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Sample records for bull semen frozen

  1. Investigation of an outbreak of infectious pustular balanoposthitis in cattle breeding bulls at a frozen semen bank.

    PubMed

    Pandey, A B; Nandi, S; Tiwari, A K; Audarya, S D; Sharma, K; Pradhan, S K; Chauhan, R S

    2014-12-01

    Infectious pustular balanoposthitis (IPB) is one of the reproductive disorders caused by bovine herpesvirus 1 (BoHV1) that can be transmitted through artificial insemination. A herd of 63 breeding bulls at a frozen semen bank in Odisha state in India experienced a suspected outbreak of IPB, with 11 bulls showing clinical signs of the infection. Clinical signs were noticed in two bulls initially and a few days after in the other nine animals. Serum samples from 53 bulls were examined for anti-BoHV1 antibodies using a virus neutralisation test (VNT) and a competitive enzyme-linked immunosorbent assay (cELISA); the remaining ten bulls were not included in the study because it was difficult to restrain them at that time. Paired serum samples were collected 21 days apart from ten clinically affected bulls (the eleventh clinically affected bull was not included in the study for the reason stated above). In the neutralisation test, the paired serum samples showed a two- to fourfold increase in anti-BoHV1 antibody titre; in the cELISA, the paired samples were also found positive for anti-BoHV1 antibodies. Serum samples from 43 in-contact bulls were collected about day 22 after the first observation of clinical infection in the herd. Among these serum samples, a total of 30 were found positive for anti-BoHV1 antibodies in the VNT and a total of 30 were found positive in cELISA. Ten samples were positive in one test but not the other and 25 tested positive in both tests. In all, 35 serum samples from in-contact bulls tested positive in either one or both of the two types of test. An overall agreement of 76.74% was found in detection of anti-BoHV1 antibodies in the two tests. Sensitivity was higher than specificity in detection of anti-BoHV1 antibodies in the serum samples. The glycoprotein C region of the genomic DNA of BoHV1 was amplified from semen samples by polymerase chain reaction. The findings from the outbreak indicate that continuous monitoring of breeding bulls at

  2. Separation of motile sperm for in vitro fertilization from frozen-thawed bull semen using progesterone induction on a microchip.

    PubMed

    Li, Jingchun; Ning, Bolin; Cao, Xinyan; Luo, Yinghua; Guo, Li; Wei, Guosheng; Liu, Shengjun; Zhang, Ying; Zhang, Aizhong; Wu, Rui; Li, Yanbing

    2016-09-01

    This study presents a novel method for the separation of motile sperm from non-progressive motile and immotile sperm and in vitro Fertilization (IVF). This separation of bull sperm was accomplished by inducing chemotaxis along a progesterone release agent in a 7.5-mm microchannel microchip composed of a biocompatible polydimethysiloxane layer and a glass gradient. The selected sperm was applied directly for IVF. In the first experiment, we tested the effect of different lengths of microchannnel (5mm, 7.5mm and 10mm) on quality parameter of separated sperm. The results showed that separated sperm using 7.5-mm microchannel chip were improved in sperm motility, swimming velocity, and beat frequency compared with other groups. In the second experiment, a medium containing sperm from swim-up method and outlet reservoir of our 7.5-mm microchannel chip was collected and mitochondrial activity of the sperm was determined by fluorescence microscopy. The sperm from the microchip had higher mitochondria activity (47.6%±6.0%) than the sperm from the swim-up method (23.6%±4.7%) (P<0.05). There were significant differences in rate of acrosome intactness between the swim-up method and the microchip (36.0%±4.1% vs. 66.8±2.1%, respectively, P<0.05). In the third experiment, we compared sperm penetration in the microchip-IVF system with a standard IVF method (droplet-IVF). The microchip-IVF group had the highest percentages of oocytes penetrated (82.2%±1.6% vs. 63.5%±2.4%) and monospermic oocytes (67.8%±3.4% vs. 42.4%±1.5%). In addition, early developmental competence of oocytes to the blastocyst stage was higher when the oocytes were inseminated in the microchip-IVF system compared with those inseminated in a standard droplet-IVF system. These results demonstrate that our microchip based on a sperm chemotaxis system is useful for motile sperm separation from frozen-thawed bull semen for IVF. Therefore, the optimized microchip system provides a good opportunity to sort

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

    PubMed

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

    2006-06-01

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

  4. Butylated hydroxytoluene can reduce oxidative stress and improve quality of frozen-thawed bull semen processed in lecithin and egg yolk based extenders.

    PubMed

    Khumran, A M; Yimer, N; Rosnina, Y; Ariff, M O; Wahid, H; Kaka, Asmatullah; Ebrahimi, M; Sarsaifi, K

    2015-12-01

    The aims of this study were to evaluate the effects of anti-oxidant butylated hydroxytoluene (BHT), when added at different concentrations into lecithin-based Bioxcell(®) (BX) and two egg-yolk-based; Tris (TY) and citrate (CE) semen extenders, on post-thaw bull sperm quality and oxidative stress. A total of 30 ejaculates from three bulls were collected using an electro ejaculator. Ejaculates were extended with one of the BX, TY and CE extenders, which contained different concentrations (0.0 - control, 0.5, 1.0, 1.5, 2.0 and 3.0mM/ml) of BHT. The extended semen samples were chilled to 4 °C, and then frozen slowly to -196 °C in 0.25 ml straws before being stored in liquid nitrogen for 2 weeks. Results showed that supplementation of BHT improved (P<0.05) general motility, progressive motility, morphology, acrosome integrity, DNA integrity and malondialdehyde of sperm at 0.5mM/ml for BX and at 1-1.5mM/ml of BHT for TY and CE when compared with the control. However, greater concentrations of 2.0 and 3.0mM/ml of BHT had a detrimental (P<0.05) effect compared with the control with all extenders evaluated. In conclusion, BHT supplementation at lesser concentrations (0.5-1.5mM/ml) could improve frozen-thawed bull sperm quality by reducing oxidative stress produced during the freezing-thawing procedures in either lecithin or egg-yolk based extenders.

  5. Dietary α-linolenic acid from flaxseed oil or eicosapentaenoic and docosahexaenoic acids from fish oil differentially alter fatty acid composition and characteristics of fresh and frozen-thawed bull semen.

    PubMed

    Moallem, Uzi; Neta, Noam; Zeron, Yoel; Zachut, Maya; Roth, Zvi

    2015-04-15

    Incorporation rates of dietary omega-3 (n-3) fatty acids (FAs) from different sources into bull plasma and sperm and the effects on physiological characteristics of fresh and frozen-thawed semen were determined. Fifteen fertile bulls were assigned to three treatment groups and supplemented for 13 weeks with encapsulated fat: (1) SFA-360 g/d per bull saturated FA; (2) FLX-450 g/d per bull providing 84.2 g/d C18:3n-3 (α-linolenic acid) from flaxseed oil; and (3) FO-450 g/d per bull providing 8.7 g/d C20:5n-3 (eicosapentaenoic acid) and 6.5 g/d C22:6n-3 (docosahexaenoic acid, DHA) from fish oil. Blood samples were taken every 2 weeks and semen was collected weekly. With respect to the FA supplements, the proportion of α-linolenic acid in plasma increased in the FLX bulls, whereas that of DHA was increased in the FO bulls, within 2 weeks. However, changes in the sperm FA fraction were first expressed in the sixth week of supplementation: in the FO and FLX bulls the DHA proportion increased (P < 0.001), whereas that of C22:5n-6 FAs (docosapentaenoic acid [DPA] n-6) decreased (P < 0.001). Sperm motility and progressive motility in fresh semen were higher (P < 0.05), and the fading rate tended to be lower in the FLX than in FO bulls (P < 0.06). Furthermore, sperm motility, progressive motility, and velocity in frozen-thawed semen were higher in FLX than in the other groups (P < 0.008). These findings indicate that the proportion of DHA in sperm can be increased at the expense of DPAn-6 by either FO or FLX supplementation, indicating de novo elongation and desaturation of short- into longer-chain n-3 FAs in testes. Furthermore, the moderate exchange of DHA and DPAn-6 in the FLX group's sperm was associated with changes in the characteristics of both fresh and frozen-thawed semen, suggesting the importance of the ratio between these two FAs for sperm structure and function.

  6. Moribund sperm in frozen-thawed semen, and sperm motion end points post-thaw and post-swim-up, are related to fertility in Holstein AI bulls.

    PubMed

    Shojaei, H; Kroetsch, T; Wilde, R; Blondin, P; Kastelic, J P; Thundathil, J C

    2012-03-15

    The objectives were to compare testicular physical characteristics and post-thaw sperm characteristics and their associations with fertility in Holstein bulls used for AI. Ten Holstein bulls (4-5 y old) were classified as either high-fertility (HF) or low-fertility (LF; n = 5 each), based on adjusted 56-d non-return rates [non-return rate (NRR); range (mean ± SD): 55.6 ± 4.6 to 71.8 ± 1.3%). Testicular physical characteristics were not significantly different between the two groups. Four ejaculates were collected from each bull and cryopreserved. Several indexes of sperm motion (based on computer-assisted sperm analysis) at post-thaw and post-swim-up were correlated with NRR. Sperm from HF bulls were in transition to a hyperactivated motility pattern, whereas those from LF bulls had only a forward progressive motility pattern. In HF vs LF bulls, there was a greater percentage of viable sperm after thawing (60.6 ± 9.7 vs 49.5 ± 8.0%, P < 0.05) and after swim-up (70.9 ± 11.0 vs 63.0 ± 8.8%, P < 0.01); these two end points were positively correlated with fertility (r = 0.45, P < 0.01 and r = 0.78; P < 0.01, respectively). Furthermore, in HF vs LF bulls, the ratio of sperm recovered after swim-up to viable sperm in post-thaw semen was higher (P < 0.001), and the proportion of moribund sperm expressed as a percentage of live sperm differed (12.6 ± 3.4 vs. 16.4 ± 3.1%, P < 0.001) and was negatively correlated (r = -0.33, P < 0.05) with fertility. In conclusion, fertility of Holstein bulls maintained in a commercial AI center was not predicted by testicular physical characteristics, but it was associated with differences in moribund sperm in the inseminate, as well as characteristics of sperm post-thaw and after swim-up.

  7. Artificial insemination of cranes with frozen semen

    USGS Publications Warehouse

    Gee, G.F.; Sexton, T.J.; Lewis, J.C.

    1979-01-01

    For the first time (1978) artificial insemination (AI) with frozen greater sandhill crane (Grus canadensis tabida) semen resulted in fertile eggs and chicks. During the 2 year (1977-78) study, 6 of 27 eggs produced were fertile. Three chicks hatched. Semen samples used for insemination were frozen and stored in liquid nitrogen for two months or less. Recent improvements in the laboratory indicated that a more effective sample can be prepared and greater fertility rates should be expected.

  8. Bulls grazing Kentucky 31 tall fescue exhibit impaired growth, semen quality, and decreased semen freezing potential

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Serum prolactin (PRL) and testosterone concentrations, body weight, body composition, semen quality, and semen freezing potential for bulls grazing the toxic tall fescue (Lolium arundinaceum [Schreb.] Darbysh. ¼ Schedonorous arundinaceum [Schreb.] Dumort.) cultivar Kentucky 31 (E+) compared with a n...

  9. The in vitro effect of leptin on semen quality of water buffalo (Bubalus bubalis) bulls.

    PubMed

    Khaki, Amir; Batavani, Rooz Ali; Najafi, Gholamreza

    2013-01-01

    The purpose of this study was to evaluate the probable effects of leptin addition in different levels to the semen extender on sperm quality (motility and motility parameters, viability, sperm membrane integrity, and DNA damage). Semen specimens were evaluated immediately after leptin addition, equilibration time and after thawing the frozen semen. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 10, 20, 50, 100, and 200 ng mL(-1) leptin. The diluted semen was kept 4 hr in refrigerator to reach to the equilibration time and then packed in 0.5 mL French straws and frozen in liquid nitrogen. Our results showed that, in the fresh semen, no significant difference was observed in all sperm quality parameters evaluated among all of the examined leptin concentrations. Addition of 10 ng mL(-1) leptin into semen extender significantly preserved sperm motility, all of the motility parameters, and viability in equilibrated semen compared to that of control group. However, in vitro addition of 200 ng mL(-1) leptin, significantly decreased theses parameters. In the frozen thawed semen, all leptin concentrations decreased sperm motility and viability, but significant decrease was observed in concentrations of 100 and 200 ng mL(-1). Adding leptin to semen extender did not have any significant influence on sperm DNA damage and sperm membrane integrity in all examined groups. These findings suggest that in vitro addition of 10 ng mL(-1) leptin could preserve sperm motility and viability in cooled semen of buffaloes.

  10. Evidence of excretion of Schmallenberg virus in bull semen

    PubMed Central

    2014-01-01

    Schmallenberg virus (SBV) is a novel orthobunyavirus, discovered in Germany in late 2011. It mainly infects cattle, sheep and goats and could lead to congenital infection, causing abortion and fetal abnormalities. SBV is transmitted by biting midges from the Culicoides genus and there is no evidence that natural infection occurs directly between ruminants. Here, we could detect SBV RNA in infected bull semen using qRT-PCR (three bulls out of seven tested positive; 29 positive semen batches out of 136). We also found that highly positive semen batches from SBV infected bulls can provoke an acute infection in IFNAR-/- mice, suggesting the potential presence of infectious virus in the semen of SBV infected bulls. PMID:24708245

  11. Cryopreservation of bull semen shipped overnight and its effect on post-thaw sperm motility, plasma membrane integrity, mitochondrial membrane potential and normal acrosomes.

    PubMed

    Anzar, M; Kroetsch, T; Boswall, L

    2011-06-01

    In the Canadian Animal Genetic Resource Program, bull semen is donated in frozen or fresh (diluted) states. This study was designed to assess the cryopreservation of diluted bull semen shipped at 4°C overnight, and to determine the post-thaw quality of shipped semen using different straw volumes and freezing rates. Semen was collected from four breeding bulls (three ejaculates per bull). Semen was diluted in Tris-citric acid-egg yolk-glycerol (TEYG) extender, cooled to 4°C and frozen as per routine (control semen). After cooling to 4°C, a part of semen was removed and shipped overnight to the research laboratory via express courier (shipped semen). Semen was packaged in 0.25 or 0.5 ml straws and frozen in a programmable freezer using three freezing rates, i.e., -10, -25 or -40°C/min. Control semen was also shipped to the research laboratory. Post-thaw sperm motility characteristics were assessed using CASA, and post-thaw sperm plasma membrane, mitochondrial membrane potential and normal acrosomes were assessed using flow cytometry. Post-thaw sperm quality was greater in shipped semen as compared to control (P<0.001). The shipped semen packaged in 0.25 ml straws had better post-thaw sperm quality than in 0.5 ml straws (P<0.001). Freezing rate had no effect on post-thaw sperm quality. In conclusion, bull semen can be shipped overnight for subsequent cryopreservation and gene banking. Overnight shipping of semen was found advantageous for bull semen cryopreservation. Semen packaging in 0.25 ml straws yielded better post-thaw quality than 0.5 ml straws.

  12. Effect of Dursban 44 on semen output of Holstein bulls.

    PubMed

    Everett, R W

    1982-09-01

    Dursban 44, an insecticide for lice control, was applied to 185 Holstein bulls 9 to 52 mo of age. These sires were in various stages of progeny testing at an artificial insemination center. Application of this product killed 7 bulls, and the remaining bulls exhibited varying severity of illness with 6 classified as very sick. This study evaluated the effect of this illness on semen production. Semen output on 40,950 ejaculates from 583 Holstein bulls collected from July 1, 1975, through March 31, 1981, was analyzed to establish normal semen production and to estimate the effect of illness caused by Dursban 44 treatment. Ejaculate number, days between collections by previous number of ejaculates, calendar months, years, and ages of bulls affected the semen output characteristics, original volume, sperm concentration, percent motile sperm, total sperm per ejaculate, percent prefreeze discards, percent postthaw sperm motility, and percent postthaw discards. Ejaculate volume, motility, total percent prefreeze discards, and percent postthaw discards were influenced negatively on the 6 very sick bulls. Percent postthaw discards were higher on all bulls treated with Dursban 44 for up to 6 mo post-treatment.

  13. A comparison of semen diluents on the in vitro and in vivo fertility of liquid bull semen.

    PubMed

    Murphy, Edel M; Murphy, Craig; O'Meara, Ciara; Dunne, Gemma; Eivers, Bernard; Lonergan, Patrick; Fair, Sean

    2017-02-01

    The aim of this study was to assess the effect of semen diluent on calving rate (CR) following artificial insemination with liquid bull semen stored for up to 3 d postcollection. In experiment 1, the effect of storing liquid semen maintained at a constant ambient temperature in 1 of 7 different diluents [Caprogen (homemade), OptiXcell, BioXcell, BullXcell, INRA96, NutriXcell, or AndroMed (all commercially available)] on total and progressive motility was assessed on d 0, 1, 2, and 3 postcollection. In experiment 2, the field fertility of liquid semen diluted in Caprogen, BioXcell, or INRA96 and inseminated on d 1, 2, or 3 postcollection was assessed in comparison to frozen-thawed semen (total of n = 19,126 inseminations). In experiment 3, the effect of storage temperature fluctuations (4 and 18°C) on total and progressive motility following dilution in Caprogen, BioXcell, and INRA96 was assessed on d 0, 1, 2, and 3 postcollection. In experiment 1, semen stored in Caprogen, BioXcell, and INRA96 resulted in the highest total and progressive motility on d 1, 2, and 3 of storage compared with OptiXcell, BullXcell, NutriXcell, and AndroMed. In experiment 2, an effect of diluent on CR was found as semen diluted in BioXcell had a lower CR on d 1, 2, and 3 of storage (46.3, 35.4, and 34.0%, respectively) in comparison with Caprogen (55.8, 52.0, and 51.9%, respectively), INRA96 (55.0, 55.1, and 52.2%, respectively), and frozen-thawed semen (59.7%). Effects were found of parity, cow fertility sub-index, as well as the number of days in milk on CR. In experiment 3, when the storage temperature of diluted semen fluctuated between 4 and 18°C, to mimic what occurs in the field (nighttime vs. daytime), BioXcell had the lowest total and progressive motility in comparison to Caprogen and INRA96. In conclusion, diluent significantly affected sperm motility when stored for up to 3 d. Semen diluted in INRA96 resulted in a similar CR to semen diluted in Caprogen and to frozen

  14. Sericin supplementation improves semen freezability of buffalo bulls by minimizing oxidative stress during cryopreservation.

    PubMed

    Kumar, Pradeep; Kumar, Dharmendra; Sikka, P; Singh, P

    2015-01-01

    The variety of mammalian cells has been successfully cryopreserved by use of the silk protein sericin due to its strong free-radical-scavenging and potent antioxidant activity. The present study was conducted to examine the protective role of sericin on buffalo spermatozoa during cryopreservation. Semen of four breeding bulls was collected twice a week using artificial vagina technique. The ejaculates of four bulls were pooled, divided into five equal fractions, diluted with the extender supplemented with different concentrations of sericin (0, 0.25, 0.5, 1.5 and 2%) and then cryopreserved. Post-thawed motility was objectively assessed by computer assisted sperm analyzer. Sperm plasma membrane integrity was assessed by hypo-osmotic swelling test (HOST). Malondialdehyde (MDA) concentration, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were determined in frozen-thawed extended seminal plasma by spectrophotometry. The extender supplemented with 0.25, 0.5 and 1% sericin resulted in the higher sperm motility and GPx acivity. Furthermore, plasma membrane integrity and SOD activity were found to be higher (P<0.05) in group supplemented with 0.25 and 0.5% sericin (P<0.05). The MDA concentration was found to be significantly lower (P<0.05) in 0.25 and 0.5% sericin treated groups than control and other treated groups. In conclusion, the supplementation of 0.25-0.5% sericin in semen extender improves frozen-thawed semen quality through protecting sperm from oxidative stress.

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

    PubMed

    Chenoweth, P J; McPherson, F J

    2016-06-01

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

  16. Hope for restoration of dead valuable bulls through cloning using donor somatic cells isolated from cryopreserved semen.

    PubMed

    Selokar, Naresh L; Saini, Monika; Palta, Prabhat; Chauhan, Manmohan S; Manik, Radheysham; Singla, Suresh K

    2014-01-01

    Somatic cells were isolated from cryopreserved semen of 4 buffalo bulls, 3 of which had died over 10 years earlier, and were established in culture. The cells expressed cytokeratin-18, keratin and vimentin indicating that they were of epithelial origin. The cells were used as nuclear donors for hand-made cloning for producing buffalo embryos. The blastocyst rate and quality, as indicated by apoptotic index, were comparable among embryos produced using cells obtained from fresh or frozen-thawed semen or those obtained from conventional cell sources such as skin. Examination of the epigenetic status revealed that the global level of H3K27me3 but not that of H3K9/14ac and H4K5ac differed significantly (P<0.05) among cloned embryos from different bulls. The relative mRNA abundance of HDAC1, DNMT1, P53 and CASPASE 3 but not that of DNMT3a differed in cells and in cloned embryos. Following transfer of 24 cloned embryos produced from fresh semen-derived cells to 12 recipients, one calf weighing 55 kg, which is now 6 months of age and is normal, was born through normal parturition. Following transfer of 20 embryos produced from frozen-thawed semen-derived cells to 10 recipients, 2 became pregnant, one of which aborted in the first trimester; the calf born was severely underweight (17 kg), and died 12 h after birth. The ability of cells derived from fresh and frozen-thawed semen to produce live offspring confirms the ability of these cells to be reprogrammed. Our findings pave the way for restoration of highly precious progeny-tested bulls, which has immense economic importance, and can also be used for restoration of endangered species.

  17. Dietary n-3 PUFAs improve fresh and post-thaw semen quality in Holstein bulls via alteration of sperm fatty acid composition.

    PubMed

    Khoshvaght, Ali; Towhidi, Armin; Zare-shahneh, Ahmad; Noruozi, Mohammad; Zhandi, Mahdi; Davachi, Navid Dadashpour; Karimi, Reza

    2016-03-15

    The goal of this study was to investigate the effect of fish oil-supplemented diet on fresh and post-thaw semen quality and sperm lipid composition in bulls. Bulls were randomly assigned to two groups (n = 6). Six bulls were used as the control group and six received the fish oil (1.2% dry matter of total diet) for 11 weeks. Semen was individually collected from each bull and frozen biweekly. Semen volume, sperm concentration, viability, progressive motility, and fatty acid profile of sperm were measured in 1st, 3rd, 5th, 7th, 9th, and 11th week of experiment. Viability, progressive motility, and fatty acid profile of post-thaw sperm were also measured in 3rd, 5th, 9th, and 11th week of experiment. Data were analyzed with using Proc GLM or MIXED (for repeated measurement data) in SAS program. The fish oil-supplemented diet increased the semen volume and sperm concentration. The fish oil-supplemented diet also altered the viability, progressive motility, and fatty acid profile of fresh and post-thaw sperm. In conclusion, feeding a fish oil-enriched diet via alteration of fatty acid profile of sperm lipid could improve in vitro quality of fresh and post-thaw sperm in Holstein bulls.

  18. Evaluation of bison (Bison bison) semen from Yellowstone National Park, Montana, USA, bulls for Brucella abortus shedding.

    PubMed

    Frey, Rebecca K; Clarke, P Ryan; McCollum, Matt P; Nol, Pauline; Johnson, Kammy R; Thompson, Brent D; Ramsey, Jennifer M; Anderson, Neil J; Rhyan, Jack C

    2013-07-01

    To determine if bison (Bison bison) bulls from Yellowstone National Park (YNP), Montana, USA, shed an infective dose of Brucella abortus in semen, 50 YNP bulls were captured on public lands in Montana during the winter and early spring (April-May) of 2010 and 2011. The bulls were immobilized, and blood and semen samples were collected for serology and Brucella culture. Thirty-five bulls (70%) were antibody-positive, and B. abortus was cultured from semen in three (9%) of the 35 antibody-positive or suspect bulls, though not at concentrations considered an infective dose. Eight bulls (six antibody-positive, two negative) had palpable lesions of the testes, epididymides, or seminal vesicles consistent with B. abortus infection. Breeding soundness exams and semen analysis suggested that antibody-positive bulls were more likely to have nonviable ejaculate (8/35; 23%) than bulls without detectable antibody (2/15; 13%).

  19. Inbreeding depression on semen quality in Austrian dual-purpose simmental bulls.

    PubMed

    Maximini, L; Fuerst-Waltl, B; Gredler, B; Baumung, R

    2011-02-01

    Using pedigree data, the inbreeding coefficients of 715 Austrian dual-purpose Simmental (Fleckvieh) bulls stationed in two artificial insemination (AI) centres in Upper and Lower Austria were calculated and incorporated in statistical models for the analysis of semen quality. Five semen quality parameters (volume, concentration, motility, number of spermatozoa per ejaculate and percentage of viable spermatozoa) of approximately 30,000 ejaculates, used in two separate data sets, were investigated. The mixed model included the fixed effects age class of the bull, bull handler, semen collector, month and year of collection, number of collection per bull and day, time interval since last collection, the linear continuous effect of the inbreeding coefficient of the bull, interactions between age class and month, and age class and interval since last collection, respectively, as well as the random effect of the bull and the random residual effect. Non-linear effects of inbreeding were significant for motility only. Despite the quite low inbreeding coefficients (mean 1.3%), all semen quality traits showed inbreeding depression, in four of the five traits significantly in at least one of the data sets. The magnitude of inbreeding depression was small, which might partly be caused by the low inbreeding levels and a potential pre-selection of the bulls in the AI centres. However, monitoring of inbreeding depression on fertility traits is recommended to avoid unrecognized deterioration of such traits.

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

    PubMed

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

    2015-01-01

    Sperm DNA damage affects the conception rate resulting from human assisted reproduction technology. The objective of this study was to adapt the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay to provide a quality parameter for bull semen based on the detection of sperm DNA damage. Fresh semen was collected from two Japanese Black bulls (A, B) several times over the course of a year, and the percentage of TUNEL-positive spermatozoa (sperm TUNEL index) was determined. Individual differences in semen were detected using the sperm TUNEL index in these bulls (P < 0.01). The sperm TUNEL index of cryopreserved semen obtained from test-mated Japanese Black (n = 30, including two bulls with a conception rate lower than 10%) and Holstein (n = 34) bulls were analyzed. The average sperm TUNEL index and conception rate resulting from artificial insemination (AI) were 4.7% and 55.7% for Japanese Black, and 4.9% and 39.5% for Holstein, respectively. A weak negative correlation between sperm TUNEL index and conception rate was observed in Holstein bulls (P < 0.05). Semen samples from six bulls with more than 10% sperm TUNEL index were studied, and these samples showed low sperm viability. However, semen resulting in a very low conception rate did not have a high sperm TUNEL index. Although it would be difficult to predict a low conception rate resulting from AI using the sperm TUNEL index alone, the index can be used as an additional parameter to provide a more comprehensive description of semen quality.

  1. Use of Hoechst 33342 stain to evaluate live fresh and frozen bull sperm by computer-assisted analysis.

    PubMed

    Tardif, A L; Farrell, P B; Trouern-Trend, V; Simkin, M E; Foote, R H

    1998-01-01

    The objective of this research was to investigate possible procedures for evaluating living bull sperm stained with Hoechst 33342 while in a simple medium and in commonly used complex egg yolk-glycerol-Tris (EYGT) and whole milk-glycerol (WMG) extenders. The two semen extenders provide good cryoprotection, but the latter one virtually obscures the sperm. To evaluate sperm motion characteristics when static nonsperm particles are present, a new Hamilton Thorne epifluorescent optical system (UV) with a strobe light was developed for potential use with DNA-stained sperm. This system permitted examination for the first time of sperm motion characteristics in milk. In Experiment 1 (four bull semen replicates with five dye concentrations and three incubation times), 2.5 microg/ml of Hoechst 33342 stained live and dead sperm sufficiently in a modified Tyrode's solution to measure all sperm characteristics without depressing motility, which was validated by using phase-contrast to analyze stained and unstained controls. In Experiments 2a and 2b, each using semen from four bulls with a 5 x 5 factorial arrangement, it was determined that 40 to 60 microg/ml of dye in EYGT or WMG, with UV illumination for 20 minutes, was optimal. There was no detrimental effect on sperm motility. In Experiment 3, analyses of two ejaculates, from each of eight bulls, confirmed that motion characteristics of sperm in EYGT and WMG were not depressed when the sperm were stained with Hoechst 33342. These experiments demonstrate that the dye concentrations and exposure times developed for use with the new epifluorescent optics facilitate evaluating bull sperm frozen in particle-filled whole milk and should be useful for sperm evaluation of a variety of species when nonsperm particulate matter may otherwise interfere.

  2. Effect of organic and inorganic selenium supplementation on semen quality and blood enzymes in buffalo bulls.

    PubMed

    El-Sharawy, Mohamed; Eid, Entsar; Darwish, Samy; Abdel-Razek, Ibrahim; Islam, Md Rashedul; Kubota, Kaiyu; Yamauchi, Nobuhiko; El-Shamaa, Ibrahim

    2016-11-21

    The present study aimed to evaluate the effect of organic and inorganic selenium (Se) supplementation on semen quality and blood serum profiles of buffalo bulls. Nine mature buffalo bulls were divided into three groups: control (non-supplemented); organic Se (10 mg Sel-Plex®/head twice weekly) and inorganic Se (10 mg sodium selenite/head twice weekly). Semen was collected twice a week for 3 months during Se supplementation. Semen properties were evaluated from fresh ejaculate. Moreover, fructose concentration, aspartate and alanine transaminase (AST and ALT) activities, total protein and total cholesterol were assayed in seminal plasma. Additionally AST, ALT, testosterone and Se levels were determined in the blood serum. Results showed that Se supplementation significantly (P < 0.05) influences the semen parameters during 3 months of treatment. Organic Se significantly (P < 0.05) increased the percentage of viable sperms compared to inorganic Se and the control group. Fructose concentration was significantly higher (P < 0.05) in the seminal plasma of organic Se-treated bulls. Serum testosterone and Se concentrations were significantly (P < 0.05) increased in the Se supplemented groups than the control group. In conclusion, Se supplementation improved the parameters of buffalo bull semen and more precisely, organic Se was more effective for the improvement of semen quality and some blood components than inorganic Se.

  3. What affects fertility of sexed bull semen more, low sperm dosage or the sorting process?

    PubMed

    Frijters, A C J; Mullaart, E; Roelofs, R M G; van Hoorne, R P; Moreno, J F; Moreno, O; Merton, J S

    2009-01-01

    Until now it has been unclear to what extent the reduced fertility with sexed semen in the dairy industry is caused by too few sperm per AI dose, or by the effect of flow cytometric sorting, which is the established procedure for sexing semen. Therefore, we evaluated the effects of low sperm numbers per dose with and without sorting on non-return rates after 56 days (NRR 56); in addition, we evaluated the effects of bulls, in order to further optimize use of sexed semen. Based on results of using sexed semen from seven Holstein bulls, an overall numerical decline of 13.6% in NRR 56 was observed (P<0.05). About two-thirds of this decline (8.6%) was due to the low dose (P<0.05), and a third (5.0%) due to the process of sorting (P<0.05). The effect of low dosage and sorting differed among bulls. We observed a sex ratio of 91.6% females for sexed semen from the first 131 calves born. Currently the best way to increase fertility of sexed semen is by closely monitoring fertility so that the highest fertility bulls are used, and by improving farm animal management. However, to make substantial progress, more in depth studies are needed on the sexing technology, especially on aspects such as sorting procedures and sperm dosage.

  4. Effects of sperm concentration at semen collection and storage period of frozen semen on dairy cow conception.

    PubMed

    Haugan, T; Gröhn, Y T; Kommisrud, E; Ropstad, E; Reksen, O

    2007-01-01

    The present study was based on data obtained from artificial inseminations (AIs) performed with cryopreserved semen from elite bulls used in the Norwegian breeding program. Semen was diluted to standardize the number of spermatozoa to 18 million per AI dose. The aim of the study was to investigate whether the net sperm concentration at semen collection and the storage period in liquid nitrogen have any effect on probability of conception in dairy cattle. We demonstrated that the natural range of sperm concentration at semen collection within some of the bulls was associated with the probability of conception. However, no primary trend among bulls was found on the effect of sperm concentration at semen collection. This appears to be due to differences among bulls in their response to the dilution ratio of seminal plasma to extender. The effect of storage time was investigated in semen that had been stored between 1000 days and 2400 days in AI straws in liquid nitrogen at the AI center. Our findings showed that use of semen with the longest storage period, i.e. 1951-2400 days, resulted in a more than one percentage point lower probability of conception than semen with a shorter storage period. In conclusion, the net sperm concentration at semen collection, which affects the dilution ratio of seminal plasma to extender, should be considered individually among bulls to achieve optimal reproductive performance. Furthermore, this study gives support to the idea that a measurable degree of damage to the spermatozoa could occur during the preservation time in liquid nitrogen.

