this study strongly suggests that postcopulatory sexual selection has an important effect on the design of the sperm head that, in turn, is important for enhancing sperm velocity, a function crucial to reaching the vicinity of the female gamete and winning fertilizations under competitive situations. © 2016 by the Society for the Study of Reproduction, Inc.

  12. Hyaluronic acid binding ability of human sperm reflects cellular maturity and fertilizing potential: selection of sperm for intracytoplasmic sperm injection.

    Science.gov (United States)

    Huszar, Gabor; Ozkavukcu, Sinan; Jakab, Attila; Celik-Ozenci, Ciler; Sati, G Leyla; Cayli, Sevil

    2006-06-01

    The current concepts of sperm biochemical markers and the central role of the HspA2 chaperone protein, a measure of sperm cellular maturity and fertilizing potential, are reviewed. Because HspA2 is a component of the synaptonemal complex, low HspA2 levels and increased frequency of chromosomal aneuploidies are related in diminished maturity sperm. We also suggest a relationship between HspA2 expression in elongating spermatids and events of late spermiogenesis, such as cytoplasmic extrusion and plasma membrane remodeling that aid the formation of the zona pellucida binding and hyaluronic acid binding sites. The presence of hyaluronic acid receptor on the plasma membrane of mature sperm, coupled with hyaluronic acid coated glass or plastic surfaces, facilitates testing of sperm function and selection of single mature sperm for intracytoplasmic sperm injection. The frequencies of sperm with chromosomal disomy are reduced approximately fourfold to fivefold in hyaluronic acid selected sperm compared with semen sperm, comparable to the increase in such abnormalities in intracytoplasmic sperm injection offspring. Hyaluronic acid binding also excludes immature sperm with cytoplasmic extrusion, persistent histones, and DNA chain breaks. Hyaluronic acid mediated sperm selection is a novel technique that is comparable to sperm zona pellucida binding. Hyaluronic acid selected sperm will also alleviate the risks related to intracytoplasmic sperm injection fertilization with sperm of diminished maturity that currently cause worldwide concern.

  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 culture when they were in direct contact with epithelial cells. Conversely, no upregulation of HSP transcription was observed when spermatozoa did not directly bind to OECs. Spermatozoa also induced a significant upregulation (P 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. © 2014 Wiley Periodicals, Inc.

  14. Human sperm immobilizing activity of aminophenyl arsenic acid and its N-substituted quinazoline, pyrimidine, and purine derivatives: protective effect of glutathione.

    Science.gov (United States)

    Uckun, Fatih M; Liu, Xing Ping; D'Cruz, Osmond J

    2002-01-01

    We examined the potential toxicity of pentavalent organic arsenicals for human sperm. We used computer-assisted sperm analysis to examine the effects of three aminophenyl arsenicals and their nine N-substituted quinazoline, pyrimidine, and purine derivatives on human sperm motility and kinematics in human semen and medium. Among the arsenicals examined, (aminophenylazo)-phenyl arsonic acid and its N-substituted pyrimidine derivative PHI-370 (2-methylthio-4-[(4'-aminophenylazo)-phenylarsonic acid] pyrimidine) exhibited rapid sperm immobilizing activity in medium with EC(50) values of 77 and 82 microM, respectively, and t(1/2) of 7 Debyes). Sperm immobilizing activity of these arsenicals was completely abrogated in the presence of seminal plasma. Furthermore, coincubation of motile sperm with PHI-370 in the presence of reduced glutathione (GSH) resulted in dose-dependent protection of sperm motility and sperm motion parameters. Coincubation of the arsenical with GSH at a molar ratio of 1:20 resulted in 95% retention of sperm progressive motility. The mean values of the other sperm movement characteristics also showed > 90% protection. These observations suggest that the rapid sperm immobilizing activity of these pentavalent arsenicals may be as a result of direct binding of the arsenical with the sperm thiol components essential for sperm motility as well as induction of oxidative damage by disruption of sperm cell's antioxidant system. Sodium arsanilate and its N-substituted pyrimidine derivative, PHI-370, are useful probes to further evaluate the mechanism of pentavalent arsanilate-induced human sperm dysfunction.

  15. Methods of sperm vitality assessment.

    Science.gov (United States)

    Moskovtsev, Sergey I; Librach, Clifford L

    2013-01-01

    Sperm vitality is a reflection of the proportion of live, membrane-intact spermatozoa determined by either dye exclusion or osmoregulatory capacity under hypo-osmotic conditions. In this chapter we address the two most common methods of sperm vitality assessment: eosin-nigrosin staining and the hypo-osmotic swelling test, both utilized in clinical Andrology laboratories.

  16. Social imaginaries, sperm and whiteness

    DEFF Research Database (Denmark)

    Andreassen, Rikke

    2017-01-01

    This article analyses narratives about so-called Viking babies and Viking sperm. Over the last few years an increasing number of British single women and lesbian couples have been creating families by becoming pregnant with Danish donor sperm, termed ‘Viking sperm’. Through analyses of British...

  17. Sperm preparation for ART

    Directory of Open Access Journals (Sweden)

    Schill Wolf-Bernhard

    2003-11-01

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

  18. How much does it cost to look like a pig in a wild boar group?

    DEFF Research Database (Denmark)

    Batocchio, Daniele; Iacolina, Laura; Canu, Antonio

    2017-01-01

    Hybridization between domestic and wild species is known to widely occur and it is reported to be oneof the major causes of the current biodiversity crisis. Despite this, poor attention has been deserved tothe behavioural ecology of hybrids, in particular in relation to their social behaviour. We...... carried out acamera trap study to assess whether phenotypically anomalous colouration in wild boar, i.e. potentiallyintrogressed with domestic pigs, affected the hierarchical structure of wild boar social groups. Chromat-ically anomalous wild boars (CAWs) were detected in 32 out of 531 wild boar videos...

  19. Viability of Frozen-Thawed Epididymal Sperm of Garut Ram Cryopreserved with Modified Tris Extender

    Directory of Open Access Journals (Sweden)

    MUHAMMAD RIZAL

    2005-06-01

    Full Text Available Sperm collected from cauda epididymis is a source of male gametes. The purposes of this study was to evaluate an quality of frozen-thawed sperm of garut ram which was collected from cauda epididymis and cryopreserved with modified Tris extender, i.e: Tris extender (control, KT, Tris extender + 60 mM lactose (LS, and Tris extender + 60 mM lactose + 0.05% glutathione (GL. Quality of collected sperm including concentration, motility, live sperm, abnormality, cytoplasmic droplet, intact acrosomal cap (IAC, and intact plasma membrane (IPM were evaluated. Results showed that mean of sperm concentration, percentages of motility, live sperm, abnormality, cytoplasmic droplet, IAC, and IPM of fresh epididymal sperm were 13,993.33 million/ml, 70.83, 82.83, 10.83, 8.5, 85.83, and 81.33%, respectively. Sperm quality after equilibration for LS and GL were significantly (P<0.05 higher than KT. Mean percentages of post thawing sperm motility, live sperm, IAC, and IPM for GL (45, 54.5, 47.83, and 48.83% were significantly (P<0.05 higher than LS (40, 49.17, 43.83, and 44.5%, and KT (35, 42.5, 39.17, and 41.5%. Mean percentages of post thawing sperm motility, live sperm, IAC, and IPM for LS were significantly (P<0.05 higher than those of KT. Hence, frozen-thawed epididymal sperm of garut ram after slaughter and cryopreserved with Tris extender + 60 mM lactose (LS and Tris extender + 60 mM lactose + 0.05% glutathione (GL possibly can be used for artificial insemination (AI or in vitro embryo production program.

  20. Comparison of detection methods for antibodies to boar spermatozoa

    African Journals Online (AJOL)

    The different methods varied in their sensitivity to determination of sperm antibody titers. The ELISA was the most sensitive of the methods. The ELISA and Protein A agglutination were the most suitable in terms of specificity and repeatability for sperm antibody detection in swine. (Tropical Veterinarian: 2003 21(1): 1-13) ...

  1. 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

    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....... Fertilization of two egg batches was attempted. Diluted sperm caused a similar percentage of fertilized eggs and a similar number of embryos and larvae, independently of storage temperature (4 or 20°C). The cryopreserved sperm resulted in a lower percentage of fertilized eggs, but embryos developed and a few...... larvae ('cryolarvae') were obtained 55 h after fertilization in one of the two egg batches. This result evidences that the tested cryopreservation protocol is applicable for eel reproduction management, although improvements will be required to enhance fertilization success...

  2. Sperm maturation in dogs: sperm profile and enzymatic antioxidant status in ejaculated and epididymal spermatozoa.

    Science.gov (United States)

    Angrimani, D S R; Lucio, C F; Veiga, G A L; Silva, L C G; Regazzi, F M; Nichi, M; Vannucchi, C I

    2014-09-01

    Spermatozoa become more susceptible to the attack of reactive oxygen species during maturation. To avoid oxidative damage, the epididymis must provide the necessary antioxidant protection. The aim of this study was to compare the canine sperm profile and the enzymatic antioxidant status of the ejaculated fractions and samples collected from the different segments of the epididymis (caput, corpus and cauda). Five adult dogs were used, and after 1-3 weeks, subsequently to bilateral orchiectomy and epididymal storage, sperm samples were collected from the different segments of the epididymis. Samples were evaluated for conventional microscopy and computer-assisted motility analysis: sperm plasma membrane permeability and the activity of the antioxidant enzymes catalase, glutathione peroxidase (GPx) and superoxide dismutase (SOD). Samples collected from the caput and corpus showed lower values for most of the motility variables evaluated, indicating different levels of immaturity. Catalase activity was observed only in ejaculated samples. Conversely, GPx activity was higher in the cauda epididymidis. Correlations were found between SOD and GPx and SOD and sperm motility in the epididymal cauda and corpus, highlighting the importance of the enzymes for the protection of spermatozoa during the transit along the epididymis. © 2013 Blackwell Verlag GmbH.

  3. Orientation of sea urchin sperms in static magnetic fields: Compared to human sperms

    Science.gov (United States)

    Sakhnini, Lama; Dairi, Maheen

    In this study we report on magnetic orientation of sea urchin and human sperms. The sea urchin and human sperms became oriented parallel to the magnetic field (1 T maximum). The human sperms were totally oriented with magnetic field at about 600 mT. However, the sea urchin sperms show different behavior due to morphological differences between them and the human sperms.

  4. Gene expression during testis development in Duroc boars

    DEFF Research Database (Denmark)

    Lervik, Siri; Kristoffersen, Anja Bråthen; Conley, Lene

    2015-01-01

    and rodents, act at specific ages and developmental stages in the boar testis. The age dependency and complexity shown for development-specific testis transcripts must be considered when selecting phenotypic parameters for genetic selection for low androstenone. The results of selection based on measurement......Androstenone is a steroid pheromone occurring in the pubertal Leydig cells. Breeding against androstenone can decrease pheromone odour in swine meat but appears to cause unwanted side effects such as delayed onset of puberty. To study causality, global gene expression in developing boar testes...... with the onset of pubertal development. With elevated steroidogenesis (weeks 16 to 27), there was an increase in the expression of genes in the MAPK pathway, STAR and its analogue STARD6. A pubertal shift in genes coding for cellular cholesterol transport was observed. Increased expression of meiotic pathways...

  5. Sperm analysis of the vas deferens fluid after a long interval of unilateral percutaneous epididymal sperm aspiration in vasectomized patients

    Directory of Open Access Journals (Sweden)

    Fernando Lorenzini

    2013-09-01

    Full Text Available Objectives Evaluation of the presence of spermatozoa in vas deferens fluid after a long interval of unilateral and homolateral percutaneous epididymal sperm aspiration (PESA in vasectomized men. When found, the spermatozoa were evaluated including concentration and motility, in order to verify the patency of the epididymal tubules. Materials and Methods Four patients, numbered in a progressive order, from one to four, with 38, 40, 48 and 51 years old and vasectomy interval of 10, 10, 25 and 11 years, respectively, whose wives did not get pregnant using intracytoplasmic sperm injection of sperm obtained by unilateral PESA and decided to try only natural conception, were submitted to intrasurgical sperm analysis of the vas deferens fluid (ISAVDF during microsurgery for reconstruction of the seminal tract. Results Time interval between PESA and ISAVDF was 13.75 ± 11.12 months (x ± s varying from 3 to 29 months. Homolateral ISAVDF and PESA showed the presence of spermatozoa. Patients 1, 2 and 4 had a high concentration of 10 x 106, 64 x 106 and 45 x 106 spermatozoa/ mL; the first two had motile sperms and patient 3 had no sperms. Conclusions Three of four patients showed spermatozoa in the vas deferens fluid after a long interval of unilateral and homolateral PESA with high concentration, including motile forms. These findings support the concept that PESA may not result in late epipidymal tubule obstruction in vasectomized patients.

  6. Proteinase inhibitors in aggregated forms of boar seminal plasma proteins

    Czech Academy of Sciences Publication Activity Database

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

    2003-01-01

    Roč. 32, - (2003), s. 99-107 ISSN 0141-8130 R&D Projects: GA ČR GA303/02/0433; GA ČR GP303/02/P069; GA MZd NJ7463 Institutional research plan: CEZ:AV0Z5052915; CEZ:MSM 113100001 Keywords : proteinase inhibitors, aggregated forms, boar seminal plasma proteins Subject RIV: CE - Biochemistry Impact factor: 1.427, year: 2003

  7. Blood parameters of boars fed diets supplemented with different ...

    African Journals Online (AJOL)

    Thirty-two cross-bred (Large White X Landrace) male weanling pigs aged 8-9 weeks and weighing between 7.37 and 7.62kg (averaging 7.49 + 0.36kg individually) were used to assess the haematological profile of boars fed varied dietary copper. Four experimental diets were formulated: control (diet 1) with no inclusion of ...