  5. Comparison of fertility of liquid or frozen semen when varying the interval from CIDR removal to insemination.

    PubMed

    Richardson, Brittany N; Larimore, Erin L; Walker, Julie A; Utt, Matthew D; DeJarnette, J Mel; Perry, George A

    2017-03-01

    Cryopreservation allows for long-term storage of semen; however, it leads to damage of sperm that may result in complete loss of viability or changes that possibly decrease sperm functionality. Liquid semen is not exposed to these stressors and may result in a longer lifespan in the female reproductive tract, thus increasing the range in timing of insemination without affecting fertility. The objective of this study was to compare fertility of liquid and frozen semen when varying the interval from CIDR removal to AI using the 7-day CO-Synch+CIDR protocol for synchronization of time of estrus. Within age group, crossbred cows (n=389) were randomly assigned to insemination at 36 or 60h after CIDR removal with either liquid or frozen semen (36L, 60L, 36F, and 60F) from one of two Angus bulls. Cows were monitored for estrous activity from CIDR removal until 60h thereafter. Cows that failed to exhibit estrus received GnRH (100μg, i.m.), and a blood sample was collected for analysis of estradiol concentration. There was no difference in pregnancy rates when liquid or frozen semen (53% and 52%) was used, but cows inseminated at 60h had a greater (P<0.01) pregnancy rate than those inseminated at 36h (72% and 31%). There was no time of AI by semen type interaction (P=0.57). Estrus was detected in 63%, 61%, 56%, and 62% of 36F, 36L, 60F, and 60L, respectively (only 5% and 1% of 36F and 36L were detected in estrus before insemination). Overall cows that exhibited estrus had a greater pregnancy rate compared with cows that did not (P<0.01; 79% compared with 24%). Among cows that did not exhibit estrus, those inseminated with liquid semen tended to have greater pregnancy rates than those inseminated with frozen semen (P=0.06). Cows that became pregnant had greater (P<0.01) concentrations of estradiol at 60h than those that did not (10.7±0.55 compared with 7.9±0.26pg/mL). In summary, there was no difference in pregnancy success between liquid and frozen semen. However, cows

  6. Royal jelly supplementation in semen extender enhances post-thaw quality and fertility of Nili-Ravi buffalo bull sperm.

    PubMed

    Shahzad, Qaisar; Mehmood, Muhammad Usman; Khan, Hamayun; ul Husna, Asma; Qadeer, Saima; Azam, Asima; Naseer, Zahid; Ahmad, Ejaz; Safdar, Muhammad; Ahmad, Mushtaq

    2016-04-01

    Two experiments were conducted to evaluate the effect of royal jelly (RJ) on post-thaw sperm quality, in vitro and in vivo fertility rate of cryopreserved buffalo bull sperm. The semen was collected from three mature regular donor buffalo bulls, ejaculates were pooled and semen evaluated initially. In Experiment 1, the ejaculates were extended in tris-citric acid diluter supplemented with different RJ concentrations (0, 0.05, 0.1, 0.2, 0.3 or 0.4%). The diluted semen was cooled to 4°C, packaged into 0.5 mL straws and frozen using standard procedure. The straws were thawed and assessed for sperm progressive motility, viability, plasma membrane, acrosome, and chromatin integrity. The results indicated that sperm progressive motility was significantly greater (P<0.05) in 0.05, 0.1, 0.2 and 0.3% RJ than 0.4% RJ supplemented and control groups. The sperm viability, plasma membrane and acrosome integrity were significantly improved (P<0.05) in 0.1% RJ supplemented group the compared to other treatment groups. In Experiment 2, cryopreserved sperm with 0.1% RJ supplementation and control (without RJ supplementation) were used to observe the in vitro fertilizing potential and in vivo fertility. In vitro fertilization method was applied to assess the cleavage rate; whereas, AI was performed in buffalo during in vivo fertility trial. The buffaloes were inseminated 12h after standing estrus and pregnancy diagnosis was performed through ultrasonography. The results revealed that the cleavage rate was higher (P<0.05) in 0.1% RJ as compared to control group. However, the pregnancy rate was similar (P>0.05) between 0.1% RJ supplemented and control groups. It is concluded that supplementation of RJ in freezing extender can improve the cryosurvival rate and in vitro fertilizing capacity of buffalo bull sperm.

  7. Assessment of motion and kinematic characteristics of frozen-thawed Sirohi goat semen using computer-assisted semen analysis

    PubMed Central

    Anand, Mukul; Yadav, Sarvajeet

    2016-01-01

    Aim: The aim was to determine the motion and kinematics characteristic of frozen-thawed spermatozoa in Sirohi goat using computer-assisted semen analysis. Materials and Methods: A study was carried out in Sirohi buck. Semen collection was made biweekly from each buck with the help of artificial vagina. A total of 12 ejaculates were collected from two bucks (six ejaculates from each buck). Freshly collected semen was pooled and later evaluated. The pooled semen sample was extended with standard glycerolated egg yolk tris extender and later subjected to a process of cryopreservation. The motion and kinematic characteristics of spermatozoa were studied during freez-thawing process. Results: Significantly (p<0.01) higher value of live percent, hypo-osmotic swelling test, and acrosomal integrity were recorded in neat semen followed by diluted and frozen thaw semen. The proportion of spermatozoa showing slow progression were the highest in the neat and diluted semen followed by rapid and non-progressively motile, while a reverse pattern was observed in the frozen thaw semen where the proportion of non-progressively motile spermatozoa were significantly (p<0.01) higher followed by slow and rapid progression. Conclusion: This study showed that the best results for motion, vitality, plasma membrane integrity, and acrosome status were obtained in the neat semen followed by diluted and frozen thaw semen. Further, the process of cryopreservation results in a shift of motility from slow to non-progressive in the post-thaw semen with a significant decrease in the path velocities when compared to neat and diluted semen. Hence, it can be concluded that freezing-thawing process reduces the motility and kinematic characters spermatozoa and may be an important factor affecting the fertilizing ability of spermatozoa resulting in poor conception rate after insemination in goats. PMID:27051209

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

    PubMed

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

    2013-05-01

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

  9. Occurrence of "Haemophilus somnus" in bovine semen and in the prepuce of bulls and steers.

    PubMed Central

    Humphrey, J D; Little, P B; Barnum, D A; Doig, P A; Stephens, L R; Thorsen, J

    1982-01-01

    Haemophilus somnus was isolated from 40 of 79 unprocessed bovine semen samples, 14 of 23 preputial washings of bulls and three of eight preputial washings of steers. The results indicate nonvenereal colonization of the male urogenital tract. It is suggested that dissemination of H. somnus from the urogenital tract may be of significance in the epizootiology of H. somnus associated diseases. PMID:7093816

  10. Rapid detection of bovine herpesvirus 1 in the semen of infected bulls by a nested polymerase chain reaction assay.

    PubMed Central

    Masri, S A; Olson, W; Nguyen, P T; Prins, S; Deregt, D

    1996-01-01

    A nested polymerase chain reaction (PCR) assay was developed for the detection of bovine herpesvirus 1 (BHV-1) in bovine semen and compared with the virus isolation method. When extended semen, commonly used in the bovine artificial insemination industry, was inoculated with BHV-1, the PCR assay detected BHV-1 DNA in semen inoculated at 0.25-2.5 TCID50 per 0.5 mL. In contrast, the lower limit of detection for virus isolation was 250 TCID50 of BHV-1 inoculated in 0.5 mL of extended semen. These methods were also used to detect BHV-1 in the semen of four bulls which were experimentally infected with BHV-1. All infected bulls demonstrated balanitis at 3 d post-inoculation (DPI) and severe balanoposthitis at 4 DPI. BHV-1 was detected in raw semen by virus isolation and PCR at 2 DPI, before balanitis was evident. For virus isolation, the last day that BHV-1 was detected during primary infection was 7 DPI for two bulls and 9 and 11 DPI for the other two bulls. In contrast, PCR detected BHV-1 in the bulls' semen until 14 or 18 DPI. For individual animals, PCR detected BHV-1 during primary infection for at least 1-10 d longer than virus isolation. Reactivation of BHV-1 from latency without the presence of visible lesions was promoted twice by two series of 5 d dexamethasone injections. For the first series of dexamethasone treatments, a positive virus isolation result was obtained on the 5th d of treatment for only one bull. In contrast, two bulls demonstrated evidence of viral reactivation on this day by PCR. All bulls shed BHV-1 in semen on d 4 after dexamethasone treatment, as evidenced by positive virus isolation and PCR results. One bull was still PCR positive 13 d later. For the second series of dexamethasone treatments, a small amount of virus was isolated from semen collected on d 3 or 4 after treatment for two bulls but not from the other two bulls. In contrast, semen samples from all bulls were PCR positive for either or both of these 2 d. In total, from 80 semen

  11. Post-thaw viability of european bison (Bison bonasus) semen frozen with extenders containing egg yolk or lipids of plant origin and examined with a heterologous in vitro fertilization assay.

    PubMed

    Pérez-Garnelo, S S; Oter, M; Borque, C; Talavera, C; Delclaux, M; Martínez-Nevado, E; Palasz, A T; De la Fuente, J

    2006-06-01

    Basic characteristics of European bison (Bison bonasus) semen were described and the efficacies of two extenders-Triladyl, containing egg yolk, and a synthetic extender, containing soybean lipids-were tested for semen cryopreservation. Seven ejaculates were collected by electroejaculation from a 10-yr-old, European bison bull. Each ejaculate was diluted at 37 degrees C to a final concentration of 200 x 10(6) sperm/ml with Triladyl or the synthetic extender. Extended semen samples were frozen according to a standard bull semen freezing protocol. After 2 wk of storage, one straw from each extender and ejaculate was thawed, and postthaw quality was evaluated by individual sperm motility and movement rate, numbers of sperm morphologic abnormalities and intact acrosomes, functional integrity of the sperm membranes determined by hypoosmotic swelling test (HOST), viability (live-dead, eosin-nigrosin stain), and a heterologous in vitro sperm penetration assay (SPA). A total of 600 in vitro-matured bovine oocytes were inseminated with 1 X 10(6) spermatozoa of Holstein semen frozen-thawed in Triladyl (control) or of European bison semen frozen in Triladyl or the synthetic extender. Nuclear status of the oocytes was determined after 18 h of sperm-oocyte coincubation. Extender had no effect on any evaluated parameters of semen after dilution and cooling (4 hr at 5 degrees C) or in postthaw individual motility, quality of movement, and sperm morphology. However, significantly (P < 0.05) higher numbers of spermatozoa with intact acrosomes, intact membranes (HOST), and viable sperm (P < 0.01) were in semen frozen in Triladyl than in the synthetic extender. Mean values for heterologous SPA for bull (control) and for bison semen frozen in the synthetic extender were very much alike-63.3+/-10.6% and 63.1 +/- 15.9%, respectively; bison semen frozen in Triladyl was lower, 43.0+/-24.2% but not significantly different. Cumulative results from a variety of viability assays of diluted

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

    PubMed Central

    Panmei, A.; Gupta, A. K.; Shivahre, P. R.; Bhakat, M.; Upadhyay, A.

    2015-01-01

    Aim: The present study was carried out on semen ejaculates of 15 Karan Fries (KF) bulls maintained at Artificial Breeding Research Centre, National Dairy Research Institute, Karnal, India with an objective to evaluate the relationship between the conventional and fluorescent based semen quality analysis of the bulls. Materials and Methods: A total of 96 ejaculates were collected from 15 KF (Holstein Friesian [HF] crossbred) bulls. Semen were evaluated for color, volume, mass activity (MA) and percentage of individual motility (IM), sperm concentration, percent live spermatozoa, hypo-osmotic swelling test and acrosome integrity, chromatin integrity, sperm viability, and membrane integrity. Data were analyzed using SPSS software package for descriptive analysis. The correlation between rankings of sires based on conventional and fluorescent semen parameters were calculated by Spearman’s rank correlation coefficient. Results: The average ejaculates volume (ml), sperm concentration (106/ml), MA, IM (%), live (%), morphological abnormalities (%), host (%), acrosome integrity (%), chromomycin A3 (CMA3) (%), SYBR-PI (%), and fluorescent isothiocyanate-peanut agglutinin (FITC-PNA) (%) were 4.57±0.36, 1162.98±97.93, 2.95±0.09, 60.8±1.22, 71.41±2.10, 9.31±1.15, 65.5±1.81, 86.6±1.59, 3.53±0.43, 65.39±2.23 and 74.47±2.53, respectively. Rank correlations were found to be significant for SYBR-PI and FITC-PNA with most of the parameters evaluated by conventional methods. Overall, among conventional criteria, IM revealed ranking of bulls almost similar to that of fluorescent criteria. Conclusion: Overview of our results indicated that, among conventional criteria, MA and IM revealed ranking of bulls almost similar to that of fluorescent criteria. PMID:27047025

  13. Effect of age and season on semen quality parameters in Sahiwal bulls.

    PubMed

    Bhakat, Mukesh; Mohanty, T K; Raina, V S; Gupta, A K; Khan, H M; Mahapatra, R K; Sarkar, M

    2011-08-01

    The objective of the study was to determine the effect of season, period, age, bull, and ejaculate on semen quality in Sahiwal bulls. Semen production records from 1996 to 2006 of 5,483 ejaculates from 46 Sahiwal bulls maintained at Artificial Breeding Complex, NDRI, Karnal, India were analyzed using least square analysis of variance by LSML software package. The overall least squares means of ejaculate volume (VOL), total volume per day (VOLD), mass activity (MA), initial motility (IM), sperm concentration per ml (SPC), and sperm concentration per ejaculate (SPCE) were 3.79 ± 0.02 ml, 5.81 ± 0.06 ml, 2.32 ± 0.01, 55.47 ± 0.001%, 766.69 ± 5.50 × 10(6)/ml and 3023.25 ± 30.15 × 10(6), respectively. All semen traits (VOL, VOLD, MA, IM and SPCE) were significantly (P < 0.01) affected by age groups, season and period, whereas season had significant effect on VOL at 5% level. During hot-humid season, highest value of VOL, VOLD, MA, IM, SPC, and SPCE were observed followed by summer and cold season. Highest value of VOL, VOLD, IM, and SPCE were observed during period-3 (2004-2006), whereas highest value of MA and SPC were observed during period-1 (1996-1999). However, lowest magnitude of MA, IM, SPC, and SPCE during period-2 (2000-2003) was observed. Ejaculate characteristics like VOL, VOLD, and SPCE increased with the increasing age of bull up to 5 years and then decreased. Significant (P < 0.01) bull to bull variation was found in VOL, VOLD, MA, IM, SPC, and SPCE traits. First ejaculate had significantly (P < 0.01) higher MA, IM, SPC, and SPCE. Hence, it could be concluded that during rainy season and period-1 and period-3 the quality of semen is quantitatively and qualitatively good. Better quality semen was obtained up to 5 years of age in Sahiwal bulls.

  14. Sperm fertility and viability following 48h of refrigeration: evaluation of different extenders for the preservation of bull semen in liquid state.

    PubMed

    Crespilho, A M; Nichi, M; Guasti, P N; Freitas-Dell'Aqua, C P; Sá Filho, M F; Maziero, R R; Dell'aqua, J A; Papa, F O

    2014-05-01

    Two experiments were conducted to compare the effectiveness of different extenders conventionally used for semen cryopreservation to maintain the viability and fertility of cooled bull semen. In Experiment 1, sperm samples obtained from 20 Nellore bulls were preserved at 5°C for 48h using two extenders containing 20% of egg yolk [Tris (TRIS-R) and Botu-Bov(®) (BB)] and another composed of 1% soy lecithin [Botu-Bov(®)-Lecithin (BB-L)] as substitutes for animal origin products. The samples were evaluated at 6, 24 and 48h for plasma and acrosomal membrane integrity, quantification of thiobarbituric acid reactive substances (ng of TBARS/10(8) cells) and sperm motility parameters by computer-assisted semen analysis (CASA). In Experiment 2, pregnancy rate (P/AI) of 973 fixed-time artificially inseminated Nellore cows were compared when cows were inseminated with conventionally cryopreserved semen in TRIS-egg yolk glycerol (TRIS-C Control, n=253) or semen cooled for 48h in TRIS-R (n=233), BB (n=247) or BB-L (n=240). Although none of the extenders used was effective on maintaining total progressive motility and cellular integrity throughout the 48-h of the refrigeration period (P<0.01), BB-L conferred greater protection against oxidative stress (P<0.05) than egg yolk-based medias. The P/AI for semen samples preserved in TRIS-C, TRIS-R, BB and BB-L were 39.92(a), 25.32(b), 26.32(b) and 33.33(ab), respectively. These results demonstrate that the three conventional extenders used for semen cryopreservation do not provide the protection required to maintain bull semen fertility under refrigeration for a 48-h period, resulting in reduced pregnancy rates. However, the use of lecithin-based medium instead of egg yolk results in greater protection against lipid peroxidation, producing P/AI results comparable to those obtained using frozen semen.

  15. Short communication: Progressive motility of frozen-thawed canine semen is highest five minutes after thawing.

    PubMed

    Karger, S; Geiser, B; Grau, M; Heuwieser, W; Arlt, S P

    2017-04-01

    Progressive motility is usually estimated by visual inspection using a light contrast microscope at X 100 immediately after semen collection or immediately after thawing frozen semen. Standard operating procedures have never been established for this test. The objective of this experiment was to examine time-dependent changes of motility after thawing cryopreserved canine semen. Semen of 35 dogs was collected, and volume, concentration, progressive motility, morphology, membrane integrity and HOS test were evaluated. For cryopreservation, CaniPRO(®) Freeze A&B was used. Semen was thawed and diluted using CaniPRO(®) culture medium. After thawing, semen was evaluated as before. In addition, every sample was evaluated for progressively motile sperm cells 0, 5, 20 and 60 min after thawing. Progressive semen motility was significantly highest five minutes after thawing.

  16. Colloid centrifugation of fresh stallion semen before cryopreservation decreased microorganism load of frozen-thawed semen without affecting seminal kinetics.

    PubMed

    Guimarães, T; Lopes, G; Pinto, M; Silva, E; Miranda, C; Correia, M J; Damásio, L; Thompson, G; Rocha, A

    2015-01-15

    Freezability of equine semen may be influenced by microorganism population of semen. The objective of this study was to verify the effect of single-layer density gradient centrifugation (SLC) of fresh semen before cryopreservation on semen's microbial load (ML) and sperm cells kinetics after freezing-thawing. For that, one ejaculate was collected from 20 healthy stallions and split into control (C) samples (cryopreserved without previous SLC) and SLC samples (subjected to SLC). Semen cryopreservation was performed according to the same protocol in both groups. Microbial load of each microorganism species and total microbial load (TML) expressed in colony-forming units (CFU/mL) as well as frozen-thawed sperm kinetics were assessed in both groups. Additional analysis of the TML was performed, subdividing the frozen-thawed samples in "suitable" (total motility ≥ 30%) and "unsuitable" (total motility < 30%) semen for freezing programs, and comparing the C and SLC groups within these subpopulations. After thawing, SLC samples had less (P < 0.05) TML (88.65 × 10(2) ± 83.8 × 10(2) CFU/mL) than C samples (155.69 × 10(2) ± 48.85 × 10(2) CFU/mL), mainly due to a reduction of Enterococcus spp. and Bacillus spp. A relationship between post-thaw motility and SLC effect on ML was noted, as only in samples with more than 30% total motility was ML reduced (P < 0.05) by SLC (from 51.33 × 10(2) ± 33.26 × 10(2) CFU/mL to 26.68 × 10(2) ± 12.39 × 10(2) CFU/mL in "suitable" frozen-thawed semen vs. 240.90 × 10(2) ± 498.20 × 10(2) to 139.30 × 10(2) ± 290.30 × 10(2) CFU/mL in "unsuitable" frozen-thawed semen). The effect of SLC on kinetics of frozen-thawed sperm cells was negligible.

  17. Effect of age and environmental factors on semen quality, glutathione peroxidase activity and oxidative parameters in Simmental bulls.

    PubMed

    Balić, I Majić; Milinković-Tur, S; Samardžija, M; Vince, S

    2012-07-15

    Taking into account that semen quality depends on animal age and climate conditions and that oxidative stress has been reported to be a common cause of infertility, the objective of this study was to monitor indicators of oxidative stress and antioxidant protection during four seasonal periods in service bulls of various age to get better insight into the significance of these factors upon evaluating service bull semen. The research was conducted over a year on 19 Simmental service bulls. Animals were divided into two groups according to age; Group I consisted of younger bulls aged two to four yrs (n=9), and Group II was comprised of older bulls aged five to ten yrs (n=10). Semen samples were obtained once in the middle of every seasonal period and blood samples for biochemical analysis were collected by jugular venipuncture immediately after ejaculate collection. The activity of total glutathione peroxidase (T-GSH-Px), selenium-dependent glutathione peroxidase (Se-GSH-Px) and selenium-independent glutathione peroxidase (non-Se-GSH-Px), together with the intensity of lipid peroxidation (thiobarbituric acid reactive substances; TBARS) and oxidative protein damage (protein carbonyl content (PCC)) were measured in seminal plasma. In samples of spermatozoa and blood serum, the activity of Se-GSH-Px and TBARS and PCC concentrations were determined. Older service bulls had significantly higher ejaculate volume in summer in comparison with younger bulls, whereas the number of spermatozoa and progressive motility percentage did not significantly vary with age. Younger animals had lower progressive motility percentage during summer than in spring, with more intensive oxidative processes observed in seminal plasma (TBARS) and spermatozoa (TBARS and PCC). Based on the results presented here, it can be concluded that younger bulls are more sensitive to elevated ambient temperatures during the summer, when intensified prooxidative processes in semen plasma and spermatozoa

  18. Relationship between in vitro sperm functional assessments, seminal plasma composition, and field fertility after AI with either non-sorted or sex-sorted bull semen.

    PubMed

    Holden, S A; Fernandez-Fuertes, B; Murphy, C; Whelan, H; O'Gorman, A; Brennan, L; Butler, S T; Lonergan, P; Fair, S

    2017-01-01

    The hypothesis of this study was that different in vitro parameters are required to predict the in vivo fertility of non-sorted (NS) and sex-sorted (SS) semen. Thus, the aim was to correlate in vitro bull sperm functional parameters (experiment 1) and seminal plasma composition (experiment 2) with pregnancy rates using 2 cohorts of bulls (NS and SS). Experiment 1: ejaculates from each bull (n = 3 ejaculates per bull; n = 6 bulls for both NS and SS) were assessed for motility, thermal stress tolerance and morphology using microscopy, and viability, osmotic resistance, mitochondrial membrane potential, and acrosome integrity using flow cytometry. Fertilizing ability was assessed using IVF. Experiment 2: ejaculates (n = 3 per bull; n = 8 and 6 bulls for NS and SS, respectively) were collected, seminal plasma harvested and frozen and later analyzed for amino acid and fatty acid composition using gas chromatography mass spectrometry. In the NS cohort of bulls, there was no correlation between pregnancy rate and any of the sperm functional parameters assessed. However, within the SS cohort, motility and viability were correlated with pregnancy rate (r = 0.84 and 0.80, respectively; P < 0.05). There was no correlation between IVF outcome and pregnancy rate in either the SS or NS cohort of bulls. In the NS cohort of bulls, concentrations of the amino acid isoleucine and the fatty acid tricosylic acid (C23:0) were correlated with pregnancy rate (r = 0.80 and 0.74, respectively; P < 0.05). Within the SS cohort of bulls, the amino acid glutamic acid and the fatty acid arachidic acid (C20:0) were correlated with pregnancy rate (r = 0.84 and 0.82, respectively; P < 0.05). In conclusion, this study suggests that different in vitro markers of fertility are required to predict the fertility of NS and SS sperm.

  19. Birth of offspring after artificial insemination of heifers with frozen-thawed, sex-sorted, re-frozen-thawed bull sperm.

    PubMed

    Underwood, S L; Bathgate, R; Maxwell, W M C; Evans, G

    2010-04-01

    from frozen-thawed semen, and with further work, may be a promising technique that will give producers access to sexed sperm from a greater range of bulls.

  20. Comparison of Biociphos-Plus and TRIS-egg yolk extender for cryopreservation of bull semen.

    PubMed

    Thun, Rico; Hurtado, Maria; Janett, F

    2002-02-01

    For optimizing routine freezing of bull semen, we examined three different cryopreservation methods using either TRIS-egg yolk-citrate extender or Biociphos-Plus. Biociphos-Plus (IMV, France) has been marketed as an extender, in which egg yolk is replaced by a sterile soybean extract to reduce the contamination risk derived from animal borne substances. We used 78 bulls of various breeds (Brown Swiss, Holstein, Simmental) between 12 and 23 months of age, and we produced a total of 800-1000 straws (0.25 ml, 20 x 10(6) spermatozoa) from each bull using three different methods. In method A, we used TRIS-egg yolk as extender and packaged at 4 degrees C. In method B, we also used TRIS-egg yolk but packaged at room temperature (RT) between 18 and 22 degrees C. In method C, Biociphos-Plus served as extender and we packaged at RT. We compared methods A, B and C by using post-thaw motility, viability, morphology and osmotic resistance as semen quality parameters. In addition, we recorded 75-day nonreturn rates (NR75) to detect the effect of extenders on fertility. With the exception of primary defects, all laboratory parameters investigated were significantly (P < 0.05) better in methods A (TRIS-egg yolk, 4 degrees C) and B (TRIS-egg yolk, RT), compared to method C (Biociphos-Plus, RT). We recorded no significant difference between methods A and B. We could not verify the differing laboratory results by fertility data (NR75). However, when we analyzed NR75 for a single breed, significant (P < 0.05) differences existed between methods A and B compared to method C in Simmental and Holstein but not in Brown Swiss. We obtained best results in Simmental using method A (69%, n = 3384), while method C (61.4%, n = 763) was superior to methods A (57.6%, n = 698) and B (57.3%, n = 737) in Holstein. After considering various factors like preparation of extender, cost of materials and ambient working temperature, we concluded from our data that bull semen processing using TRIS-egg yolk

  1. Relationships between feed efficiency, scrotal circumference, and semen quality traits in yearling bulls.

    PubMed

    Hafla, A N; Lancaster, P A; Carstens, G E; Forrest, D W; Fox, J T; Forbes, T D A; Davis, M E; Randel, R D; Holloway, J W

    2012-11-01

    A meta-analysis was conducted to examine phenotypic relationships between feed efficiency, scrotal circumference, and semen quality traits in yearling bulls. Data evaluated were obtained from 5 postweaning trials involving Angus (n = 92), Bonsmara (n = 62), and Santa Gertrudis (n = 50) bulls fed diets that ranged from 1.70 to 2.85 Mcal ME/kg DM. After an adaptation period of 24 to 28 d, feed intake was measured daily, and BW was measured at 7- or 14-d intervals during the 70- to 77-d trials. Ultrasound carcass traits (12th-rib back fat thickness, BF; LM area, LMA) and scrotal circumference (SC) were measured at the start and end of each trial. Semen samples were collected by electroejaculation within 51 d of the end of the trials when the age of bulls averaged from 365 to 444 d and were evaluated for progressive sperm motility and morphology. Residual feed intake (RFI) was calculated as the difference between actual DMI and expected DMI from linear regression of DMI on ADG and midtest BW(0.75), with trial, trial by ADG, and trial by midtest BW(0.75) as random effects. Across all studies, bulls with low RFI phenotypes (<0.5 SD below the mean RFI of 0) consumed 20% less DM and had 10% less BF but had similar ADG, SC, and semen quality traits compared with high-RFI bulls (>0.5 SD above the mean RFI of 0). Gain to feed ratio was strongly correlated with ADG (0.60) and weakly correlated with initial BW (-0.17) and DMI (-0.26). Residual feed intake was not correlated with ADG, initial age, or BW but was correlated with DMI (0.71), G:F (-0.70), and BF (0.20). Initial SC (-0.20), gain in SC (-0.28), and percent normal sperm (-0.17) were correlated with G:F, but only sperm morphology was found to be weakly associated with RFI (0.13). These data suggest that RFI is not phenotypically associated with SC or sperm motility but is weakly associated with sperm morphology.

  2. Effect of species, breed, and age on bacterial load in bovine and bubaline semen

    PubMed Central

    Sannat, Chandrahas; Nair, Ajit; Sahu, S. B.; Sahasrabudhe, S. A.; Kumar, Ashish; Gupta, Amit Kumar; Shende, R. K.

    2015-01-01

    Aim: The present study was conducted to investigate the effect of species, breed and age on bacterial load in fresh and frozen semen of Cattle and Buffalo bull. Materials and Methods: Present study covered 56 cow and 10 buffalo bulls stationed at Central Semen Station Anjora, Durg (Chhattisgarh). Impact of breeds on bacterial load in semen was assessed using six breeds of cattle viz. Sahiwal, Gir, Red Sindhi, Tharparkar, Jersey and Holstein Friesian (HF) cross. Cow bulls were categorized into four different groups based on their age (<4 years, 4-5 years, 5-6 years and > 6 years) to study variation among age groups. Bacterial load was measured in fresh and frozen semen samples from these bulls using the standard plate count (SPC) method and count was expressed as colony forming unit (CFU) per ml of semen. Results: Higher bacterial load was reported in fresh (2.36 × 104 ± 1943 CFU/ml) and frozen (1.00 × 10 ± 90 CFU/ml) semen of cow bulls as compared to buffalo bulls (1.95 × 104 ± 2882 and 7.75 × 102 ± 160 CFU/ml in fresh and frozen semen, respectively). Jersey bull showed significantly higher bacterial count (p < 0.05) both in fresh (4.07 × 104 ± 13927 CFU/ml) and frozen (1.92 × 103 ± 178 CFU/ml) semen followed by HF cross, Sahiwal, Gir, Red Sindhi and Tharparkar bull. Bulls aged < 4 years and more than 6 years yielded increased bacterial load in their semen. Although a minor variation was reported between species and among age groups, no significant differences were measured. Conclusion: Bacterial load in semen did not differ significantly between species and age groups; however significant variation was reported among different breeds. Bulls of Jersey breed showed significantly higher bacterial load in semen as compared to the crossbred and indigenous bull. PMID:27047115

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

    PubMed

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

    2014-10-01

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

  4. A High Percentage of Beef Bull Pictures in Semen Catalogues Have Feet and Lower Legs that Are Not Visible

    PubMed Central

    Franks, Marcy K.; Grandin, Temple

    2015-01-01

    Simple Summary When cattle breeders purchase semen from a website, the only way they can visually appraise a bull’s conformation is by looking at his photograph. Correct foot and leg structure is important to help reduce lameness. Only 19.4% of the bull pictures on four major websites had fully visible feet and lower legs. A possible explanation for this may be deliberate covering of feet and legs with photo editing software to cover up conformation defects. Visibility of feet and lower legs would help semen buyers avoid bulls with obvious feet or leg problems. Abstract A total of 1379 beef bull pictures were surveyed to determine visibility of feet and legs from four American semen company websites. Five different breeds were represented: Angus, Red Angus, Hereford (polled and horned), Simmental, and Charolais. In addition to visibility, data on other variables were collected to establish frequencies and correlations. These included breed, color, material that obscured visibility, such as grass, picture taken at livestock show or outside, semen company, photographer, video, and age of bull. A foot and leg visibility score was given to each bull picture. Only 19.4% of the pictures had fully visible feet and legs. Both the hooves and dewclaws were hidden on 32.5% of the pictures. Correlation between bull’s birthdate and the first four visibility scores was statistically significant (P < 0.0001). As age increased the feet and legs were more likely to be visible in the bull’s picture. This may possibly be due to greater availability of both photo editing software and digital photography. One positive finding was that 6% of the bulls had a video of the bull walking which completely showed his feet and legs. PMID:26479372

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

    PubMed

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

    2004-01-01

    In this study the concentration of copper, iron, zinc, cadmium, lead, and nickel in bull and ram semen and relation of these metals to spermatozoa morphology was investigated. Analysis by atomic absorption spectrophotometry showed that copper concentration was significantly higher (p<0.0001) in ram semen in comparison with bull semen. The zinc concentration was higher in bull semen in comparison with ram semen. The iron and cadmium concentrations in the semen were similar. Higher concentration of lead was found in ram semen. Higher levels of nickel were found in ram semen in comparison with bulls. In bull semen 11.79+/-4.88% of pathological spermatozoa was found. Higher occurrence of pathological spermatozoa was in ram semen (17.17+/-3.76) in comparison with the semen of bulls. Separated tail, tail torso, and knob twisted tail were the most frequent forms of pathological spermatozoa in both species. Correlation analysis in bulls showed high positive relation between iron and zinc (r = 0.72), nickel and separated tail (r = 0.76), separated tail and tail torso (r = 0.71), tail torso and total number of pathological spermatozoa (r=0.72), and between tail ball and total number of pathological spermatozoa (r = 0.78). In rams high positive correlation between cadmium and lead (r=0.98), nickel and separated tail (r=0.77), separated tail and total number of pathological spermatozoa (r=0.69), knob twisted tail and retention of cytoplasmic drop (r=0.78), and between knob twisted tail and other pathological spermatozoa (r = 0.71) was found. High negative correlation in ram semen was observed between copper and nickel (r=0.71), copper and separated tail (r=0.70), and between iron and tail torso (r=0.67). The results suggest that the studied metals have a direct effect on spermatozoa quality.

  6. The use of beef bull semen reduced the risk of abortion in Neospora-seropositive dairy cows.

    PubMed

    López-Gatius, F; Santolaria, P; Yániz, J L; Garbayo, J M; Almería, S

    2005-03-01

    There is an evidence that the epidemiology of neosporosis differs in dairy and beef cattle, such that beef cattle carry a lower risk of abortion. The aim of the present study was to establish whether artificial insemination using semen from beef bulls could reduce the risk of abortion in dairy cows seropositive for the Neospora caninum parasite. Our study was based on yearly serological screening for neosporosis and on the confirmation of Neospora infection in aborted fetuses in two high-producing dairy herds with a mean 28% seroprevalence of N. caninum antibodies. The study population comprised of 273 pregnancies in seropositive animals: 156 pregnancies monitored after insemination using Holstein-Friesian semen and 117 after insemination using beef bull semen. Abortion rates for these animals were 28.2% (77 of 273), 34.6% (54 of 156) and 19.7% (23 of 117). Logistic regression analysis indicated no significant effects of lactation number and previous abortion on the abortion rate. Based on the odds ratio, a 1-unit increase in the Neospora antibody titre yielded a 1.01-fold increase in the abortion rate. The likelihood of abortion was two times higher for cows in one of the two herds and 2.8 times lower (one of 0.36) for pregnant cows inseminated with beef bull semen rather than Holstein-Friesian semen. Our results indicate that the use of beef bull semen can reduce the risk of abortion in dairy cows, and suggest that annual screening for neosporosis, specifically the antibody titre to the protozoan, could be an useful predictor of abortion risk in reproductive health programmes.

  7. Genome-wide association study for semen volume and total number of sperm in Holstein-Friesian bulls.

    PubMed

    Hering, D M; Oleński, K; Ruść, A; Kaminski, S

    2014-12-30

    In artificial insemination industry bulls producing high volume of semen with relatively high concentration of sperm are very desirable since they ensure stable production of commercial straws especially in case of top bulls. The aim of the study was to screen the entire bull genome to identify markers and candidate genes underlying semen volume (SV) and total number of sperm (TNS) in ejaculate produced by Holstein-Friesian bulls. Data on semen production were retrieved from records of AI center and included a population of 877 Holstein-Friesian bulls. Each bull was genotyped using the Illumina BovineSNP50 BeadChip. Genome-wide association analysis was performed with the use of GoldenHelix SVS7 software. An additive model for Linear Regression Analysis was used to estimate the effect of SNP marker for SV and TNS. After Bonferroni correction, 3 markers located on chromosome 22 reached the highest significance (rs41625599, rs41584616, rs42012507) for both traits. In the vicinity of these significant markers 3 genes are located (DCP1A, SFMBT1, TMEM110). Moreover, marker rs110109069 located on chromosome 25 was significantly associated with TNS and marker rs42438348 located on chromosome 10 has been found to be associated with SV. Some additional candidate genes were suggested to be potentially involved in analyzed traits (GALC, PRKCD, PHF7, TLR9, SPATA7). Identifying SNPs associated with the lower total number of sperm may be very useful for early recognition of a young sire as less suitable for effective semen production in artificial insemination centers.

  8. Successful preservation of capercaillie (Tetrao urogallus L.) semen in liquid and frozen states.

    PubMed

    Kowalczyk, Artur; Łukaszewicz, Ewa; Rzońca, Zenon

    2012-03-15

    Experiments on semen collection and preservation were undertaken by Wrocław University of Environmental and Life Sciences and Forestry Wisła, Poland to assist in the protection of the capercaillie (Tetrao urogallus L.) and to create an ex situ in vitro cryobank. Semen was collected from 11 captive-bred males, using dorsoabdominal massage. Ejaculates once obtained were diluted 3-fold at room temperature with EK diluent and then a number of them were stored at 4 °C for 18, 24, and 48 hours, while the remaining ejaculates were equilibrated with 6% dimethylacetamide and frozen by pipetting, drop-by-drop directly onto a liquid nitrogen surface. Frozen pellets were thawed at 60 °C in a water bath after 4 to 28 mo of storage. In total, 103 individually collected ejaculates (54 stored as liquid and 49 frozen in liquid nitrogen) were of appropriate value for further processing. The volume of ejaculates varied from 30 to 240 μL; spermatozoa concentration from 70 × 10(6) mL(-1) to 1950 × 10(6) mL(-1). The total amount of live spermatozoa in the fresh semen varied from 85.3% to 99.0%, of which from 41.1% to 85.3% were morphologically normal. Among morphologically abnormal forms, bulb-head (5.6% to 36.0%) and midpiece deformations (1.3% to 16.6%) were the most frequent. Dilution and semen storage up to 24 h at 4 °C did not affect the semen quality, as far as motility and sperm morphology are concerned. A significant (P < 0.05) decrease in total live (94.9 vs. 91.7%) and live normal cells (66.4 vs. 56.7%) was observed after 48 h. About 30% to 40% of spermatozoa remained motile. Cryopreservation significantly decreased (P < 0.05) the total number of live and live normal spermatozoa however, in relation to the fresh semen, their average content was 44.1% and 37.4%, respectively. Significant (P < 0.05) individual differences were observed in the quality of the fresh, liquid stored and the frozen-thawed semen assessed in terms of spermatozoa motility and morphology. After a

  9. Effect of seminal plasma vesicular structures in canine frozen-thawed semen.

    PubMed

    Goericke-Pesch, S; Hauck, S; Failing, K; Wehrend, A

    2015-12-01

    Membrane vesicles (MVs) in the ejaculate have been identified in various species and are considered to affect membrane fluidity due to their characteristic molecular composition. Addition of MV to human frozen semen has been shown to improve post-thaw motility. Similarly, a beneficial effect has been suggested for frozen equine semen. As post-thaw canine semen quality varies widely between dogs, the aim of our study was to test for the effect of addition of canine MV on post-thaw semen quality in dogs. Semen samples from 10 male dogs were purified from MV and prepared for freezing. In experiment 1, three groups were compared: sperm frozen (1) with MV (S1); (2) without MV, but MV added immediately after thawing (S2); and (3) without MV (C). Semen analysis included computer-assisted sperm analysis of motility parameters immediately after thawing (t0), after 10 (t10) and 30 minutes (t30), % living sperm, % membrane intact, % morphologically normal sperm (all t0 and t30). Computer-assisted sperm analysis motility distance and velocity parameters (all P < 0.05) and % living sperm (P < 0.001) were significantly affected by treatment with a temporary increase of distance and velocity parameters at t0 to t10, but a significant decrease of the aforementioned parameters at t30 in samples with MV. In experiment 2, different MV protein concentrations added after thawing were compared: 0.05 mg, 0.1 mg, and 0.2 mg/mL. Computer-assisted sperm motility analysis was performed at t0, t10, and t30. No differences between MV concentrations were identified, only a significant interaction between effect of treatment and time for progressive motility (P < 0.01). Our study identified a short-term beneficial effect of canine MV on post-thaw distance and velocity parameters, whereas at t30 progressive motility, motility parameters and % living sperm were reduced in samples with MV compared to C. The results point to species-specific differences regarding the MV effect on frozen

  10. Cryopreservation of bull semen: Evolution from egg yolk based to soybean based extenders.

    PubMed

    Layek, S S; Mohanty, T K; Kumaresan, A; Parks, J E

    2016-09-01

    Since the inception of bovine semen cryopreservation, egg yolk and milk based extenders have been used to protect sperm from the detrimental effects of cooling and freezing. In recent years, demand for alternatives to conventional commercial extenders has arisen as the risk of introducing exotic diseases through transporting egg yolk based products has been recognized. Egg yolk can also interfere with sperm evaluation and the presence of particulate material in the extender may reduce fertility. Soybeans contain lecithin, a phospholipid fraction that can substitute for high molecular weight lipoprotein and phospholipids from egg yolk and prevent or ameliorate damage to the sperm plasma membrane that occurs during extension, cooling, and cryopreservation. Soy lecithin based extenders have been evaluated for processing and freezing bovine semen, although extender from soybean milk has not been studied as extensively. Commercially available soy lecithin based extenders are used increasingly but remain under scrutiny and are not universally accepted. With these observations in mind, this review is intended to examine effects of conventional cryopreservation procedures, methods of assessment, and potential for developing soybean extract as an acceptable alternative to traditional egg yolk and milk based extenders for bull sperm cryopreservation.

  11. Toxoplasma gondii transmission by artificial insemination in sheep with experimentally contaminated frozen semen.

    PubMed

    Consalter, Angélica; Silva, Andressa F; Frazão-Teixeira, Edwards; Matos, Luis F; de Oliveira, Francisco C R; Leite, Juliana S; Silva, Franciele B F; Ferreira, Ana M R

    2017-03-01

    Toxoplasma gondii is a parasite considered one of the major causes of reproductive problems in sheep. Furthermore, the presence of the agent in ram semen urges the possibility of sexual transmission in this species. The aim of this study was to evaluate if ram's frozen semen spiked with T. gondii tachyzoites would be able to cause infection in sheep by laparoscopic artificial insemination (AI). Nine ewes tested seronegative to anti-T. gondii antibodies by the modified agglutination test (MAT) were superovulated and inseminated to collect embryos. Animals were divided into two groups: G1 (n = 5), ewes inseminated with semen containing 4 × 10(7) tachyzoites; and G2 (n = 4), ewes inseminated with tachyzoite-free semen (control group). To confirm infection, ewe's blood samples were collected on days -14, -7, 0, 7, 14, 21, 28, 35, 49 and 57 after AI for analysis by MAT and PCR. Tissue samples of these ewes were also collected for histopathology and immunohistochemistry (IHC). Seven days after AI, all ewes of group G1 had specific antibodies to T. gondii, while those of G2 were negative. Toxoplasma gondii DNA was detected in the blood of one ewe and parasites were observed in tissues of all five animals inseminated with contaminated semen, indicating that semen freezing protocol does not affect T. gondii transmission by artificial insemination in sheep.

  12. Strategies to improve the fertility of fresh and frozen donkey semen.

    PubMed

    de Oliveira, José Victor; Oliveira, Pedro Victor de Luna Freire; Melo e Oña, Cely Marini; Guasti, Priscilla Nascimento; Monteiro, Gabriel Augusto; Sancler da Silva, Yamê Fabres Robaina; Papa, Patrícia de Mello; Alvarenga, Marco Antônio; Dell'Aqua Junior, Jose Antonio; Papa, Frederico Ozanam

    2016-04-15

    Fertility rates of donkey semen in jennies are lower compared to mares. The aims of this study were to evaluate different sperm cryopreservation methods and insemination strategies to improve the fertility of donkey semen in jennies. Three experiments were performed: (1) the comparison of two freezing methods of donkey semen (conventional method and automated method); (2) the determination of a suitable insemination dose of fresh donkey semen for jennies and mares; and (3) the influence of the semen deposition site on fertility of jennies inseminated with frozen donkey semen. For experiment 1, no differences were observed in total motility, angular velocity, curvilinear velocity, straight-line velocity, and plasma membrane integrity between samples frozen with the conventional (Styrofoam box) and the automated method (TK 4000C). However, the automated method provided higher values of progressive motility and rapid cells in frozen-thawed samples in comparison with the conventional method (P < 0.05). For experiment 2, mares were bred using 500 × 10(6) fresh sperm (M); and jennies using 1 × 10(9) (J1) or 500 × 10(6) fresh sperm (J5). Pregnancy rates in M, J1, and J5 were 93% (14/15), 73% (11/15), and 40% (6/15), respectively. When using different insemination doses, 500 × 10(6) or 1 × 10(9) sperm, no significant difference was observed in pregnancy rates of mares (M, 14/15) and jennies (J1, 11/15). Furthermore, there was no significant difference between the two insemination doses in jennies. However, with an insemination dose of 500 × 10(6) fresh sperm, the pregnancy rates were significantly higher in mares (M, 14/15) than in jennies (J5, 6/15; P < 0.05). For experiment 3, the inseminations were carried out in the uterine body (UB) or in the uterine horn of jennies with frozen-thawed donkey semen. No pregnancies were achieved with inseminations performed in the UB (0/12). The pregnancy rate for uterine horn group was 28.26% (13/46) and thus

  13. Artificial insemination of red deer (Cervus elaphus) with frozen-thawed wapiti semen.

    PubMed

    Haigh, J C; Bowen, G

    1991-09-01

    Semen collected from wapiti (Cervus elaphus) in Canada in 1983 was frozen in two extenders. In 1988, the semen was used to inseminate 200 red deer hinds on 2 farms in New Zealand. Oestrus was synchronized in the hinds with progesterone-impregnated intravaginal devices (CIDR); 200 iu pregnant mares' serum gonadotrophin was given to each hind on Day 11. The CIDRs were removed on Day 12 at 20/h, as the numbers of the hinds were recorded. On Day 14, 54-56 h after CIDR removal, the hinds were brought into the yards in the same batches and laparoscopically inseminated. Semen from three sires was used. The overall conception rate was 51%. Gestation length ranged from 239 to 247 days. One hind was lost at calving, 3 calves had to be hand raised and there were 2 neonatal calf deaths.

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

    PubMed

    Pan, Qing; Ju, Zhihua; Huang, Jinming; Zhang, Yan; Qi, Chao; Gao, Qin; Zhou, Lei; Li, Qiuling; Wang, Lingling; Zhong, Jifeng; Liu, Mei; Wang, Changfa

    2013-01-01

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

  15. Comparison of different methods for assessment of sperm concentration and membrane integrity with bull semen.

    PubMed

    Anzar, Muhammad; Kroetsch, Tom; Buhr, Mary M

    2009-01-01

    Assessing semen quality is crucially important for the exploitation of genetically superior sires in an artificial insemination (AI) program. In this study, we compare modern and conventional techniques to estimate bovine sperm concentration and membrane integrity. First, the NucleoCounter SP-100 was validated for sperm concentration and provided statistically reliable and repeatable estimates among aliquots and replicates of 25 fresh ejaculates. Sperm concentrations in 78 ejaculates were then determined with hemacytometer, flow cytometer, and NucleoCounter SP-100 and were significantly correlated (P < .001), with regression coefficients among these 3 techniques close to 1 (P < .01). However, the sperm concentration determined by hemacytometer was lower (P < .01) than by flow cytometer and NucleoCounter SP-100. Forty frozen-thawed semen samples were then assessed for sperm concentration and membrane integrity with hemacytometer, flow cytometer and NucleoCounter SP-100. Significant relationships were found for sperm concentration determined by hemacytometer and NucleoCounter SP-100 and for sperm membrane integrity determined by flow cytometer and NucleoCounter SP-100 (P < .01). Finally, the standard curves of sperm concentrations in 6 spectrophotometers, comparing optical density against counts drawn by hemacytometer and NucleoCounter SP-100 (n = 94 fresh ejaculates) showed different (P < .01) intercepts and regression coefficients (linear, quadratic, cubic). It was calculated that a breeding station can improve its production potential by 13% with the use of NucleoCounter SP-100 instead of hemacytometer for calibration of spectrophotometers. Flow cytometer and NucleoCounter SP-100 can be used with equal confidence to estimate sperm concentration and membrane integrity in domestic animals and human semen.

  16. Quantitative Assessment of the Risk of Release of Foot-and-Mouth Disease Virus via Export of Bull Semen from Israel.

    PubMed

    Meyer, A; Zamir, L; Ben Yair Gilboa, A; Gelman, B; Pfeiffer, D U; Vergne, T

    2017-03-23

    Various foot-and-mouth disease (FMD) virus strains circulate in the Middle East, causing frequent episodes of FMD outbreaks among Israeli livestock. Since the virus is highly resistant in semen, artificial insemination with contaminated bull semen may lead to the infection of the receiver cow. As a non-FMD-free country with vaccination, Israel is currently engaged in trading bull semen only with countries of the same status. The purpose of this study was to assess the risk of release of FMD virus through export of bull semen in order to estimate the risk for FMD-free countries considering purchasing Israeli bull semen. A stochastic risk assessment model was used to estimate this risk, defined as the annual likelihood of exporting at least one ejaculate of bull semen contaminated with viable FMD virus. A total of 45 scenarios were assessed to account for uncertainty and variability around specific parameter estimates and to evaluate the effect of various mitigation measures, such as performing a preexport test on semen ejaculates. Under the most plausible scenario, the annual likelihood of exporting bull semen contaminated with FMD virus had a median of 1.3 * 10(-7) for an export of 100 ejaculates per year. This corresponds to one infected ejaculate exported every 7 million years. Under the worst-case scenario, the median of the risk rose to 7.9 * 10(-5) , which is equivalent to the export of one infected ejaculate every 12,000 years. Sensitivity analysis indicated that the most influential parameter is the probability of viral excretion in infected bulls.

  17. Differences in ability of jennies and mares to conceive with cooled and frozen semen containing glycerol or not.

    PubMed

    Vidament, Marianne; Vincent, Pierrick; Martin, François-Xavier; Magistrini, Michele; Blesbois, Elisabeth

    2009-05-01

    A suitable method for the cryopreservation of donkey semen would be very valuable for the ex situ management of genetic diversity in this species. This report uses a variety of observation and trials to evaluate the effect of cryoprotectants in per-cycle pregnancy rates (PC) in equids females (jennies (donkey) and mares (horse)). This was explored by (1) comparing the results of insemination of jennies and mares with cooled or frozen donkey semen, (2) examining the possible toxic effect of the cryoprotectant (CPA) glycerol in these two species and (3) studying alternative solutions. Donkey and horse semen was either used immediately, or cooled according to some steps of the pre-freezing procedure or frozen and thawed. The pre-freezing procedure included semen dilution, centrifugation, resuspension in milk or in INRA82+2% egg yolk+various % CPA (expressed as final concentrations in extended semen (v/v)) and then cooling to 4 degrees C. PC was similar in mares and jennies inseminated with donkey semen cooled to 4 degrees C in milk. However, the PC was significantly higher in mares than in jennies when donkey semen was frozen with 2.2% glycerol (36%, n=50 cycles vs. 11%, n=38 cycles; P<0.01). Increasing the concentrations of glycerol (0, 2.2, 3.5, 4.8%) before cooling stallion semen resulted in a progressive decrease in mare PC (87, 53, 53, 13% (n=15 cycles for each concentration); P<0.0001). The addition of 2.2% glycerol before cooling donkey semen decreased the PC measured in jennies to 0. The replacement of glycerol by 2% dimethylformamide increased the fertility obtained in jennies with cooled donkey semen (PC: 67%, n=12 cycles) but did not increase the fertility obtained with frozen-thawed donkey semen (PC: 11%, n=28 cycles with dimethylformamide vs. 0%, n=16 cycles with glycerol). In conclusion, this study clearly shows that the ability of jennies to conceive after AI with donkey frozen semen is lower than that of mares. Glycerol affects the fertility of donkey

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

    PubMed

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

    2016-07-01

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

  19. Intravaginal artificial insemination in bitches using frozen/thawed semen after dilution in powdered coconut water (ACP-106c).

    PubMed

    Uchoa, D C; Silva, T F P; Mota Filho, A C; Silva, L D M

    2012-12-01

    The aim of this study was to evaluate powdered coconut water extender (ACP-106c; ACP Serviços Tecnológicos Ltda, ACP Biotecnologia, Fortaleza, Ceará, Brazil) as a diluent for freezing dog semen and the fertility after vaginal insemination of semen frozen therein. Ten ejaculates were collected from five dogs, evaluated fresh, diluted in ACP-106c, 10% egg yolk and 6% glycerol, cooled and frozen. In the first phase of the study, straws with frozen semen were thawed and immediately subjected to the same analysis as the fresh semen and, in addition, to Computer-Assisted Semen Analysis (CASA). In phase 2, 10 bitches that had been subjected to natural breeding during a preceding oestrous cycle were vaginally inseminated with thawed semen that had been re-diluted in ACP-106c. After thawing, a mean of 77% sperm motility was obtained through subjective analysis and 77.3% through CASA. Following artificial insemination, a 60% pregnancy rate was observed, resulting in a 50% parturition rate and a mean litter size of 3.4 (SEM 0.6), with 47.1% males and 52.9% females. ACP-106c can be successfully used for freezing canine semen, and vaginal deposition of such semen yields similar pregnancy rates to those reported in other studies.

  20. Estimation of genetic parameters and effects of cytoplasmic line on scrotal circumference and semen quality traits in Angus bulls.

    PubMed

    Garmyn, A J; Moser, D W; Christmas, R A; Minick Bormann, J

    2011-03-01

    The purpose of this study was to estimate the heritability of scrotal circumference (SC) and semen traits, genetic correlations between SC and semen quality traits, and the effect of cytoplasmic line on SC and semen traits. Breeding soundness exam (BSE) data were collected on registered Angus bulls at 4 ranches over 7 yr. The American Angus Association provided historical pedigree information to estimate the effect of cytoplasmic line on SC and semen quality traits. After editing, the evaluated data set contained 1,281 bulls with breeding soundness exam data that traced back to 100 founder dams. Data were analyzed using a 2-trait animal model to obtain heritability, genetic correlation between SC and semen quality traits, as well as the effect of cytoplasmic line as a random effect for SC, percent motility (MOT), percent primary abnormalities (PRIM), percent secondary abnormalities (SEC), and percent total abnormalities (TOT) using multiple-trait derivative-free REML. Fixed effects included source ranch and collection year, and test age was used as a covariate. Estimates of heritability for SC, MOT, PRIM, SEC, and TOT were 0.46, 0.05, 0.27, 0.23, and 0.25, respectively. Genetic correlations between SC and MOT, PRIM, SEC, and TOT were 0.36, -0.19, -0.11, and -0.23, respectively. The proportions of phenotypic variance accounted for by cytoplasmic line for SC, MOT, PRIM, SEC, and TOT were <0.001, 0.013, 0.023, 0.002, and <0.001, respectively. Genetic correlations between SC and semen quality traits were low to moderate and favorable. Cytoplasmic line may have a marginal effect on MOT and PRIM, but is likely not a significant source of variation for SC, SEC, or TOT.

  1. Development of extender and techniques for frozen turkey semen. 1. Development.

    PubMed

    Graham, E F; Nelson, D S; Schmehl, M K

    1982-03-01

    An extender for turkey semen with a frozen-thawed recovery of greater than or equal to 50% motile spermatozoa and a vigorous swirl was developed. The effect of glucose, a comparison of 10 different sugars in the extender, the use of sodium acetate and potassium acetate, the ratio of dimethylsulfoxide to ethylene glycol, the percent total cryoprotectant, equilibration time, and osmotic pressure were all tested. Replacement of approximately half of the extender with an iso-osmolar glucose solution yielded higher percentages of motile spermatozoa with both fresh and frozen-thawed semen samples. Replacement of glucose with other carbohydrates did not enhance recovery of frozen-thawed semen. The proportion of sodium and potassium acetates to TESNaK2 in the extender had no effect on motility. Approximately equal proportions of ethylene glycol to dimethylsulfoxide, with a final concentration of 11.2% after dilution, produced at or near optimal recovery of spermatozoal motility after freezing. Twenty to 90 min equilibration times before freezing yielded significantly higher recovery of motile sperm postthaw than shorter periods. A wide range of extender osmotic pressures were compatible with recovery of spermatozoal motility postthaw. The final extender consisted of 4.70 g sodium acetate, 3.39 g potassium acetate 9.23 g TES, .353 g sodium hydroxide, .492 g potassium hydroxide, 32.00 g glucose, H2O q.s. 1000 ml, 370 mOsm/kg, pH 7.2. Ethylene glycol and dimethylsulfoxide were added (1:1 ratio) for a final concentration of 11.2% cryoprotectant after dilution. Semen was held undiluted for 6 min, extended 1:4 at 22 C, and equilibrated for 30 min at 0 C before pellet freezing.

  2. Effects of package, extender, and light on stored frozen bull spermatozoa.

    PubMed

    Coulter, G H; Foote, R H

    1977-09-01

    Spermatozoa were analyzed when fresh, freshly frozen, and stored at --196 C for 6 and 18 mo after processing in egg yolk-citrate and Promine-D extender with varying exposure to light and packaging in straws or glass ampules. Over all treatments there were decreases with time in spermatozoal motility, oxygen uptake, and cellular glutamic oxaloacetic transaminase due to freezing-thawing. Percent progressive motility decreased from spermatozoa freshly frozen (36.5%) to those stored for 6 mo (33.6%). Oxygen uptake in freshly frozen samples was 8.4 microliter per 10(8) cells per h. In frozen semen stored for 6 and 18 mo corresponding values were 12.2 and 8.3 microliter. Packaging methods did not have a significant effect on oxygen uptake, but package interacted with storage. Egg yolk-citrate extender supported greater progressive spermatoal motility than did promine-D. Extender interacted with storage for all criteria. Visible light radiation reduced progressive spermatozoal motility from 40.2 to 38.0% and oxygen uptake from 11.8 to 10.8 microliter per 10(8) cells per h. Light interacted with storage time. Correlations between all criteria and storage times were not important practically.

  3. Comparative fertility of freshly collected vs frozen-thawed semen with laparoscopic oviductal artificial insemination in domestic cats.

    PubMed

    Lambo, C A; Grahn, R A; Lyons, L A; Bateman, H l; Newsom, J; Swanson, W F

    2012-12-01

    Artificial insemination (AI) is potentially invaluable as an adjunct to natural breeding for the conservation management of non-domestic felid populations. The efficacy of AI, however, must be substantially improved for applied use, especially when using frozen semen. Our recent advances in using laparoscopic oviductal AI (LO-AI) with low sperm numbers and freezing of cat semen in a soy lecithin-based cryoprotectant medium suggest that combining these two approaches might improve pregnancy outcomes with frozen-thawed spermatozoa. In this study, our objectives were to (i) assess the effect of two gonadotropin dosages (100 vs 150 IU eCG) on ovarian response in domestic cats and (ii) compare the relative fertility of frozen-thawed and fresh semen in vivo following LO-AI. All 16 females ovulated after gonadotropin treatment and were inseminated with fresh semen from one male and frozen-thawed semen from a second male. There were no differences between gonadotropin dosages in CL number, pregnancy percentage or litter size. Half (8/16) of the females conceived, with seven females giving birth to a total of 36 offspring. Paternity analysis showed that more kittens resulted from LO-AI with fresh (28/36, 78%) than frozen-thawed (8/36, 22%) semen, possibly due to impaired motility and longevity of thawed sperm. These results demonstrated that viable offspring can be produced by AI using semen frozen in a soy lecithin-based medium. Insemination with greater numbers of frozen-thawed spermatozoa, combined with further refinement of cat sperm cryopreservation methods, may be necessary to optimize pregnancy success with LO-AI in domestic and nondomestic cats.

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

    PubMed

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

    2016-08-01

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

  5. Comparing ethylene glycol with glycerol and with or without dithiothreitol and sucrose for cryopreservation of bull semen in egg-yolk containing extenders.

    PubMed

    Büyükleblebici, Serhat; Tuncer, Pürhan Barbaros; Bucak, Mustafa N; Taşdemir, Umut; Eken, Ayşe; Büyükleblebici, Olga; Durmaz, Emre; Sarıözkan, Serpil; Endirlik, Burcu Ü

    2014-08-01

    There are few studies performed for investigating the roles of different ratio and cryoprotectants with dithiothreitol or sucrose on sperm motility characteristics and antioxidant capacities of post-thawed bull spermatozoa. The objectives of this study were to compare glycerol (G) and ethylene glycol (EG) at different concentrations as cryoprotectants and dithiothreitol (D) or sucrose (S) (with/without) as antioxidants in Tris extender for cryopreservation of bull semen. Twenty-four ejaculates obtained from three bulls were included in the study. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and diluted using both of the dithiothreitol 5mM or sucrose 25 mM, and control (without additives) was cooled to 4 °C and frozen in 0.25-ml French straws. when compared to control, different doses cryoprotectants and antioxidants addition no significantly increased the percentages of post-thaw sperm progressive and motitilities, acrosome abnormality and plasma membrane integrity (P>0.05). However, EG3+S yielded the greatest percentages of the total abnormality (P<0.05). As regard to antioxidant activities G7 and EG5 led to lowest MDA activity with or without D or S but, these results were not supported to the GPx activity (P<0.01). The sperm motion characteristics such as VAP, VCL, ALH and BCF gave significantly different results (P<0.05). When compared the DNA integrity, different doses cryoprotectants without antioxidants addition significantly increased the percentages of the tail intensity and tail moment (P<0.05). There were no significant differences observed in non-return rates among all treatment groups (P>0.05).

  6. Influence of cool storage before freezing on the quality of frozen-thawed semen samples in dogs.

    PubMed

    Santana, M; Batista, M; Alamo, D; González, F; Niño, T; Cabrera, F; Gracia, A

    2013-02-01

    The aim of this study was to determinate the semen quality of frozen-thawed samples that were chilled for up to 2 days before freezing. The ejaculates (n = 18) from six dogs were collected, pooled and divided into six aliquots. The first aliquot (C, control) was frozen in liquid nitrogen using a conventional protocol to reach a final concentration of 100 × 10(6) spermatozoa/ml, 20% egg yolk and 5% glycerol. The remaining five aliquots were diluted with a chilled extender (Tris-glucose and 20% egg yolk) and cooled at 4 °C as follows: R1, the semen was cooled for 1 h; R6, the semen was cooled for 6 h; R12, the semen was cooled for 12 h; R24, the semen was cooled for 24 h and R48, the semen was cooled for 48 h. After the chilling period, a second extender was added (Tris-glucose, 20% egg yolk, 10% glycerol and Equex at 1%) to reach a final composition similar to aliquot C, and then, the semen samples (R1, R6, R12, R24 and R48) were frozen in liquid nitrogen. The post-thaw sperm quality was assessed in 30 straws from each experimental group. After freezing-thawing, the total sperm motility (approximately 60-70%) in the semen chilled for up to 48 h did not show any differences from the samples frozen by the conventional cryopreservation method (63.2%). No significant differences were detected in the percentages of abnormal sperm cells among the fresh semen, the control group and the frozen samples after the different cooling times. Finally, the post-thaw percentages of damaged acrosomes showed a very uniform distribution, with mean values ranging between 7% and 10.5%. The results clearly demonstrated that cooling the semen up to 48 h before freezing did not produce a decrease in the semen quality when was compared with semen frozen by a traditional procedure.

  7. Separation of motile spermatozoa from frozen-thawed buffalo semen: swim-up vs filtration procedures.

    PubMed

    Mustafa, G; Anzar, M; Arslan, M

    1998-07-15

    Three experiments were conducted to maximize the recovery rate of motile spermatozoa from frozen-thawed buffalo semen. In Experiment 1, the swim-up of motile spermatozoa was performed in the presence or absence of HEPES in TALP medium and CO2 in the environment. The recovery rate of motile spermatozoa in TALP medium (control), TALP + HEPES + CO2, TALP + HEPES and TALP + CO2 was 15, 18, 12 and 10%, respectively (P > 0.05), with sperm motility at 87, 89, 90 and 90%, respectively (P > 0.05). In Experiment 2, the pH of TALP medium was adjusted to 7.0, 7.5, 8.0, 8.5 and 9.0, and swim-up procedure was performed in the presence of HEPES and CO2. The recovery rate of motile spermatozoa at different pH was 14, 20, 24, 27 and 16%, respectively (P < 0.05). Motility of separated spermatozoa was 88, 91, 90, 89 and 90%, respectively (P > 0.05). In Experiment 3, the efficiency of ion-exchange filtration and Swim-up procedure in separating motile spermatozoa from frozen-thawed buffalo semen was compared. The recovery rate of motile spermatozoa was 95% in filtration procedure and 33% in swim-up procedure (P < 0.005). In all experiments, normal acrosomes did not vary due to treatments (P > 0.05). In conclusion, HEPES and CO2 had no significant effect on swim-up of buffalo spermatozoa. The pH 8.5 of TALP improved the recovery rate of motile spermatozoa in swim-up procedure. The ion-exchange filtration was found superior to swim-up procedure in harvesting maximum number of motile spermatozoa from frozen-thawed buffalo semen (95 vs 33%; P < 0.001).