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

    Czech Academy of Sciences Publication Activity Database

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

    2009-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Janko Mrkun

    2014-01-01

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

  10. Dietary fatty acids influence sperm quality and function.

    Science.gov (United States)

    Ferramosca, A; Moscatelli, N; Di Giacomo, M; Zara, V

    2017-05-01

    Recently, obesity has been linked to male infertility. In animal models the administration of a high-fat diet caused a reduction in sperm quality, by impairing gamete energy metabolism. The aim of this study was to investigate a possible effect of dietary fatty acids supplementation in the modulation of sperm energy metabolism and, in turn, in the improvement of sperm quality in rats fed a high-fat diet. Sexually mature male Sprague-Dawley rats were divided into four groups and fed for 4 weeks a standard diet (control group), a high-fat diet (enriched in 35% of fat and 15% sucrose), a high-fat diet supplemented with 2.5% olive oil (a source of monounsaturated fatty acids) or a high-fat diet supplemented with 2.5% krill oil (a source of n-3 polyunsaturated fatty acids). Liver and adipose tissue weight, plasma glucose, insulin and lipid concentrations were determined. Activities of enzymes involved in sperm energetic metabolism were evaluated by spectrophotometric assays. Sperm mitochondrial respiratory efficiency was also assayed. The obtained results suggest that olive oil partially counteracts the negative effects of a high-fat diet on sperm quality, by increasing gamete motility, by reducing oxidative stress and slightly improving mitochondrial respiration efficiency. On the other hand, krill oil determines an increase in sperm concentration and motility, an increase in the activities of lactate dehydrogenase, Krebs cycle enzymes and respiratory chain complexes; a parallel increase in the cellular levels of ATP and a reduction in oxidative damage were also observed. These results suggest that dietary fatty acids are able to positively influence sperm quality and function. © 2017 American Society of Andrology and European Academy of Andrology.

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

    Science.gov (United States)

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

    2013-02-01

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

  12. THE EVOLUTION OF SPERM SIZE IN BIRDS.

    Science.gov (United States)

    Briskie, James V; Montgomerie, Robert; Birkhead, Tim R

    1997-06-01

    Sperm size varies enormously among species, but the reasons for this variation remain obscure. Since it has been suggested that swimming velocity increases with sperm length, earlier studies proposed longer (and therefore faster) sperm are advantageous under conditions of intense sperm competition. Nonetheless, previous work has been equivocal, perhaps because the intensity of sperm competition was measured indirectly. DNA profiling now provides a more direct measure of the number of offspring sired by extrapair males, and thus a more direct method of assessing the potential for sperm competition. Using a sample of 21 species of passerine birds for which DNA profiling data were available, we found a positive relation between sperm length and the degree of extrapair paternity. A path analysis, however, revealed that this relationship arises only indirectly through the positive relationship between the rate of extrapair paternity and length of sperm storage tubules (SSTs) in the female. As sperm length is correlated positively with SST length, an increase in the intensity of sperm competition leads to an increase in sperm length only through its effect on SST length. Why females vary SST length with the intensity of sperm competition is not clear, but one possibility is that it increases female control over how sperm are used in fertilization. Males, in turn, may respond on an evolutionary time scale to changes in SST size by increasing sperm length to prevent displacement from rival sperm. Previous theoretical analyses predicting that sperm size should decrease as sperm competition becomes more intense were not supported by our findings. We suggest that future models of sperm-size evolution consider not only the role of sperm competition, but also how female control and manipulation of ejaculates after insemination selects for different sperm morphologies. © 1997 The Society for the Study of Evolution.

  13. Sperm structure and sperm transfer in Pseudopythina subsinuata (Bivalvia, Galeommatoidea)

    DEFF Research Database (Denmark)

    Jespersen, Åse

    2009-01-01

    to the elongate cells. Most females contain one to three "sperm trees", structures consisting of a short stem and numerous branches. They are firmly implanted in the abfrontal part of the gill filament and protrude into the posterior part of the suprabranchial (brooding) chamber. Implantation of the trees causes...... the receptacle and thereupon fuse. A similar process is known in the allied P. tsurumaru, but the resulting structure ("sperm-carrying body") is not attached to the gills....

  14. Sperm Proteome Maturation in the Mouse Epididymis.

    Science.gov (United States)

    Skerget, Sheri; Rosenow, Matthew A; Petritis, Konstantinos; Karr, Timothy L

    2015-01-01

    In mammals, transit through the epididymis, which involves the acquisition, loss and modification of proteins, is required to confer motility and fertilization competency to sperm. The overall dynamics of maturation is poorly understood, and a systems level understanding of the complex maturation process will provide valuable new information about changes occurring during epididymal transport. We report the proteomes of sperm collected from the caput, corpus and cauda segments of the mouse epididymis, identifying 1536, 1720 and 1234 proteins respectively. This study identified 765 proteins that are present in sperm obtained from all three segments. We identified 1766 proteins that are potentially added (732) or removed (1034) from sperm during epididymal transit. Phenotypic analyses of the caput, corpus and cauda sperm proteomes identified 60 proteins that have known sperm phenotypes when mutated, or absent from sperm. Our analysis indicates that as much as one-third of proteins with known sperm phenotypes are added to sperm during epididymal transit. GO analyses revealed that cauda sperm are enriched for specific functions including sperm-egg recognition and motility, consistent with the observation that sperm acquire motility and fertilization competency during transit through the epididymis. In addition, GO analyses revealed that the immunity protein profile of sperm changes during sperm maturation. Finally, we identified components of the 26S proteasome, the immunoproteasome, and a proteasome activator in mature sperm.

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

    Science.gov (United States)

    2011-01-01

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

  16. Advances in sperm retrieval techniques in azoospermic men: A systematic review

    Directory of Open Access Journals (Sweden)

    Rupin Shah

    2018-03-01

    Full Text Available Objective: To evaluate various methods of operative sperm retrieval in men with non-obstructive azoospermia (NOA and to determine the optimal surgical approach in terms of effectiveness, morbidity, and complications. Materials and methods: PubMed and Cochrane databases were searched to identify five recent reviews and meta-analyses evaluating outcomes for sperm retrieval in men with NOA. Results and Conclusion: Micro-TESE is the most efficient method for retrieving sperm but requires special expertise and can be traumatic for the testes. Conventional biopsies are twice more likely to retrieve sperm than fine-needle aspiration. Testicular aspiration performed by multiple passes into the testis is traumatic and is not efficient for sperm retrieval. Needle-aspiration biopsy and open real-time testicular mapping by the single seminiferous tubule technique can offer less traumatic methods for sperm retrieval, which can be tried before proceeding to micro-TESE. The first attempt at sperm retrieval is the best chance the patient has and should combine various techniques sequentially to give the highest chance of success with the least morbidity. Keywords: Non-obstructive azoospermia, Operative sperm retrieval, Testicular failure, Testicular sperm extraction

  17. Fatty acid content in epididymal fluid and spermatozoa during sperm maturation in dogs.

    Science.gov (United States)

    Ramos Angrimani, Daniel S; Nichi, Marcilio; Losano, João Diego A; Lucio, Cristina F; Lima Veiga, Gisele A; Franco, Márcia V M Junqueira; Vannucchi, Camila I

    2017-01-01

    During sperm maturation, there is a reorganization of fatty acids from plasmatic membrane of the spermatozoa, which allows higher membrane integrity and acquisition of sperm motility. However, the fatty acid profile during sperm maturation remains unclear in dogs. Thus, the aim of this study was to identify the fatty acids from the epididymal spermatozoa and plasma during the sperm maturation, and observed changes in the motility and plasmatic membrane parameters. Twenty one adult dogs were used, subsequently to bilateral orchiectomy and epididymal storage, sperm samples were collected from the different segments of the epididymis. Samples were evaluated for conventional microscopy, computer-assisted motility analysis, sperm plasma membrane permeability and the fatty acid analysis (lipids were extracted, transmethylated and analyzed by chromatography). Caput and corpus sperm showed lower values for the motility variables evaluated and plasmatic membrane integrity, indicating different levels of the fatty acids organization. Saturated, monounsaturated and polyunsaturated fatty acids were in higher concentrations in the spermatozoa from epididymis cauda. Highlighting the presence of caprylic, stearic and docosahexaenoic acids. These findings demonstrate the influence of the fatty acid profile during sperm maturation, assigning physical and chemical changes in sperm cells, essential for fertilization.

  18. Boar effects and their relations to fertility and litter size in sows

    African Journals Online (AJOL)

    User

    Boar effects and their relations to fertility and litter size in sows. D.O. Umesiobi. Department of Agriculture, School of Agriculture and Environmental Sciences, Central University of Technology,. Free State, Private Bag X20539, Bloemfontein 9300, South Africa. Abstract. Twenty Large White boars and 60 sows were used in ...

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

    Directory of Open Access Journals (Sweden)

    Jaime Gosálvez

    2014-06-01

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

  20. Colloid single-layer centrifugation improves post-thaw donkey (Equus asinus) sperm quality and is related to ejaculate freezability.

    Science.gov (United States)

    Ortiz, I; Dorado, J; Acha, D; Gálvez, M J; Urbano, M; Hidalgo, M

    2015-01-01

    The aim of this study was to determine whether colloid single-layer centrifugation (SLC) improves post-thaw donkey sperm quality and if this potential enhancement is related to ejaculate freezability. Semen from Andalusian donkeys was frozen following a standard protocol. SLC was performed on frozen-thawed semen and post-thaw sperm parameters were compared with uncentrifuged samples. Sperm quality was estimated by integrating in a single value sperm motility (assessed by computer-assisted sperm analysis), morphology and viability (evaluated under brightfield or fluorescence microscopy). Sperm freezability was calculated as the relationship between sperm quality obtained before freezing and after thawing. Ejaculates were classified into low, medium and high freezability groups using the 25th and 75th percentiles as thresholds. All sperm parameters were significantly (P<0.01) higher in SLC-selected samples in comparison to uncentrifuged frozen-thawed semen and several kinematic parameters were even higher than those obtained in fresh semen. The increment of sperm parameters after SLC selection was correlated with ejaculate freezability, obtaining the highest values after SLC in semen samples with low freezability. We concluded that, based on the sperm-quality parameters evaluated, SLC can be a suitable procedure to improve post-thaw sperm quality of cryopreserved donkey semen, in particular for those ejaculates with low freezability.

  1. 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. Copyright © 2017. Published by Elsevier Inc.

  2. Influence of different anaesthetic protocols over the sperm quality on the fresh, chilled (4°C) and frozen-thawed epididymal sperm samples in domestic dogs.

    Science.gov (United States)

    Batista, M; Vilar, J; Rosario, I; Terradas, E

    2016-10-01

    This study assessed the influence of three different anaesthetic protocols on semen quality obtained from the epididymis. Sixty male dogs undergoing to routine sterilization were assigned to three anaesthetic protocols: thiopental group (TG, n = 20), propofol group (PG, n = 20) and ketamine-dexmedetomidine group (KDG, n = 20). Immediately after orchidectomy, the cauda epididymides and vas deferent ducts were isolated and then a retrograde flushing was performed to collect spermatozoa. In experiment 1, after the initial evaluation of the semen (sperm concentration, sperm motility and the percentages of live spermatozoa, abnormal spermatozoa and acrosome membrane integrity), semen samples were diluted in Tris-glucose-egg yolk extender and chilled for 48 hr, and the sperm motility was assessed at 6, 24 and 48 hr. In experiment 2, semen samples were diluted in Tris-glucose-egg yolk extender and chilled for 24 hr, and then samples were frozen in two extenders with different glycerol concentrations, to reach a final concentration of 50-100 × 10(6) spermatozoa ml(-1) , 20% egg yolk, 0.5% Equex and 4% and 5% glycerol, respectively. Mean values of total sperm concentration, sperm viability and the percentages of intact acrosome and abnormal spermatozoa were not significantly different between experimental groups, and therefore, the anaesthetic protocols assessed did not affect sperm parameters mentioned above. However, our study confirmed a detrimental effect of the use of thiopental (TG) over the total sperm motility (p sperm motility (p epididymal sperm samples. The anaesthetic protocols including the application of propofol or ketamine-dexmedetomidine can be used to recover sperm in domestic canids without significant changes in sperm quality compared when semen is collected routinely and these techniques could be applicable to endangered wild canids. © 2016 Blackwell Verlag GmbH.

  3. Spermiogram and sperm head morphometry assessed by multivariate cluster analysis results during adolescence (12-18 years and the effect of varicocele

    Directory of Open Access Journals (Sweden)

    Fernando Vásquez

    2016-01-01

    Full Text Available This work evaluates sperm head morphometric characteristics in adolescents from 12 to 18 years of age, and the effect of varicocele. Volunteers between 150 and 224 months of age (mean 191, n = 87, who had reached oigarche by 12 years old, were recruited in the area of Barranquilla, Colombia. Morphometric analysis of sperm heads was performed with principal component (PC and discriminant analysis. Combining seminal fluid and sperm parameters provided five PCs: two related to sperm morphometry, one to sperm motility, and two to seminal fluid components. Discriminant analysis on the morphometric results of varicocele and nonvaricocele groups did not provide a useful classification matrix. Of the semen-related PCs, the most explanatory (40% was related to sperm motility. Two PCs, including sperm head elongation and size, were sufficient to evaluate sperm morphometric characteristics. Most of the morphometric variables were correlated with age, with an increase in size and decrease in the elongation of the sperm head. For head size, the entire sperm population could be divided into two morphometric subpopulations, SP1 and SP2, which did not change during adolescence. In general, for varicocele individuals, SP1 had larger and more elongated sperm heads than SP2, which had smaller and more elongated heads than in nonvaricocele men. In summary, sperm head morphometry assessed by CASA-Morph and multivariate cluster analysis provides a better comprehension of the ejaculate structure and possibly sperm function. Morphometric analysis provides much more information than data obtained from conventional semen analysis.