  8. Effect of ketoprofen treatment on the uterine inflammatory response after AI of jennies with frozen semen.

    PubMed

    Vilés, K; Rabanal, R; Rodríguez-Prado, M; Miró, J

    2013-04-15

    Artificial insemination (AI) involving the placing of frozen-thawed semen directly into the jenny uterine body is associated with very low pregnancy rates. This might be because of an exacerbation of the acute response of the endometrium to sperm, as seen in mares with persistent induced mating endometritis. Pregnancy rates can be increased in such mares, however, by including anti-inflammatory treatments in the insemination protocol (Bucca S, Carli A, Buckley T, Dolci G, Fogarty U. The use of dexamethasone administered to mares at breeding time in the modulation of persistent mating induced endometritis. Theriogenology 2008;70:1093-100; Rojer H, Aurich C. Treatment of persistent mating-induced endometritis in mares with the non-steroid anti-inflammatory drug vedaprofen. Reprod Domest Anim 2010;45:e458-60). To investigate the endometritis caused by the use of frozen-thawed semen in jennies, and to assess the response to ketoprofen treatment, endometrial cytological samples and biopsies from six healthy jennies were examined in a crossover design experiment. Samples were taken from jennies in estrus (E; control) and at 6 hours after AI with or without ketoprofen (+K and -K, respectively). Ketoprofen was administered iv 24 hours before and for 4 days after insemination (total = 2.2 mg/kg/24 hours for 5 days). All animals showed a severe inflammatory response to semen deposition. Polymorphonuclear neutrophil numbers in the cytological smears and biopsies differed significantly between the +K and E animals. No significant differences were recorded, however, between the +K and -K treatments. Eosinophils were observed in all sample types from all groups; these cells appear to be a feature of the normal jenny endometrium. Slight fibrosis was observed in some biopsies, but no significant relationship with inflammation was found. Intense cyclooxygenase-2 (COX-2) immunohistochemical labeling was detected in the -K biopsies. Less intense labeling was seen in those of the +K

  9. Chicks produced in the Magellanic penguin (Spheniscus magellanicus) after cloacal insemination of frozen-thawed semen.

    PubMed

    O'Brien, Justine Kellie; Steinman, Karen J; Montano, Gisele A; Dubach, Jean M; Robeck, Todd R

    2016-07-01

    The in vitro and in vivo functionality of cryopreserved spermatozoa was examined over two breeding seasons in a zoological colony of Magellanic penguins (Spheniscus magellanicus). Frozen-thawed semen was inseminated into five anesthetized females, over a total of eight egg production cycles, with a different male used for each artificial insemination (AI) within each season. Females were maintained within the colony in cordoned nest sites to prevent copulation with their paired male, and were inseminated every 3-10 days until the first oviposition. Semen frozen from seven males using a straw method retained 39.8%, 25.7%, 74.0%, and 52.1% of its initial total motility, progressive motility, average path velocity, and plasma membrane integrity, respectively. Normal morphology of motile cells was reduced (P < 0.05) during freeze-thawing from 76.7% immediately prior to freezing to 65.3% post-thawing. Conceptive females received 1.6 ± 0.2 inseminations before the first oviposition, with 19.2 ± 1.6 × 10(6) motile, morphologically normal spermatozoa per insemination. Overall fertility was 53.3% (8/15 eggs), hatchability was 50.0% (4/8), and genetic analyses confirmed that all embryos and hatchlings were sired by the AI male. Fertile eggs were laid at 4.0-12.1 days after AI, indicating that frozen-thawed spermatozoa resided in the female reproductive tract for up to ∼7.2 days prior to fertilization. Results demonstrate that frozen-thawed Magellanic penguin spermatozoa are fully functional in vivo and support the use of genome banking and AI as tools for managing the sustainability of zoological penguin populations. Zoo Biol. 35:326-338, 2016. © 2016 Wiley Periodicals, Inc.

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

    SciTech Connect

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

    1987-01-01

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

  11. Effect of heterologous seminal plasma and semen extenders on motility of frozen-thawed ram spermatozoa.

    PubMed

    Mataveia, G A; Terblanche, S J; Nöthling, J O; Gerber, D

    2010-09-01

    Ram seminal plasma increases the fertility of frozen-thawed ram spermatozoa deposited into the cervix. The aim of the current study was to compare the effect of ram seminal plasma to that of bull seminal plasma, dog prostatic fluid, protein-free TALP TrilEq (Triladyl with 0.5 mt of Equex STM paste added to each 100 mt) and heat-treated skim milk on longevity and percentages of progressively motile and aberrantly motile frozen-thawed ram spermatozoa. Three ejaculates from each of 6 rams were extended in TrilEq, pooled and frozen in straws as a single batch per ram. One hundred and eight straws (3 straws from each ram for each fluid) were thawed in random order. Once thawed, a straw was emptied into a tube with 0.85 ml of the appropriate fluid at 37 degrees C and kept at that temperature for 6 h. Motility was assessed at x200 magnification immediately (time zero) and 2, 4 and 6 h after thawing. Progressive motility decreased from each time to the next (P < 0.05) and was 39.0 % (0 h), 26.0 % (2 h), 19.6 % (4 h) and 12.6 % (6 h); SEM 1.24, n = 108 for each group. Ram seminal plasma resulted in higher progressive motility than bull seminal plasma, lower than milk, and similar to the other fluids. Ram seminal plasma resulted in lower aberrant motility than protein-free TALP and similar aberrant motility to other fluids. The effect of ram seminal plasma and dog prostatic fluid was very similar. The effect of ram seminal plasma on the fertility of frozen-thawed ram spermatozoa deposited into the cervix is not due an exceptionally beneficial effect on the motility of spermatozoa.

  12. Effect of semen collection by transrectal massage of accessory sexual glands or artificial vagina on the outcome of breeding soundness examinations of Italian yearling beef bulls.

    PubMed

    Sylla, Lakamy; Palombi, Claudio; Stradaioli, Giuseppe; Vagniluca, Antonio; Monaci, Maurizio

    2015-03-15

    Although semen quality is one of the major traits that influence breeding soundness examination outcomes in bulls, field conditions occasionally do not allow for the collection of semen samples by means of an artificial vagina. The aims of the present study were to report the results of a large number of semen collections that were performed via the transrectal massage (TRM) of the accessory sexual glands of Italian yearling beef bulls and compare this semen collection method to the artificial vagina (AV) method in term of breeding soundness examination outcomes; furthermore, we determined whether the breed affected the semen characteristics. In the TRM group (n = 475), the semen samples were collected via TRM of the accessory sexual glands, and in the AV group (n = 502), the AV method was used. In the TRM group, semen samples were obtained from 81.3% of the bulls and penile protrusion was observed in 87.6% of the animals during semen collection. The sperm concentrations (920.5 ± 439.0 vs. 281.0 ± 259.8 × 10(6)/mL) and the percentages of total abnormal spermatozoa (22.8 ± 15.0 vs. 18.8 ± 12.9) were significantly higher in the AV group than those in the TRM group. The percentage of bulls that did not meet the minimum requirement for normal cells (≥70%) was 6.2% higher in the AV group than that in the TRM group (P < 0.05). Moreover, the samples collected from Chianina bulls by TRM exhibited a lower percentage of motile sperm and a higher percentage of abnormal spermatozoa when compared with the other two breeds. The major drawbacks of the TRM technique were the inability to conduct complete evaluation of the libido and mating ability of the yearling bulls, a significant reduction of the number of spermatozoa collected, and an increase in the variability of the semen characteristics due to breed. In conclusion, despite the drawbacks, TRM guarantees that semen evaluation can be conducted in cases in which the semen samples cannot be collected

  13. Dimethylformamide as a cryoprotectant for canine semen diluted and frozen in ACP-106C.

    PubMed

    Mota Filho, A C; Teles, C H A; Jucá, R P; Cardoso, J F S; Uchoa, D C; Campello, C C; Silva, A R; Silva, L D M

    2011-10-15

    The objective was to assess the effect of adding various concentrations of dimethylformamide on characteristics of canine semen diluted in powdered coconut water (ACP-106C; ACP Biotecnologia, Fortaleza, CE, Brazil) and frozen at -196°C. Fifteen ejaculates were collected by manual stimulation from five adult Boxer dogs. The sperm-rich fraction was diluted in ACP-106C (ACP Biotecnologia) containing 10% egg yolk and divided into four aliquots. The cryoprotectants used for each aliquot were 6% glycerol (control group; CG) or 2%, 4%, or 6% dimethylformamide (DF2, DF4, and DF6, respectively). After thawing, total motility (mean ± SEM) for CG (58.4 ± 24.6) was higher (P < 0.05) than that of the other groups (2% dimethylformamide, 24.4 ± 12.3; 4% dimethylformamide, 26.5 ± 16.1; and 6% dimethylformamide, 21.7 ± 17.9). Furthermore, there was a greater percentage of fast, average, and slow moving sperm (assessed with computer-aided semen analysis; CASA) in CG in comparison with the other three groups. Therefore, based on concentrations tested in this study, dimethylformamide, together with ACP-106C (ACP Biotecnologia) and 10% egg yolk as a diluent, yielded unsatisfactory in vitro results for freezing canine semen.

  14. Intrauterine insemination of farmed fallow deer (Dama dama) with frozen-thawed semen via laparoscopy.

    PubMed

    Asher, G W; Kraemer, D C; Magyar, S J; Brunner, M; Moerbe, R; Giaquinto, M

    1990-09-01

    Estrus and ovulation of mature fallow does (n=155) on two North American farms were synchronized by intravaginal silastic devices containing 0.3 g progesterone (CIDR-type G) for 14 d. Each of 151 does received laparoscopic intrauterine inseminations of either 50x10(6) (n=125) or 25x10(6) (n=26) frozen-thawed spermatozoa, 65 to 68 h after CIDR device withdrawal. Four does received intrauterine inseminations per vaginam of 50x10(6) spermatozoa 68 to 69 hours after CIDR device withdrawal. Semen from crossbred Dama dama damaxDama dama mesopotamica sires was collected in New Zealand by electroejaculation. The overall pregnancy rate to artificial insemination, as assessed by rectal ultrasonography at Day 45, was 67.7%. The pregnancy rates for does receiving laparoscopic inseminations were 58.2% (Texas; 50x10(6) spermatozoa; n=79 does); 80.8% (Texas; 25x10(6) spermatozoa; n=26 does) and 76.1% (New York; 50x10(6) spermatozoa; n=46 does). Three of the four does receiving intrauterine inseminations per vaginam became pregnant to the frozen-thawed semen.

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

    PubMed

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

    2015-10-01

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

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

    PubMed

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

    2011-03-01

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

  17. Effect of various concentrations of caffeine, pentoxifylline, and kallikrein on hyperactivation of frozen bovine semen.

    PubMed

    Barakat, Ibrahim A H; Danfour, Mohamed A; Galewan, Fatma A M; Dkhil, Mohamed A

    2015-01-01

    Caffeine, pentoxifylline, and kallikrein are substances that affect the efficiency of sperms in the fertilization process; however, they have not been adequately studied. The present study aimed to examine the influence of caffeine, kallikrein, and pentoxifylline on sperm motility in bovine as well as investigate their optimum concentrations for increasing the movement of sperms in bovine. Frozen bovine sperms were thawed in universal IVF medium supplemented with 1, 5, and 10 mM caffeine or pentoxifylline or 1, 4, and 8 U/mL kallikrein and were then incubated for 30 min. Treated semen parameters were analyzed using a computer assisted semen analyzer (CASA). Data analysis showed that the mean values concerning progression and motility of sperm increased in caffeine and pentoxifylline treatments when compared with the kallikrein group. The obtained results revealed that kallikrein is not necessary for the improvement of bovine sperm motility. Additionally, our results revealed that 5 mM from caffeine was the best concentration added to the medium, followed by 1 or 5 mM from pentoxifylline. Therefore, it is concluded from the present study that caffeine has hyperactivation efficacy at 5 mM concentration compared to other treatments.

  18. Effect of different levels and sources of zinc supplementation on quantitative and qualitative semen attributes and serum testosterone level in crossbred cattle (Bos indicus x Bos taurus) bulls.

    PubMed

    Kumar, Nishant; Verma, Ramesh Prashad; Singh, Lallan Prasad; Varshney, Vijay Prakash; Dass, Ram Sharan

    2006-01-01

    An experiment was conducted on 16 crossbred bulls (about 2 years of age, 316.2+/-0.77 kg average body weight), divided into groups I, II, III and IV to study the effect of different levels of Zn supplementation from inorganic and organic sources on semen quality. The animals in the first 3 groups were supplemented with 0, 35 and 70 ppm Zn from Zn sulfate, respectively and the animals in-group IV were supplemented with 35 ppm Zn as Zn propionate. Semen collection and evaluation was done in the first month (to assess semen quality at the start of the experiment) and 7th, 8th and 9th month of experimental feeding to evaluate the effect of supplemental Zn on semen attributes. We gave 6 months for Zn feeding, so that 3 sperm cycles of spermatogenesis had passed and the collected semen reflected the complete effect of Zn supplementation. Six ejaculates from each bull were collected and evaluated for semen quantitative (ejaculate volume, sperm concentration and sperm number per ejaculate) and qualitative characteristics (semen pH, mass motility, individual motility, sperm livability percent and abnormal sperm percent, percent intact acrosome, bovine cervical mucus penetration test, hypo-osmotic sperm swelling test) and activity of seminal plasma enzymes i.e., alkaline phosphatase, acid phosphatase, GOT and GPT. Testosterone level in the blood serum of crossbred bulls was also estimated. Mean values of semen quantitative and qualitative characteristics at the start of the experiment were statistically non significant (P > 0.05) in all the crossbred cattle bulls, however, there were statistically significant differences among the bulls of different groups after 6 months of zinc supplementation. Mean ejaculate volume (mL) was 2.37, 4.70, 5.86 and 6.38, respectively in groups I to IV, indicating a statistically significant (P < 0.05) higher semen volume in Zn-supplemented groups as compared to the control group of bulls. Similarly, sperm concentration (million.mL(-1)), live

  19. Isolation of bacteria in semen and evaluation of antibiotics in extender for cryopreservation of buffalo (Bubalus bubalis) bull spermatozoa.

    PubMed

    Andrabi, S M H; Khan, L A; Shahab, M

    2016-12-01

    This study was designed to investigate the occurrence of bacterial species in water buffalo semen at the time of collection/processing and to assess the efficacy of some selected antibiotics (GTLS; gentamycin, tylosin and linco-spectin or SP; streptomycin and penicillin) in cryodiluent on bacterial control and quality including in vivo fertility of buffalo spermatozoa. For this purpose, four experiments were conducted. In experiment 1, a total of 11 bacterial species were isolated from buffalo ejaculates. In experiment 2, total aerobic bacterial counts at post dilution and thawing were lower (P < 0.05) in GTLS than in SP or control. The majority of the bacterial isolates from ejaculates were more susceptible to GTLS than SP. In experiment 3, sperm acrosome integrity was higher (P < 0.05) in GTLS and SP compared to control. In experiment 4, the in vivo fertility results for GTLS were higher (P < 0.05) than that for SP. In conclusion, a number of bacterial species were isolated from the bubaline semen, which requires an efficient control before its use in artificial insemination program. The GTLS combination of antibiotics may be incorporated into a freezing extender/protocol without compromising the post-thaw quality and in vivo fertility of buffalo bull spermatozoa.

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

    PubMed

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

    2009-07-01

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

  1. Effect on post-cryopreserved semen characteristics of Holstein bulls of adding combinations of vitamin C and either catalase or reduced glutathione to Tris extender.

    PubMed

    Eidan, Sajeda M

    2016-04-01

    This study was undertaken to investigate the influence of adding combinations of vitamin C to Tris extender with either catalase or reduced glutathione on post-cryopreserved semen characteristics of Holstein bulls for different preservation periods (cooling at 5°C, 48 h, 1, 2 and 3 months post cryopreservation, PC). Seven Holstein bulls of 2.5-3 years of age were used in this experiment. Semen was collected via artificial vagina in one ejaculate per bull per week for the 7 week experimental period. Pooled semen was equally divided into three treatments using Tris extender. Combinations of vitamin C (2.5mM) were added with either catalase (100 IU/ml, T2) or reduced glutathione (2mM, T3) to Tris extender and comparisons in response were made with the control group (Tris extender, T1). Individual sperm motility (IM), viability (V), plasma membrane integrity (PMI), and acrosome integrity (AI) were assessed during all periods of the study along with Malondialdehyde (MDA) concentrations and freezing ability. The IM was greater (P ≤ 0.01) in the T2 as compared with the T1 group at all periods of the study. Furthermore, the IM were greater (P ≤ 0.01) in the T3 as compared with the T1 group at the 48 h time period and at 3 months PC. The V, PMI and AI were greater (P ≤ 0.01) in T2 and T3 as compared with the T1 group at all the experimental periods. The MDA was greater (P ≤ 0.01) in the T2 as compared with the T1 group at 3 months PC. In conclusion, there was improved semen quality if semen of Holstein bulls was collected and stored in combinations of vitamin C with either catalase (T2) or reduced glutathione (T3) being added to Tris extender.

  2. Laparoscopic intra-uterine insemination of fallow deer with frozen-thawed or fresh semen after synchronisation with CIDR devices.

    PubMed

    Asher, G W; Morrow, C J; Jabbour, H N; Mulley, R C; Veldhuizen, F A; Langridge, M

    1992-03-01

    This study investigated the efficacy of fixed-time laparoscopic intra-uterine insemination of farmed fallow deer (Dama dama) with frozen-thawed or fresh semen. In the trials with frozen-thawed semen, a total of 547 mature non-lactating does across five New Zealand farms were used. For oestrous synchronisation and artificial insemination, a standard control regimen was applied to at least 30% of the does on each farm, involving the insertion of single CIDR type-G devices intravaginally for 14 days, deposition of 50 x 10(6) frozen-thawed spermatozoa at 65 hours after withdrawal of the CIDR device and the continuous presence of vasectomised bucks from the insertion of the CIDR device until 10 days after insemination. Various aspects of this protocol were changed for the remaining does on each farm, including inseminations at 60 or 70 hours, the absence of vasectomised bucks, insemination with 25 x 10(6) or 10 x 10(6) spermatozoa, synchronisation with CIDR type-S devices and synchronisation with prostaglandin. The conception rate, based on rectal ultrasonography at 45 days after insemination, was 67% across all treatments (n=547). Corrected conception rates (+/-s.e.), calculated following between-farm adjustments, were 67+/- 3% for the control regimen, 67+/- 9% and 73 +/- 8% for inseminations at 60 and 70 hours respectively, 61 +/- 9% for absence of bucks, 80 +/- 8% and 74 +/- 9% for inseminations with 25 x 10(6) and 10 x 10(6) spermatozoa respectively, 62 +/- 10% for CIDR type-S device synchronisation, and 49 +/- 10% for prostaglandin synchronisation. Despite apparent differences, none of the treatments resulted in adjusted conception rates that were significantly different from the control regimen (P>0.01). In the trials with fresh semen, 216 does in the USA were inseminated at 69-71 hours after withdrawal of the CIDR device using either cryopreserved semen from New Zealand (n=158; 25 x 10(6) spermatozoa per inseminate) or fresh semen (n=58; 7.5 x10(6) to 20 x 10

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  4. Effects of pipothiazine palmitate on handling stress and on the characteristics of semen collected by electroejaculation in bison (Bison bison) bulls.

    PubMed

    Toosi, B M; Gratton, G; McCorkell, R B; Wynne-Edwards, K E; Woodbury, M R; Lessard, C

    2013-04-01

    Handling North American bison can pose risk to the handler and evoke stress in the animal. Moreover, this induced stress might affect qualities of semen collected by electroejaculation. The objective of this study was to investigate if a long acting neuroleptic tranquilizer (LAN) would reduce the stress of bison and thereby improve the quality of electroejaculated semen. Eight experimental replicates were conducted between May and November. In each replicate, the same six bison bulls were randomly assigned into LAN-treated (n=3) and non-treated control (n=3) groups. Pipothiazine palmitate (Piportil L4) was administered intramuscularly as a single dose of 100 mg in replicates 1-4 or 200 mg in replicates 5-8. Within each replicate, semen was collected by electroejaculation at 4, 6, 11 and 13 days post treatment. Behavioral parameters, sperm morphology and motility parameters were analyzed. A blood sample was collected before each electroejaculation and serum concentrations of testosterone, cortisol and corticosterone were determined. Treatment bulls with 100 mg of Piportil L4 reduced the restraint time and the struggling of bison bulls during handling compared to the control group (P<0.05). Semen motility parameters and serum concentrations of testosterone, cortisol and corticosterone were not significantly affected when 100mg of the LAN was administered (P>0.05). However, giving 200 mg of Piportil L4 reduced the restraint time of bison bulls and the duration of semen collection (P<0.05). Also, this treatment improved total and progressive sperm motilities when compared to the respective controls (P<0.05). Interestingly, serum concentration of corticosterone, as an endocrine stress indicator, was decreased after administration of 200mg of Pipothiazine palmitate, while testosterone concentrations were increased compared to those values in untreated control bulls (corticosterone: 0.10±0.01 compared with 0.15±0.02 ng/mL; testosterone: 9.11±1.68 compared with 5.33±0

  5. Effect of spermatozoal concentration and number on fertility of frozen equine semen.

    PubMed

    Leipold, S D; Graham, J K; Squires, E L; McCue, P M; Brinsko, S P; Vanderwall, D K

    1998-06-01

    Information on the number of motile spermatozoa needed to maximize pregnancy rates for frozen-thawed stallion semen is limited. Furthermore, concentration of spermatozoa per 0.5-mL straw has been shown to affect post-thaw motility (7). The objectives of this study were 1) to compare the effect of increasing the concentration of spermatozoa in 0.5-mL straws from 400 to 1,600 x 10(6) spermatozoa/mL on pregnancy rate of mares, and 2) to determine whether increasing the insemination dose from approximately 320 to 800 million progressively motile spermatozoa after thawing would increase pregnancy rates. Several ejaculates from each of 5 stallions were frozen in a skim milk-egg yolk based freezing medium at 2 spermatozoal concentrations in 0.5-mL polyvinyl-chloride straws. Half of each ejaculate was frozen at 400 x 10(6) cells/mL and half at 1,600 x 10(6) cells/mL. Insemination doses were based on post-thaw spermatozoal motility and contained approximately 320 x 10(6) (320 to 400) motile spermatozoa or approximately 800 x 10(6) (800 to 900) motile spermatozoa. Sixty-three mares were assigned to 1 of 4 spermatozoal treatments (1--low spermatozoal number, low concentration; 2--low spermatozoal number, high concentration; 3--high spermatozoal number, low concentration; 4--high spermatozoal number, high concentration) and were inseminated daily. Post-thaw spermatozoal motility was similar for cells frozen at both spermatozoal concentrations (P > 0.1). One-cycle pregnancy rates were 15, 40, 28 and 33%, respectively, for Treatments 1, 2, 3 and 4. Packaging spermatozoa at the high concentration tended to increase pregnancy rates vs packaging at the low concentration (37 vs 22%; P = 0.095). Furthermore, when the lower spermatozoal number was used, there tended (P < 0.1) to be a higher pregnancy rate if spermatozoa were packaged at the higher concentration. There was no increase in pregnancy rates when higher numbers of motile spermatozoa were inseminated (27 vs 31%; P > 0

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Reproductive performance is of paramount importance to the cattle industry. Since recent progress has been achieved by optimizing estrus and ovulation synchronization protocols in cows, improvements are desired to increase the fertility of bulls enrolled in artificial insemination (AI) programs. Thi...

  7. Localization and functional modification of L-type voltage-gated calcium channels in equine spermatozoa from fresh and frozen semen.

    PubMed

    Albrizio, M; Moramarco, A M; Nicassio, M; Micera, E; Zarrilli, A; Lacalandra, G M

    2015-02-01

    It is well known that insemination of cryopreserved semen always results in lower fertility when compared with fresh semen, but there is an increased interest and demand for frozen equine semen by the major breeder associations because of the utility arising from semen already "on hand" at breeding time. In this article, we report that equine sperm cells express L-type voltage-gated calcium channels; their localization is restricted to sperm neck and to the principal piece of the tail in both fresh and frozen-thawed spermatozoa. We also studied the causes of cryoinjury at the membrane level focusing on the function of L-type calcium channels. We report that in cryopreserved spermatozoa the mean basal value of [Ca(2+)]i is higher than that of spermatozoa from fresh semen (447.130 vs. 288.3 nM; P < 0.001) and L-type channels function differently in response to their agonist and antagonist in relation to semen condition (fresh or frozen-thawed). We found that on addition of agonist to the culture medium, the increase in intracellular calcium concentrations ([Ca(2+)]i) was greater in frozen semen than in fresh semen (Δ[Ca(2+)]i = 124.59 vs. 16.04 nM; P < 0.001), whereas after the addition of antagonist the decrease in [Ca(2+)]i was lower in frozen semen than in fresh semen (Δ[Ca(2+)]i = 32.5 vs. 82.5 nM; P < 0.001). In this article, we also discuss the impact of cryopreservation on sperm physiology.

  8. Improved fertility in gilts and sows after artificial insemination of frozen-thawed boar semen by supplementation of semen extender with caffeine and CaCl2.

    PubMed

    Yamaguchi, Shoichiro; Funahashi, Hiroaki; Murakami, Tetsuya

    2009-12-01

    Supplementation of semen extender with caffeine and CaCl(2) for artificial insemination (AI) of fresh spermatozoa has been demonstrated to reduce recruitment of uterine polymorphonuclear leukocytes (PMNs) and the activity of phagocytosis. Here, we determined if addition of caffeine and CaCl(2) to semen extender improves the fertility of frozen-thawed boar semen. In experiment 1, gilts were cervically inseminated twice with frozen-thawed boar spermatozoa (25 x 10(8) cells per dose) suspended in Modena solution (n=7) or modified Beltsville Thawing Solution supplemented with caffeine and CaCl(2) (BCC, n=7). The gilts were slaughtered 4 h later, and their oviducts and uterine horns plus the body of the uterus were flushed to recover PMNs and non-phagocytosed spermatozoa. There was no difference in the total number of uterine PMNs between gilts inseminated with Modena solution and those inseminated with BCC (3.8 x 10(8) vs. 1.5 x 10(8) cells, respectively); however, the total number of uterine spermatozoa was higher when gilts were inseminated with BCC (40.6 x 10(6) cells) compared with those inseminated with Modena solution (1.4 x 10(6) cells, P<0.05). In experiment 2, gilts and sows were subjected to intrauterine insemination twice with frozen-thawed spermatozoa suspended (25 x 10(8) sperm per dose) in Modena (n=21) or BCC (n=21). The overall pregnancy and farrowing rates were higher in females inseminated with BCC (71.4 and 61.9%, respectively) compared with those inseminated with Modena solution (38.1 and 28.6%, respectively, P<0.05). However, no significant difference in litter size of piglets was observed between treatments (7.2 +/- 1.6 piglets for Modena solution vs. 8.2 +/- 0.9 piglets for BCC solution). In conclusion, we demonstrated that use of BCC solution for frozen-thawed boar semen produced better pregnancy and farrowing rates following AI than Modena solution, probably by reducing the phagocytosis of spermatozoa.

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

    PubMed

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

    2009-01-15

    In order to determine zinc and catalase content of seminal plasma in the buffalo and to study their associations with the semen characteristics, 54 semen samples were collected from 10 buffalo bulls; semen volume and sperm concentration, gross and progressive motility and viability were evaluated, seminal plasma was then harvested by centrifugation and its zinc content was estimated by atomic absorption spectrophotometer and its catalase activity determined by using a commercial kit. The zinc content of the seminal plasma (Mean +/- SEM) was recorded as 154.40 +/- 1.74 mg L(-1), while, the mean catalase value was 32.00 +/- 0.42 U mL(-1). The mean zinc values was highly correlated with sperm progressive motility and viability and with catalase values (p = 0.000 for all) and also was associated with gross motility (p = 0.020) and negatively with abnormal morphology (p = 0.049). The catalase values were highly associated with sperm progressive motility, viability and zinc content (p = 0.000 for all) and was associated with sperm gross motility (p = 0.024). For further clarification of these correlations, the samples were categorized in three groups of excellent (Ex, >90% motile, n = 33), good (Go, 80-89% motile, n = 15) and moderate (Mo, <79% motile, n = 6) according to their percentage of sperm motility. The mean progressive motility in Ex group was 92.54 +/- 0.51%, in Go group was 81.66 +/- 0.62% and in Mo group was 71.66 +/- 1.05%. The mean zinc and catalase values were recorded as 161.07 +/- 1.63 mg L(-1) and 33.41 +/- 0.34 U mL(-1) in Ex, 146.70 +/- 1.91 mg L(-1) and 31.01 +/- 0.67 in Go and 136.42 +/- 4.97 mg L(-1) and 26.51 +/- 0.87 U mL(-1) in Mo groups. The mean zinc value in Ex group was highly associated with sperm motility, viability and catalase values, in Go group was associated with catalase values and highly associated with sperm abnormal morphology and in Mo group it was highly associations with catalase values only. The mean catalase value in Ex group

  10. Quantification of leptin in seminal plasma of buffalo bulls and its correlation with antioxidant status, conventional and computer-assisted sperm analysis (CASA) semen variables.

    PubMed

    Kumar, Pradeep; Saini, Monika; Kumar, Dharmendra; Jan, M H; Swami, Dheer Singh; Sharma, R K

    2016-03-01

    The present study is the first to quantify leptin in seminal plasma of buffalo and investigate its relationship with seminal attributes. Ten ejaculates each from 10 Murrah buffalo bulls were collected. Semen quality variables such as semen volume, sperm concentration, sperm abnormalities, membrane integrity, antioxidant enzyme activities (superoxide dismutase, catalase and total antioxidant capacity), malondialdehyde (MDA) concentration, as well as sperm kinetics and motility variables were evaluated. The leptin concentration in serum and seminal plasma were estimated by the ELISA method. Bulls were classified in two groups on the basis of sperm concentration with Group I having >800 million sperm/mL and Group II <500 million sperm/mL. Greater (P<0.05) mean sperm abnormalities, seminal leptin concentrations and MDA concentrations were recorded in Group II than Group I. The seminal leptin was positively correlated with sperm abnormalities and MDA concentration while being negatively correlated with sperm concentration, but there was no correlation with sperm kinetic and motility variables, sperm membrane integrity and seminal plasma antioxidant enzyme activity. Thus, the data suggest that seminal leptin has a role in spermatogenesis and can be used as a marker for spermatogenesis to predict the capacity of buffalo bulls for semen production.

  11. Comparison of cryoprotective effects of lycopene and cysteamine in different cryoprotectants on bull semen and fertility results.

    PubMed

    Tuncer, P B; Büyükleblebici, S; Eken, A; Taşdemir, U; Durmaz, E; Büyükleblebici, O; Çoşkun, E

    2014-10-01

    The objectives of this study were to compare glycerol and ethylene glycol at different concentrations as cryoprotectants and lycopene or cysteamine (with/without) as antioxidants in Tris extender for bull semen. Twenty-four ejaculates were obtained from three bulls. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and added using both of the cysteamine 5 mm or lycopene 500 μg/ml, and control (without additives). The addition of 7% glycerol with cysteamine, 5% ethylene glycol with cysteamine and 3% ethylene glycol with cysteamine groups gave the lowest CASA motility than the other groups. However, 7% glycerol and 7% glycerol with lycopene resulted in a better rate of CASA progressive motility compared with that of other groups. Generally, all the lycopene groups signed better protective effects on acrosome and total morphology than the other groups. Glycerol 7% and 3% ethylene glycol with lycopene groups yielded to slight higher percentages of membrane integrity assessed by HOST than that of the other groups, but 7% glycerol with cysteamine and 3% ethylene glycol with cysteamine showed the worst percentages of membrane integrity. Glycerol 7% and 5% glycerol with lycopene gave rise to a higher value of VAP, VSL and VCL compared with that of the other groups. On the contrary, adding to 5% glycerol with cysteamine showed negative effect for VAP, VSL, VCL and ALH values. All cryoprotectant groups with lycopene decreased chromatin damage than the other groups. Ethylene glycol 3% led to lower non-return rates of inseminated cows. However, this result was not considered to be statistically important.

  12. Infertility and venereal disease in cattle inseminated with semen containing bovine herpesvirus type 5.

    PubMed

    Kirkland, P D; Poynting, A J; Gu, X; Davis, R J

    2009-07-25

    A group of 20 cows and heifers experienced poor conception rates and probable ovarian dysfunction after being artificially inseminated. When first examined, some showed signs of vulvovaginitis, with pustular, ulcerative lesions consistent with a herpesvirus infection. They had had no contact with bulls during the current breeding season. Bovine herpesvirus type 5 (BHV-5) was isolated from samples of frozen semen from the batch that had been used for the artificial insemination programme. BHV-5 was also isolated from the semen of a second apparently healthy bull during routine screening of its semen.

  13. Longitudinal study of the detection of Bluetongue virus in bull semen and comparison of real-time polymerase chain reaction assays.

    PubMed

    Gu, Xingnian; Davis, Rodney J; Walsh, Susan J; Melville, Lorna F; Kirkland, Peter D

    2014-01-01

    Infection with Bluetongue virus (BTV) is a significant impediment to the global movement of bovine semen. Repeat testing of blood from donor animals is specified in the World Organization for Animal Health (OIE) Manual for the export of semen from regions where BTV may be present. Screening of blood or semen samples has usually been carried out by virus isolation (VI) either by inoculation of chicken embryos followed by passage onto insect and mammalian cell cultures or in vivo inoculation of sheep followed by serology to detect seroconversion. Direct testing of semen for BTV would enable earlier release of semen samples and avoid repeat testing of the donor, as well as provide an option for releasing batches of semen that were collected without certification of the donor. Quantitative (real-time) reverse transcription polymerase chain reaction (qRT-PCR) assays overcome most of the limitations of other methods and have the potential to provide higher sensitivity. The present study compared 5 qRT-PCR assays, including 2 commercially available kits, for the detection of BTV in semen serially collected from 8 bulls over a period of 90 days after experimental infection. The results of the study show that at least one of the qRT-PCR assays is extremely reproducible and has both very high sensitivity and specificity to reliably detect all available serotypes. The preferred qRT-PCR gave consistently superior results to VI, sheep inoculation, and conventional RT-PCR. Therefore, the assay can be recommended for the screening of bovine semen for freedom from BTV.

  14. Glutathione supplementation to semen extender improves the quality of frozen-thawed canine spermatozoa for transcervical insemination.

    PubMed

    Ogata, Kazuko; Sasaki, Aiko; Kato, Yuka; Takeda, Arisa; Wakabayashi, Mikio; Sarentonglaga, Borjigin; Yamaguchi, Mio; Hara, Asuka; Fukumori, Rika; Nagao, Yoshikazu

    2015-01-01

    The present study was conducted to evaluate whether supplementation of semen extender with glutathione (GSH) can maintain the quality of frozen-thawed canine spermatozoa. Eighteen ejaculates were obtained from 5 dogs and placed in extender (20% egg yolk, Tris, citric acid, lactose, raffinose, antibiotics and 6.5% glycerol) containing 0 (control), 2.5, 5, 7.5 or 10 mM GSH. The samples were cooled to 4 C and then frozen in liquid nitrogen vapor. Motility parameters of the sperm were evaluated at 0, 1, 2, 3, 4, 12 and 24 h after thawing. Sperm motility was higher in the 5 mM GSH group than in the control or 2.5 and 10 mM GSH groups; this effect was observed at 1 to 24 h after thawing (P < 0.05). The 5 mM GSH group had a higher sperm viability index at 12 and 24 h after thawing compared with the other groups (P < 0.05). Acrosome integrity, evaluated at 4 h after thawing, was greater in two of the GSH-treated groups (5 and 10 mM) compared with the control. Lipid peroxidation (LP) levels immediately after thawing were lower in the 5 and 10 mM GSH groups compared with the control, while those at 12 h after thawing did not differ significantly. Frozen-thawed semen in the 5 mM GSH group was used for transcervical insemination of 4 bitches, resulting in delivery of 5 puppies from 2 bitches. These results indicate that supplementation of semen extender with 5 mM GSH was effective in improving motility, longevity and acrosomal integrity and inhibiting LP levels in post-thaw canine spermatozoa, without any adverse impacts on full-term development after transcervical insemination.

  15. The effect of season on spermatozoa motility, plasma membrane and acrosome integrity in fresh and frozen-thawed semen from Xinong Saanen bucks.

    PubMed

    Wang, W; Luo, J; Sun, S; Xi, L; Gao, Q; Haile, A B; Shi, H; Zhang, W; Shi, H

    2015-02-01

    The aim of this study was to evaluate whether the season of ejaculate collection influences seminal quality parameters of pre- and post-freeze-thawing in Xinong Saanen bucks. Ejaculates were collected from eight bucks throughout the four seasons (spring, summer, autumn and winter) in a 12 months' time period, identified in the Northern Hemisphere. Semen samples were evaluated by the combinations of conventional and Computer-Assisted Sperm Analysis (CASA) when fresh and after frozen-thawed, respectively. The results clearly demonstrated that season of ejaculate collection influenced (p < 0.05) fresh semen quality. Highest semen quality was observed during autumn. On the contrary, undesirable indices (significantly lower, p < 0.05) were observed in winter as compared with the other remaining seasons. CASA has clearly shown the influences of seasonal variations on semen motility parameters. Furthermore, season of ejaculate collection was also found to influence sperm freezability. Semen characteristics after frozen-thawed followed a similar pattern with that of fresh ejaculate except in spring. The results revealed that sperm quality was higher (p < 0.01) in summer and autumn than in spring and winter. In conclusion, seasonal variation influences semen quality in Xinong Saanen bucks. In addition to summer and autumn, fresh ejaculates in spring can also be successfully used for AI. Sperm from ejaculates collected during summer and autumn are more suitable for cryopreservation. Hence, it is possible to increase the efficiency of goat breeding by manipulating the seasonal variations of semen quality for immediate AI and/or cryopreservation.

  16. Embryo production and possible species preservation by nuclear transfer of somatic cells isolated from bovine semen.

    PubMed

    Liu, Jie; Westhusin, Mark; Long, Charles; Johnson, Gregory; Burghardt, Robert; Kraemer, Duane

    2010-12-01

    Somatic cells in semen are a potential source of nuclei for nuclear transfer to produce genetically identical animals; this is especially important when an animal has died and the only viable genetic material available is frozen semen. Usefulness of somatic cells obtained from fresh (cultured) and frozen (isolated, not cultured) bovine semen for nuclear transfer was evaluated. Twelve ejaculates were collected from nine bulls representing three breeds: Charolais, Brahman, and crossbred Rodeo bull. All samples were processed immediately and cell growth was obtained from seven of the twelve ejaculates (58.3%). Cells from three bulls (with the best growth rates) were evaluated by optical microscopy and used in cloning experiments. In culture, these cells exhibited classic epithelial morphology and expressed cytokeratin and vimentin, indicating they were of epithelial origin. When cells from the three bulls were used as donor cells, 15.9% (18/113), 34.5% (29/84), and 14.4% (13/90) of the fused embryos developed into blastocysts, respectively. Of the blastocyst stage embryos, 38.9% (7/18), 72.4% (21/29), and 61.5% (8/13) hatched, respectively. Somatic cells isolated (not cultured) from frozen bovine semen were also used in the cloning experiments. Although cleavage occurred, no compact morulae or blastocysts were obtained. In conclusion, epithelial cell growth was obtained from fresh bovine ejaculates with relatively high efficiency. Somatic cells from semen can be used as nucleus donors to produce cloned blastocyst-stage embryos.

  17. A retrospective clinical study of endoscopic-assisted transcervical insemination in the bitch with frozen-thawed dog semen.

    PubMed

    Mason, S J

    2016-11-24

    Since the conclusion of data collation from previously published work, a further 352 inseminations using frozen-thawed dog semen by endoscopic-assisted transcervical insemination (EIU) have been performed by the author. Insemination was performed on the second day in which crenulation of the anterior vagina was detected in conjunction with a progesterone concentration of >10 ng/ml. All semen samples were analysed for total number of sperm, total motility and progressive motility using computer-assisted semen analysis (CASA). The insemination dose was based on the progressively motile normal spermatozoa (PMNS). Insemination was performed on all bitches as previously described using a ureterorenoscope. Additional extender was inseminated subsequent to the semen to expand and fill the uterus. The semen and additional extender were inseminated slowly over a period of 15-20 min. Pregnancy was determined by B-mode ultrasound equipped with a 7.5-MHz probe whilst standing and/or via the whelping rate. The number of sperm inseminated ranged from 9 × 10(6) PMNS to 519 × 10(6) PMNS, with progressive motility values ranging between 20% and 80%. The overall pregnancy rate was 68% (238/352). When stratified by PMNS, pregnancy rates were as follows: >150 × 10(6) PMNS - 76% (110/145), 100-150 × 10(6) - 68% (87/128) and <100 × 10(6) PMNS - 52% (41/79). Pregnancy rate was significantly higher when >150 × 10(6) PMNS (p = .003) or 100-150 ×10(6) PMNS (p = .027) were inseminated compared to <100 × 10(6) PMNS. These data are concordant with previous reports recommending the insemination of >150 × 10(6) PMNS to maximize pregnancy rate. These results indicate that one optimally timed EIU insemination results in similar pregnancy rates to previous publications of one optimally timed, or two or more non-optimally timed inseminations using the Norwegian catheter.

  18. Sperm distribution and fertilization after unilateral and bilateral laparoscopic artificial insemination with frozen-thawed goat semen.

    PubMed

    Anakkul, Nitira; Suwimonteerabutr, Junpen; Tharasanit, Theerawat; Khunmanee, Sarawanee; Diloksumpan, Paweena; Berg, Debra K; Techakumphu, Mongkol

    2014-11-01

    Generally, laparoscopic artificial insemination (LAI) provides a higher success rate than of cervical insemination in goats. However, the sperm distribution after LAI in goats remains unknown, particularly when frozen-thawed semen is used. This study evaluated the distribution of frozen-thawed goat spermatozoa after LAI and compared the effects of sperm numbers and deposition sites (unilateral and bilateral sites) on pregnancy rate. In experiment 1, the frozen-thawed spermatozoa were stained either with CellTracker Green CMFDA (CT-Green) or CellTracker Red CMPTX (CT-Red), and in vitro evaluations of viability and motility were performed. In experiment 2, the labeled spermatozoa were deposited via LAI into the left (CT-Green) and right (CT-Red) uterine horns (n = 4). After ovariohysterectomy (6 hours after insemination), the distributions of green- and red-colored spermatozoa were assessed via tissue section, flushing, and the oviductal contents were also collected. Experiment 3 was designed to test the pregnancy rates in a group of 120 does after LAI using different numbers of spermatozoa (60 and 120 × 10(6) sperm per LAI) and different deposition sites. The results demonstrated that the fluorochromes used in this study did not impair sperm motility or viability. Frozen-thawed goat spermatozoa can migrate transuterinally after LAI, as evidenced by the observations of both CT-Green- and CT-Red-labeled spermatozoa in both uterine horns. Lower numbers of spermatozoa (60 × 10(6)) that are inseminated unilaterally (either ipsilateral or contralateral to the site of ovulation) can efficiently be used for LAI in goats (with a 56.67% pregnancy rate).

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

    PubMed

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

    2016-07-01

    The micromolar calcium-activated neutral protease gene (CAPN1) is a physiological candidate gene for sperm motility. However, the molecular mechanisms involved in regulating the expression of the CAPN1 gene in bulls remain unknown. In this study, we investigated the expression pattern of CAPN1 in testis, epididymis, and sperm at the RNA and protein levels by qRT-PCR, western blot, immunohistochemistry, and immunofluorescence assay. Results revealed that the expression of CAPN1 levels was higher in the sperm head compared with that in other tissues. Moreover, we identified a novel single-nucleotide polymorphism (g.-1256 A>C, ss 1917715340) in the noncanonical core promoter of the CAPN1 gene between base g.-1306 and g.-1012. Additionally, we observed greater sperm motility in bulls with the genotype CC than in those with the genotype AA (P<0.01), indicating that different genotypes were associated with the bovine semen trait. Furthermore, a higher fluorescence intensity of the C allele than that of the A allele at g. -1256 A>C was revealed by transient transfection in MLTC-1 cells and luciferase report assay. Finally, CAPN1 was highly expressed in the spermatozoa with the CC genotype compared with that with the AA genotype by qRT-PCR. This study is the first report on genetic variant g.-1256 A>C in the promoter region of CAPN1 gene association with the semen quality of Chinese Holstein bulls by influencing its expression. g.-1256 A>C can be a functional molecular marker in cattle breeding.

  20. Effects of semen extenders and storage temperatures on characteristics of frozen-thawed Bryde's (Balaenoptera edeni) whale spermatozoa.

    PubMed

    Hiwasa, Mami; Suzuki, Yo; Watanabe, Hiroyuki; Bhuiyan, Mohammad Musharraf Uddin; Matsuoka, Kohji; Fujise, Yoshihiro; Ishikawa, Hajime; Ohsumi, Seiji; Fukui, Yutaka

    2009-12-01

    The present study investigated effects of three semen extenders and storage temperatures on post-thaw characteristics of Bryde's whale spermatozoa. Spermatozoa were collected from the vasa deferens of three mature Bryde's whales captured during the Japanese whale research in the north-west Pacific (May to August 2007) after death. The three semen extenders used for freezing were 1) a commercialized synthetic extender (AndroMed: AM), 2) Tris-based + 10% bovine serum albumin (BSA) and 3) Tris-based + egg yolk (EY). The sperm samples from the three whales were frozen with the three extenders, and the post-thaw spermatozoa were stored at three different temperatures (35 C; 20-25 C, room temperature; and 5 C) for 0, 6, 12, 24, 48 and 96 h. At each time-point, total and progressive motility (PM), viability (live or dead), the hypo-osmotic test, defective acrosomes and malformation were examined. Immediately after thawing, AM resulted in similar recovery rates (60.4 and 83.3%) in 2 of the 3 whales examined and had comparable post-thaw recovery rates to those obtained using the EY and BSA extenders. Immediately after thawing, the proportion of PM in EY (17.6%) was higher (P<0.05) than that in BSA (15.0%). In the hypo-osmotic test, the proportions of AM (26.0%) and BSA (25.2%) were higher (P<0.05) than that of EY (17.3 %). The three extenders had similar viabilities (36.7, 37.9 and 32.1%, respectively), but the viability of BSA was higher (P<0.05) than that of EY. The present study showed that a synthetic semen extender, AndroMed, could be used for cryopreservation of whale spermatozoa in addition to Tris-based extenders containing bovine serum albumin or egg yolk. Storage of the post-thaw Bryde's whale spermatozoa was better at 5 C than at room temperature or 35 C. The frozen-thawed Bryde's whale spermatozoa maintained their motility and viability for at least two days at room temperature and for four days at 5 C.

  1. l-Carnitine in rooster semen cryopreservation: Flow cytometric, biochemical and motion findings for frozen-thawed sperm.

    PubMed

    Fattah, A; Sharafi, M; Masoudi, R; Shahverdi, A; Esmaeili, V; Najafi, A

    2017-02-01

    Rooster semen cryopreservation is not efficient for artificial insemination in breeder flocks. l-Carnitine (LC) has been evaluated for effectiveness in cryopreservation media on the characteristics of rooster sperm after freeze-thawing. Motility characteristics, membrane functionality, abnormal morphology, apoptotic like changes, mitochondria activity and lipid peroxidation of rooster sperms were assessed after freeze-thawing with different concentrations of LC in Beltsville medium. Semen samples were collected from 12 roosters, twice a week, and diluted in the extenders that contained different concentrations of LC. Supplementation of Beltsevile with 1 and 2 mM LC was found to result in higher total motility (68.2± 1.7% and 69.1± 1.7%, respectively), progressive motility (28.4± 1.6%, 29.8± 1.6%), membrane functionality (76.2± 1.9% and 75.9± 1.9%), viability (58.2 ± 1.1%, 59.1 ± 1.1%) and lower significant of lipid peroxidation (2.53 ± 0.08 nmol/ml, 2.49 ± 0.08 nmol/ml) compared to control group containing no LC. Lower motility, progressive motility, and viability were observed in frozen-thawed sperm in extender containing 8 mM LC (35.8± 1.7%, 9.6± 1.2% and 27.1 ± 1.2%, respectively) compared to control. Morphology and mitochondrial activity were not affected by different concentrations of LC. Our results showed that supplementation of Beltsville extender with 1 and 2 mM LC significantly improved the quality of rooster sperm quality after freeze-thawing.

  2. Functional SNPs of INCENP Affect Semen Quality by Alternative Splicing Mode and Binding Affinity with the Target Bta-miR-378 in Chinese Holstein Bulls

    PubMed Central

    Zhang, Yan; Jiang, Qiang; Huang, Jinming; Ju, Zhihua; Wang, Xiuge; Zhong, Jifeng; Wang, Changfa

    2016-01-01

    Inner centromere protein (INCENP) plays an important role in mitosis and meiosis as the main member of chromosomal passenger protein complex (CPC). To investigate the functional markers of the INCENP gene associated with semen quality, the single nucleotide polymorphisms (SNPs) g.19970 A>G and g.34078 T>G were identified and analyzed. The new splice variant INCENP-TV is characterized by the deletion of exon 12. The g.19970 A>G in the exonic splicing enhancer (ESE) motif region results in an aberrant splice variant by constructing two minigene expression vectors using the pSPL3 exon capturing vector and transfecting vectors into MLTC-1 cells. INCENP-TV was more highly expressed than INCENP-reference in adult bull testes. The g.34078 T>G located in the binding region of bta-miR-378 could affect the expression of INCENP, which was verified by luciferase assay. To analyze comprehensively the correlation of SNPs with sperm quality, haplotype combinations constructed by g.19970 A>G and g.34078 T>G, as well as g.-692 C>T and g.-556 G>T reported in our previous studies, were analyzed. The bulls with H1H12 and H2H2 exhibited a higher ejaculate volume than those with H2H10 and H9H12, respectively (P < 0.05). Bulls with H11H11 and H2H10 exhibited higher initial sperm motility than those with H2H2 (P < 0.05). The expression levels of INCENP in bulls with H1H12 and H11H11 were significantly higher than those in bulls with H9H12 (P < 0.05), as determined by qRT-PCR. Findings suggest that g.19970 A>G and g.34078 T>G in INCENP both of which appear to change the molecular and biological characteristics of the mRNA transcribed from the locus may serve as a biomarkers of male bovine fertility by affecting alternative splicing mode and binding affinity with the target bta-miR-378. PMID:27669152

  3. Effect of separating bull semen into X and Y chromosome-bearing fractions on the sex ratio of resulting embryos.

    PubMed Central

    Hagele, W C; Hare, W C; Singh, E L; Grylls, J L; Abt, D A

    1984-01-01

    Seventy-six, day 12 to day 15 bovine embryos, collected from 14 donors which had been inseminated with either X or Y chromosome-bearing spermatozoa fractions of semen separated by a thermal convection counterstreaming sedimentation and forced convection galvanization process, were processed for sexing by chromosomal analysis. Fifty-seven embryos were sexed; 20 from Y chromosome-bearing and 37 from X chromosome-bearing fractions of semen. Statistical analysis of the sexing data indicated that there was no significant difference in the male: female ratio for donors receiving male fractions compared to those receiving female fractions. The Y chromosome-bearing fractions produced a male: female ratio that was indistinguishable from the expected 1:1 ratio. However, the X chromosome-bearing fractions of semen produced a highly significant deviation from the expected 1:1 ratio towards the male. PMID:6478299

  4. Effect of artificial insemination protocol and dose of frozen/thawed stallion semen on pregnancy results in mares.

    PubMed

    Govaere, J L J; Hoogewijs, M K; De Schauwer, C; De Vliegher, S; Van Soom, A; Duchateau, L; de Kruif, A

    2014-06-01

    Deep intra-uterine insemination is commonly accepted as a routine procedure for artificial insemination in horses. The motives and principles of deep insemination are well described, but the equipment used may differ. In this trial, the efficiency of two different insemination pipettes for deep intra-uterine insemination in the mare was compared with insemination into the uterine body using commercially available frozen-thawed semen of two stallions of proven fertility. These inseminations were performed using two different doses. The semi-flexible Minitube pipette was compared with a newly designed insemination device with a more flexible telescopic insemination catheter (Ghent device). The semi-flexible Minitube pipette performed better than the newly designed insemination device with respect to pregnancy outcome (p = 0.008). The superiority of deep horn insemination over uterine body insemination was reflected by the better pregnancy rates obtained after deep insemination using the same low doses (30.6% better pregnancy rates) (p = 0.0123).

  5. Conception rates in European fallow does (Dama dama dama) following intrauterine insemination with frozen-thawed semen from Mesopotamian fallow (Dama dama mesopotamica) and crossbred (Dama dama dama x Dama dama mesopotamica) bucks.

    PubMed

    Mylrea, G E; Evans, G; English, A W

    1991-09-01

    Ninety eight parous fallow does received laparoscopic intrauterine insemination of frozen-thawed semen at one of 2 fixed intervals following oestrus synchronisation treatment. Semen was collected from a Mesopotamian (Dama dama mesopotamica) and a crossbred (F1) (Dama dama dama x Dama dama mesopotamica) fallow buck. Does were inseminated at either 56 or 66 hours after the removal of an intravaginal controlled internal drug releasing device. Eighty eight does received a single straw of frozen-thawed semen containing a total of 50 x 10(6) spermatozoa, while the remaining 10 received split straws containing 25 x 10(6) spermatozoa. Overall, the use of F1 semen containing 50 x 10(6) spermatozoa resulted in a 68% (17/25) conception rate compared with the Mesopotamian semen, which resulted in a 41% (26/63) conception rate. Conceptions were also achieved using 25 x 10(6) spermatozoa of either Mesopotamian or F1 semen (3/8 versus 2/2, respectively). Overall, the conception rate was higher for F1 than Mesopotamian semen (P less than 0.025) and there was a significant interaction with time of insemination (P less than 0.05); for F1 semen there was no difference in conception rate at the 2 insemination times, but for Mesopotamian semen conception was significantly higher (P less than 0.005) following insemination at 66 hours than at 56 hours.

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

    PubMed

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

    2012-01-01

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

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

    PubMed Central

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

    2012-01-01

    The aim of the present study was to investigate the effect of in vitro supplementation of selenium on fresh and frozen spermatozoa quality of buffalo (Bubalus bubalis) bulls. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 0.5, 1, 2, 4 and 8 µg mL-1 sodium selenite and the sperm motility and viability were evaluated at 0 (T0) (immediately after dilution), 60 (T1) and 120 (T2) min after diluting semen. In the second step, semen samples were diluted with tris-egg yolk-glycerol extender containing the same amounts of sodium selenite, cooled to 4 ˚C, equilibrated and semen parameters (motility, viability, membrane integrity and DNA damage) were estimated. Then, the semen was packed in 0.5 mL French straws and frozen in liquid nitrogen. Later, the semen was thawed and analyzed for the same parameters, as well as total antioxidant capacity. Results showed that addition of 1 and 2 µgmL-1 selenium to the semen extender significantly increased the sperm motility of fresh and equilibrated semen compared to the control without affecting other parameters. However, in frozen-thawed semen, extenders containing 1 and 2 µg mL-1 selenium significantly improved sperm motility, viability, membrane integrity and semen total antioxidant capacity and also resulted in lower DNA damaged sperms. In this study selenium supplementation of semen extender of 4 and 8 µg mL-1 had deleterious effects on sperm parameters as early as the samples were prepared for freezing. PMID:25653769

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

    PubMed

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

    2015-12-01

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

  9. Implementation of flow cytometry for quality control in four Danish bull studs.

    PubMed

    Christensen, P; Hansen, C; Liboriussen, T; Lehn-Jensen, H

    2005-02-01

    A flow cytometric method for simultaneous determination of sperm concentration and viability has recently been developed. In 2001, four Danish bull studs purchased flow cytometers and eight technicians were trained for routine analysis of raw and frozen-thawed semen. After initial training of the technicians, an experiment was carried out to document the precision of the system. The aim was also to assess if flow cytometric determination of sperm concentration could result in a more uniform production of semen doses. Results of this experiment showed high precision in the determination of sperm concentration and coefficients of variation were 3.5 and 2.4% for raw and frozen-thawed semen, respectively. Sperm viability was also assessed with high precision and coefficients of variation were 0.9% for raw semen and 1.7% for frozen-thawed semen. Furthermore, the experiment showed that package of semen doses after flow cytometric determination of sperm concentration in the raw semen results in a significantly smaller variation in the number of sperm per dose. In the second experiment, frozen semen was exchanged between the participating studs and were analysed by flow cytometry as well as by microscopic assessments by the eight technicians. Results show that the average correlation between technicians were 0.38 for motility assessments while flow cytometric agreement between technicians was significantly higher (average correlation was 0.86 for sperm viability and 0.92 for sperm concentration). The experiment also showed very high agreements between assessments within lab technician (correlations r=0.98 (sperm concentration) and r=0.99 (sperm viability)). Experiment 3 revealed that straws from the same batch varies in both concentration and viability. It is concluded that flow cytometric determination of sperm concentration and viability can be used to improve semen assessment by AI studs and result in a better quality control.

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

    PubMed

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

    2016-07-15

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

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

    PubMed

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

    2011-06-01

    The hypothalamic gonadotropin-releasing hormone receptor (GnRHR) plays an essential physiological role in reproductive function, which triggers the synthesis and release of luteinizing hormone and follicle stimulating hormone in the pituitary. The objective of this study was to investigate the effects of polymorphisms of GnRHR gene on the quality of fresh and frozen semen in Holstein bulls. The PCR-RFLP method was applied to detect G286A and T340C transitions determining MboII and BspMI polymorphisms, respectively, in the exon I of bovine GnRHR gene and evaluated its associations with sperm quality traits in 131 Holstein bulls. In polymorphic locus 286, bulls with the GA genotype had significantly higher sperm motility in frozen semen (FMOT) than GG genotype (P < 0.01). In polymorphic locus 340, bulls with heterozygote CT genotype had significantly higher sperm motility (MOT), semen volume per ejaculate (VOL), and lower abnormal spermatozoa rate (ASR) than homozygote TT genotype (P < 0.05). Bulls contained one A allele or C allele had a favorable, positive effect on sperm quality traits. These results indicate that GnRHR gene can be a potential marker for improving sperm quality traits, and imply that bulls with GA or CT genotype should be selected in breeding program.

  12. Is caprine arthritis encephalitis virus (CAEV) transmitted vertically to early embryo development stages (morulae or blastocyst) via in vitro infected frozen semen?

    PubMed

    Al Ahmad, M Z Ali; Chebloune, Y; Chatagnon, G; Pellerin, J L; Fieni, F

    2012-05-01

    The aim of this study was to determine, in vivo, whether in vitro infected cryopreserved caprine sperm is capable of transmitting caprine arthritis-encephalitis virus (CAEV) vertically to early embryo development stages via artificial insemination with in vitro infected semen. Sperm was collected from CAEV-free bucks by electroejaculation. Half of each ejaculate was inoculated with CAEV-pBSCA at a viral concentration of 10(4) TCID(50)/mL. The second half of each ejaculate was used as a negative control. The semen was then frozen. On Day 13 of superovulation treatment, 14 CAEV-free does were inseminated directly into the uterus under endoscopic control with thawed infected semen. Six CAEV-free does, used as a negative control, were inseminated intrauterine with thawed CAEV-free sperm, and eight CAEV-free does were mated with naturally infected bucks. Polymerase chain reaction (PCR) was used to detect CAEV proviral-DNA in the embryos at the D7 stage, in the embryo washing media, and in the uterine secretions of recipient does. At Day 7, all the harvested embryos were PCR-negative for CAEV proviral-DNA; however, CAEV proviral-DNA was detected in 8/14 uterine smears, and 9/14 flushing media taken from does inseminated with infected sperm, and in 1/8 uterine swabs taken from the does mated with infected bucks. The results of this study confirm that (i) artificial insemination with infected semen or mating with infected bucks may result in the transmission of CAEV to the does genital tack seven days after insemination, and (ii) irrespective of the medical status of the semen or the recipient doe, it is possible to obtain CAEV-free early embryos usable for embryo transfer.

  13. Antioxidative effects of cysteamine, hyaluronan and fetuin on post-thaw semen quality, DNA integrity and oxidative stress parameters in the Brown Swiss bull.

    PubMed

    Sarıözkan, S; Tuncer, P B; Büyükleblebici, S; Bucak, M N; Cantürk, F; Eken, A

    2015-03-01

    The aim of this study was to compare the effectiveness of antioxidants including cysteamine (2.5, 7.5 mm), hyaluronan (0.25, 1 mg ml(-1) ) and fetuin (5, 10 mg ml(-1) ) in the freezing of Brown Swiss bull semen. The best percentages of CASA motilities were achieved with 10 mg ml(-1) of fetuin and 2.5 mm of cysteamine. For sperm morphology, 10 mg ml(-1) of fetuin and 2.5 mm of cysteamine had better protective effects (P < 0.001). The results of hypo-osmotic swelling test showed that the percentage values of membrane integrity in all the groups, excluding that supplemented with 5 mg ml(-1) of fetuin, were higher than those of the control group (P < 0.001). Results obtained for the DNA damage of sperm cells demonstrated that 0.25 mg ml(-1) of hyaluronan, and 2.5 and 7.5 mm of cysteamine led to lower rates of spermatozoa with damaged DNA, compared with the control group (P < 0.001). The maintenance of superoxide dismutase and glutathione peroxidase antioxidant activities following freeze-thawing with 2.5 and 7.5 mm of cysteamine and 10 mg ml(-1) of fetuin was demonstrated to be at a higher level in comparison with the control group (P < 0.001). Malondialdehyde formation was found to be lower in the groups supplemented with 0.25 mg ml(-1) of hyaluronan and 7.5 mm of cysteamine after the freeze-thawing process (P < 0.001).

  14. Effect of seminal plasma removal before cryopreservation of bovine semen obtained by electroejaculation on semen quality and in vitro fertility.

    PubMed

    Campanholi, Suzane Peres; Monteiro, Fabio Morato; Ribeiro Dias, Erika Aline; Mercadante, Maria Eugênia Zerlotti; de Paz, Claudia Cristina Paro; Dell'Aqua Junior, José Antonio; Papa, Frederico Ozanam; Dell'Aqua, Camila de Paula Freitas; Vantini, Roberta; Garcia, Joaquim Mansano

    2017-02-01

    Cryopreservation of bull semen is a common biotechnology procedure in cattle breeding. However, when the ejaculate is obtained by electroejaculation, wide variation is observed in the sperm/seminal plasma (SP) ratio that can affect the freezability of semen in this species. The removal of SP may improve the quality of frozen bull semen. The objective of this study was to evaluate the effect of SP removal from the ejaculate on the cryopreservation of semen from 38 Nellore bulls collected by electroejaculation. After collection, the ejaculate was divided into three aliquots: (1) control (N) diluted to a concentration of 60 × 10(6) spermatozoa/mL and frozen with SP; (2) centrifugation (C) at ×600g for 10 minutes and the pellet resuspended and frozen at the same concentration as N; and (3) filtration (F) through SpermFilter and sperm recovered and frozen at the same concentration as N. After thawing, sperm kinetics, plasma and acrosome membrane integrity, mitochondrial membrane potential, oxidative stress, and in vitro fertility were evaluated. Statistical analysis was performed using the SAS 9.2 package, and differences were considered significant when P < 0.05. Higher average path velocity and straight-line velocity were observed in the groups submitted to SP removal compared to the control group (P < 0.01). In contrast, filtered samples exhibited higher beat cross frequency, straightness, and linearity compared to the other groups. Plasma membrane integrity was reduced when SP was removed, but lower oxidative stress was observed in groups C and F (34.91 ± 2.95% and 31.63 ± 2.95%, respectively) compared to group N (57.39 ± 2.95%). However, the percentage of hatched blastocysts was similar in the N and F groups (21.22 ± 1.05% and 24.00 ± 1.05%, respectively) and higher compared to group C (18.83 ± 1.05%). In conclusion, removal of SP by centrifugation for bull semen freezing reduced the rate of in vitro-produced embryos, whereas filtration of

  15. Effects of caffeine on sperm characteristics after thawing and inflammatory response in the uterus after artificial insemination with frozen-thawed boar semen.