  4. EFFECT OF ORGANIC AND INORGANIC MINERAL SUPPLEMENTATION ON SEMINAL QUALITY OF BOARS EXPOSED TO HEAT STRESS EFEITO DA SUPLEMENTAÇÃO DE MINERAIS ORGÂNICOS E INORGÂNICOS NA QUALIDADE DO SÊMEN DE SUÍNOS SUBMETIDOS A ESTRESSE TÉRMICO

    Directory of Open Access Journals (Sweden)

    Daiane Donin Spessatto

    2009-12-01

    Full Text Available

    Seasonal high temperatures or inadequate nutrition can decrease reproductive efficiency in boars, especially through a reduction in spermatozoal number / ejaculate and percentage of normal spermatozoa. The aim of this study was to evaluate the effect of organic and inorganic trace mineral supplementation on seminal quality in boars exposed to high environmental temperatures. The experiment was conducted in Southern Brazil, west of Parana State. Boars (2 years of age were divided into three groups to receive: inorganic (GIn, n=4 and organic (GOr, n=4 mineral supplementation and a lactation diet (GCo, n=5. Inorganic and organic diets contained a premix of inorganic and organic trace minerals, respectively, with the same quantity of each trace mineral, based on NRC (NRC, 1998 for boars. The lactation diet was based on NRC for lactating sows and contained a higher level of inorganic trace minerals, protein and metabolic energy. Maximum mean environmental temperatures were higher than the normal thermal comfort temperature for boars (26oC during the experimental period, and were associated with a reduction in semen quality. Results are expressed as mean ± SEM. The semen volume of Inorganic and Organic diet groups were higher than Lactation group animals (345.7 ± 92.6 mL and 338.4 ± 67.8 mL versus 302.5 ± 81.4 mL, respectively; P=0.02. Boars in the Organic diet group had higher sperm concentration when compared to the Inorganic diet group (233.5 ± 76.7 X 106 sptz/mL versus 181.2 ± 77.3 X 106 sptz/mL, respectively; P=0.006. Percentage of normal spermatozoa, averaged higher in the Organic group than both Inorganic and Lactation groups (93.31 ± 5.20% versus 78.48 ± 12.15% and 82.59 ± 17.27%, respectively; P=0.00021. High temperatures (>34.5 oC reduced normal spermatozoa number in all groups, but with significant differences only in the Inorganic and Lactation groups (P=0.03. Organic minerals were benefic to seminal quality, reducing some of

  5. Polyclonal VDAC3 antibody decreases human sperm motility: a novel approach to male contraception

    Directory of Open Access Journals (Sweden)

    Asmarinah Asmarinah

    2011-02-01

    Full Text Available Background: Voltage dependent anion channels (VDAC mediate transport of anions, cations and ATP which play an important role in sperm motility. This study was aimed to examine the effect of polyclonal VDAC3 antiserum to human sperm motility.Methods: Polyclonal VDAC3 antiserum used in this study was produced in rabbits by immunization of VDAC3-specific synthetic peptides.  Preimmunserum was collected before immunization and used for control experiment. Recognition of VDAC3 antiserum to antigen in human sperm was performed by western blot. Thirty sperm samples obtained from fertile men which had high quality of sperm motility were washed and collected by Percoll gradient. Sperm motility was assessed by means of evaluation of sperm velocity (seconds per 0.1 mm distance and the number of unmoved sperm (million per ml which were observed 0 minute, 30 minutes and 60 minutes after addition of VDAC3 antiserum and preimmunserum as a control. Both data were analyzed by SPSS 13.0 software.Results: VDAC3 antiserum recognized VDAC3 protein in human sperm. Statistical analysis demonstrated that there were increasing numbers of unmoved spermatozoa after addition of anti-VDAC3 antiserum in vitro for 60 minutes observation compared with preimmunserum (control. We found also that sperm velocity decreased signifi cantly after giving anti-VDAC3 antiserum in vitro for 0 minute, 30 minutes, and 60 minutes compared with pre-immunee serum (control.Conclusion: VDAC3 antiserum can decrease motility of human sperm. and may provide a novel principle of male contraception in the future. (Med J Indones 2011; 20:5-10Keywords: VDAC3 antiserum, sperm, motility, contraception

  6. Activation of free sperm and dissociation of sperm bundles (spermatozeugmata) of an endangered viviparous fish, Xenotoca eiseni.

    Science.gov (United States)

    Liu, Yue; Yang, Huiping; Torres, Leticia; Tiersch, Terrence R

    2018-04-01

    Knowledge of sperm motility activation for viviparous fishes has been limited to study of several species in Poeciliidae, and the dissociation of sperm bundles is even less understood. The goal of this study was to use the endangered Redtail Splitfin (Xenotoca eiseni) as a model to investigate the activation of sperm from viviparous fishes by study of free sperm and spermatozeugmata (unencapsulated sperm bundles). The specific objectives were to evaluate the effects of: (1) osmotic pressure and refrigerated storage (4 °C) on activation of free sperm, (2) osmotic pressure, ions, and pH on dissociation of spermatozeugmata, and (3) CaCl 2 concentration and pH on sperm membrane integrity. Free sperm were activated in Ca 2+ -free Hanks' balanced salt solution at 81-516 mOsmol/kg. The highest motility (19 ± 6%) was at 305 mOsmol/kg and swim remained for 84 h. Glucose (300-700 mOsmol/kg), NaCl (50-600 mOsmol/kg), and KCl, MgCl 2 , and MnCl 2 at 5-160 mM activated sperm within spermatozeugmata, but did not dissociate spermatozeugmata. CaCl 2 at 5-160 mM dissociated spermatozeugmata within 10 min. Solutions of NaCl-NaOH at pH 11.6 to 12.4 dissociated spermatozeugmata within 1 min. The percentage of viable cells had no significant differences (P = 0.2033) among different concentrations of CaCl 2 , but it was lower (P < 0.0001) at pH 12.5 than at pH between 7.0 and 12.0. Overall, this study provided a foundation for quality evaluation of sperm and spermatozeugmata from livebearing fishes, and for development of germplasm repositories for imperiled goodeids. Copyright © 2018 Elsevier Inc. All rights reserved.

  7. Cryopreservation of yamú (Brycon amazonicus) sperm for large scale fertilization

    DEFF Research Database (Denmark)

    Velasco-Santamaría, Yohana M.; Medina-Robles, Mauricio; Cruz-Casallas, Pablo E.

    2006-01-01

    . Sperm was diluted (1:4) in a solution of glucose, egg yolk and dimethyl sulfoxide (DMSO). Sperm concentration was determined using a Neubauer chamber, and motility evaluated after activation with 1% NaHCO3. In the laboratory, four sizes of straw (0.5, 1.8, 2.5 and 4.0 mL) and two thawing temperatures...

  8. Challenges in Development of Sperm Repositories for Biomedical Fishes: Quality Control in Small-Bodied Species.

    Science.gov (United States)

    Torres, Leticia; Liu, Yue; Guitreau, Amy; Yang, Huiping; Tiersch, Terrence R

    2017-12-01

    Quality control (QC) is essential for reproducible and efficient functioning of germplasm repositories. However, many biomedical fish models present significant QC challenges due to small body sizes (<5 cm) and miniscule sperm volumes (<5 μL). Using minimal volumes of sperm, we used Zebrafish to evaluate common QC endpoints as surrogates for fertilization success along sequential steps of cryopreservation. First, concentrations of calibration bead suspensions were evaluated with a Makler ® counting chamber by using different sample volumes and mixing methods. For sperm analysis, samples were initially diluted at a 1:30 ratio with Hanks' balanced salt solution (HBSS). Motility was evaluated by using different ratios of sperm and activation medium, and membrane integrity was analyzed with flow cytometry at different concentrations. Concentration and sperm motility could be confidently estimated by using volumes as small as 1 μL, whereas membrane integrity required a minimum of 2 μL (at 1 × 10 6 cells/mL). Thus, <5 μL of sperm suspension (after dilution to 30-150 μL with HBSS) was required to evaluate sperm quality by using three endpoints. Sperm quality assessment using a combination of complementary endpoints enhances QC efforts during cryopreservation, increasing reliability and reproducibility, and reducing waste of time and resources.

  9. Development of Domestic Cat Embryo Produced by Preserved Sperms

    Directory of Open Access Journals (Sweden)

    KARTINI ERIANI

    2008-12-01

    Full Text Available The ability to mature and fertilize oocytes of endangered species may allow us to sustain genetic and global biodiversity. Epididymis sperms may be the last chance to ensure preservation of genetic materials after injury or death of a valuable animal. Studies have been conducted to determine wether both epididymis sperms and oocytes can be used to produce viable embryos and offspring. The purpose of this study was to determine how long cats sperms contained in epididymis were remain motile and had intact membranes when preserved at 4 ° C, and to determine whether such those preserved sperms are able to fertilize oocytes. Epididymis was preserved immediately in phosphate buffer saline at 4 ° C for 1, 3, and 6 days. The observation of sperm quality and viability after preservation was performed by vital staining acrosom and Hoechst-Propidium Iodine. Biological functions of sperms were evaluated by in vitro culture technique for fertilization, micro fertilization and embryonic development rate in CR1aa medium. The results showed that average motility of sperms collected from ductus deferens, cauda and corpus epididymis decreased not significantly (P > 0.05 from 0, 1, 3, and 6 days of preservation times (from 83.0%, 80.2%, 79.0%; 80.9%, 75.0%, 75.5%; 52.0%, 63.2%, 55.0% to 34.6%, 34.6%, 33.3%, respectively. The general results showed that sperms from epididymis preserved for 1, 3, and 6 days can be used for IVF. The rate of embryonal cleavage produced by IVF technique using sperms collected from epididymis preserved for 1-, 3- and 6-days were 33.3, 26.7, and 20.0%, respectively and significantly different (p < 0.05 from that of controll (50.0%. In conclusion, sperms contained in epididyimis preserved at 4 ° C in PBS (Phospate Buffer Saline for 1-6 days can be used to IVF and in vitro production of cat embryos.

  10. Sperm cleanup and centrifugation processing for cryopreservation.

    Science.gov (United States)

    Sieme, Harald; Oldenhof, Harriëtte

    2015-01-01

    Fertility rates with artificial insemination are highest with good-quality sperm samples. Therefore, nonviable sperm, cellular debris, and seminal plasma are preferably removed from semen samples prior to use or for preservation. Such compounds are sources where reactive oxygen species are generated during storage or upon cryopreservation, impairing sperm function. In this chapter we describe methods to remove seminal plasma and cellular debris from sperm samples, and for selecting morphologically normal motile sperm. The methods that are described here include: ordinary centrifugation, sperm swim-up, glass wool and Sephadex filtration/adherence, and single-layer as well as discontinuous two-layer iodixanol density gradient centrifugation.

  11. Social genetic effects for growth in pigs differ between boars and gilts.

    Science.gov (United States)

    Nielsen, Hanne M; Ask, Birgitte; Madsen, Per

    2018-02-01

    Average daily gain (ADG) in pigs is affected by the so-called social (or indirect) genetic effects (SGE). However, SGE may differ between sexes because boars grow faster than gilts and their social behaviours differ. We hypothesized that direct genetic effects (DGE) and SGE for ADG in pigs differ between boars and gilts and that accounting for these differences will improve the predictive ability of a social genetic effects model (SGM). Our data consisted of ADG from 30 to 94 kg for 32,212 uncastrated males (boars) and 48,252 gilts that were raised in sex-specific pens. Data were analyzed using a univariate model with sex as a fixed effect and a bivariate model with ADG in boars and gilts as separate traits using both a classical animal model (CM) and a SGM. With the univariate model, the heritability for ADG was 0.22 ± 0.01 for the CM, while the estimate of the total heritable variance (T 2 ) was 0.23 ± 0.01 with the SGM. With the bivariate model, the genetic variance for SGE was higher for boars (13.8 ± 5.8) than for gilts (9.3 ± 3.9). For the bivariate model, T 2 was 0.32 ± 0.02 for boars and 0.27 ± 0.01 for gilts. Estimates of the genetic correlations between DGE (0.88 ± 0.02) and SGE (0.30 ± 0.30) for boars versus gilts indicated that ADG in boars and gilts are different traits. Moreover, the estimate of the genetic correlation between DGE and SGE indicated presence of genetic effects of competition among gilts but not among boars. Compared to a CM, the univariate SGM improved predictive ability significantly only for gilts and the bivariate SGM improved predictive ability significantly for both boars and gilts. We found significant genetic variances of SGE for ADG. The covariance between DGE and SGE was much more negative for gilts than for boars when applying the bivariate model. Because the estimate of the genetic correlation for ADG between gilts and boars differed significantly from 1 and the predictive ability for boars and gilts

  12. Structural Classification of Wild Boar (Sus scrofa) Vocalizations.

    Science.gov (United States)

    Garcia, Maxime; Gingras, Bruno; Bowling, Daniel L; Herbst, Christian T; Boeckle, Markus; Locatelli, Yann; Fitch, W Tecumseh

    2016-04-01

    Determining whether a species' vocal communication system is graded or discrete requires definition of its vocal repertoire. In this context, research on domestic pig ( Sus scrofa domesticus ) vocalizations, for example, has led to significant advances in our understanding of communicative functions. Despite their close relation to domestic pigs, little is known about wild boar ( Sus scrofa) vocalizations. The few existing studies, conducted in the 1970s, relied on visual inspections of spectrograms to quantify acoustic parameters and lacked statistical analysis. Here, we use objective signal processing techniques and advanced statistical approaches to classify 616 calls recorded from semi-free ranging animals. Based on four spectral and temporal acoustic parameters-quartile Q25, duration, spectral flux, and spectral flatness-extracted from a multivariate analysis, we refine and extend the conclusions drawn from previous work and present a statistically validated classification of the wild boar vocal repertoire into four call types: grunts, grunt-squeals, squeals, and trumpets. While the majority of calls could be sorted into these categories using objective criteria, we also found evidence supporting a graded interpretation of some wild boar vocalizations as acoustically continuous, with the extremes representing discrete call types. The use of objective criteria based on modern techniques and statistics in respect to acoustic continuity advances our understanding of vocal variation. Integrating our findings with recent studies on domestic pig vocal behavior and emotions, we emphasize the importance of grunt-squeals for acoustic approaches to animal welfare and underline the need of further research investigating the role of domestication on animal vocal communication.

  13. First isolation and characterization of Brucella microti from wild boar.

    Science.gov (United States)

    Rónai, Zsuzsanna; Kreizinger, Zsuzsa; Dán, Ádám; Drees, Kevin; Foster, Jeffrey T; Bányai, Krisztián; Marton, Szilvia; Szeredi, Levente; Jánosi, Szilárd; Gyuranecz, Miklós

    2015-07-11

    Brucella microti was first isolated from common vole (Microtus arvalis) in the Czech Republic in Central Europe in 2007. As B. microti is the only Brucella species known to live in soil, its distribution, ecology, zoonotic potential, and genomic organization is of particular interest. The present paper is the first to report the isolation of B. microti from a wild boar (Sus scrofa), which is also the first isolation of this bacterial species in Hungary. The B. microti isolate was cultured, after enrichment in Brucella-selective broth, from the submandibular lymph node of a female wild boar that was taken by hunters in Hungary near the Austrian border in September 2014. Histological and immunohistological examinations of the lymph node sections with B. abortus-, B. suis- and B. canis-specific sera gave negative results. The isolate did not require CO2 for growth, was oxidase, catalase, and urease positive, H2S negative, grew well in the presence of 20 μg/ml basic fuchsin and thionin, and had brownish pigmentation after three days of incubation. It gave strong positive agglutination with anti-A and anti-M but had a negative reaction with anti-R monospecific sera. The API 20 NE test identified it as Ochrobactrum anthropi with 99.9% identity, and it showed B. microti-specific banding pattern in the Bruce- and Suis-ladder multiplex PCR systems. Whole genome re-sequencing identified 30 SNPs in orthologous loci when compared to the B. microti reference genome available in GenBank, and the MLVA analysis yielded a unique profile. Given that the female wild boar did not develop any clinical disease, we hypothesize that this host species only harboured the bacterium, serving as a possible reservoir capable of maintaining and spreading this pathogen. The infectious source could have been either a rodent, a carcass that had been eaten or infection occurred via the boar rooting in soil. The low number of discovered SNPs suggests an unexpectedly high level of genetic homogeneity

  14. Mannan-binding proteins from boar seminal plasma

    Czech Academy of Sciences Publication Activity Database

    Jelínková-Slavíčková, Petra; Liberda, J.; Maňásková, Pavla; Ryšlavá, H.; Jonáková, Věra; Tichá, M.

    2004-01-01

    Roč. 62, 1-2 (2004), s. 167-182 ISSN 0165-0378. [Congress of the European Society for Reproductive & Developmental Immunology /4./. Rhodes, 04.06.2003-06.06.2003] R&D Projects: GA ČR GA303/02/0433; GA ČR GP303/02/P069; GA MŠk VS96141; GA MZd NJ7463 Institutional research plan: CEZ:AV0Z5052915 Keywords : boar seminal plasma proteins * mannan-binding proteins * oviductal epithelium Subject RIV: CE - Biochemistry Impact factor: 2.726, year: 2004

  15. Exposure of extensively farmed wild boars (Sus scrofa scrofa) to selected pig pathogens in Greece.

    Science.gov (United States)

    Marinou, K A; Papatsiros, V G; Gkotsopoulos, E K; Odatzoglou, P K; Athanasiou, L V

    2015-06-01

    Increased density and distribution of wild boar populations are likely to promote interactions and transmission of certain pathogens, not only among wild boar but also from wild boar to livestock or humans and vice versa. The purpose of this study was to determine seroprevalence against seven selected pathogens in wild boar living in four different areas in Greece. In total, 359 serum samples were collected from extensively farmed wild boar (Sus scrofa scrofa) originating from four distinct geographical areas throughout Greece from April 2012 to August 2013. Samples were tested for antibodies to Actinobacillus pleuropneumoniae, African swine fever virus (ASFV), Aujeszky's disease virus (ADV), classical swine fever virus (CSFV), Erysipelothrix rhusiopathiae, Mycoplasma hyopneumoniae and porcine reproductive and respiratory syndrome virus (PRRSV). Prevalence was compared among the four regions using Fisher's exact test. Low overall seropositivities of 2.4% and 5.6% were detected for E. rhusiopathiae and PRRSV, respectively, higher ones for ADV (32.0%) and the highest (72.5% and 90.5%) for M. hyopneumoniae and A. pleuropneumoniae, respectively. All sera tested were found negative for antibodies directed against CSFV and ASFV. This is the first report of exposure of wild boars to selected pig pathogens in Greece. These results are indicative of the circulation of these pathogens in Greece with the exception of CSFV and ASFV and suggestive of the potential role of wild boars on their maintenance and transmission to their domestic counterparts and vice versa.

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

    OpenAIRE

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

    2014-01-01

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

  17. Hunting of roe deer and wild boar in Germany: Is non-lead ammunition suitable for hunting?

    Science.gov (United States)

    Martin, Annett; Gremse, Carl; Selhorst, Thomas; Bandick, Niels; Müller-Graf, Christine; Greiner, Matthias; Lahrssen-Wiederholt, Monika

    2017-01-01

    Non-lead hunting ammunition is an alternative to bullets that contain lead. The use of lead ammunition can result in severe contamination of game meat, thus posing a health risk to consumers. With any kind of ammunition for hunting, the terminal effectiveness of bullets is an animal welfare issue. Doubts about the effectiveness of non-lead bullets for a humane kill of game animals in hunting have been discussed. The length of the escape distance after the shot has been used previously as an indicator for bullet performance. The object of this study was to determine how the bullet material (lead or non-lead) influences the observed escape distances. 1,234 records of the shooting of roe deer (Capreolus capreolus) and 825 records of the shooting of wild boar (Sus scrofa) were evaluated. As the bullet material cannot be regarded as the sole cause of variability of escape distances, interactions of other potential influencing variables like shot placement, shooting distance, were analyzed using conditional regression trees and two-part hurdle models. The length of the escape distance is not influenced by the use of lead or non-lead ammunition with either roe deer or wild boar. With roe deer, the length of the escape distance is influenced significantly by the shot placement and the type of hunting. Increasing shooting distances increased the length of the escape distance. With wild boar, shot placement and the age of the animals were found to be a significant influencing factor on the length of the escape distance. The length of the escape distance can be used as an indicator for adequate bullet effectiveness for humane killings of game animals in hunting.Non-lead bullets already exist which have an equally reliable killing effect as lead bullets.

  18. Female sperm storage in reptiles.

    Science.gov (United States)

    Sever, David M; Hamlett, William C

    2002-02-01

    Internal fertilization and oviparity most likely are symplesiomorphies for modern reptiles, and viviparity has evolved independently numerous times in Sauria and Serpentes. Oviducal sperm storage is known in females of all taxa except Amphisbaenia. However, in Rhynchocephalia and Crocodilia, sperm storage is poorly studied, and specialized sperm storage tubules (Ssts) are unknown. We use the molecular phylogenetic hypothesis [(Chelonia+Archosauria) (Squamata)] to trace evolution of sperm storage characters. Ssts arose independently in Chelonia and Squamata. Turtles possess albumen-secreting glands in the anterior half of the oviduct (the tuba or isthmus), and the most distal of these glands also serve as Ssts; in addition, some turtles possess Ssts in the adjacent segment of the oviduct, the uterus. Squamates lack albumen-secreting glands, and the ancestral state is possession of Ssts in the posterior infundibulum (uterine tube). Secondarily, iguanids have evolved vaginal Ssts. In this paper, we present the first ultrastructural observations on vaginal Ssts in lizards, using Anolis sagrei (Polychrotidae). Proximally, the neck of these simple tubular glands continues the alternation of ciliated and secretory cells lining the lumen of the vagina. However, the epithelial cells of the distal sperm storage area are neither secretory nor ciliated. The Ssts of Anolis are more similar to those of birds more than to infundibular receptacles in snakes and lizards. Copyright 2002 Wiley-Liss, Inc.

  19. Camera traps and activity signs to estimate wild boar density and derive abundance indices.

    Science.gov (United States)

    Massei, Giovanna; Coats, Julia; Lambert, Mark Simon; Pietravalle, Stephane; Gill, Robin; Cowan, Dave

    2018-04-01

    Populations of wild boar and feral pigs are increasing worldwide, in parallel with their significant environmental and economic impact. Reliable methods of monitoring trends and estimating abundance are needed to measure the effects of interventions on population size. The main aims of this study, carried out in five English woodlands were: (i) to compare wild boar abundance indices obtained from camera trap surveys and from activity signs; and (ii) to assess the precision of density estimates in relation to different densities of camera traps. For each woodland, we calculated a passive activity index (PAI) based on camera trap surveys, rooting activity and wild boar trails on transects, and estimated absolute densities based on camera trap surveys. PAIs obtained using different methods showed similar patterns. We found significant between-year differences in abundance of wild boar using PAIs based on camera trap surveys and on trails on transects, but not on signs of rooting on transects. The density of wild boar from camera trap surveys varied between 0.7 and 7 animals/km 2 . Increasing the density of camera traps above nine per km 2 did not increase the precision of the estimate of wild boar density. PAIs based on number of wild boar trails and on camera trap data appear to be more sensitive to changes in population size than PAIs based on signs of rooting. For wild boar densities similar to those recorded in this study, nine camera traps per km 2 are sufficient to estimate the mean density of wild boar. © 2017 Crown copyright. Pest Management Science © 2017 Society of Chemical Industry. © 2017 Crown copyright. Pest Management Science © 2017 Society of Chemical Industry.

  20. Psychosocial counselling of identifiable sperm donors

    NARCIS (Netherlands)

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

    2016-01-01

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

  1. Psychosocial counselling of identifiable sperm donors

    NARCIS (Netherlands)

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

    2016-01-01

    What do identifiable sperm donors feel about psychosocial counselling? Identifiable sperm donors found it important that psychosocial counselling focused on emotional consequences and on rules and regulations and they expected to have access to psychosocial counselling at the time that

  2. Variation in sperm morphology among Afrotropical sunbirds

    Czech Academy of Sciences Publication Activity Database

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

    2016-01-01

    Roč. 158, č. 1 (2016), s. 155-166 ISSN 0019-1019 Institutional support: RVO:68081766 Keywords : comparative analysis * Nectariniidae * phylogenetic signal * sperm competition * sperm size Subject RIV: EG - Zoology Impact factor: 2.279, year: 2016

  3. Delta opioid receptor on equine sperm cells: subcellular localization and involvement in sperm motility analyzed by computer assisted sperm analyzer (CASA

    Directory of Open Access Journals (Sweden)

    Lacalandra Giovanni M

    2010-06-01

    Full Text Available Abstract Background Opioid receptors and endogenous opioid peptides act not only in the control of nociceptive pathways, indeed several reports demonstrate the effects of opiates on sperm cell motility and morphology suggesting the importance of these receptors in the modulation of reproduction in mammals. In this study we investigated the expression of delta opioid receptors on equine spermatozoa by western blot/indirect immunofluorescence and its relationship with sperm cell physiology. Methods We analyzed viability, motility, capacitation, acrosome reaction and mitochondrial activity in the presence of naltrindole and DPDPE by means of a computer assisted sperm analyzer and a fluorescent confocal microscope. The evaluation of viability, capacitation and acrosome reaction was carried out by the double CTC/Hoechst staining, whereas mitochondrial activity was assessed by means of MitoTracker Orange dye. Results We showed that in equine sperm cells, delta opioid receptor is expressed as a doublet of 65 and 50 kDa molecular mass and is localized in the mid piece of tail; we also demonstrated that naltrindole, a delta opioid receptor antagonist, could be utilized in modulating several physiological parameters of the equine spermatozoon in a dose-dependent way. We also found that low concentrations of the antagonist increase sperm motility whereas high concentrations show the opposite effect. Moreover low concentrations hamper capacitation, acrosome reaction and viability even if the percentage of cells with active mitochondria seems to be increased; the opposite effect is exerted at high concentrations. We have also observed that the delta opioid receptor agonist DPDPE is scarcely involved in affecting the same parameters at the employed concentrations. Conclusions The results described in this paper add new important details in the comprehension of the mammalian sperm physiology and suggest new insights for improving reproduction and for

  4. Profound Transcriptomic Differences Found between Sperm Samples from Sperm Donors vs. Patients Undergoing Assisted Reproduction Techniques Tends to Disappear after Swim-up Sperm Preparation Technique

    Directory of Open Access Journals (Sweden)

    Marcos Meseguer

    2010-01-01

    Full Text Available Background: Although spermatozoa delivers its RNA to oocytes at fertilization, its biologicalrole is not well characterized. Our purpose was to identify the genes differentially and exclusivelyexpressed in sperm samples both before and after the swim-up process in control donors andinfertile males with the purpose to identify their functional significance in male fertility.Materials and Methods: This was a nested case-control study. Ten sperm samples were obtainedfrom infertile patients [n=5 (two aliquots each from five samples; one before the swim-upprocess and one after] and donors [n=5 (two aliquots from five samples, one before the swim-upprocess and one after]. Oligonucleotide microarrays were employed to study the genome-wideexpression of pooled samples from infertile patients vs. donors. A total of four microarrays wereperfomed: two with sperm sample aliquots before swim-up and two with sperm samples aliquotsafter swim-up, from both the case and control groups. The results were evaluated to detect whichgenes expressed differentially [fold change (FC>5 and p<0.05] and which genes were exclusiveto each of the groups, both before and after swim-up.Results: Profound differences were detected between the fresh sperm samples of donors vs.infertile patients with respect to both differentially and exclusively expressed genes. Neverthelessthese differences seemed to decrease after the swim-up selection process.Conclusion: There are important differences between the expression profiles of sperm samplesof fertile donors vs. infertile patients who require assisted reproduction techniques (ART. Thesedifferences are potential forecasters of fertility success, although their reliability needs to beexplored further.