    PubMed

    Yamaguchi, S; Suzuki, C; Noguchi, M; Kasa, S; Mori, M; Isozaki, Y; Ueda, S; Funahashi, H; Kikuchi, K; Nagai, T; Yoshioka, K

    2013-01-01

    We previously reported that AI with frozen-thawed boar semen supplemented with caffeine increased the number of uterine sperm by inhibiting migration of polymorphonuclear leukocytes (PMNs) into the uterine lumen, and also improved fertility of gilts and sows. The objective of the present study was to determine the effects of the addition of caffeine to a thawing solution on postthaw sperm quality and uterine inflammatory response after AI with frozen-thawed boar semen. Incubation of frozen-thawed sperm in Modena solution supplemented with 10 mM caffeine for 90 minutes improved (P < 0.05) percentages of progressive motility, straightness, and linearity of sperm movement compared with no caffeine, without causing damage to plasma or acrosomal membranes. Gilts inseminated once with 2 × 10(9) frozen-thawed sperm suspended in Modena solution with or without caffeine, and gilts that did not receive AI, were slaughtered 4 hours later. Uteri were recovered for analysis of number of uterine PMNs and mRNA expression (quantitative reverse transcription polymerase chain reaction) of tumor necrosis factor-α, interleukin (IL)-1β, IL-6, IL-8, monocyte chemoattractant protein-1, and cyclooxygenase 2 in the endometrium. Caffeine decreased (P < 0.05) both the number of total uterine PMNs and expression of IL-8 mRNA in the endometrium after AI. The amount of IL-8 and cyclooxygenase 2 mRNA after AI in the absence of caffeine were higher than samples from gilts that did not receive AI (P < 0.05), whereas there were no significant differences between treatments in expression levels of tumor necrosis factor-α, IL-1β, IL-6, or monocyte chemoattractant protein-1 mRNA. Pregnancy rate in sows inseminated with sperm supplemented with caffeine (16 of 23; 70%) tended (P < 0.1) to exceed that without caffeine (12 of 26; 46%), but litter size was not affected. In conclusion, the addition of caffeine to the thawing solution inhibited migration of uterine PMNs, probably by downregulating IL-8

  16. Addition of superoxide dismutase mimics during cooling process prevents oxidative stress and improves semen quality parameters in frozen/thawed ram spermatozoa.

    PubMed

    Santiani, Alexei; Evangelista, Shirley; Sepúlveda, Néstor; Risopatrón, Jennie; Villegas, Juana; Sánchez, Raúl

    2014-10-01

    High levels of reactive oxygen species (ROS), which may be related to reduced semen quality, are detected during semen cryopreservation in some species. The objectives of this study were to measure the oxidative stress during ram semen cryopreservation and to evaluate the effect of adding 2 antioxidant mimics of superoxide dismutase (Tempo and Tempol) during the cooling process on sperm motility, viability, acrosomal integrity, capacitation status, ROS levels, and lipid peroxidation in frozen and/or thawed ram spermatozoa. Measuring of ROS levels during the cooling process at 35, 25, 15, and 5 °C and after freezing and/or thawing showed a directly proportional increase (P < 0.05) when temperatures were lowering. Adding antioxidants at 10 °C confered a higher motility and sperm viability after cryopreservation in comparison with adding at 35 °C or at 35 °C/5 °C. After freezing and/or thawing, sperm motility was significantly higher (P < 0.05) in Tempo and Tempol 1 mM than that in control group. Percentage of capacitated spermatozoa was lower (P < 0.05) in Tempo and Tempol 1 mM in comparison with that in control group. In addition, ROS levels and lipid peroxidation in group Tempo 1 mM were lower (P < 0.05) than those in control group. These results demonstrate that ram spermatozoa are exposed to oxidative stress during the cooling process, specifically when maintained at 5 °C and that lipid peroxidation induced by high levels of ROS decreases sperm motility and induces premature sperm capacitation. In contrast, the addition of Tempo or Tempol at 0.5 to 1 mM during the cooling process (10 °C) protects ram spermatozoa from oxidative stress.

  17. Designing of an artificial neural network model to evaluate the association of three combined Y-specific microsatellite loci on the actual and predicted postthaw motility in crossbred bull semen.

    PubMed

    Deb, Rajib; Singh, Umesh; Raja, Thirvvothur Venkatesan; Kumar, Sushil; Tyagi, Shrikant; Alyethodi, Rafeeque R; Alex, Rani; Sengar, Gyanendra; Sharma, Sheetal

    2015-06-01

    The freezing of bull semen significantly hamper the motility of sperm which reduces the conception rate in dairy cattle. The prediction of postthaw motility (PTM) before freezing will be useful to take the decision on discarding or freezing of the germplasm. The artificial neural network (ANN) methodology found to be useful in prediction and classification problems related to animal science, and hence, the present study was undertaken to compare the efficiency of ANN in prediction of PTM on the basis of the number of ejaculates, volume, and concentration of sperms. The combined effect of Y-specific microsatellite alleles on the actual and predicted PTM was also studied. The results revealed that the prediction accuracy of PTM based on the semen quality parameters was comparatively lower because of higher variability in the data set. The ANN gave better prediction accuracy (34.88%) than the multiple regression analysis models (32.04%). The root mean square error was lower for ANN (8.4353) than that in the multiple regression analysis (8.6168). The haplotype or combined effect of microsatellite alleles on actual and predicted PTM was found to be highly significant (P < 0.01). On the basis of results, it was concluded that the ANN methodology can be used for prediction of PTM in crossbred bulls.

  18. Embryo transfer and sex determination following superovulated hinds inseminated with frozen-thawed sex-sorted Y sperm or unsorted semen in Wapiti (Cervus elaphus songaricus).

    PubMed

    Gao, Q H; Wang, H E; Zeng, W B; Wei, H J; Han, C M; Du, H Z; Zhang, Z G; Li, X M

    2011-07-01

    The aim of this study was to evaluate embryo production in superovulated wapiti hinds inseminated with either Y-sorted or unsorted semen. Eighteen hinds were allocated to three treatment groups: AI following multiple ovulation (CIDR/FSH) with 10×10(6) Y-sorted frozen-thawed semen (Y group, n=6), or 10×10(6) and 100×10(6) unsorted frozen-thawed semen for the unsorted (n=6) and the control group (n=6). The embryos from the sixth day following insemination were collected and classified. Fifteen embryos from the unsorted or the control group, and four embryos from the Y group were sex determinated based on DNA analysis of the amelogenin gene. Twenty-one embryos from the Y group and 42 embryos from the unsorted or the control group were transferred into 21 and 42 synchronized recipients via standard procedures on 6th day post estrus, respectively. There were no significant differences in the number of recovered eggs, transferable embryos, degenerated embryos or unfertilized oocytes per hind among the three groups of the control (9.2±3.6, 4.7±1.9, 3.0±2.0, 1.5±1.4), the unsorted (8.2±1.9, 4.8±0.7, 1.7±1.0, 1.7±1.0) and the Y group (8.8±4.2, 4.2±1.8, 2.2±1.2, 2.5±2.1), respectively (P>0.05). The sex ratio of embryos from the Y group (4M/0F) was significantly (P<0.05) distinct from that of the unsorted and control group (8M/7F). The sex ratio of the offspring from sexed embryos (8M/0F) was deviated significantly (P<0.05) from that of the non-sexed embryos (11M/9F). In conclusion, the results suggested that the male embryos of predicted sex can be achieved with AI of sex-sorted cryopreserved sperm. PCR amplification using the amelogenin gene primers can be applied to DNA analysis of micro samples from wapiti embryo biopsies for sex identification. The male offspring can be produced after transferred with the male embryos of predicted sex.

  19. The possibility of obtaining intergeneric hybrids via White Kołuda (Anser anser L.) goose insemination with fresh and frozen-thawed Canada goose (Branta canadensis L.) gander semen.

    PubMed

    Kowalczyk, Artur; Łukaszewicz, Ewa

    2012-02-01

    The objective of the present experiments was to produce the intergeneric hybrids of domesticated and wild goose via artificial insemination with fresh and frozen-thawed semen. The experiments were carried out during two successive goose reproductive seasons, on eight five-year-old Canada Goose (Branta canadensis L.) males used as semen donors and 16 two-year-old White Kołuda geese designated to fertility tests. Pooled semen was collected twice a week by the dorso-abdominal massage. In freshly collected semen, ejaculate volume, color, consistency, degree of fecal or blood contamination, spermatozoa concentration, motility, and morphology were evaluated. Part of the semen collected in the first year of the experiment (Experiment 1) was used for geese insemination with fresh semen, while the remainder was frozen. In Experiment 2 all samples were subjected exclusively to freezing procedure. Geese were inseminated once a week with fresh semen in a dose of 80 μl or 160 μl, and twice a week with frozen-thawed semen in a dose of 80 μl (160 μl per wk) or 100 μl (200 μl per wk). Eggs were set weekly and incubated up to hatching. The volume of ejaculates varied from 0.100 to 0.470 ml; spermatozoa concentration from 140 to 310 million ml(-1); progressive movement was observed in 40 to 60% of spermatozoa; the percentage of total live spermatozoa ranged from 69.3 to 92.0%, the highest percentage (34.0-68.3) was represented by live normal spermatozoa and those with bulb-head (13.3-41.0). Cryopreservation caused a decrease in percentage of motile cells to 30%; total live spermatozoa contribution by 27.2%p, including those live normal by 15.9%p (in relation to the fresh semen), bulb-head spermatozoa by 10.9%p, and increase (by 5.9%p) in number of spermatozoa with other deformations. Goose insemination 1×/week with fresh semen containing about 10.3 million live normal spermatozoa resulted in 66.7% of fertile eggs and with dose higher by 2.8 million spermatozoa (on average

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

    PubMed Central

    2012-01-01

    Background Proximal cytoplasmic droplets (PCDs), a remnant of germ cell cytoplasm, are common non-specific morphological defects in bovine semen. This study evaluated the effect of higher percentages of PCDs on the quality of frozen-thawed bovine semen, embryo production and early embryo development. Methods Three ejaculates from each of five (group 1: PCD ≤ 1%, control) and eight adult Bos indicus bulls (group 2: PCD ≥ 24%) were analysed. Semen samples were examined for: post-thaw motility, vigour of movement, concentration, sperm morphology, slow thermoresistance test (STT), membrane integrity, acrosome status, mitochondrial function using fluorescent probes association (FITC-PSA, PI and JC-1) and sperm chromatin integrity using acridine orange assay. Two bulls from group 2, with 28.5% and 48.5% PCD, respectively, and three bulls from the control group, each with 0% PCD, were selected for IVF (in vitro fertilisation). Results Semen analyses revealed a significant correlation (P < 0.01) between increased rates of PCD and sperm quality traits. Nevertheless, no differences were observed in sperm motility and vigour either before or after the STT or in the percentage of intact acrosomes (analysed by differential interference contrast microscopy (DIC) after STT), but membrane integrity, acrosome status (evaluated with FITC-PSA staining method after thawing) and mitochondrial function were reduced, when compared with group 1 (P < 0.05). The higher incidence of PCD was positively correlated to chromatin damage, especially after three hours of incubation at 37°C. IVF showed similar results for bull C2 (group 1, control) and bull P2 (group 2, group with higher PCDs). Conclusion Higher PCD levels influenced spermatozoa quality traits. IVF and embryo development data showed that cleavage, blastocyst formation and blastocyst hatching may have been influenced by the interaction of morphology traits and individual bull effects. PMID:22240071

  1. PCK1 is negatively regulated by bta-miR-26a, and a single-nucleotide polymorphism in the 3' untranslated region is involved in semen quality and longevity of Holstein bulls.

    PubMed

    Huang, Jinming; Guo, Fang; Zhang, Zebin; Zhang, Yuanpei; Wang, Xiuge; Ju, Zhihua; Yang, Chunhong; Wang, Changfa; Hou, Minghai; Zhong, Jifeng

    2016-03-01

    Phosphoenolpyruvate carboxykinase 1 (PCK1) is a multi-functional enzyme that plays important roles in physiological processes, including reproduction. We previously reported that the PCK1 transcript has five splice variants; PCK1-AS4, which lacks exon 5, is enriched in the testis of Holstein bulls. In the present study, we profiled select PCK1 transcript variants in the testis, epididymus, and semen of high- and low-performance bulls, and examined the possibility that microRNAs may be involved in single nucleotide polymorphism (SNP)-mediated modulation of PCK1 expression. PCK1-AS4 abundance is not significantly different between high- and low-performance bulls. Luciferase reporter assays, however, showed that bovine PCK1 expression is repressed by bta-miR-26a in HepG2 hepatocyte cells. One SNP (c. + 2183 G > T) at the miRNA-binding site of PCK1 does not influence PCK1 expression, but is associated with elevated ejaculation volume, fresh sperm motility, and genomic estimated breeding value of longevity, as well as with reduced values of composite index and calving ease. Collectively, the identified 3'-untranslated-region SNP variant highlights the importance of PCK1 in the fecundity of Holstein bulls, and implicates a role for bta-miR-26a in regulating PCK1 abundance. Further study is needed to assess the effects of other genetic variants in 5'-flanking region and exons of PCK1 on enzyme levels in the testis and sperm. Mol. Reprod. Dev. 83: 217-225, 2016. © 2016 Wiley Periodicals, Inc.

  2. Different concentrations of cysteamine and ergothioneine improve microscopic and oxidative parameters in ram semen frozen with a soybean lecithin extender.

    PubMed

    Najafi, Abozar; Kia, Hossein Daghigh; Mohammadi, Hossein; Najafi, Mir Hossein; Zanganeh, Zaynab; Sharafi, Mohsen; Martinez-Pastor, Felipe; Adeldust, Hamideh

    2014-08-01

    The aim of this study was to evaluate the effects of ergothioneine and cysteamine as antioxidant supplements in a soybean lecithin extender for freezing ram semen. Twenty-four ejaculates were collected from four rams and diluted with extenders (1.5% soybean lecithin, 7% glycerol) containing no supplements (control) and cysteamine or ergothioneine (2, 4, 6 or 8mM). Motility by CASA, viability, plasma membrane functionality (HOS test), total abnormality, lipid peroxidation, glutathione peroxidase (GPx) activity and capacitation status (CTC staining) were assessed after thawing. Using 6mM of either antioxidant improved total motility. Cysteamine at 6mM and ergothioneine at 4 and 6mM improved viability and reduced lipid peroxidation (malondialdehyde concentration). Both antioxidants improved membrane functionality significantly, except at 8mM. Progressive motility, kinematic parameters, GPx activity, capacitation status and sperm abnormalities were not influenced by the antioxidant supplements. In conclusion, cysteamine at 6mM and ergothioneine at 4 or 6mM seem to improve the post-thawing quality of ram semen cryopreserved in a soybean lecithin extender.

  3. Scrotal insulation and its relationship to abnormal morphology, chromatin protamination and nuclear shape of spermatozoa in Holstein-Friesian and Belgian Blue bulls.

    PubMed

    Rahman, Mohammad Bozlur; Vandaele, Leen; Rijsselaere, Tom; Maes, Dominiek; Hoogewijs, Maarten; Frijters, Adrie; Noordman, Jakomien; Granados, Ana; Dernelle, Eric; Shamsuddin, Mohammed; Parrish, John J; Van Soom, Ann

    2011-10-15

    The objectives of this study were to identify the stages of spermatogenesis susceptible to elevated testicular temperature in terms of sperm motility, viability, morphology, chromatin protamination and nuclear shape. The latter two valuable parameters are not included in routine semen analysis. Scrotal insulation (SI) was applied for 48 h in 2 Holstein-Friesian (HF) and 2 Belgian Blue (BB) bulls and semen was collected at 7 d intervals along with semen collection of a non-insulated bull of each breed. Semen samples were frozen and assigned to 4 groups: period 1 (preinsulation) = -7 d and 0 d, where 0 d = initiation of SI after semen collection; period 2 = 7 d (sperm presumed in the epididymis during SI); period 3 = 14 d to 42 d (cells presumed at spermiogenesis and meiosis stages during SI); period 4 = 49 d to 63 d (cells presumed at spermatocytogenesis stage during SI). The percentages of progressively motile and viable spermatozoa as assessed by computer-assisted sperm analysis (CASA) and fluorescence microscopy, respectively were decreased whereas abnormal sperm heads, nuclear vacuoles and tail defects were increased at period 3 (P < 0.05) compared to period 1, 2 or 4 in SI bulls of both HF and BB breeds. Protamine deficient spermatozoa as observed by chromomycin A(3) (CMA(3)) staining were more present (P < 0.05) at period 2 and 3 in both breeds compared to period 1 or 4. Sperm nuclear shape as determined by Fourier harmonic amplitude (FHA) was most affected by heat stress during period 3 (P < 0.01) and a higher response was observed in BB bulls than HF bulls. In conclusion, sperm cells at the spermiogenic and meiotic stages of development are more susceptible to heat stress. The lack of chromatin protamination is the most pertinent result of heat stress, together with subtle changes in sperm head shape, which can be detected by FHA but not by conventional semen analysis.

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

    PubMed Central

    Allouche, Lynda; Madani, Toufik; Mechmeche, Mohamed; Clement, Laetitia; Bouchemal, Allaoua

    2017-01-01

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

  5. Applications and cost benefits of sexed semen in pasture-based dairy production systems.

    PubMed

    Butler, S T; Hutchinson, I A; Cromie, A R; Shalloo, L

    2014-05-01

    Sexed semen technology is now commercially available in many countries around the world, and is primarily used in dairy cattle breeding. Sperm are sorted by flow cytometry on the basis of a 4% difference in DNA content between sperm containing X and Y chromosomes. Despite reliably producing a 90% gender bias, the fertility of the sexed semen product is compromised compared with conventional semen. The negative implications of the reduced fertility of sexed semen are amplified in seasonal systems of dairy production, as the importance of fertility is greater in these systems compared with year-round calving systems. A review of the literature indicates that conception rates (CR) to 1st service with frozen-thawed sexed semen are ~75% to 80% of those achieved with conventional frozen-thawed semen. Preliminary results from a large-scale field trial carried out in Ireland in 2013 suggest that significant improvements in the performance of sexed semen have been made, with CR of 87% of those achieved with conventional semen. The improved fertility of a sexed semen product that delivers a 90% gender bias has considerable implications for the future of breeding management in pasture-based dairy production systems. Sexed semen may facilitate faster, more profitable dairy herd expansion by increasing the number of dairy heifer replacements born. Biosecurity can be improved by maintaining a closed herd during the period of herd expansion. In a non-expansion scenario, sexed semen may be used to increase the value of beef output from the dairy herd. The replacement heifer requirements for a herd could be met by using sexed semen in the 1st 3 weeks of the breeding season, with the remaining animals bred to beef sires, increasing the sale value over that of a dairy bull calf. Alternatively, very short gestation sires could be used to shorten the calving interval. Market prices have a considerable effect on the economics of sexed semen use, and widespread use of sexed semen should

  6. In vitro development of bovine embryos cultured with stem cell factor or insulin-like growth factor-I following IVF with semen of two bulls having different field fertility.

    PubMed

    Dhali, A; Anchamparuthy, V M; Butler, S P; Pearson, R E; Gwazdauskas, F C

    2009-12-01

    The usefulness of IVF as a potential tool to evaluate the field fertility of bulls is equivocal and growth factor addition to culture media research is needed to delineate components needed for providing defined environments for embryos. The overall aim was to evaluate the in vitro development of embryos derived using a serum supplemented and serum-free production systems and semen from two bulls of different field fertility. The study was conducted to determine the combinatorial effect of stem cell factor (SCF) and/or insulin-like growth factor-I (IGF-I) in culture on subsequent embryo development in cattle. Oocytes were aspirated separately from >or=3 to <3mm follicles to test different follicle size populations and were matured in TCM-199 supplemented with LH, FSH, estradiol and BSA (Fraction V). Matured oocytes were fertilized in BSA supplemented synthetic oviductal fluid (SOF)-IVF medium. Presumptive zygotes were cultured for 8d (in humidified 5% CO(2) at 38.5 degrees C) in BSA supplemented SOF-in vitro culture (IVC) medium. SOF-IVC medium was supplemented with fetal bovine serum (4%), IGF-I (100ng/mL), SCF (50ng/mL) or IGF-I (100ng/mL)+SCF (50ng/mL). The development competence of embryos did not differ between the bulls and among the culture environments. Nevertheless, there was an effect of follicle size on cleavage rate (P<0.05) and a greater cleavage rate resulted from oocytes aspirated from >or=3mm follicles (71.0+/-1.5%) compared to those collected from <3mm follicles (64.8+/-1.6%). The overall cleavage rate (%); blastocyst formation (%); and expanded/hatched blastocyst formation (%) were 68.2+/-1.5 and 67.7+/-1.7; 29.4+/-1.4 and 28.6+/-1.5; and 18.6+/-1.2 and 18.5+/-1.1, respectively, for the bull of above and below average field fertility. The results indicate that follicle size for oocyte aspiration is effective for determining IVC success and that IVF may not discriminate among bulls of different field fertility.

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

    PubMed

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

    2017-02-07

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

  8. Initial analysis of sperm DNA methylome in Holstein bulls

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  9. Ascorbic acid, catalase and chlorpromazine reduce cryopreservation-induced damages to crossbred bull spermatozoa.

    PubMed

    Paudel, K P; Kumar, S; Meur, S K; Kumaresan, A

    2010-04-01

    The present study evaluated the effectiveness of ascorbic acid, catalase, chlorpromazine and their combinations in reducing the cryodamages to crossbred bull (Bos taurus x Bos indicus) spermatozoa. A total of 32 ejaculates (eight each from four bulls) were diluted in Tris-citric acid-fructose-egg yolk-glycerol extender. Each ejaculate was split into six parts (five treatment and one control). Treatment groups included 10 mm ascorbic acid, 0.1 mm chlorpromazine, 200 IU/ml catalase, 10 mm ascorbic acid + 0.1 mm chlorpromazine or 200 IU/ml catalase + 0.1 mm chlorpromazine in the extender. Fluorescent probes (Fluorescein isothiocyanate--Pisum sativum agglutinin + Propidium iodide) were used for the assessment of spermatozoa viability and acrosomal status. The proportion of acrosome intact live (AIL), acrosome intact dead, acrosome reacted live and acrosome reacted dead sperm was assessed in fresh, equilibrated and frozen-thawed semen. The functional status of the sperm was assessed using hypo-osmotic sperm swelling test (HOSST). Activities of acrosin and hyaluronidase enzyme were also determined. Lipid peroxidation level was assayed based on the melonaldehyde (MDA) production. In cryopreserved semen, the values of AIL spermatozoa, HOSST response, hyaluronidase and acrosin activity were reduced by 53%, 47%, 34% and 54%, respectively from their initial values in fresh semen. However, MDA level was threefold higher in the frozen-thawed sperm compared with fresh sperm. Significant (p < 0.05) improvement in motility, viability, HOSST response, retention of hyaluonidase and acrosin and reduction in MDA was recorded in ascorbic acid, catalase, ascorbic acid + chlorpromazine and catalase + chlorpromazine incorporated groups. The percentage of AIL sperm was significantly (p < 0.05) higher in ascorbic acid, catalase and ascorbic acid + chlorpromazine incorporated groups compared with the control. Chlorpromazine alone did not improve the post-thaw semen quality but when combined

  10. Comparison of efficiency between two artificial insemination methods using frozen-thawed semen in domestic cat (Felis catus): artificial insemination in domestic cats.

    PubMed

    Villaverde, Ana Izabel Silva Balbin; Melo, Cely Marini; Martin, Ian; Ferreira, Tatiana Henriques; Papa, Frederico Ozanam; Taconeli, Cesar Augusto; Lopes, Maria Denise

    2009-09-01

    The aim of this study was to compare the efficiency of the intravaginal (IVAI) vs. intrauterine artificial insemination (IUAI) using frozen-thawed sperm in the domestic cat. Semen was collected from two tom cats using an artificial vagina and samples were assessed for motility (computer-assisted sperm analysis (CASA)), sperm morphology and plasma membrane integrity. After dilution with TRIS/OEP/YOLK (4% of glycerol), sperm samples were loaded into 0.25 mL straws (25 x 10(6)motile sperm/straw), incubated at 5 degrees C for 20 min and cryopreserved over liquid nitrogen (LN(2)) vapor for 15 min and then immersed in LN(2). For each AI, four straws from the same male were thawed (12s at 46 degrees C) and centrifuged at 250 x g for 8 min to pellet the sperm. The supernatant was discarded and sperm pellet resuspended with the remaining liquid, approximately 100 microL, and analyzed as described above. Queens were treated with a single im injection of 100 IU eCG to induce ovarian follicular development. Final oocyte maturation and ovulation was induced with 100 IU hCG given im at 82-84 h after eCG administration. Thirty hours after hCG administration, females were inseminated either intrauterine (n=8 queens) or intravaginally (n=8 queens), using thawed sperm from a single male. Although a pronounced decrease in sperm motility, acrosome and plasma membrane integrity was observed in sperm samples from both cats, a pregnancy rate of 75% was achieved when using the intrauterine AI method compared with 0% pregnancy when inseminated intravaginally.

  11. Transcriptome analysis of bull semen with extreme nonreturn rate: use of suppression-subtractive hybridization to identify functional markers for fertility.

    PubMed

    Lalancette, C; Thibault, C; Bachand, I; Caron, N; Bissonnette, N

    2008-04-01

    Spermatozoa are terminally differentiated cells produced during the complex process of spermatogenesis. Although the role of their residual RNA content is still being debated, this transcriptome may represent a fingerprint of spermatogenesis quality. In the present study, we undertook differential transcript profiling of spermatozoa from fertile bulls with extreme nonreturn rates (NRRs): a low-fertile group, and a high-fertile group. Using the suppression-subtractive hybridization technique in combination with macroarray analysis, we also identified novel genes. Both extreme NRR index groups retained redundant identity, such as ribosomal and mitochondrial sequences, at a statistically significant level. An elevated number of 12S, 18S, and Large Chain R rRNA gene copies were found in low-fertile bulls and validated in spermatozoa by quantitative RT-PCR for a small cohort of bulls with known fertility index. Whereas the high-NRR library exhibited a large proportion (29%) of transcripts associated with known functions (e.g., metabolism, signal transduction, translation, glycosylation, and protein degradation), only 10% of the low-NRR sequences did. This difference is also conveyed by two other categories: 17% Bovine Genome and 48% unknown in the high-NRR library, compared with 3% and 80%, respectively, in the low-NRR library. Some of the unknown transcripts are similar to expressed sequence tags detected in the male reproductive organ of certain plants and retain homology to a putative human protein. Whereas the individual transcriptome profiles may be useful in fertility assessment, these findings also suggest cross-species conservation, could contribute to a better understanding of spermatogenesis, and provide new insights regarding idiopathic infertility.

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

    PubMed

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

    2016-01-01

    In artificial insemination, poor quality of semen unsuitable for cryopreservation and susceptibility of spermatozoa to cryodamage in crossbred bulls have been a matter of concern. Present study was designed to identify the testicular cytology indices that might be used to predict the semen quality and cryotolerance of spermatozoa in bulls. Based on the ejaculate rejection rate and sperm cryotolerance, bulls (Holstein Friesian X Tharparkar crossbred) were classified into either good (producing good quality semen with spermatozoa having good cryotolerance; n = 4) or poor (producing poor quality semen with spermatozoa having poor cryotolerance; n = 4). Testicular cytology was studied in all the 8 bulls using fine needle aspiration technique. Testicular cytology of good bulls and poor bulls differed significantly. The proportion of Sertoli cells was significantly higher in good bulls (25.3 ± 1.6) compared to poor bulls (11.0 ± 0.8). The Sertoli cell index was 46.1 ± 5.0 in good bulls while it was only 13.8 ± 1.3 in poor bulls. The cut off values, as determined using Receiver Operating Characteristics analysis, indicate that the bulls having testicular cytogram comprising of < 15.5% Sertoli cells, < 24.3 Sertoli cell index and > 4.0 spermatogenic cells to Sertoli cell ratio might be a poor bull in terms of semen quality and cryotolerance of spermatozoa. The proportion of Sertoli cells in the testicular cytology had positive (P < 0.05) relationship with semen quality and cryotolerance of spermatozoa.

  13. Avian artificial insemination and semen preservation

    USGS Publications Warehouse

    Gee, G.F.; Risser, Arthur C.; Todd, Frank S.

    1983-01-01

    Summary: Artificial insemination is a practical propagation tool that has been successful with a variety of birds. Cooperative, massage, and electroejaculation and modifications of these three basic methods of semen collection are described for a variety of birds. Semen color and consistency and sperm number, moti!ity, and morphology, as discussed, are useful indicators of semen quality, but the most reliable test of semen quality is the production of fertile eggs. Successful cryogenic preservation of avian semen with DMSO or glycerol as the cryoprotectant has been possible. Although the methods for preservation require special equipment, use of frozen semen requires only simple insemination supplies

  14. Cryopreservation of American kestrel semen with dimethylsulfoxide

    USGS Publications Warehouse

    Gee, G.F.; Morrell, C.A.; Franson, J. Christian; Pattee, Oliver H.

    1993-01-01

    Semen samples from 15 male American Kestrels (Falco sparverius) were frozen in dimethyl sulfoxide (DMSO). The semen was thawed 1-14 mo later and used to inseminate six females during three breeding seasons. Kestrels inseminated with thawed semen containing 4% DMSO produced only infertile eggs (N = 14). Kestrels inseminated with thawed semen containing 6%, 8%, or 10% DMSO produced fertile eggs (N = 14) and live chicks (N = 6). Progressive motility of spermatozoa in thawed semen containing 10% DMSO was less (44 ? 6%) than in thawed semen containing 6% (62 ? 10%) or 8% (61 ? 1%) DMSO.

  15. Relationship between the magnitude of the inbreeding coefficient and milk traits in Holstein and Jersey dairy bull semen used in Brazil.

    PubMed

    Soares, M P; Gaya, L G; Lorentz, L H; Batistel, F; Rovadoscki, G A; Ticiani, E; Zabot, V; Di Domenico, Q; Madureira, A P; Pértile, S F N

    2011-09-06

    Artificial insemination has been used to improve production in Brazilian dairy cattle; however, this can lead to problems due to increased inbreeding. To evaluate the effect of the magnitude of inbreeding coefficients on predicted transmitting abilities (PTAs) for milk traits of Holstein and Jersey breeds, data on 392 Holstein and 92 Jersey sires used in Brazil were tabulated. The second-degree polynomial equations and points of maximum or minimal response were estimated to establish the regression equation of the variables as a function of the inbreeding coefficients. The mean inbreeding coefficient of the Holstein bulls was 5.10%; this did not significantly affect the PTA for percent milk fat, protein percentage and protein (P = 0.479, 0.058 and 0.087, respectively). However, the PTAs for milk yield and fat decreased significantly after reaching inbreeding coefficients of 6.43 (P = 0.034) and 5.75 (P = 0.007), respectively. The mean inbreeding coefficient of Jersey bulls was 6.45%; the PTAs for milk yield, fat and protein, in pounds, decreased significantly after reaching inbreeding coefficients of 15.04, 9.83 and 12.82% (P < 0.001, P = 0.002, and P = 0.001, respectively). The linear regression was only significant for fat and protein percentages in the Jersey breed (P = 0.002 and P = 0.005, respectively). The PTAs of Holstein sires were more affected by smaller magnitudes of inbreeding coefficients than those of Jersey sires. It is necessary to monitor the inbreeding coefficients of sires used for artificial insemination in breeding schemes in Brazil, since the low genetic variability of the available sires may lead to reduced production.

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

    PubMed

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

    2016-12-01

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

  17. Buffalo (Bubalus bubalis) SCNT embryos produced from somatic cells isolated from frozen-thawed semen: effect of trichostatin A on the in vitro and in vivo developmental potential, quality and epigenetic status.

    PubMed

    Selokar, Naresh L; Saini, Monika; Agrawal, Himanshu; Palta, Prabhat; Chauhan, Manmohan S; Manik, Radheysham; Singla, Suresh K

    2016-08-01

    This study examined the effects of trichostatin A (TSA) treatment of reconstructed buffalo embryos, produced by hand-made cloning using somatic cells isolated from over a decade old frozen-thawed semen, on their in vitro and in vivo developmental competence, quality and epigenetic status. Following treatment of reconstructed embryos with TSA (0, 50 or 75 nM) for 10 h prior to culture, the cleavage (100.0 ± 0, 94.5 ± 2.3 and 96.1 ± 1.2%, respectively) and blastocyst rate (50.6 ± 2.3, 48.4 ± 2.7 and 48.1 ± 2.6%, respectively), total cell number (275 ± 17.4, 289 ± 30.1 and 317 ± 24.2, respectively) and apoptotic index (5.6 ± 0.7, 3.4 ± 0.9 and 4.5 ± 1.4, respectively) were not significantly different among the three groups. However, TSA treatment increased (P < 0.05) the global level of H4K5ac and decreased (P < 0.05) that of H3K27me3 in blastocysts whereas the global level of H3K18ac was not affected significantly. Transfer of embryos treated with 75 nM TSA (n = 10) to recipients resulted in two pregnancies (20%), one out of which was aborted in the second and the other in the third trimester whereas transfer of control embryos (n = 20) or those treated with 50 nM TSA (n = 12) did not result in any pregnancy. In conclusion, these results suggest that TSA treatment of cloned buffalo embryos produced using somatic cells isolated from frozen-thawed semen improved their epigenetic status but not the in vitro developmental potential and offspring rate.