  5. Heat shock protein 90 has roles in intracellular calcium homeostasis, protein tyrosine phosphorylation regulation, and progesterone-responsive sperm function in human sperm.

    Science.gov (United States)

    Li, Kun; Xue, Yamei; Chen, Aijun; Jiang, Youfang; Xie, Haifeng; Shi, Qixian; Zhang, Songying; Ni, Ya

    2014-01-01

    Heat shock protein 90 plays critical roles in client protein maturation, signal transduction, protein folding and degradation, and morphological evolution; however, its function in human sperm is not fully understood. Therefore, our objective in this study was to elucidate the mechanism by which heat shock protein 90 exerts its effects on human sperm function. By performing indirect immunofluorescence staining, we found that heat shock protein 90 was localized primarily in the neck, midpiece, and tail regions of human sperm, and that its expression increased with increasing incubation time under capacitation conditions. Geldanamycin, a specific inhibitor of heat shock protein 90, was shown to inhibit this increase in heat shock protein 90 expression in western blotting analyses. Using a multifunctional microplate reader to examine Fluo-3 AM-loaded sperm, we observed for the first time that inhibition of heat shock protein 90 by using geldanamycin significantly decreased intracellular calcium concentrations during capacitation. Moreover, western blot analysis showed that geldanamycin enhanced tyrosine phosphorylation of several proteins, including heat shock protein 90, in a dose-dependent manner. The effects of geldanamycin on human sperm function in the absence or presence of progesterone was evaluated by performing chlortetracycline staining and by using a computer-assisted sperm analyzer. We found that geldanamycin alone did not affect sperm capacitation, hyperactivation, and motility, but did so in the presence of progesterone. Taken together, these data suggest that heat shock protein 90, which increases in expression in human sperm during capacitation, has roles in intracellular calcium homeostasis, protein tyrosine phosphorylation regulation, and progesterone-stimulated sperm function. In this study, we provide new insights into the roles of heat shock protein 90 in sperm function.

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

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past two decades, Denmark has developed into an important destination for fertility travelers in need of donor sperm. A variety of different clinics providing sperm with different modes of clinical care have developed, and an increasing number of their customers come from abroad. One...... of the reasons why Danish fertility clinics have enabled women from Denmark and abroad to turn to sperm donation is, that two of the largest sperm banks in Europe are Danish. In contrast to many other countries where sperm is in demand, the Danish sperm banks are able to provide enough sperm for the national...... market as well as clinics worldwide. Furthermore, one of the sperm banks market sperm for self-insemination and deliver it to private customers by UPS. Although sperm donation is an old and low technology compared to other reproductive technologies, this development has taken place at the same time...

  7. Social genetic effects for growth in pigs differ between boars and gilts

    DEFF Research Database (Denmark)

    Nielsen, Hanne M.; Ask, Birgitte; Madsen, Per

    2018-01-01

    between boars and gilts and that accounting for these differences will improve the predictive ability of a social genetic effects model (SGM). Our data consisted of ADG from 30 to 94 kg for 32,212 uncastrated males (boars) and 48,252 gilts that were raised in sex-specific pens. Data were analyzed using......Average daily gain (ADG) in pigs is affected by the so-called social (or indirect) genetic effects (SGE). However, SGE may differ between sexes because boars grow faster than gilts and their social behaviours differ. We hypothesized that direct genetic effects (DGE) and SGE for ADG in pigs differ...

  8. Social genetic effects for growth in pigs differ between boars and gilts

    DEFF Research Database (Denmark)

    Nielsen, Hanne M.; Ask, Birgitte; Madsen, Per

    2018-01-01

    Average daily gain (ADG) in pigs is affected by the so-called social (or indirect) genetic effects (SGE). However, SGE may differ between sexes because boars grow faster than gilts and their social behaviours differ. We hypothesized that direct genetic effects (DGE) and SGE for ADG in pigs differ...... between boars and gilts and that accounting for these differences will improve the predictive ability of a social genetic effects model (SGM). Our data consisted of ADG from 30 to 94 kg for 32,212 uncastrated males (boars) and 48,252 gilts that were raised in sex-specific pens. Data were analyzed using...

  9. Gold-standard and improved framework for sperm head segmentation.

    Science.gov (United States)

    Chang, Violeta; Saavedra, Jose M; Castañeda, Victor; Sarabia, Luis; Hitschfeld, Nancy; Härtel, Steffen

    2014-11-01

    Semen analysis is the first step in the evaluation of an infertile couple. Within this process, an accurate and objective morphological analysis becomes more critical as it is based on the correct detection and segmentation of human sperm components. In this paper, we present an improved two-stage framework for detection and segmentation of human sperm head characteristics (including acrosome and nucleus) that uses three different color spaces. The first stage detects regions of interest that define sperm heads, using k-means, then candidate heads are refined using mathematical morphology. In the second stage, we work on each region of interest to segment accurately the sperm head as well as nucleus and acrosome, using clustering and histogram statistical analysis techniques. Our proposal is also characterized by being fully automatic, where a user intervention is not required. Our experimental evaluation shows that our proposed method outperforms the state-of-the-art. This is supported by the results of different evaluation metrics. In addition, we propose a gold-standard built with the cooperation of a referent expert in the field, aiming to compare methods for detecting and segmenting sperm cells. Our results achieve notable improvement getting above 98% in the sperm head detection process at the expense of having significantly fewer false positives obtained by the state-of-the-art method. Our results also show an accurate head, acrosome and nucleus segmentation achieving over 80% overlapping against hand-segmented gold-standard. Our method achieves higher Dice coefficient, lower Hausdorff distance and less dispersion with respect to the results achieved by the state-of-the-art method. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  10. Bioenergetics of mammalian sperm capacitation.

    Science.gov (United States)

    Ferramosca, Alessandra; Zara, Vincenzo

    2014-01-01

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

  11. Microfluidic single sperm entrapment and analysis

    NARCIS (Netherlands)

    de Wagenaar, B.; Berendsen, Johanna Theodora Wilhelmina; Berendsen, J.T.W.; Bomer, Johan G.; Olthuis, Wouter; van den Berg, Albert; Segerink, Loes Irene

    2015-01-01

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

  12. Sperm quality differences between the rainy and dry seasons in captive black-handed spider monkeys (Ateles geoffroyi).

    Science.gov (United States)

    Hernández-López, Leonor; Parra, Gerardo Cerezo; Cerda-Molina, Ana Lilia; Pérez-Bolaños, Stephanella C; Díaz Sánchez, Vicente; Mondragón-Ceballos, Ricardo

    2002-05-01

    The present work provides an assessment of sperm measures (concentration, motility, viability, etc.) of three black-handed spider monkeys (Ateles geoffroyi) during the rainy and dry seasons in Mexico City, as well as an evaluation of the between-subjects variability of sperm quality. Twenty samples obtained by rectal electroejaculation and digested with trypsin were evaluated. The results showed that during the dry season (n = 9) the semen samples were of better quality than those obtained during the rainy season (n = 11). The individual animals showed differences in sperm concentration, although there were no differences in sperm quality. Copyright 2002 Wiley-Liss, Inc.

  13. Effect of Vitrification on Sperm Parameters and Apoptosis in Fertile Men

    Directory of Open Access Journals (Sweden)

    M Adib

    2011-01-01

    Full Text Available Introduction & Objective: Today, cryopreservation of the human sperm is a common technique for treating infertility. It has been indicated that cryopreservation by different methods decrease the sperm motility and viability in fertile men, but still effect of freezing of the sperm by vitrification method have not been evaluated on sperm parameters and apoptosis. The aim of this study was to evaluate the effect of vitrification of sperm of fertile men on different sperm parameters (motility, morphology, viability and count and apoptosis after thawing. Materials & Methods: In this experimental study which was conducted at Yazd Infertility Research and Clinical Center in 2009, seventeen semen samples were collected by masturbation from people who came to this centre. Semen analysis was performed according to WHO standards. Smear was provided from these samples and fixed for TUNEL staining. Some samples were directly cryopreserved by cryoloope in liquid nitrogen and stored at least for Seven days. After thawing, samples were evaluated for sperm parameters. The collected data was analyzed by the SPSS software using paired T-test and Willcoxon statistical test. Results: The progressive movement of sperm was significantly decreased by vitrification. Also significant decrease in viability and morphology of the sperm and increase in the rate of apoptosis was observed after vitrification. The amount of apoptosis had negatively correlated with normal parameters of spermatozoa (especially progressive motility and viability. Conclusion: These results indicated that vitrification is harmful for sperm parameters and of apoptosis rate in fertile men. However, the apoptosis rate was lower compared to other freezing methods.

  14. Solea senegalensis sperm cryopreservation: New insights on sperm quality.

    Directory of Open Access Journals (Sweden)

    Marta F Riesco

    Full Text Available Cryopreservation of Senegalese sole sperm can represent an alternative to overcome some reproductive problems of this species. However, it is important to guarantee the safe use of cryopreserved sperm by selecting an appropriate protocol according to a high demand quality need to be ensured. It has been demonstrated that traditional assays such as motility and viability do not provide enough information to identify specific damage caused by cryopreservation process (freezing and thawing. Specific tests, including lipid peroxidation and DNA damage, should be performed. In the present study, motility and lipid peroxidation were performed as specific tests allowing us to discard cryopreservation conditions such as methanol as internal cryoprotectant and bovine serum albumin as external cryoprotectant. In addition, a caspase 3/7 detection by flow cytometry was performed to analyze apoptosis activity in the best selected conditions. Moreover, new highly sensitive tests based on transcript number detection have recently been described in fish sperm cryopreservation. For this reason, a transcript level detection assay was performed on certain oxidative and chaperone genes related to fertilization ability and embryo development (hsp70, hsp90BB, hsp90AA, gpx to select the best cryopreservation conditions. DMSO+ egg yolk proved to be the best cryoprotectant combination in terms of transcript level. This study describes an optimized cryopreservation protocol for Solea senegalensis sperm demonstrating for the first time that transcript degradation is the most sensitive predictor of cell status in this species after cryopreservation.

  15. Cryopreservation of sperm bundles (spermatozeugmata) from endangered livebearing goodeids.

    Science.gov (United States)

    Liu, Yue; Torres, Leticia; Tiersch, Terrence R

    2018-04-14

    More than half of fishes in the family Goodeidae are considered to be endangered, threatened, or vulnerable. Sperm cryopreservation is an effective tool for conserving genetic resources of imperiled populations, but development of protocols with livebearing fishes faces numerous challenges including the natural packaging of sperm into bundles. In this study the cryopreservation of sperm bundles (spermatozeugmata) of three goodeids species was evaluated. Sperm quality was evaluated by activation with NaCl-NaOH solution (at 300 mOsmol/kg and pH 11.8), and analysis of dissociable bundles and dissociation duration. Using Redtail Splitfin (Xenotoca eiseni) as a model, the effects of cryoprotectants (dimethyl sulfoxide, methanol, and glycerol) with different concentrations (5-15% v/v %), equilibration exposure times (1-60 min), cooling rates (5-40 °C/min), concentrations (4 × 10 4 -4 × 10 6 bundles/ml), buffers (HBSS, PBS and NaCl), and buffer osmolalities (200-400 mOsmol/kg) were investigated. After cooling and thawing, sperm bundles maintained their packed form. A specific protocol was developed (10% dimethyl sulfoxide, 20-min equilibration, 10 °C/min cooling rate, 4 × 10 6 bundles/ml, and 300 mOsmol/kg HBSS). This protocol yielded 89 ± 5% of post-thaw dissociable bundles with 209 ± 10 s of dissociation duration for X. eiseni, 96 ± 9% with 814 ± 14 s for Blackfin Goodea (Goodea atripinni), and 66 ± 2% with 726 ± 25 s for Striped Goodeid (Ataeniobius toweri). This is the first study of cryopreservation of sperm within bundles for livebearing fishes and provides a basis for establishment of germplasm repositories for goodeids and other livebearers. Copyright © 2018. Published by Elsevier Inc.

  16. Epididymal contraction and sperm parameters are affected by clonidine.

    Science.gov (United States)

    da Silva Júnior, E D; de Souza, B P; Vilela, V V; Rodrigues, J Q D; Nichi, M; de Agostini Losano, J D; Dalmazzo, A; Barnabe, V H; Jurkiewicz, A; Jurkiewicz, N H

    2014-11-01

    The use of clonidine, a selective agonist of α2-adrenoceptors, is related to the fertility impairment. Thus, it has been described that changes in the epididymal function are related to the loss of fertility. Therefore, this study was sought to further evaluate the effects of clonidine in the rat distal cauda epididymis contractions and its consequence in the sperm parameters. The in vitro effects of clonidine in the isolated distal cauda epididymis were evaluated by pharmacological experiments. The consecutive contractile responses for clonidine in distal cauda epididymis showed desensitization. The noradrenaline-induced contractions were desensitized after in vitro clonidine pre-treatment (10(-5) M for 10 min). Clonidine was unable to alter the noradrenaline contractions if the in vitro pre-treatment was made in the presence of idazoxan (α2-adrenoceptor antagonist), whereas prazosin (α1-adrenoceptor antagonist) was ineffective. Moreover, the in vitro clonidine pre-treatment increased frequency and amplitude of spontaneous contraction of distal cauda epididymis. In addition, to induce in vivo desensitization of α2-adrenoceptors, male Wistar rats were treated with crescent doses of clonidine and distal cauda of epididymis contraction and sperm parameters were analyzed. The in vivo treatment with clonidine diminished the potency of the contractions induced by adrenergic agonists and augmented the frequency and amplitude of spontaneous contraction of distal cauda epididymis. This treatment also altered the sperm transit time in epididymis, epididymal sperm reserves, sperm lipid peroxidation, and antioxidant enzymes activity. The results suggest that clonidine was able to affect the sperm quantity and quality by decreasing the transit time related to the increase in the frequency and amplitude of spontaneous contractions in epididymis, although the contractions induced by adrenergic agonists were desensitized. © 2014 American Society of Andrology and European

  17. Development of sperm vitrification protocols for freshwater fish (Eurasian perch, Perca fluviatilis) and marine fish (European eel, Anguilla anguilla).

    Science.gov (United States)

    Kása, Eszter; Bernáth, Gergely; Kollár, Tímea; Żarski, Daniel; Lujić, Jelena; Marinović, Zoran; Bokor, Zoltán; Hegyi, Árpád; Urbányi, Béla; Vílchez, M Carmen; Morini, Marina; Peñaranda, David S; Pérez, Luz; Asturiano, Juan F; Horváth, Ákos

    2017-05-01

    Vitrification was successfully applied to the sperm of two fish species, the freshwater Eurasian perch (Perca fluviatilis) and marine European eel (Anguilla anguilla). Sperm was collected, diluted in species-specific non-activating media and cryoprotectants and vitrified by plunging directly into liquid nitrogen without pre-cooling in its vapor. Progressive motility of fresh and vitrified-thawed sperm was evaluated with computer-assisted sperm analysis (CASA). Additional sperm quality parameters such as sperm head morphometry parameters (in case of European eel) and fertilizing capacity (in case of Eurasian perch) were carried out to test the effectiveness of vitrification. The vitrification method for Eurasian perch sperm resulting the highest post-thaw motility (14±1.6%) was as follows: 1:5 dilution ratio, Tanaka extender, 30% cryoprotectant (15% methanol+15% propylene-glycol), cooling device: Cryotop, 2μl droplets, and for European eel sperm: dilution ratio 1:1, with 40% cryoprotectant (20% MeOH and 20% PG), and 10% FBS, cooling device: Cryotop, with 2μl of sperm suspension. Viable embryos were produced by fertilization with vitrified Eurasian perch sperm (neurulation: 2.54±1.67%). According to the ASMA analysis, no significant decrease in head area and perimeter of vitrified European eel spermatozoa were found when compared to fresh spermatozoa. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. The Effect of Orally Administered L-carnitine on Testis Tissue Sperm Parameters and Daily Sperm Production in Adult Mice

    OpenAIRE

    Zohre Zare; Hossein Eimani; Moslem Mohammadi; Mahmood Mofid; Hossein Dashtnavard

    2010-01-01

    Introduction: The purpose of this study was to evaluate body and testis weight, testis tissue,counts, motility, viability, morphology, and chromatin quality of epididymal sperm, aswell as the testicular spermatid number (TSN) per gram of testis, and daily sperm production(DSP) in L-carnitine treated mice.Materials and Methods: In the present study, adult male NMRI mice (mean age of 4weeks) were administered L-carnitine by gavage for two weeks. The experimental groupsreceived 1mg L-carnitine/1...

  19. Out-of-season sperm cryopreserved in different media of the Amazonian freshwater fish pirapitinga (Piaractus brachypomus).

    Science.gov (United States)

    Nascimento, A F; Maria, A N; Pessoa, N O; Carvalho, M A M; Viveiros, A T M

    2010-04-01

    The pirapitinga (Piaractus brachypomus) is a freshwater fish that inhabits the Amazon and Orinoco River basins. The use of cryopreserved sperm has been considered to facilitate procedures during the artificial reproduction. The aim of the present study was to develop a freezing protocol for pirapitinga sperm collected outside the spawning season. Sperm samples were diluted in four freezing media prepared by a combination of two extenders (glucose and BTS-Beltsville Thawing Solution) and two cryoprotectant agents (DMSO and methylglycol) loaded into 0.5-mL straws, frozen in a nitrogen-vapor shipping dewar (dry-shipper) and stored in liquid nitrogen at -196 degrees C. Post-thaw sperm motility was evaluated both subjectively using a light microscope and by a computer-assisted sperm analyzer (CASA). Curvilinear, average path and straight-line velocities were also determined. There were no differences (P>0.05) in post-thaw sperm motility between evaluations performed subjectively and using the CASA. Sperm samples cryopreserved in glucose-methylglycol yielded the greatest post-thaw sperm motility (81%) and fastest sperm velocities when compared to the samples frozen in the other three media (P<0.05). Out-of-season sperm cryopreserved in glucose and methylglycol under the conditions described above is of high quality and can therefore be used to facilitate artificial reproduction procedures, as only females will need handling for hormonal induction and gamete collection during the spawning season. Although the CASA system provides precise data on sperm motility, the subjective evaluation is practical and can be conducted by well-trained personnel at commercial fish farms as an acceptable evaluation of sperm quality. Copyright 2009 Elsevier B.V. All rights reserved.

  20. Validation of image cytometry for sperm concentration measurement

    DEFF Research Database (Denmark)

    Egeberg Palme, Dorte L; Johannsen, Trine Holm; Petersen, Jørgen Holm

    2017-01-01

    Sperm concentration is an essential parameter in the diagnostic evaluation of men from infertile couples. It is usually determined by manual counting using a hemocytometer, and is therefore both laborious and subjective. We have earlier shown that a newly developed image cytometry (IC) method may...

  1. Clinical Application of Sperm Chromatin Structure Assessment in Andrology Patients

    NARCIS (Netherlands)

    M. Smit (Marij)

    2011-01-01

    textabstractInfertility, defined as the inability to conceive spontaneously within one year, is a common medical problem. Traditionally, fertility investigations initially focus on the evaluation of ovulation and tubal patency in females, and on assessment of sperm quantity and quality in males. In

  2. Effects of curcumin on sperm parameters abnormalities induced by ...

    African Journals Online (AJOL)

    Morphine, which is commonly used for the treatment of severe pain, gastrointestinal tract and kidneys. Curcumin petals consist of, glycosides, flavonoids, and anthocyanin. The study aims at evaluating curcumin effect and morphine on sperm parameters, testis tissue and serum testosterone level in rat. In this experimental ...

  3. Testicular Morphometry and Sperm Quality of Rabbit Bucks Fed ...

    African Journals Online (AJOL)

    Twenty rabbit bucks of mixed breeds, aged four to five weeks which weighed between 627.4g to 631.5g were used to evaluate the effect of Moringa oleifera leaf meal (MOLM) on testicular morphometry and sperm quality. Five bucks were randomly assigned to each of the four diets containing MOLM at 0%. 5%, 10% and ...

  4. Haematological and sperm count changes following Exposure to ...

    African Journals Online (AJOL)

    The plants Hyptis suaveolens, Clome viscosa and Urena lobata are herbs commonly found growing in pasture and also used as medicinal plant in Nigeria. They were suspected to be toxic following a pilot toxicity study, and their toxic effects were thus evaluated on haematological parameters and sperm count of albino rats.

  5. Pentoxifylline effects on capacitation and fertility of stallion epididymal sperm

    NARCIS (Netherlands)

    Guasti, P. N.; Monteiro, G. A.; Maziero, R. R.D.; Carmo, M. T.; Dell'Aqua, J. A.; Crespilho, A. M.; Rifai, E. A.; Papa, F. O.

    2017-01-01

    The aims of this study were to determinate whether pentoxifylline (PTX) increases the motion parameters of fresh and frozen-thawed equine epididymal spermatozoa, to evaluate the tyrosine phosphorylation of frozen-thawed epididymal sperm in the presence of PTX and to determine whether the

  6. The bioaccumulation of lead in the organs of roe deer (Capreolus capreolus L.), red deer (Cervus elaphus L.), and wild boar (Sus scrofa L.) from Poland.

    Science.gov (United States)

    Bąkowska, Małgorzata; Pilarczyk, Bogumiła; Tomza-Marciniak, Agnieszka; Udała, Jan; Pilarczyk, Renata

    2016-07-01

    The aim of this study was to evaluate the level of lead (Pb) in the livers and kidneys of free-living animals from Poland, with regard to the differences in tissue Pb content between the species. The research material consisted of liver and kidney samples collected from roe deer (Capreolus capreolus), red deer (Cervus elaphus), and wild boar (Sus scrofa) that had been hunted in 16 voivodeships of Poland. The concentration of lead had been measured using inductively coupled plasma optical emission spectrometry (ICP-OES) method. The results show that differences in lead concentration in the organs depended on the geographic location. In roe deer and red deer, the highest mean lead concentrations in the livers and kidneys, observed in the central region of Poland, were twice as high as the lowest concentration of Pb in these animals from the northeastern region of the country. In wild boar, the highest mean concentration of Pb was noted in the livers of animals from the central region of Poland and in the kidneys of animals from the northwestern region, while the lowest lead concentrations in both organs were typical for wild boar from the southeast part of the country. Our results show that areas located in the center and in the north of Poland carry most of the burden of lead bioaccumulation.

  7. Effect of purine nucleoside analogue-acyclovir on the sperm parameters and testosterone production in rats.

    Science.gov (United States)

    Movahed, Elham; Sadrkhanlou, Rajabali; Ahmadi, Abbas; Nejati, Vahid; Zamani, Zahra

    2013-04-01

    Acyclovir (ACV), a synthetic purine nucleoside analogue derived from guanosine, is known to be toxic to gonads and the aim of this study was to evaluate the effect of ACV on the sperm parameters and testosterone production in rat. In this experimental study, forty adult male Wistar rats (220 ± 20 g) were randomly divided into five groups (n=8 for each group). One group served as control and one group served as sham control [distilled water was intraperitoneally (i.p.) injected]. ACV was administered intraperitoneally in the drug treatment groups (4, 16 and 48 mg/kg/day) for 15 days. Eighteen days after the last injection, rats were sacrificed by CO2 inhalation. After that, cauda epididymides were removed surgically. At the end, sperm concentrations in the cauda epididymis, sperm motility, morphology, viability, chromatin quality and DNA integrity were analyzed. Serum testosterone concentrations were determined. The results showed that ACV did not affect sperm count, but decreased sperm motility and sperm viability at 16 and 48 mg/kg dose-levels. Sperm abnormalities increased at 48 mg/kg dose-level of ACV. Further, ACV significantly increases DNA damage at 16 and 48 mg/kg dose-levels and chromatin abnormality at all doses. Besides, a significant decrease in serum testosterone concentrations was observed at 16 and 48 mg/ kg doses. The present results highly support the idea that ACV induces testicular toxicity by adverse effects on the sperm parameters and serum level of testosterone in male rats.

  8. Spermicide, cryptic female choice and the evolution of sperm form and function.

    Science.gov (United States)

    Holman, L; Snook, R R

    2006-09-01

    Sperm competition and cryptic female choice profoundly affect sperm morphology, producing diversity within both species and individuals. One type of within-individual sperm variation is sperm heteromorphism, in which each male produces two or more distinct types of sperm simultaneously, only one of which is typically fertile (the "eusperm"). The adaptive significance of nonfertile "parasperm" types is poorly understood, although numerous sperm-heteromorphic species are known from many disparate taxa. This paper examines in detail two female-centred hypotheses for the evolution and maintenance of this unconventional sperm production strategy. First, we use game theoretical models to establish that parasperm may function to protect eusperm from female-generated spermicide, and to elucidate the predictions of this idea. Second, we expand on the relatively undeveloped idea that parasperm are used by females as a criterion for cryptic female choice, and discuss the predictions generated by this idea compared to other hypotheses proposed to explain sperm heteromorphism. We critically evaluate both hypotheses, suggest ways in which they could be tested, and propose taxa in which they could be important.

  9. Infection with Paragonimus westermani of boar-hunting dogs in Western Japan maintained via artificial feeding with wild boar meat by hunters.

    Science.gov (United States)

    Irie, Takao; Yamaguchi, Yohei; Doanh, Pham Ngoc; Guo, Zhi Hong; Habe, Shigehisa; Horii, Yoichiro; Nonaka, Nariaki

    2017-08-18

    Infection of boar-hunting dogs with Paragonimus westermani was investigated in Western Japan. Blood and rectal feces were collected from 441 dogs in the three districts (205 in Kinki, 131 in Chugoku and 105 in Shikoku District). In a screening ELISA for serum antibody against P. westermani antigen, 195 dogs (44.2%) showed positive reaction. In the 195 dogs, 8 dogs were found excreting P. westermani eggs after molecular analysis of fecal eggs, and additional 7 were identified serologically for the parasite infection because of their stronger reactivity against P. westermani antigen than against antigens of other species of Paragonimus. A spatial analysis showed that all of the P. westermani infections were found in Kinki and Chugoku Districts. In this area, dogs' experience of being fed with raw boar meat showed high odds ratio (3.35) to the sero-positivity in the screening ELISA, and the frequency of such experiences was significantly higher in sero-positive dogs. While clear relationship was not obtained between predation of boars by dogs during hunting and their sero-positivity. Therefore, it is suggested that human activity of feeding with wild boar meat is the risk factor for P. westermani infection in boar-hunting dogs. Considering that hunting dogs could play as a major definitive host and maintain the present distribution of P. westermani in Western Japan, control measures for the infection in hunting dogs, such as prohibition of raw meat feeding and regular deworming, should be undertaken.