  18. Fertility aspects in yearling beef bulls grazing endophyte-infected tall fescue pastures.

    PubMed

    Schuenemann, G M; Edwards, J L; Hopkins, F M; Rohrbach, N R; Adair, H S; Scenna, F N; Waller, J C; Oliver, J W; Saxton, A M; Schrick, F N

    2005-01-01

    During a 2-year study, yearling beef bulls were used to determine the effects of grazing on endophyte-infected tall fescue on endocrine profiles, semen quality and fertilisation potential. Bulls were allotted to graze tall fescue pastures infected with Neotyphodium coenophialum (E+; n = 20 per year) or Jesup/MaxQ (Pennington Seed, Atlanta, GA, USA; NTE; n = 10 per year). Bulls were grouped by scrotal circumference (SC), bodyweight (BW), breed composites and age to graze tall fescue pastures from mid-November until the end of June (within each year). Blood samples, BW, SC and rectal temperatures (RT) were collected every 14 days. Semen was collected from bulls every 60 days by electroejaculation and evaluated for motility and morphology. The developmental competence of oocytes fertilised in vitro with semen from respective treatments was determined. Bulls grazing E+ pastures had decreased BW gain (P < 0.01), increased overall RT (P < 0.01) and decreased prolactin (P < 0.01) compared with animals grazing NTE pastures. Neither percentage of normal sperm morphology nor motility differed between bulls grazed on the two pasture types. Semen from E+ bulls demonstrated decreased cleavage rates (P = 0.02) compared with semen from NTE bulls. However, development of cleaved embryos to the eight-cell and blastocyst stages did not differ between the two groups. In conclusion, semen from bulls grazing E+ tall fescue resulted in decreased cleavage rates in vitro, which may lower reproductive performance owing to reduced fertilisation ability.

  19. Sperm macromolecules associated with bull fertility.

    PubMed

    Kaya, Abdullah; Memili, Erdoğan

    2016-06-01

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

  20. In vitro assessment of sperm from bulls of high and low field fertility.

    PubMed

    Al Naib, A; Hanrahan, J P; Lonergan, P; Fair, S

    2011-07-01

    The aim of this study was to investigate the reasons for differences in field fertility of bulls following insemination with frozen-thawed semen. The study was carried out in two separate parts over two years and comparisons were made between 5 high and 4 low fertility Holstein Friesian bulls as determined by their either 90 day non-return rate (Year 1) or calving rate (Year 2). Two high fertility Limousin bulls were included in Year 1 for comparative purposes. The ability of sperm from each bull to penetrate artificial mucus was assessed (Year 1 = 7 replicates; Year 2 = 5 replicates). Glass capillary tubes (2 per bull per replicate) were filled with artificial mucus and incubated with sperm stained in 1% Hoechst 33342 for 30 min at 37 °C. The number of sperm were subsequently counted at 10 mm intervals along the tube between 40 and 80 mm markers. Sperm mitochondrial activity of each bull was assessed by the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assay (4 replicates in each year). Sperm were incubated with MTT for 1 h at 37 °C following which the absorbance of formazan was read using a spectrophotometer. Sperm viability after thawing was assessed for each bull using a live/dead sperm viability kit (Year 1 = 3 replicates; Year 2 = 4 replicates). A minimum of 250 cells were assessed per bull in each replicate and classified as either live or dead. Finally, the ability of sperm to fertilize oocytes in vitro and their ability to develop to blastocyst stage embryos were assessed (5 replicates in each year involving 220 to 306 oocytes per bull). Data transformation to normalize residuals was required for mucus sperm penetration (square root) and IVF (cleavage and blastocyst rate) results (arcsin). The mean number of sperm counted at each 10 mm mark between 40 and 80 mm was higher in the high fertility (56.0; 95% CI 39.5 to 75.3) compared to the low fertility (42.9; 95% CI 29.3 to 59.1) Holstein Friesian bulls but the difference did not

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

    PubMed

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

    2014-09-01

    Amino acids seem to be crucial components for semen freezing extender due to antioxidant properties. Therefore, this study aimed to assess motility parameters, membrane integrity, intracellular reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and DNA damage to detect the optimum concentrations of cysteine and glutamine for buffalo semen cryopreservation. Twenty ejaculates of four buffalo bulls were diluted in tris-egg yolk extender and divided into seven equal groups consisting of cysteine (5, 7.5 and 10 mmol), glutamine (10, 15 and 20 mmol) and no additive. Supplementation of 5 and 7.5 mmol cysteine and 15 mmol glutamine in cryopreservation extender significantly increased post-thaw motility and plasma membrane integrity of spermatozoa with significant reduction in intracellular ROS when compared with control groups (P < 0.05). Cysteine at 7.5 mmol concentration elevated progressive motility and MMP, compared with control (P < 0.05). No significant differences were observed for motion patterns and DNA damage of frozen-thawed buffalo spermatozoa in extender containing amino acids. The findings of this study showed that supplementation of 7.5 mmol cysteine and 15 mmol glutamine in semen cryopreservation extender has more potential to decrease intracellular ROS, and subsequently elevate motility and membrane integrity of buffalo frozen-thawed spermatozoa.

  2. Catalase supplementation on thawed bull spermatozoa abolishes the detrimental effect of oxidative stress on motility and DNA integrity.

    PubMed

    Fernández-Santos, M R; Domínguez-Rebolledo, A E; Esteso, M C; Garde, J J; Martínez-Pastor, F

    2009-08-01

    The potential protective effect of catalase supplementation during in vitro culture of frozen/thawed bull spermatozoa was investigated. Frozen/thawed semen collected from three fighting bulls was diluted in phosphate buffered saline (PBS) and incubated at 37 degrees C under different experimental conditions: Control, Catalase (CAT) (200 U/mL), Oxidant (OXI) (100 microm Fe(2+)/1 mm ascorbate), and Catalase + Oxidant (CAT/OXI). We assessed sperm motility, acrosomal integrity, viability and chromatin status (SCSA) at 0, 2 and 6 h of incubation. Our results showed that catalase abolished the effect of the oxidant, protecting spermatozoa against reactive oxygen species, and improving both sperm motility and chromatin status during incubation. The OXI treatment significantly reduced the percentage of motile sperm after 6 h of incubation. The statistical model also showed that there were differences in sperm motility between CAT/OXI (20.8 +/- 2.9%) and OXI (11.6 +/- 7.6%) (p < 0.001). There were no significant effects of OXI on sperm viability, acrosomal status or proportion of abnormal tails. %DFI (spermatozoa with moderate or high DNA Fragmentation Index) was significantly higher on OXI (p < 0.001). Catalase prevented DNA fragmentation even in the presence of the oxidant (%DFI: 30.3 +/- 0.8% OXI vs. 17.4 +/- 0.7% CAT/OXI). We conclude that catalase supplementation after thawing could protect bull spermatozoa against oxidative stress, and it could improve media used for processing thawed spermatozoa.

  3. Effect of single-layer centrifugation or washing on frozen-thawed donkey semen quality: Do they have the same effect regardless of the quality of the sample?

    PubMed

    Ortiz, I; Dorado, J; Morrell, J M; Crespo, F; Gosálvez, J; Gálvez, M J; Acha, D; Hidalgo, M

    2015-07-15

    The aims of this study were to determine the sperm quality of frozen-thawed donkey sperm samples after single-layer centrifugation (SLC) using Androcoll-E in comparison to sperm washing or no centrifugation and to determine if the effect on the sperm quality after SLC or sperm washing depends on the quality of the sample. Frozen-thawed sperm samples from Andalusian donkeys were divided into three aliquots, and they were processed using three different techniques after thawing: uncentrifuged diluted control (UDC), sperm washing (SW), and SLC. Afterward, sperm quality index was estimated by integrating all parameters (total and progressive sperm motility, membrane integrity, and DNA fragmentation) in a single value. The relationship between the sperm quality of thawed UDC samples and the effect on sperm parameters in SW and SLC-selected samples was assessed. Sperm quality index was significantly higher (P < 0.001) in SLC (0.8 ± 0.0) samples than that in UDC (0.6 ± 0.0) and SW (0.6 ± 0.0) samples, regardless of the sperm quality index after thawing of the sperm sample. In conclusion, SLC of frozen-thawed donkey spermatozoa using Androcoll-E-Small can be a suitable procedure for selecting frozen-thawed donkey sperm with better quality, in particular in those samples where an improvement in motility is needed.

  4. Ultrastructure and fertilizing ability of Limousin bull sperm after storage in CEP-2 extender with and without egg yolk.

    PubMed

    Ducha, Nur; Susilawati, T; Aulanni'am; Wahyuningsih, Sri; Pangestu, Mulyoto

    2012-10-15

    Sperm can change physiology and structure during storage in refrigerator temperature or frozen temperature that caused by cold shock or free radical. The aim of this study to evaluate ultrastructure and fertilizing ability of Limousin bull sperm after storage in cauda epididymal plasma-based (CEP-2) extender with or without 20% egg yolk concentration at refrigerator temperature. Semen sample collected from three Limousin bull were diluted with CEP-2 with 20% egg yolk and CEP-2 without egg yolk, cooled and stored at 4-5 degrees C during eight days. Sperm ultrastructure were observed with scanning electron microscopy (SEM). Fertilizing ability of Limousin bull sperm were assessed on cleavage rate of embryo using in vitro fertilization method. The percentage data were transformed into arcsine before being analysis with ANOVA and Duncan's multiple comparison test. The result of study showed morphologically normal sperm after storage in CEP-2 with 20% egg yolk, whereas in CEP-2 without egg yolk morphologically abnormal sperm especially neck was fractured and head was destroyed. Fertilizing ability of Limousin bull sperm were significantly higher in CEP-2 extender with egg yolk 20% (74.29 +/- 4.95%; p < 0.05) than without egg yolk (30.00 +/- 12.02%; p < 0.05). Egg yolk 20% in CEP-2 extender protected ultrastructure and fertilizing ability after storage during eight days.

  5. Repression of common bull sperm flora and in vitro impairment of sperm motility with Pseudomonas aeruginosa introduced by contaminated lubricant.

    PubMed

    Smole, I; Thomann, A; Frey, J; Perreten, V

    2010-08-01

    Semen collected from clinically healthy bulls at an artificial insemination centre was examined for bacterial diversity. While bacteria that are normally present in the common flora of bovine semen were absent, such as Mycoplasma sp., Proteus sp. and Corynebacterium sp., all semen samples contained an unusually high number of Pseudomonas aeruginosa strains. Analysis via pulsed field gel electrophoresis demonstrated that one particular P. aeruginosa strain, present in a sealed bottle of lubricant, was widespread in bull semen. This strain was shown to secrete substances that inhibited both the growth of bacteria constituting the normal bull sperm flora and the motility of spermatozoa in vitro. This study demonstrated that commercially available lubricants might contain bacteria that can spread amongst breeding bulls and affect the quality of semen. Bacteriological controls and species' identification are necessary at several production levels, including lubricants and extenders, to ensure high semen quality and avoid the spread of pathogens.

  6. Quantification of damage at different stages of cryopreservation of endangered North American bison (Bison bison) semen and the effects of extender and freeze rate on post-thaw sperm quality.

    PubMed

    Hussain, S A; Lessard, C; Anzar, M

    2011-12-01

    Semen cryopreservation is an important technique for the banking of animal germplasm from endangered species and exploitation of genetically superior sires through artificial insemination. Being a member of bovidae family, bison semen has poor freezing ability as compared to dairy and beef bulls' semen. This study was designed to quantify the damage to bison sperm at different stages of cryopreservation, and to determine the effects of extender (commercial Triladyl(®) vs. custom made tris-citric acid [TCA]) and freeze rate (-10, -25 and -40°C/min) on post-thaw quality of bison semen. Semen was collected from five bison bulls (three woods and two plains) via electroejaculation. In Experiment 1, semen was diluted in Triladyl® extender and frozen with freeze rate -10°C/min. Sperm motility characteristics were recorded in fresh, diluted, cooled (4°C) and freeze-thawed semen using computer-assisted sperm analyzer (CASA). In Experiment 2, semen was diluted in Triladyl® or TCA extender, and frozen with three different freeze rates, i.e. -10, -25 or -40°C/min. Thawing was performed at 37°C for 60s. Post-thaw sperm motility characteristics were assessed using CASA, and sperm structural characteristics (plasma membrane, mitochondrial membrane potential and acrosomes) were evaluated using flow cytometer, at 0 and 3h while incubating semen at 37°C. In Experiment 1, total and progressive motilities did not differ among pre-freeze stages of cryopreservation (P>0.05). However, sperm total and progressive motilities declined (P<0.001) in freeze-thawed semen by 35% and 42%, respectively, compared to after cooling (pre-freeze) semen. In Experiment 2, Triladyl®, as compared to TCA, yielded greater (P<0.05) post-thaw sperm total motility (41% compared to 36%) and progressive motility (34% compared to 29%) at 0h, respectively. The percent change in post-thaw sperm total and progressive motilities, VAP, VCL, VSL, IPM-high ΔΨm and IPM-IACR during 3h incubation at 37°C, was

  7. Applications of sexed semen in cattle production.

    PubMed

    Hohenboken, W D

    1999-12-01

    Sexed semen will contribute to increased profitability of dairy and beef cattle production in a variety of ways. It could be used to produce offspring of the desired sex from a particular mating to take advantage of differences in value of males and females for specific marketing purposes. Commercial dairy farmers, those who produce and market milk, could use sexed semen to produce replacement daughters from genetically superior cows and beef crossbred sons from the remainder of their cow population. To increase the rate of response to selection, seedstock dairy cattle breeders could produce bulls for progeny testing from a smaller number of elite dams by using sexed semen to ensure that all of them produced a son. Using sexed semen could then reduce the cost of progeny testing those bulls, because fewer matings would be necessary to produce any required number of daughters. Commercial beef cattle farmers, producing animals for eventual slaughter, could use sexed semen to capitalize on the higher value of male than female offspring for meat production. They could also use sexed semen to produce specialized, genetically superior replacement heifers from as small a proportion of the herd as possible. This would allow the remainder of the herd to produce male calves from bulls or breeds with superior genetic merit for growth, feed conversion efficiency, and carcass merit. Single-sex, bred-heifer systems, in which each female is sold for slaughter soon after weaning her replacement daughter, would be possible with the use of X-chromosome-sorted semen. Use of sexed semen would make terminal crossbreeding systems more efficient and sustainable in beef cattle. Fewer females would be required to produce specialized maternal crossbred daughters, and more could be devoted to producing highly efficient, terminal crossbred sons.

  8. Cysteamine supplementation revealed detrimental effect on cryosurvival of buffalo sperm based on computer-assisted semen analysis and oxidative parameters.

    PubMed

    Swami, Dheer Singh; Kumar, Pradeep; Malik, R K; Saini, Monika; Kumar, Dharmendra; Jan, M H

    2017-02-01

    The aim of this study was to investigate the effect of addition of cysteamine to the semen extender on post-thaw semen quality. A total of 30 ejaculates were collected from six bulls. Each ejaculate was divided into five equal parts and diluted to final concentration of 80 million sperms/mL using Optixcell(®)(IMV, France) semen extender supplemented with different concentrations of cysteamine (0, 0.75, 1.25, 2.5 and 5mM) and cryopreserved. In the frozen-thawed samples, the VAP, VSL, VCL ALH and sperm motility of control samples was greater (P<0.05) than cysteamine treated samples. The sperm abnormality and malondialdehyde (MDA) concentration were found highest in 5mM cysteamine treated samples. The cysteamine treated samples travelled significantly less distance in cervical mucus as compared to control. Further, cysteamine decreased acrosomal integrity of sperm. In incubation test, control samples showed better sperm motility as compared to treatment groups. Further, cysteamine supplementation decreased the total antioxidants and increased the MDA concentration of sperm. From the study, we hypothesized that cysteamine cannot stimulate synthesis of glutathione (GSH) intracellularly in sperm to combat free radicals because during the maturation, sperm lost its cytoplasm which is necessary for biochemical reaction in which cysteamine reacts with cystine to form a mixed disulfide which taken up by cells and split into cysteine in the cytoplasm. Synthesis of GSH depends on the availability of cysteine. In conclusion, the results of our study strongly emphasize that cysteamine would not be a suitable additive in extender for freezing buffalo bull semen.

  9. Evaluation of bull fertility in dairy and beef cattle using cow field data.

    PubMed

    Berry, D P; Evans, R D; Mc Parland, S

    2011-01-01

    A successful outcome to a given service is a combination of both male and female fertility. Despite this, most national evaluations for fertility are generally confined to female fertility with evaluations for male fertility commonly undertaken by individual breeding organisations and generally not made public. The objective of this study was to define a pertinent male fertility trait for seasonal calving production systems, and to develop a multiple regression mixed model that may be used to evaluate male fertility at a national level. The data included in the study after editing consisted of 361,412 artificial inseminations from 206,683 cow-lactations (134,911 cows) in 2,843 commercial dairy and beef herds. Fixed effects associated with whether a successful pregnancy ensued (pregnant = 1) or not (pregnant = 0) from a given service were year by month of service, day of the week, days since calving, cow parity, level of calving difficulty experienced, whether or not the previous calving was associated with perinatal mortality, and age of the service bull at the date of insemination. Non-additive genetic effects such as heterosis and recombination loss as well as inbreeding level of the service bull, dam or mating were not associated with a successful pregnancy; there was no difference in pregnancy rate between fresh or frozen semen. Random effects included in the model were the additive genetic effect of the cow, as well as a within lactation and across lactation permanent environmental effect of the cow; pedigree group effects based on cow breed were also included via the relationship matrix. Temporal differences in the AI technician and service bull were also included as random effects. A difference in five percentage units in male fertility was evident between the average effects of different dairy and beef breeds. The correlation between raw pregnancy rates for bulls with more than 100 services (n = 431) and service bull solutions from the mixed model analysis

  10. Use of internal artificial vaginas for breeding soundness evaluation in range bulls: an alternative for electroejaculation allowing observation of sex drive and mating ability.

    PubMed

    Barth, Albert D; Arteaga, Andres A; Brito, Leonardo F C; Palmer, Colin W

    2004-09-01

    The objective of this study was to test the efficacy of an inexpensive, reusable internal artificial vagina (IAV) developed for breeding soundness evaluation of range beef bulls. In addition, sexual behavior during semen collection by IAV was compared to behavior during pasture breeding. Breeding soundness exams (BSEs) were conducted on 165 bulls in two consecutive years (96 in Year 1 and 69 in Year 2). In Year 1, BSEs were done twice in all bulls, once by a conventional protocol using electroejaculation (EEJ), followed by the IAV method, one week later. In Year 2, all BSEs were done on one day; 69 bulls by the IAV method followed by EEJ in 21 bulls that failed to serve the IAV. For semen collection using an IAV, mount cows were restrained in breeding crates and an IAV was inserted into the vagina just beyond the depth of the vestibular sphincter. After each copulation, the IAV was replaced for the next bull to be tested. Semen collection by IAV was successful for all bulls that mounted and penetrated cows during the testing period (54.3 and 69.6% of the bulls served the cows with IAVs in Year 1 and 2, respectively). Semen was collected successfully by EEJ from all bulls in both years. Differences were observed between semen collection methods in semen volume and percentage of sperm staining alive; however, the differences were opposite in Year 1 and 2 and, therefore, were probably due to natural variations in time and within bull rather than the method of semen collection. Semen collection by IAV allowed the detection of problems that prevented copulation in 8 bulls (4.8%) that were determined to be satisfactory potential breeders when semen was collected by EEJ. In Year 1, breeding observations were made at pasture for 15 bulls that served, and 15 bulls that did not serve cows with an IAV. Bulls that did not serve the IAV during the test period had fewer mounts, attempts to mount, and completed services at pasture than bulls that had served the IAV, indicating

  11. Cryogenic preservation of semen from the Aleutian Canada goose (Branta canadensis leucopareia)

    USGS Publications Warehouse

    Gee, G.F.; Sexton, T.J.

    1990-01-01

    Aleutian Canada geese (Branta canadensis leucopareia) were inseminated with frozen-thawed semen containing 6% or 7% dimethylsulfoxide (DMSO) resulting in 32 fertile eggs and 17 goslings; with 7% DMSO, 19 of 31 eggs were fertile. Beltsville Poultry Semen Extender (BPSE), adjusted to 270 ? 30 mOs and 7.5 ? 0.4 pH, was used to dilute semen samples and the DMSO before cryopreservation. About half of the live spermatozoa in the fresh semen (92.9 ? 2.5% live cells, laboratory studies; 87.3 ? 7.3%, insemination trials) survived the freeze-thaw process (46.7 ? 7.8%, laboratory; 33.3 ? 17.8%, insemination trials). Samples of frozen-thawed semen contained a greater percentage of bent spermatozoa (27.1 ? 8.4% of live cells) than fresh semen (14.4 ? 3.0% of live cells). Fecal- and urate-contaminated semen (a common problem when collecting goose semen) reduced the sperm motility score from 3.2 ? 0.6 to 2.7? 0.7 and number of live spermatozoa in frozen-thawed semen from 49 ? 9% to 24 ?18%. Other variables examined that had less of an effect on semen quality included semen extenders, semen holding temperature, dilution and equilibration, relationship between hour of semen collection and level of semen contamination, and the relationship between season and sperm concentration.

  12. The sequential appearance of sperm abnormalities after scrotal insulation or dexamethasone treatment in bulls.

    PubMed Central

    Barth, A D; Bowman, P A

    1994-01-01

    Scrotal insulation and dexamethasone treatment were used as a model to compare the effect of testicular heating and stress on spermatogenesis. Insulation was applied to the scrotum of eight bulls (insulated) for a period of four days, eight bulls were treated daily for seven days with 20 mg dexamethasone injected intramuscularly, and four bulls were untreated controls. Semen from four bulls in each group was collected and evaluated over a six-week period after treatment. Blood samples for testosterone analysis were taken hourly for eight hours at the beginning and the end of the six-week period from the control bulls and before and after treatment from the four insulated and four dexamethasone-treated bulls that were not used for semen collection. At the end of the last blood sampling period, the four bulls in each group were castrated for the collection of testicular tissue for the determination of testosterone concentrations. Basal, peak episodic, and mean serum testosterone concentrations among control bulls, pre and postinsulated bulls, and pretreatment samples of dexamethasone-treated bulls were not different (p > 0.05); however, bulls that had received dexamethasone treatments had significantly lower basal, peak episodic, and mean testosterone concentrations (p < 0.05). Tissue concentrations of testosterone in control, insulated, and dexamethasone-treated bulls were not significantly different but tended to be lower in dexamethasone-treated bulls (p > 0.13). The spermiograms of the control bulls varied insignificantly over the six-week sampling period; however, there was a marked increase in sperm defects in insulated and dexamethasone-treated bulls. The types of sperm defects and the temporal relationships of rises and declines of sperm defects were quite similar for both treatments. All bulls recovered to approximately pretreatment levels of sperm defects by six weeks after the initiation of treatment. Results indicate that two of the most common types of

  13. Effect of supplemental trace mineral level and form on peripubertal bulls

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objectives were to determine if different supplemental trace mineral levels and /or forms (sulfate and metal amino acid complexes) influence age at puberty, semen quality, endocrine status and scrotal circumference in peripubertal bulls. Fifty crossbred, peripubertal bulls were blocked by age (...

  14. Bull breeding soundness evaluation in Southern Africa.

    PubMed

    Irons, P C; Nöthling, J O; Bertschinger, H J

    2007-10-01

    The motivation for and process leading up to the publication of a new bull breeding soundness certification standard endorsed by the South African Veterinary Association is described. The veterinary certificate of bull breeding soundness and explanatory notes and minimum standards are shown. The first component of the certificate is a declaration by the veterinarian that the bull complies with the minimum standards set for examinations for the selected purpose, these being for use as a natural service sire, as a donor of semen for distribution, and for insurance purposes. This is followed by the details of the bull and owner, and a list of the recommended examinations and tests for the bull with provision for which were performed. Certificates are available in book form with the explanatory notes and minimum standards on the reverse, and a carbon copy which remains in the book. The clarity and ease of completion of the document are regarded as being positive features. Bulls are either classified as breeding sound or not, with no actual parameters indicated on the document and no certificate issued for those which do not meet the set criteria. Contact details of the parties involved are shown on the certificate to allow for communication as a means of avoiding disputes.

  15. Seasonal and cryopreservation impacts on semen quality in boars

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  16. Retained functional integrity of bull spermatozoa after double freezing and thawing using PureSperm density gradient centrifugation.

    PubMed

    Maxwell, W M C; Parrilla, I; Caballero, I; Garcia, E; Roca, J; Martinez, E A; Vazquez, J M; Rath, D

    2007-10-01

    The main aim of this study was to compare the motility and functional integrity of bull spermatozoa after single and double freezing and thawing. The viability and morphological integrity of spermatozoa selected by PureSperm density gradient centrifugation after cryopreservation of bovine semen in two commercial extenders (Experiment 1) and the function of bull spermatozoa before and after a second freezing and thawing assisted by PureSperm selection (Experiment 2) were examined. On average, 35.8 +/- 12.1% of sperm loaded onto the PureSperm density gradient were recovered after centrifugation. In Experiment 1, post-thaw motility and acrosome integrity were higher for spermatozoa frozen in Tris-egg yolk extender than in AndroMed, whether the assessments were made immediately after thawing [80.4 +/- 12.7 vs 47.6 +/- 19.0% motile and 78.8 +/- 8.3 vs 50.1 +/- 19.5% normal apical ridge (NAR), p < 0.05] or after preparation on the gradient (83.3 +/- 8.6 vs 69.4 +/- 15.9% motile and 89.5 +/- 7.2 vs 69.1 +/- 11.4% NAR, p < 0.05). For semen frozen in Tris-egg yolk extender, selection on the PureSperm gradient did not influence total motility but significantly improved the proportion of acrosome-intact spermatozoa. After the gradient, both the total motility and percentage of normal acrosomes increased for spermatozoa frozen in AndroMed (Minitüb Tiefenbach, Germany). In Experiment 2, there was no difference in sperm motility after the first and second freeze-thawing (82.9 +/- 12.7 vs 68.8 +/- 18.7%). However, the proportion of acrosome-intact spermatozoa was significantly improved by selection through the PureSperm gradient, whether measured by phase contrast microscopy (78.9 +/- 9.7 vs 90.4 +/- 4.0% NAR, p < 0.05) or flow cytometry (53.4 +/- 11.7 vs 76.3 +/- 6.0% viable acrosome-intact spermatozoa, p < 0.001). The improvement in the percentage of spermatozoa with normal acrosomes was maintained after resuspension in the cooling extender and cooling to 4 degrees C (88

  17. Fertilization rates and in vitro embryo production using sexed or non-sexed semen selected with a silane-coated silica colloid or Percoll.

    PubMed

    Rodríguez Villamil, P; Wei, H; Moreira, G; Caccia, M; Fernandez Taranco, M; Bó, G A

    2012-07-01

    The aim of this study was to evaluate sperm fertilization rates and in vitro embryo development rates for sexed and non-sexed semen selected using a silane-coated silica colloid method (Isolate) or Percoll. Frozen/thawed, sexed and unsexed semen samples from four Holstein bulls were randomly allocated to one of two different density gradient selection methods. Sperm quality (motility, concentration, morphology and membrane integrity) were evaluated and compared before and after sperm selection. Sperm motility and morphology improved (P < 0.005) after the sperm selection process with no differences between the two methods. For non-sexed semen, Percoll gradient increased the mean (± SEM) percentage of sperm recovered (57.3 ± 2.8) compared to Isolate (46.0 ± 1.8; P < 0.01). However, membrane integrity was higher after Isolate than Percoll (sexed semen: 41.0 ± 0.6 vs. 38.8 ± 0.8 and non-sexed semen 60.8 ± 1.6 vs. 58.8 ± 0.5; P < 0.05). The percentage of blastocysts produced was higher when either sexed or non-sexed semen was selected by Isolate (14.0 ± 1.0; 22.0 ± 1.1) than by Percoll (10.5 ± 1.5; 17.0 ± 2.1, respectively; P < 0.05). In summary, Isolate was a more effective method for the recovery of high quality sperm for in vitro fertilization embryo production.

  18. Sexed-semen usage for Holstein AI in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The dairy industry has used sexed-semen to reduce the birth of undesirable bull calves for over a decade. While the efficacy of sexed-semen has been determined experimentally, we sought to tabulate statistics on the generalized use of the technology in the US dairy herd and determine its effectivene...

  19. Identification of single nucleotide polymorphism in protein phosphatase 1 regulatory subunit 11 gene in Murrah bulls

    PubMed Central

    Jain, Varsha; Patel, Brijesh; Umar, Farhat Paul; Ajithakumar, H. M.; Gurjar, Suraj K.; Gupta, I. D.; Verma, Archana

    2017-01-01

    Aim: This study was conducted with the objective to identify single nucleotide polymorphism (SNP) in protein phosphatase 1 regulatory subunit 11 (PPP1R11) gene in Murrah bulls. Materials and Methods: Genomic DNA was isolated by phenol–chloroform extraction method from the frozen semen samples of 65 Murrah bulls maintained at Artificial Breeding Research Centre, ICAR-National Dairy Research Institute, Karnal. The quality and concentration of DNA was checked by spectrophotometer reading and agarose gel electrophoresis. The target region of PPP1R11 gene was amplified using four sets of primer designed based on Bos taurus reference sequence. The amplified products were sequenced and aligned using Clustal Omega for identification of SNPs. Animals were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using EcoNI restriction enzyme. Results: The sequences in the NCBI accession number NW_005785016.1 for Bubalus bubalis were compared and aligned with the edited sequences of Murrah bulls with Clustal Omega software. A total of 10 SNPs were found, out of which 1 at 5’UTR, 3 at intron 1, and 6 at intron 2 region. PCR-RFLP using restriction enzyme EcoNI revealed only AA genotype indicating monomorphism in PPP1R11 gene of all Murrah animals included in the study. Conclusion: A total of 10 SNPs were found. PCR-RFLP revealed only AA genotype indicating monomorphism in PPP1R11 gene of all Murrah animals included in the study, due to which association analysis with conception rate was not feasible. PMID:28344410

  20. A comparison of ABAcard(®) p30 and RSID™-Semen test kits for forensic semen identification.

    PubMed

    Boward, Emily S; Wilson, Stacey L

    2013-11-01

    The screening and confirmatory tests available to a forensic laboratory allow evidence to be examined for the presence of bodily fluids. With the majority of evidence being submitted involving sexual assaults, it is important to have confirmatory tests for the identification of semen that are straightforward, quick, and reliable. The purpose of this study was to compare two commonly used semen identification kits utilized by forensic laboratories: ABAcard(®) p30 and Rapid Stain Identification of Human Semen (RSID™-Semen). These kits were assessed with aged semen stains, fresh and frozen post-vasectomy semen, post-coital samples collected on different substrates, post-vasectomy semen mixed with blood, saliva, and urine, a series of swabs collected at increasing time intervals after sexual intercourse, and multiple non-semen samples. The test kits were compared on the basis of sensitivity, specificity, and the cost and time effectiveness of each protocol. Overall, both semen identification tests performed well in the studies. Both kits proved specificity for identifying semen, however the ABAcard(®) p30 test surpassed the RSID™-Semen test in sensitivity, cost per test, and simplified test protocol.

  1. Maintaining semen quality by improving cold chain equipment used in cattle artificial insemination

    PubMed Central

    Lieberman, Daniel; McClure, Elizabeth; Harston, Stephen; Madan, Damian

    2016-01-01

    Artificial insemination of dairy cattle is a common practice in the developing world that can improve farmer incomes and food security. Maintaining the fertilizing potential of frozen semen as it is manipulated, transported and stored is crucial to the success of this process. Here we describe simple technological improvements to protect semen from inadvertent thermal fluctuations that occur when users mishandle semen using standard equipment. We show that when frozen semen is mishandled, characteristics of semen biology associated with fertility are negatively affected. We describe several design modifications and results from thermal performance tests of several improved prototypes. Finally, we compare semen that has been mishandled in standard and improved equipment. The data suggest that our canister improvements can better maintain characteristics of semen biology that correlate with fertility when it is mishandled. PMID:27313137

  2. Maintaining semen quality by improving cold chain equipment used in cattle artificial insemination

    NASA Astrophysics Data System (ADS)

    Lieberman, Daniel; McClure, Elizabeth; Harston, Stephen; Madan, Damian

    2016-06-01

    Artificial insemination of dairy cattle is a common practice in the developing world that can improve farmer incomes and food security. Maintaining the fertilizing potential of frozen semen as it is manipulated, transported and stored is crucial to the success of this process. Here we describe simple technological improvements to protect semen from inadvertent thermal fluctuations that occur when users mishandle semen using standard equipment. We show that when frozen semen is mishandled, characteristics of semen biology associated with fertility are negatively affected. We describe several design modifications and results from thermal performance tests of several improved prototypes. Finally, we compare semen that has been mishandled in standard and improved equipment. The data suggest that our canister improvements can better maintain characteristics of semen biology that correlate with fertility when it is mishandled.