  10. Infection with Paragonimus westermani of boar-hunting dogs in Western Japan maintained via artificial feeding with wild boar meat by hunters

    Science.gov (United States)

    IRIE, Takao; YAMAGUCHI, Yohei; DOANH, Pham Ngoc; GUO, Zhi Hong; HABE, Shigehisa; HORII, Yoichiro; NONAKA, Nariaki

    2017-01-01

    Infection of boar-hunting dogs with Paragonimus westermani was investigated in Western Japan. Blood and rectal feces were collected from 441 dogs in the three districts (205 in Kinki, 131 in Chugoku and 105 in Shikoku District). In a screening ELISA for serum antibody against P. westermani antigen, 195 dogs (44.2%) showed positive reaction. In the 195 dogs, 8 dogs were found excreting P. westermani eggs after molecular analysis of fecal eggs, and additional 7 were identified serologically for the parasite infection because of their stronger reactivity against P. westermani antigen than against antigens of other species of Paragonimus. A spatial analysis showed that all of the P. westermani infections were found in Kinki and Chugoku Districts. In this area, dogs’ experience of being fed with raw boar meat showed high odds ratio (3.35) to the sero-positivity in the screening ELISA, and the frequency of such experiences was significantly higher in sero-positive dogs. While clear relationship was not obtained between predation of boars by dogs during hunting and their sero-positivity. Therefore, it is suggested that human activity of feeding with wild boar meat is the risk factor for P. westermani infection in boar-hunting dogs. Considering that hunting dogs could play as a major definitive host and maintain the present distribution of P. westermani in Western Japan, control measures for the infection in hunting dogs, such as prohibition of raw meat feeding and regular deworming, should be undertaken. PMID:28717056

  11. Towards the identification of reliable sperm biomarkers for male infertility: A sperm proteomic approach.

    Science.gov (United States)

    Intasqui, P; Agarwal, A; Sharma, R; Samanta, L; Bertolla, R P

    2018-04-01

    Male infertility evaluation is mainly based on semen analysis. Thus, identification of additional diagnostic methods is valuable. The aim of this study was to analyse the sperm proteome of infertile men to identify the underlying mechanisms and reliable diagnostic biomarkers. This cross-sectional study consisted of 16 infertile men and seven proven fertile men. An LC-MS/MS approach was performed in five pooled samples of each group (proven fertile men, primary infertility and secondary infertility). Differentially expressed proteins were used for functional enrichment analyses, and the most central proteins involved in altered functions in both infertile groups and the testis-specific proteins were validated using Western blotting and immunocytochemistry. In total, 1,305 sperm proteins were identified, of which 102 were underexpressed and 15 were overexpressed proteins in both infertile groups. Underexpressed proteins were mostly related to protein post-translational modification and folding, especially BAG6, HSPA2 and SPA17. Validation analysis revealed an underexpression of BAG6 in infertile men, whereas HSPA2 and SPA17 expressions did not differ between the groups. No differences were observed in the sperm localisation of these proteins. An overexpression of HIST1H2BA-a testis-specific protein-was observed in both proteomic approaches. Therefore, BAG6 and HIST1H2BA are potential candidates for male infertility biomarkers. © 2017 Blackwell Verlag GmbH.

  12. Frozen-thawed rhesus sperm retain normal morphology and highly progressive motility but exhibit sharply reduced efficiency in penetrating cervical mucus and hyualuronic acid gel

    Science.gov (United States)

    Tollner, Theodore L.; Dong, Qiaoxiang; VandeVoort, Catherine A.

    2011-01-01

    The preservation of the genetic diversity of captive populations of rhesus monkeys is critical to the future of biomedical research. Cryopreservation of rhesus macaque sperm is relatively simple to perform, yields high post-thaw motility, and theoretically, provides via artificial insemination (AI) a way to easily transfer genetics among colonies of animals. In the interest of optimizing semen cryopreservation methods for use with vaginal AI, we evaluated the ability of frozen-thawed rhesus sperm to penetrate periovulatory cervical mucus (CM). Motile sperm concentration of pre–freeze (“fresh”) and post-thawed (“thawed”) samples from 5 different males were normalized for both computer assisted sperm motion analysis and CM penetration experiments. Sperm samples were deposited into slide chambers containing CM or gel composed of hyaluronic acid (HA) as a surrogate for CM and numbers of sperm were recorded as they entered a video field a preset distance from the sperm suspension-CM (or HA) interface. Fresh and thawed sperm were dried on glass slides, “Pap”-stained, and assessed for changes in head dimensions and head and flagellar shape. While retaining better than 80% of fresh sperm progressive motility, thawed sperm from the same ejaculate retained on average only 18.6% of the CM penetration ability. Experiments using HA gel yielded similar results only with reduced experimental error and thus improved detection of treatment differences. Neither the percentage of abnormal forms nor head dimensions differed between fresh and thawed sperm. While findings suggests that sperm-CM interaction is a prominent factor in previous failures of vaginal AI with cryopreserved macaque sperm, neither sperm motility nor morphology appears to account for changes in the ability of cryopreserved sperm to penetrate CM. Our data points to a previously unidentified manifestation of cryodamage which may have implications for assessment of sperm function beyond the cervix and

  13. Evaluation of serum estradiol/ progestrone ratio on day of embryo transfer and it’s effect on intra-cytoplasmic sperm induction outcome

    Directory of Open Access Journals (Sweden)

    Masomeh Hagshafiha

    2013-07-01

    Full Text Available Background : One of the important problems in fertilization in vitro (IVF is failure of implantation. This could be the result of estrogen and progesterone effects in endometrial acceptance during ovulation stimulation. Although progesterone has a vital role in primary phase of pregnancy, but the estradiol role in luteal phase is unknown. The aim of this study is assessment of the ratio of estradiol to progesterone in embryo transfer day on Intracytoplasmic sperm injection (ICSI outcomes. Methods : This is a cohort study. The subjects were 311 infertile women referred to Urmia Kosar infertility clinic & Urmia reproductive health research center who treated with ICSI method between August-Jan 2011 . Five cc blood was drawn for determine of estradiol and progesterone in transfer day. Transfer occurred after a variable in vitro culture period ranging from 48 to 72 hours after ovulation induction. Chemiluminescent ELICA the level of mention hormones was used to determine the ratio of the progesterone level to the estradiol serum level and was compared based on treatment outcomes. Results : A total of 311 patients, 115 (37% were pregnant and happened abortion were 18 (5.8% . The mean ratio of estradiol to progesterone in transfer day in two groups of miscarriage/ non miscarriage and pregnant/ non pregnant was 32.26±23.86 , 28.17±26.5 and 28.58±2.4 , 36.09±4.39 respectively. There is no significant difference between two groups in regard of estradiol on progesterone ratio (P=0.5, P= 0.2 . Conclusion: The results of this prospective cohort study show that there is no effect of estradiol to progesterone ratio on day of embryo transfer, successful pregnancy in ICSI cycles and abortion rate following of ICSI .

  14. Comparative Bacteriological Study of Two Wild Boar Populations in Sierra Morena (Ja�n, Spain

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    Antonio NOTARIO

    2010-12-01

    Full Text Available The aim of this study was to investigate the presence of various bacterial species in the wild boar populations of Lugar Nuevo and Selladores-Contadero woodlands from Sierra Morena (Spain. Bacteriological analyses were carried out on a total of 229 wild boar individuals hunted in the period 2000-2003 in eleven experimental plots which are representative for the different biotopes of the area. The following species were detected: Brucella ovis, Clostridium sp., Corynebacterium sp., Streptococcus pneumoniae, Escherichia coli, Mycobacterium bovis, Mycobacterium avium subsp. paratuberculosis, Mannheimia haemolytica, Pasteurella multocida, Pseudomonas aeruginosa, Chlamydophila psittaci, Staphylococcus aureus and Staphylococcus sp. and the bacterial prevalence was estimated for each of them. The results provide useful indications of the health status of wild boar in both locations and highlight the potential of the wild boar populations to act as biological reservoirs of certain microorganisms that can be passed onto other vertebrate wild animals and humans.

  15. First record of wild boar infected with Trichinella pseudospiralis in Poland

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    Bilska-Zając Ewa

    2016-06-01

    Full Text Available Introduction: The paper describes identification of Trichinella species isolated from wild boars (Sus scrofa in the most popular hunting region of the West Pomeranian Province of Poland.

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

    DEFF Research Database (Denmark)

    Lacour, Sandrine A.; Heckmann, Aurelie; Mace, Pauline

    2013-01-01

    Freeze-tolerance of encapsulated Trichinella muscle larvae (ML) is mainly determined by Trichinella species, but is also influenced by host species, the age of the infection and the storage time and temperature of the infected meat. Moreover, the freeze-tolerance of the encapsulated species appears...... 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 degrees C for 19,30 and 56h, and for 1-8 weeks. Larvae were recovered by artificial digestion. Their mobilities were recorded using Saisam (R) image analysis software and their infectivities were...

  17. Seroepidemiologic survey for Chlamydia suis in wild boar (Sus scrofa) populations in Italy.

    Science.gov (United States)

    Di Francesco, Antonietta; Donati, Manuela; Morandi, Federico; Renzi, Maria; Masia, Marco Antonio; Ostanello, Fabio; Salvatore, Daniela; Cevenini, Roberto; Baldelli, Raffaella

    2011-07-01

    We used serology to estimate the prevalence of exposure to chlamydiae in Italian populations of wild boars (Sus scrofa). Sera from 173 hunter-killed wild boars harvested during the 2006-2009 hunting seasons in three Italian regions were tested for antibodies to Chlamydia suis, Chlamydophila pecorum, Chlamydophila abortus, and Chlamydophila psittaci by the microimmunofluorescence test. Antibody titers to chlamydiae ≥ 1:32 were detected in 110 of the 173 samples tested (63.6%). Specific reactivity could be assessed only in 44 sera with antibody titers to C. suis that were two- to threefold higher than antibody titers against the other chlamydial species; the other 66 sera had similar reactivity against all the chlamydia species tested. Antibody to C. suis was detected in sera from wild boar populations with rare or no known contact with domestic pigs. These results suggest that the wild boar could be a chlamydia reservoir and may acquire chlamydiae independent of contacts with the domestic pig.

  18. Boar taint - the effects of selected candidate genes associated with androstenone and skatole levels - a review

    Czech Academy of Sciences Publication Activity Database

    Zadinová, K.; Stupka, R.; Stratil, Antonín; Čítek, J.; Vehovský, K.; Urbanová, D.

    2016-01-01

    Roč. 34, č. 2 (2016), s. 107-128 ISSN 0860-4037 Institutional support: RVO:67985904 Keywords : boar teint * and rostenone * skatole Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 0.725, year: 2016

  19. Pre-birth sense of smell in the wild boar: the ontogeny of the olfactory mucosa.

    Science.gov (United States)

    Fulgione, Domenico; Trapanese, Martina; Buglione, Maria; Rippa, Daniela; Polese, Gianluca; Maresca, Viviana; Maselli, Valeria

    2017-08-01

    Animals recognize their surrounding environments through the sense of smell by detecting thousands of chemical odorants. Wild boars (Sus scrofa) completely depend on their ability to recognize chemical odorants: to detect food, during scavenging and searching partners, during breeding periods and to avoid potential predators. Wild piglets must be prepared for the chemical universe that they will enter after birth, and they show intense neuronal activity in the olfactory mucosa. With this in mind, we investigated the morpho-functional embryonic development of the olfactory mucosa in the wild boar (in five stages before birth). Using mRNA expression analysis of olfactory marker protein and neuropeptide Y, involved in the function of olfactory sensory neurons, we show early activation of the appropriate genes in the wild boar. We hypothesize olfactory pre-birth development in wild boar is highly adaptive. Copyright © 2017 Elsevier GmbH. All rights reserved.

  20. Role of sperm surface proteins in reproduction

    Czech Academy of Sciences Publication Activity Database

    Jonáková, Věra; Maňásková, Pavla; Davidová, Nina; Tichá, M.; Pěknicová, Jana

    2009-01-01

    Roč. 30, Supplement (2009), s. 63-64 ISSN 0196-3635. [9th International Congress of Andrology. 07.03.2009-10.03.2009, Barcelona] R&D Projects: GA MŠk(CZ) 1M06011; GA ČR(CZ) GA523/08/H064; GA ČR(CZ) GA303/06/0895 Institutional research plan: CEZ:AV0Z50520701 Keywords : boar seminal plasma proteins * spermadhesins * proteinase inhibitor * DQH * boar spermatozoa Subject RIV: CE - Biochemistry

  1. Sperm vacuoles negatively affect outcomes in intracytoplasmic morphologically selected sperm injection in terms of pregnancy, implantation, and live-birth rates.

    Science.gov (United States)

    Greco, Ermanno; Scarselli, Filomena; Fabozzi, Gemma; Colasante, Alessandro; Zavaglia, Daniela; Alviggi, Erminia; Litwicka, Katarzyna; Varricchio, Maria Teresa; Minasi, Maria Giulia; Tesarik, Jan

    2013-08-01

    To retrospectively evaluate whether sperm vacuoles influence clinical results, with a particular focus on live-birth rates, in 101 intracytoplasmic morphologically selected sperm injection (IMSI) cycles. Retrospective, observational study. Medical center. A total of 101 couples with at least two failed intracytoplasmic sperm injection (ICSI) attempts and impaired sperm morphology. Patients divided into two groups according to sperm morphology and vacuolization pattern: group A comprising patients with good quality spermatozoa (type I and/or type II spermatozoa) (n = 63 patients); group B comprising patients with low quality spermatozoa (type III and/or IV spermatozoa) (n = 38 patients). Fertilization rate, embryo quality, pregnancy, implantation, and live-birth rates. No statistically significant differences were observed between group A and B with regard to "early" assisted reproduction outcomes (fertilization rate and embryo quality). However, the "late" outcomes (pregnancy, implantation, and live-birth rates) were statistically significantly higher in group A. These results confirm a correlation between sperm vacuoles and a negative IMSI outcome, suggesting that sperm vacuoles are related to the late paternal effect. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

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

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    Kriangsak Mengumphan

    2010-10-01

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

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

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    Nasim Charehjooy

    2014-03-01

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

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

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    Ottó Bencsik

    2014-09-01

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

  5. Cat fertilization by mouse sperm injection.

    Science.gov (United States)

    Jin, Yong-Xun; Cui, Xiang-Shun; Yu, Xian-Feng; Lee, Sung-Hyun; Wang, Qing-Ling; Gao, Wei-Wei; Xu, Yong-Nan; Sun, Shao-Chen; Kong, Il-Keun; Kim, Nam-Hyung

    2012-11-01

    Interspecies intracytoplasmic sperm injection has been carried out to understand species-specific differences in oocyte environments and sperm components during fertilization. While sperm aster organization during cat fertilization requires a paternally derived centriole, mouse and hamster fertilization occur within the maternal centrosomal components. To address the questions of where sperm aster assembly occurs and whether complete fertilization is achieved in cat oocytes by interspecies sperm, we studied the fertilization processes of cat oocytes following the injection of cat, mouse, or hamster sperm. Male and female pronuclear formations were not different in the cat oocytes at 6 h following cat, mouse or hamster sperm injection. Microtubule asters were seen in all oocytes following intracytoplasmic injection of cat, mouse or hamster sperm. Immunocytochemical staining with a histone H3-m2K9 antibody revealed that mouse sperm chromatin is incorporated normally with cat egg chromatin, and that the cat eggs fertilized with mouse sperm enter metaphase and become normal 2-cell stage embryos. These results suggest that sperm aster formation is maternally dependent, and that fertilization processes and cleavage occur in a non-species specific manner in cat oocytes.

  6. Etiologies of sperm oxidative stress

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    Parvin Sabeti

    2016-04-01

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

  7. Comparison of three methods to evaluate wild boar diet

    Czech Academy of Sciences Publication Activity Database

    Zeman, J.; Hrbek, J.; Drimaj, J.; Kudláček, T.; Kamler, J.; Plhal, R.; Heroldová, Marta

    2016-01-01

    Roč. 65, č. 3 (2016), s. 221-224 ISSN 0139-7893 Institutional support: RVO:68081766 Keywords : Sus scrofa * food composition * diet analysis * stomach * faeces Subject RIV: EH - Ecology, Behaviour Impact factor: 0.739, year: 2016

  8. The Comparison Analysis of the Main Chemical Composition Parameters of Wild Boar Meat and Pork

    OpenAIRE

    Erika Bodnárné Skobrák; Károly Bodnár; Edit Mikóné Jónás; János Gundel; András Jávor

    2011-01-01

    The chemical composition of wild boar meat from wild boar parks with various habitats and different feeding facilities was analysed. Samples were collected from m. serratus anterior – during the winter hunting period. The protein, the total fat, the saturated and unsaturated fatty acid content were measured and also and those elements which have great importance on human alimentation. The results were compared with other author’s data of the same parameters of pork. The aim of the study was t...

  9. Hormonal and behavioral correlates of emotional states in sexually trained boars.

    Science.gov (United States)

    Bishop, J D; Malven, P V; Singleton, W L; Weesner, G D

    1999-12-01

    Physiological and behavioral traits of sexually mature boars were compared between episodes of copulation and sexual frustration in order to determine reliable indicators of the differences in emotional states. Ten boars, approximately 6 mo of age, were trained to mount a stationary artificial sow (ArtSow) and to ejaculate when digital pressure was applied to the extended penis. This method of semen collection is the typical procedure of the industry. All 10 boars used in this study were fully trained to this procedure before the onset of the study. Each boar was subjected to trials in which one of the following two treatments was applied. In the control (CTRL) treatment, boars were treated the same as during their training (i.e., allowed to complete ejaculation). In the frustration (FRUS) treatment, boars were allowed to mount the ArtSow, but because no manual pressure was applied to the extended penis, ejaculation never occurred. Blood was collected via indwelling catheters before onset of the trial, during exposure to the ArtSow, and after returning to their home pen. Concentrations of testosterone, cortisol, and beta-endorphin were quantified. Behavior of the boars was recorded during exposure to the ArtSow and for 30 min after return to their home pen. Relative to preexposure levels, serum cortisol increased (Pcortisol did not allow us to distinguish between the excitement of copulation and the negative affect associated with sexual frustration, whereas increases in serum beta-endorphin and motor activity seemed to be indicators of the negative emotional state of sexual frustration in trained boars.

  10. Variation in mitochondrial DNA of Vietnamese pigs: Relationships with Asian domestic pigs and Ryukyu wild boars

    OpenAIRE

    HONGO, Hitomi; ISHIGURO, Naotaka; WATANOBE, Takuma; SHIGEHARA, Nobuo; ANEZAKI, Tomoko; LONG, Vu The; BINH, Dang Vu; TIEN, Nguyen Trong; NAM, Nguyen Huu; 一美, 本郷

    2002-01-01

    Mitochondrial DNA (mtDNA) sequences (574 bp) of 30 Vietnamese pigs (large and small) were examined and compared with those of 61 haplotypes from wild boars and domestic pigs from various locations in Asia. The large Vietnamese pigs had genetic links to Ryukyu wild boars in southern Japan. The small Vietnamese pigs were closely related to other East Asian domestic pigs. These results indicate that Vietnamese pigs are genetically diverse and may be descendents of wild and domestic pigs from oth...

  11. Males and females contribute unequally to offspring genetic diversity in the polygynandrous mating system of wild boar.

    Directory of Open Access Journals (Sweden)

    Javier Pérez-González

    Full Text Available The maintenance of genetic diversity across generations depends on both the number of reproducing males and females. Variance in reproductive success, multiple paternity and litter size can all affect the relative contributions of male and female parents to genetic variation of progeny. The mating system of the wild boar (Sus scrofa has been described as polygynous, although evidence of multiple paternity in litters has been found. Using 14 microsatellite markers, we evaluated the contribution of males and females to genetic variation in the next generation in independent wild boar populations from the Iberian Peninsula and Hungary. Genetic contributions of males and females were obtained by distinguishing the paternal and maternal genetic component inherited by the progeny. We found that the paternally inherited genetic component of progeny was more diverse than the maternally inherited component. Simulations showed that this finding might be due to a sampling bias. However, after controlling for the bias by fitting both the genetic diversity in the adult population and the number of reproductive individuals in the models, paternally inherited genotypes remained more diverse than those inherited maternally. Our results suggest new insights into how promiscuous mating systems can help maintain genetic variation.

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

    Science.gov (United States)

    Takami, Yasuoki; Sota, Teiji

    2007-07-01

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

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

    Science.gov (United States)

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

    2013-04-22

    Sperm production is a key male reproductive trait and an important parameter in sperm competition theory. Under sperm competition, paternity success is predicted to depend strongly on male allocation to sperm production. Furthermore, because sperm production is inherently costly, individuals should economize in sperm expenditure, and conditional adjustment of the copulation frequency according to their sperm availability may be expected. However, experimental studies showing effects of sperm production on mating behaviour and paternity success have so far been scarce, mainly because sperm production is difficult to manipulate directly in animals. Here, we used phenotypic engineering to manipulate sperm-production rate, by employing dose-dependent RNA interference (RNAi) of a spermatogenesis-specific gene, macbol1, in the free-living flatworm Macrostomum lignano. We demonstrate (i) that our novel dose-dependent RNAi approach allows us to induce high variability in sperm-production rate; (ii) that a reduced sperm-production rate is associated with a decreased copulation frequency, suggesting conditional adjustment of mating behaviour; and (iii) that both sperm production and copulation frequency are important determinants of paternity success in a competitive situation, as predicted by sperm competition theory. Our study clearly documents the potential of phenotypic engineering via dose-dependent RNAi to test quantitative predictions of evolutionary theory.

  14. The Persistence of Sperm and the Development of Time Since Intercourse (TSI) Guidelines in Sexual Assault Cases at Forensic Science Ireland, Dublin, Ireland.

    Science.gov (United States)

    Casey, David G; Domijan, Katarina; MacNeill, Sarah; Rizet, Damien; O'Connell, Declan; Ryan, Jennifer

    2017-05-01

    The persistence of sperm using confirmatory microscopic analysis, the persistence of sperm with tails, time since intercourse (TSI) analysis, and results from the acid phosphatase (AP) reaction from approximately 5581 swabs taken from circa 1450 sexual assault cases are presented. The observed proportions of sperm in the vagina and anus declines significantly after 48 h TSI, and sperm on oral swabs were observed up to 15 h TSI. The AP reaction as a predictor of sperm on intimate swabs is questioned. All AP reaction times gave a low true positive rate; 23% of sperm-positive swabs gave a negative AP reaction time. We show the AP reaction is an unsafe and an unreliable predictor of sperm on intimate swabs. We propose that TSI not AP informs precase assessment and the evaluative approach for sexual assault cases. To help inform an evaluative approach, TSI guidelines are presented. © 2016 American Academy of Forensic Sciences.

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

    Directory of Open Access Journals (Sweden)

    Rubina Merchant

    2011-01-01

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

  16. Hepatitis E Virus in Sylvatic and Captive Wild Boar from Portugal.

    Science.gov (United States)

    Mesquita, J R; Oliveira, R M S; Coelho, C; Vieira-Pinto, M; Nascimento, M S J

    2016-10-01

    Hepatitis E virus (HEV) is a zoonotic agent today considered a major Public Health issue in industrialized countries. HEV strains belonging to zoonotic genotype 3 are widely present in swine, being today considered important reservoirs for human disease. Unlike in swine, only scarce data are available on the circulation of HEV in wild boar. This study describes the detection and molecular characterization of HEV in livers from sylvatic wild boar hunted in Portugal and destined for consumption. Additionally, the detection of HEV in stools of a confined wild boar population also destined for consumption is also described. A total of 80 liver samples collected during the hunting season of 2011/2012 and 40 stools collected in February 2012 from a wild boar breeding farm in Portugal were tested by a nested broad-spectrum RT-PCR assay targeting open reading frame (ORF) 1. Twenty livers (25.0%) and 4 stools (10%) were positive for HEV. Phylogenetic analysis showed that all strains clustered with sequences classified as HEV genotype 3 subgenotype e. To our knowledge, this is the first report documenting the occurrence and molecular analysis of HEV in sylvatic and captive wild boar destined for human consumption in Portugal. This report demonstrates for the first time the circulation of HEV in wildlife reservoirs of Portugal adding knowledge to the epidemiology of HEV in wild boar populations. © 2014 Blackwell Verlag GmbH.

  17. Expression patterns and role of SDF-1/CXCR4 axis in boar spermatogonial stem cells.

    Science.gov (United States)

    Park, Hyun Jung; Lee, Won-Yong; Kim, Jin Hoi; Park, Chankyu; Song, Hyuk

    2018-03-15

    The signaling of chemokine stromal cell-derived factor (SDF)-1 and its receptor C-X-C motif chemokine receptor 4 (CXCR4) is involved in the cellular proliferation, survival, and migration of various cell types. Although SDF-1/CXCR4 has been implicated in the maintenance of the spermatogonial population during mouse testis development, their expression patterns and functions in boar testis remain unclear. In the present study, the expression pattern of SDF-1 and CXCR4 was determined during pre-pubertal and post-pubertal stage boar testes and in vitro cultured porcine spermatogonial stem cells (pSSCs). The role of these proteins in colony formation in cultured pSSCs was also investigated. Interestingly, SDF-1 expression was observed in PGP 9.5-positve spermatogonia in all developing stages of boar testis; however, CXCR4 expression was only detected in spermatogonia from 5-day-old boar testis. In addition, SDF-1 and CXCR4 expression was observed in cultured pSSCs from 5-day-old boar testes, and inhibition of the CXCR4 receptor signaling pathway by AMD3100 significantly decreased the colony formation of pSSCs. These results suggest that SDF-1 and CXCR4 are useful markers for detecting stage-specific spermatogonia in boar testis. Our results reveal the role of the SDF-1/CXCR4 axis in pSSC in vitro culture. Copyright © 2018. Published by Elsevier Inc.

  18. Zoonotic tick-borne bacteria among wild boars (Sus scrofa in Central Italy

    Directory of Open Access Journals (Sweden)

    Valentina Virginia Ebani

    2017-03-01

    Full Text Available The objective of this investigation was to estimate the occurrence of infections by the three zoonotic bacteria Anaplasma phagocytophilum (A. phagocytophilum, Borrelia burgdorferi sensu lato (B. burgdorferi s.l. and Coxiella burnetii in wild boars (Sus scrofa in Central Italy. The spleen samples from 100 hunted wild boars were submitted to DNA extraction and PCR assays were carried out to detect the three agents. One (1% animal was positive for A. phagocytophilum, and three (3% for B. burgdorferi s.l. No positive reactions were observed for Coxiella burnetii. Wild boars did not seem to play an important role in the epidemiology of the three investigated agents. However, the detection of A. phagocytophilum and B. burgdorferi s.l. in the spleen of the tested animals showed that wild boars can harbor these pathogens, thus ticked that feeding on infected wild boars are likely to become infected, too, which represents a source of infection for other animals and humans. This is the first detection of A. phagocytophilum in wild boars in Italy.

  19. Contemporary Genetic Structure, Phylogeography and Past Demographic Processes of Wild Boar Sus scrofa Population in Central and Eastern Europe

    OpenAIRE

    Kusza, Szilvia; Podgórski, Tomasz; Scandura, Massimo; Borowik, Tomasz; Jávor, András; Sidorovich, Vadim E.; Bunevich, Aleksei N.; Kolesnikov, Mikhail; Jędrzejewska, Bogumiła

    2014-01-01

    The wild boar (Sus scrofa) is one of the most widely distributed mammals in Europe. Its demography was affected by various events in the past and today populations are increasing throughout Europe. We examined genetic diversity, structure and population dynamics of wild boar in Central and Eastern Europe. MtDNA control region (664 bp) was sequenced in 254 wild boar from six countries (Poland, Hungary, Belarus, Ukraine, Moldova and the European part of Russia). We detected 16 haplotypes, all k...

  20. In Romania, exposure to Toxoplasma gondii occurs twice as often in swine raised for familial consumption as in hunted wild boar, but occurs rarely, if ever, among fattening pigs raised in confinement.

    Science.gov (United States)

    Paştiu, Anamaria Ioana; Györke, Adriana; Blaga, Radu; Mircean, Viorica; Rosenthal, Benjamin Martin; Cozma, Vasile

    2013-06-01

    A wide range of swine husbandry practices p