  3. Semen collection using phantom in dromedary camel.

    PubMed

    Ziapour, S; Niasari-Naslaji, A; Mirtavousi, M; Keshavarz, M; Kalantari, A; Adel, H

    2014-12-10

    Semen collection is relatively long, unsafe, and tedious procedure in dromedary camel. The innovation of safe, hygienic, and simple approach to collect semen could make great progress in development of AI program in this species. This study investigated two methods of semen collection using phantom and artificial vagina in dromedary camel. Semen was collected using phantom (n = 4 bulls; 26 collections) and artificial vagina (n = 6 bulls; 11 collections) and diluted with INRA96 at the ratio of 1:10. The duration of semen collection, semen parameters, and morphometric features of sperm were evaluated. For specimen collected through phantom and AV, the respected duration of semen collection (411.2 ± 48.19 vs 326 ± 37.05 sec), volume (6.6 ± 0.87 vs 6 ± 1.57 ml), osmolarity (328 ± 1.6 vs 319.4 ± 3.21 mOsm/kg H2O), pH (7.7 ± 0.06 vs 7.9 ± 0.16) of semen, concentration (161.4 ± 44.05 × 10(6)/mL vs 160.2 ± 58.42 × 10(6)/mL), total motility (84.1 ± 1.89 vs 78.3 ± 3.97%), progressive forward motility (45.5 ± 3.69 vs 44.3 ± 6.41%), live percentage (72.2 ± 3.11 vs 76 ± 2.53%), and plasma membrane integrity (61.5 ± 2.49 vs 58.9 ± 4.19%) of sperm were similar (P > 0.05). The number of specimens contaminated with visible particles was greater using AV (72.7%) compared to phantom (0%; P < 0.05). Total length, head, middle-piece, and tail length of sperm were 45.9 ± 0.1, 5.6 ± 0.01, 7 ± 0.02, and 34.2 ± 0.16 μm, respectively. The frequency of abnormal sperm was 13.28% among which coiled tail, detached head, and proximal protoplasmic droplets had greater incidence. In conclusion, phantom could be considered as a suitable approach to collect semen due to simplicity, safety, and lack of specimen contamination in dromedary camel.

  4. Association of oxidative status and semen characteristics with seminal plasma proteins of buffalo semen

    PubMed Central

    Sharma, L.; Pandey, V.; Nigam, R.; Saxena, A.; Swain, D. K.; Yadav, B.

    2016-01-01

    To study the influence of season on oxidative status of buffalo semen and their association with semen characteristics and seminal plasma proteins, ejaculates were collected twice a week in winter, summer and rainy seasons from six adult Bhadawari buffalo bulls. The neat semen was analyzed for semen characteristics immediately after collection and oxidative status viz. lipid peroxidation (LPO), catalase (CAT), super oxide dismutase (SOD) activity, and total protein (TP) were estimated in seminal plasma. The protein profiling was carried out by one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The significant effect of season was observed on TP, SOD activity and % protein fractions of seminal plasma proteins of buffalo bulls. The TP values showed positive correlation with ejaculate volume (EV), sperm concentration (SC), and % live-dead (LD) and negative correlation with progressive motility (PM), and hypo-osmotic swelling test (HOST). The SOD activity showed positive correlation with PM, LD, HOST and % acrosoamal integrity (AI). Besides that, results showed correlation of TP with 6.5, 38 and 66 kDa proteins, LPO with 70, 72, 84 and 86 kDa proteins, CAT with 70 kDa and 86 kDa proteins, and SOD with 6.5, 24.5, 44.5, 70 and 72 kDa proteins. In conclusion, this study indicated that TP and SOD activity of seminal plasma of buffalo bulls were influenced by season and were found to be associated with some of the semen characteristics and expression of seminal plasma proteins. PMID:28224004

  5. Effects of In Vitro Zinc Sulphate Additive to The Semen Extender on Water Buffalo (Bubalusbubalis) Spermatozoa before and after Freezing

    PubMed Central

    Dorostkar, Kamran; Alavi Shoushtari, Sayed Mortaza; Khaki, Amir

    2014-01-01

    Background The objective of the study was to investigate the effects of in vitro zinc sulphate additive to semen extender on sperm parameters (progressive motility, viability, membrane integrity and DNA stability) after cryopreservation. Materials and Methods In this Prospective longitudinal laboratory study, semen samples of 5 buffalo bulls of 3-5 years old were collected at 5 different occasions from Iran, Urmia during summer and autumn 2011, 25 samples were used in each treatment. Sperm progressive motility, viability and abnormal morphology were measured before and at 0.5 (T0), 1(T1) and 2(T2) hours after diluting semen(1:10 v/v) in Tris-citric acid based extender (without egg yolk and glycerol) at 37˚C containing none (control group), 0.072, 0.144, 0.288, 0.576 and 1.152 mg/L zinc sulphate to investigate dose and time effects. Next, a Tris-citric acid-egg yolk-glycerol extender (20% egg yolk and 7% glycerol) containing the same amount of zinc sulphate was prepared, diluted semen (1:10 v/v) was cooled and kept into a refrigerated chamber (4˚C) for 4 hours to equilibrate. Sperm progressive motility, viability, abnormal morphology, membrane integrity and DNA damage were estimated.The equilibrated semen was loaded in 0.5 ml French straws and frozen in liquid nitrogen. Later, the frozen semen was thawed and the same parameters as well as total antioxidant capacity (TAC) of the frozen-thawed semen were determined. Results The results showed that zinc sulphate additive at the rate of 0.288 mg/L gave a higher protection of sperm progressive motility (53.7 ± 1.8% vs. 40.5 ± 1.7%), viability (70.8 ± 1.8% vs. 60.1 ± 1.5%), membrane integrity (67.3 ± 1.6% vs. 56.6 ± 1.7%), DNA stability (10.1 ± 0.47% vs. 11.8 ± 0.33% damaged DNA) through the process of dilution, equilibration and freeze-thawing and caused a higher TAC level (81 ± 3.3% vs. 63 ± 3.2 µmol/L) after freez-thawing compared to the control group. Adding 0.576 and 1.152 mg/L zinc sulphate, however

  6. Update on sexed semen technology in cattle.

    PubMed

    Seidel, G E

    2014-05-01

    The technology in current use for sexing sperm represents remarkable feats of engineering. These flow cytometer/cell sorters can make over 30 000 consecutive evaluations of individual sperm each second for each nozzle and sort the sperm into three containers: X-sperm, Y-sperm and unsexable plus dead sperm. Even at these speeds it is not economical to package sperm at standard numbers per inseminate. However, with excellent management, pregnancy rates in cattle with 2 million sexed sperm per insemination dose are about 80% of those with conventional semen at normal sperm doses. This lowered fertility, in part due to damage to sperm during sorting, plus the extra cost of sexed semen limits the applications that are economically feasible. Even so, on the order of 2 million doses of bovine semen are sexed annually in the United States. The main application is for dairy heifers to have heifer calves, either for herd expansion or for sale as replacements, often for eventual export. Breeders of purebred cattle often use sexed semen for specific matings; thawing and then sexing frozen semen and immediately using the few resulting sexed sperm for in vitro fertilization is done with increasing frequency. Beef cattle producers are starting to use sexed semen to produce crossbred female replacements. Proprietary improvements in sperm sexing procedures, implemented in 2013, are claimed to improve fertility between 4 and 6 percentage points, or about 10%.

  7. Frozen Frozen CO2

    NASA Technical Reports Server (NTRS)

    2005-01-01

    2 October 2005 This Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) image shows a view of frozen carbon dioxide in the south polar residual cap of Mars. Much of the south polar residual cap exhibits terrain that resembles stacks of sliced Swiss cheese, but this portion of the cap lacks the typical, circular depressions that characterize much of the region. Carbon dioxide on Mars freezes at a temperature of around 148 Kelvins, which is -125oC or about -193oF.

    Location near: 87.2oS, 28.4oW Image width: width: 3 km (1.9 mi) Illumination from: upper left Season: Southern Spring

  8. The effects of addition of omega-3, 6, 9 fatty acids on the quality of bovine chilled and frozen-thawed sperm.

    PubMed

    Kandelousi, M A Sheikholeslami; Arshami, J; Naserian, A A; Abavisani, A

    2013-01-01

    This study was aimed to investigate the effects of omega-3, 6, 9 fatty acids on the characteristics of bovine chilled and frozen-thawed semen. For this purpose, oil containing different levels of omega-3, 6, 9 fatty acids were added to semen extender. To emulsify the oil in semen extender, polyethylene glycol (PEG) was added as a suitable solvent and the solution was finally sonicated. Five proven Holstein bulls were randomly selected and their ejaculates were collected using an artificial vagina. Groups were designed as control, treatments 1, 2, 3 and 4. The control group contained only the basic extender (Tris-citrate buffer, egg yolk and glycerol) without any additives. In treatment 1, only 5% PEG was added to the diluent; while in treatments 2, 3 and 4 different concentrations of omega-3, 6, 9 fatty acids (1.0, 2.5 and 5.0%) in combination with PEG were added to the basic extender. After dilution, the semen samples were packaged into 0.5 ml straws, a process that was followed by cooling the semen straws. Motility, viability and morphology of semen samples were evaluated after 24 and 48 h of storage in refrigerator (5 °C) or after one month of storage in the liquid nitrogen. Immotility was increased and all the other parameters including motility, viability and morphology were significantly decreased in all the groups compared with fresh samples during cold storage and freezing-thawing. Our results demonstrated the following: 1) PEG has significant detrimental effects, especially on the sperm motility; 2) addition of omega-3, 6, 9 fatty acids could not improve the sperm motility in chilled storage condition and after freezing-thawing; and 3) omega-3, 6, 9 fatty acidscould not also attenuate the other deleterious effects of PEG. In conclusion, our findings reveal that addition of these fatty acids to the semen extender does not enhance the resistance of the bovine sperm membrane to cooling and freezing-thawing and that further studies are required to find

  9. Risk of Salmonella transmission via cryopreserved semen in turkey flocks.

    PubMed

    Iaffaldano, N; Reale, A; Sorrentino, E; Coppola, R; Di Iorio, M; Rosato, M P

    2010-09-01

    To investigate the possibility to carry pathogen bacteria in turkey flocks via cryopreserved semen, research was carried out 1) to investigate the microbial contamination of fresh and frozen thawed turkey semen and 2) to evaluate the effect of the freezing-thawing process on the survival of 3 serovars of Salmonella spp. experimentally inoculated in turkey semen. Five pools of semen diluted 4-fold were cooled, added with 8% of dimethylacetamide as a cryoprotectant, and aliquots of 80 muL were directly plunged into liquid nitrogen to form frozen pellets. Mesophilic viable counts, total and fecal coliforms, Enterobacteriaceae, enterococci, Campylobacter spp., and Salmonella spp. were investigated on fresh and thawed samples. Further, 5 pools of diluted semen were each divided into 3 subsamples, inoculated with 7.8 +/- 0.2 log cfu.mL(-1) of Salmonella Liverpool, Salmonella Montevideo, and Salmonella Braenderup, respectively, and cryopreserved before to assess the postthaw viability of Salmonella spp. strains. Fresh semen was highly contaminated by all of the saprophytic bacteria investigated and the cryopreservation process reduced the amount of mesophilic viable count and total coliforms (P < 0.05) and fecal coliforms, Enterobacteriaceae, and enterococci (P < 0.01) by about 1 log cfu.mL(-1). Conversely, neither Campylobacter spp. nor Salmonella spp. were found as endogenous bacteria in semen. In the inoculated semen, both Salmonella Liverpool, Salmonella Montevideo, and Salmonella Braenderup colonies were recovered postthaw, showing a significant reduction of 2.03 +/- 0.28, 3.08 +/- 0.22, and 2.72 +/- 0.23 log cfu.mL(-1), respectively, compared with the fresh semen (P < 0.001). In conclusion, the cryopreservation process allowed us to obtain a low reduction of microbial count both in endogenous saprophytic bacteria and artificially inoculated Salmonella spp. strains; therefore, the possibility of Samonella spp. transmission to flocks through the use of infected

  10. Expected net present value of pure and mixed sexed semen artificial insemination strategies in dairy heifers.

    PubMed

    Olynk, N J; Wolf, C A

    2007-05-01

    Sexed semen has been a long-anticipated tool for dairy farmers to obtain more heifer calves, but challenges exist for integrating sexed semen into commercial dairy farm reproduction programs. The decreased conception rates (CR) experienced with sexed semen make virgin heifers better suited for insemination with sexed semen than lactating dairy cows. This research sought to identify when various sexed semen breeding strategies provided higher expected net present value (NPV) than conventional artificial insemination (AI) breeding schemes, indicating which breeding scheme is advisable under various scenarios. Budgets were developed to calculate the expected NPV of various AI breeding strategies incorporating conventional (non-sexed) and sexed semen. In the base budgets, heifer and bull calf values were held constant at $500 and $110, respectively. The percentage of heifers expected to be born after breeding with conventional and sexed semen used was 49.2 and 90%, respectively. Breeding costs per AI were held constant at $15.00 per AI for conventional semen and $45.00 per AI for sexed semen of approximately the same genetic value. Conventional semen CR of 58 and 65% were used, and an AI submission rate was set at 100%. Breeding strategies with sexed semen were assessed for breakeven heifer calf values and sexed semen costs to obtain a NPV equal to that achieved with conventional semen. Breakeven heifer calf values for pure sexed semen strategies with a constant 58 and 65% base CR in which sexed semen achieved 53% of the base CR are $732.11 and $664.26, respectively. Breakeven sexed semen costs per AI of $17.16 and $22.39, compared with $45.00 per AI, were obtained to obtain a NPV equal to that obtained with pure conventional semen for base CR of 58 and 65%, respectively. The strategy employing purely sexed semen, with base CR of both 58 and 65%, yielded a lower NPV than purely conventional semen in all but the best-case scenario in which sexed semen provides 90% of

  11. Reproduction in nondomestic birds: Physiology, semen collection, artificial insemination and cryopreservation

    USGS Publications Warehouse

    Gee, G.F.; Bertschinger, H.; Donoghue, A.M.; Blanco, J.; Soley, J.

    2004-01-01

    Pioneering work by Quinn and Burrows in the late 1930s led to successful artificial insemination (AI) programs in the domestic poultry industry. A variety of species specific modifications to the Quinn and Burrows massage technique made AI possible in nondomestic birds. Massage semen collection and insemination techniques span the entire range of species from sparrows to ostriches. Also, cooperative semen collection and electroejaculation have found limited use in some nondomestic species. Artificial insemination produces good fertility, often exceeding fertility levels in naturally copulating populations. However, aviculturists should explore other ways to improve fertility before resorting to AI. Artificial insemination is labor intensive and may pose risks to nondomestic birds as well as handlers associated with capture and insemination. Semen collection and AI makes semen cryopreservation and germ plasma preservation possible. Yet, semen cryopreservation techniques need improvement before fertility with frozen-thawed semen will equal fertility from AI with fresh semen.

  12. Lycopene and resveratrol improve post-thaw bull sperm parameters: sperm motility, mitochondrial activity and DNA integrity.

    PubMed

    Bucak, M N; Ataman, M B; Başpınar, N; Uysal, O; Taşpınar, M; Bilgili, A; Öztürk, C; Güngör, Ş; İnanç, M E; Akal, E

    2015-06-01

    We focussed on evaluating the protective effect of lycopene and resveratrol on post-thaw bull sperm and oxidative stress parameters. Nine ejaculates for each bull were used in the study. Each ejaculate, splitted into three equal aliquots and diluted at 37 °C with base extenders containing lycopene (1 × 10(-3)  g ml(-1) ) and resveratrol (1 mm), and no antioxidant (control), was cooled to 5 °C and then frozen. Frozen straws were thawed in a water bath for evaluation. The supplementation of the semen extender with lycopene and resveratrol increased the percentages of post-thawed computer-assisted sperm analysis (CASA) motility (55.8 ± 3.8 and 61.9 ± 4.0%) and progressive motility (38 ± 2.4 and 37 ± 8.8), compared with the controls (50.7 ± 2.65 and 33.3 ± 3.74%, respectively, P < 0.05). Resveratrol provided a higher ALH (4.3 ± 0.1), in comparison with the control (3.9 ± 0.3, P < 0.05). The supplementation of the semen extender with lycopene and resveratrol produced a higher mitochondrial activity (24.6 ± 2.9 and 30.1 ± 6.5% respectively), compared with that of the control (11.8 ± 9.5%, P < 0.05). It was determined that both antioxidants resulted in a lower percentage of sperm with damaged DNA than that of the control (P < 0.05). Sperm motion characteristics except for ALH, acrosome integrity, sperm viability and oxidative stress parameters were not affected by the adding of lycopene and resveratrol.

  13. Cryopreservation of canine semen after cold storage in a Neopor box: effect of extender, centrifugation and storage time.

    PubMed

    Hidalgo, M; Portero, J M; Demyda-Peyrás, S; Ortiz, I; Dorado, J

    2014-07-05

    The aim of this work was to assess the combined effect of sperm centrifugation, semen extender and storage time before freezing on post-thaw sperm quality and freezability on chilled stored canine semen in a Neopor box. Sperm parameters evaluated were total and progressive sperm motility by Computer-Assisted Sperm Analysis (CASA) and sperm viability and acrosome integrity using a triple fluorescent stain. Sperm quality and freezability indexes were also studied. First, the effect of centrifugation and two commercial extenders from Minitübe (Biladyl A and CaniPRO Freeze A) was evaluated in chilled semen after 24 and 45 hours of cold storage. No significant differences were observed between treatments in almost all the sperm parameters assessed. Secondly, chilled semen was frozen after 24 and 45 hours of cold storage in a Neopor box. The best results were obtained when semen was centrifuged, chilled with CaniPRO Freeze A and then frozen after 24 hours of cold storage, showing no differences in both post-thaw sperm quality and freezability in comparison with semen immediately frozen after collection. In conclusion, dog semen centrifuged after collection and extended with CaniPRO Freeze can be frozen after 24 hours of cold storage in a Neopor box, obtaining similar results to semen immediately frozen after collection.

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

    PubMed

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

    2015-06-01

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

  15. Red Bull Stratos Presentation

    NASA Video Gallery

    Red Bull Stratos High Performance Director Andy Walshe & Technical Project Director Art Thompson share the Stratos story with JSC. Supported by a team of experts, Felix Baumgartner reached 128,100 ...

  16. Electroejaculation increased vocalization and plasma concentrations of cortisol and progesterone, but not substance P, in beef bulls.

    PubMed

    Whitlock, B K; Coffman, E A; Coetzee, J F; Daniel, J A

    2012-09-01

    Electroejaculation is a reliable method of obtaining a semen sample for a bull breeding soundness examination, but is sometimes regarded as painful. Substance P is a neuropeptide involved in the integration of pain, stress, and anxiety. We hypothesized that substance P is a measure of pain in bulls following electroejaculation. The specific objective was to compare vocalization and plasma concentrations of cortisol, progesterone, and substance P immunoreactivity in bulls following electroejaculation. Nine Angus bulls (501.9 ± 14.3 kg) were used. Blood samples were collected at -60, -30, 0, 2, 10, 20, 30, 45, 60, 75, 90, 120 min relative to treatment. At Time 0, bulls were subject to electroejaculation, rectal probe insertion without electroejaculation, or no manipulation. Treatments were administered contemporaneously to three bulls. Treatments were repeated weekly until each bull had received each treatment in a 3 × 3 Latin square design. More bulls (P = 0.0147) in the electroejaculation group vocalized (5 of 9 bulls; 55.6%) when compared to controls (0 of 9 bulls; 0%). Mean plasma cortisol and progesterone concentration following electroejaculation in bulls were higher (P < 0.05) than concentrations in probed and control bulls through the 45 min sample. However, mean plasma substance P concentration following electroejaculation in bulls (77.2 ± 17.2 pg/mL) was not different (P = 0.6264) from probed (79.1 ± 17.2 pg/mL) or control bulls (93.4 ± 17.2 pg/mL). A significant increase in vocalization and plasma cortisol and progesterone concentrations in bulls following electroejaculation was likely owing to acute stress. However, the lack of a difference in plasma concentrations of substance P after electroejaculation was interpreted as a lack of pain associated with nociception.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    PubMed

    Persson, Ylva; Söderquist, Lennart; Ekman, Stina

    2007-11-05

    The lame sire, unsound for breeding, can cause substantial economic loss due to reduced pregnancies in the beef-producing herd. To test the hypothesis that joint disorder is a possible cause of infertility in beef sires, right and left hind limb bones from 34 beef sires were examined postmortem to identify lesions in the femorotibial, femoropatellar (stifle), tarsocrural, talocalcaneus, and proximal intertarsal (tarsal) joints. The bulls were slaughtered during or after the breeding season due to poor fertility results. Aliquots of the cauda epididymal contents taken postmortem from 26 bulls were used for sperm morphology evaluation. As a control, hind limbs (but no semen samples) from 11 beef bulls with good fertility results were included. Almost all infertile bulls (30/34) had lesions in at least one joint. Twenty-eight bulls (28/30, 93%) had lesions in the stifle joint, and 24 (24/28, 86%) of these were bilateral. Fourteen bulls (14/30, 47%) had lesions in the tarsal joint, and 10 (10/14, 71%) of these were bilateral. Four bulls (4/34, 12%) had no lesions, three bulls (3/34, 9%) had mild osteoarthritis (OA), 5 (5/34, 15%) moderate OA, 17 (17/34, 50%) severe OA and 5 (5/34, 15%) deformed OA. Almost all OA lesions (97%) were characterized as lesions secondary to osteochondrosis dissecans. All the bulls with satisfactory sperm morphology (n = 12/34) had joint lesions, with mostly severe or deformed bilateral lesions (83%). Consequently, the most likely cause of infertility in these 12 bulls was joint disease. Almost all control bulls (10/11) had OA lesions, but most of them were graded as mild (55%) or moderate (36%). None of the control bulls had severe lesions or deformed OA. We suggest that joint lesions should be taken into consideration as a contributory cause of reproductive failure in beef sires without symptoms of lameness.

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

    PubMed

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

    2016-11-01

    The effects of l-cysteine in extender on antioxidant enzymes profile during cryopreservation, post-thaw quality parameters and in vivo fertility of Nili-Ravi buffalo bull spermatozoa were studied. Semen samples from 4 buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of l-cysteine (0.0, 0.5, 1.0, 2.0 and 3.0 mm) and frozen in 0.5-ml French straws. The antioxidative enzymes [catalase, super oxide dismutase and total glutathione (peroxidase and reductase)] were significantly higher (P < 0.05) at pre-freezing and post-thawing in extender containing 2.0 mm l-cysteine as compared to other groups. Post-thaw total motility (%), progressive motility (%), rapid velocity (%), average path velocity (μm s(-1) ), straight line velocity (μm s(-1) ), curvilinear velocity (μm s(-1) ), beat cross frequency (Hz), viable spermatozoa with intact plasmalemma (%), acrosome and DNA integrity (%) were higher with the addition of 2.0 mm l-cysteine as compared to other groups (P < 0.05). The fertility rates (59 versus 43%) were higher (P < 0.05) in buffaloes inseminated with doses containing 2.0 mm of l-cysteine than in the control. In conclusion, the addition of 2.0 mm l-cysteine in extender improved the antioxidant enzymes profile, post-thaw quality and in vivo fertility of Nili-Ravi buffalo bull spermatozoa.

  20. Protective effects of l-carnitine on astheno- and normozoospermic human semen samples during cryopreservation.

    PubMed

    Zhang, Wei; Li, Feng; Cao, Haifeng; Li, Chuyan; Du, Congqi; Yao, Lingnv; Mao, Huan; Lin, Wenqin

    2016-04-01

    This study was conducted to determine the effects of l-carnitine (LC), as an antioxidant, in preventing spermatozoa damage during the freezing-thawing process in both astheno- and normozoospermic human semen samples. Seventy semen samples (37 asthenozoospermic and 33 normozoospermic) were involved in this study. Cryopreservation medium supplemented with 1.0 g/l LC was mixed with semen at a ratio of 1:1 (v/v). Controls were cryopreserved with freezing medium only. Assessment of motility, viability (VIA), mitochondrial membrane potential (MMP) and DNA fragmentation index (DFI) were performed on aliquots of fresh semen, frozen-thawed control and frozen-thawed LC treated samples. Supplementation of the cryopreservation medium with LC induced a significant improvement in post-thaw sperm parameters in both the asthenozoospermic and normozoospermic semen samples, compared with those of the control, regarding sperm fast forward motility, forward motility, total motility and VIA. LC showed better protective effects towards asthenozoospermia for DFI (F = 115.85, P < 0.01) and VIA (F = 67.14, P < 0.01) than did normozoospermic semen samples. We conclude that supplementation with LC prior to the cryopreservation process reduced spermatozoa cryodamage in both asthenozoospermic and normozoospermic semen samples. LC had better protective effects for asthenozoospermic human semen samples. Future research should focus on the molecular mechanism for and the different protective effects of LC between asthenozoospermic and normozoospermic semen samples during cryopreservation.

  1. A successful new approach to honeybee semen cryopreservation.

    PubMed

    Wegener, Jakob; May, Tanja; Kamp, Günter; Bienefeld, Kaspar

    2014-10-01

    Honeybee biodiversity is under massive threat, and improved methods for gamete cryopreservation could be a precious tool for both the in situ- and ex situ-conservation of subspecies and ecotypes. Recent cryoprotocols for drone semen have improved the viability and fertility of frozen-thawed semen by using increased diluent:semen-ratios, but there is still much room for progress. As semen cryopreserved after dilution often appeared hyperactive, we speculated that the disruption of sperm-sperm interactions during dilution and cryopreservation could reduce the fertile lifespan of the cells. We therefore developed protocols to reduce admixture, or abolish it altogether by dialyzing semen against a hypertonic solution of cryoprotectant. Additionally, we tested methods to reduce the cryoprotectant concentration after thawing. Insemination of queens with semen cryopreserved after dialysis yielded 49%, 59% and 79% female (= stemming from fertilized eggs) pupae in three separate experiments, and the numbers of sperm found in the spermathecae of the queens were significantly higher than those previously reported. Post-thaw dilution and reconcentration of semen for cryoprotectant removal reduced fertility, but sizeable proportions of female brood were still produced. Workers stemming from cryopreserved semen did not differ from bees stemming from untreated semen with regard to indicators of fluctuating asymmetry, but were slightly heavier. Cryopreservation after dialysis tended to increase the proportion of cells with DNA-nicks, as measured by the TUNEL-assay, but this increase appears small when compared to the baseline variations of this indicator. Overall, we conclude that cryoprotectant-addition through dialysis can improve the quality of cryopreserved drone semen. Testing of offspring for vitality and genetic integrity should continue.

  2. Trehalose improves semen antioxidant enzymes activity, post-thaw quality, and fertility in Nili Ravi buffaloes (Bubalus bubalis).

    PubMed

    Iqbal, Sajid; Andrabi, Syed Murtaza Hassan; Riaz, Amjad; Durrani, Aneela Zameer; Ahmad, Nasim

    2016-03-15

    Our objectives were to study the effect of trehalose in extender on (1) antioxidant enzymes profile during cryopreservation (after dilution, before freezing, and after thawing), (2) in vitro quality (after thawing), and (3) in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa. Semen samples (n = 20) from four buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of trehalose (0.0, 15, 30, 45, and 60 mM) and frozen in French straws. At post dilution, profile of sperm catalase (U/mL) was higher (P < 0.05) in extenders containing 15, 30, and 45 mM of trehalose as compared to control. Although profiles of superoxide dismutase (U/mL) and total glutathione (μM) were higher (P < 0.05) in extenders containing 15 and 30 mM of trehalose as compared to control. At prefreezing, sperm catalase, superoxide dismutase, and total glutathione profiles were higher (P < 0.05) in all the treatment groups as compared to control. At post thawing, the profiles of catalase and total glutathione were higher (P < 0.05) in extender containing 30-mM trehalose as compared to other treatment groups and control. Whereas, profile of superoxide dismutase was higher (P < 0.05) in extenders containing 30, 45, and 60 mM of trehalose as compared to control and 15mM group. Post thaw total sperm motility (%) was higher (P < 0.05) in extender containing 30-mM trehalose as compared to control and 15 and 60-mM groups. Although sperm progressive motility (%), rapid velocity (%), average path velocity (μm/s), straight line velocity (μm/s), curvilinear velocity (μm/s), plasma membrane (structural and functional, %), acrosome (%), and DNA (%) integrity were higher (P < 0.05) in extender containing 30 mM trehalose as compared to other treatment groups and control. The fertility rates (61% vs. 43%) were higher (P < 0.05) in buffaloes inseminated with semen doses cryopreserved in extender containing 30 mM of trehalose than the control. It is

  3. Artificial insemination and cryopreservation of semen from nondomestic birds

    USGS Publications Warehouse

    Gee, G.F.; Bakst, M.R.; Wishart, G.J.

    1995-01-01

    Studies of Al and cryopreservation of semen from nondomestic birds began because of the increased emphasis on conservation of avian species threatened with extinction. Over the years, aviculturists have developed techniques for Al and cryopreservation of semen obtained from a variety of birds ranging from passerines to Andean condors. Generally, for each new species, we develop a practical semen collection technique and then evaluate the semen. A commercial semen extender (Beltsville Poultry Semen Extender) is modified and used to dilute the semen and provide support for the sperm during the freezing process (the pH and osmolality of the extender is adjusted to reflect the pH and osmolality of the semen being frozen). We find that the freezing schedule developed by Sexton (1977), which utilizes dimethylsulfoxide (DMS0) as cryoprotectant, works well for many species. We cool the sample sequentially in an ethanol bath, in liquid nitrogen vapor, and lastly in liquid nitrogen. Although we have experimented with a variety of freezing protocols, we prefer a 15-min equilibration period in DMSO at 5 C. We begin the freezing process by cooling at -1 C/min from 5 to -20 C in the ethanol bath. The samples are transferred into a vapor tank at a location just above liquid nitrogen and frozen at -50 C/min to -80 C. To complete the freezing process, the samples are plunged into the liquid nitrogen in the bottom of the vapor tank. The samples remain in liquid nitrogen until they are thawed just before insemination. If necessary, the freezing equipment can be transported in a van to remote locations.

  4. Cryopreservation of turkey semen: effect of breeding line and freezing method on post-thaw sperm quality, fertilization, and hatching

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryopreservation methods for poultry semen are not reliable for germplasm preservation, especially for turkeys, where fertility rates from frozen/thawed semen are particularly low. The objective was to evaluate cryopreservation methods for effectiveness in promoting cryosurvival and post-thaw funct...

  5. The utility of nanowater for ram semen cryopreservation.

    PubMed

    Murawski, Maciej; Schwarz, Tomasz; Grygier, Joanna; Patkowski, Krzysztof; Oszczęda, Zdzisław; Jelkin, Igor; Kosiek, Anna; Gruszecki, Tomasz M; Szymanowska, Anna; Skrzypek, Tomasz; Zieba, Dorota A; Bartlewski, Pawel M

    2015-05-01

    Nanowater (NW; water declusterized in the low-temperature plasma reactor) has specific physicochemical properties that could increase semen viability after freezing and hence fertility after artificial insemination (AI) procedures. The main goal of this study was to evaluate ram semen quality after freezing in the media containing NW. Ejaculates from 10 rams were divided into two equal parts, diluted in a commercially available semen extender (Triladyl®; MiniTüb GmbH, Tiefenbach, Germany) prepared with deionized water (DW) or NW, and then frozen in liquid nitrogen. Semen samples were examined for sperm motility and morphology using the sperm class analyzer system and light microscopy. Cryo-scanning electron microscopy (cryo-SEM) was employed to determine the size of extracellular water crystals in frozen semen samples. Survival time at room temperature, aspartate aminotransferase (AspAT) and alkaline phosphatase (ALP) concentrations post-thawing as well as conception/lambing rates after laparoscopic intrauterine AI of 120 ewes were also determined. There were no significant differences between DW and NW groups in sperm progressive motility (26.4 ± 12.2 and 30.8 ± 12.4%) or survival time (266.6 ± 61.3 and 270.9 ± 76.7 min) after thawing and no differences in the percentages of spermatozoa with various morphological defects before or after freezing. There were, however, differences (P < 0.05) in AspAT (DW: 187.1 ± 160.4 vs. NW: 152.7 ± 118.3 U/l) and ALP concentrations (DW: 2198.3 ± 1810.5 vs. NW: 1612.1 ± 1144.8 U/l) in semen samples post-thawing. Extracellular water crystals were larger (P < 0.05) in ejaculates frozen in NW-containing media. Ultrasonographic examinations on day 40 post-AI revealed higher (P < 0.05) conception rates in ewes inseminated with NW (78.3%) compared with DW semen (58.3%), and the percentages of ewes that carried lambs to term were 73.3% and 45.0% in NW and DW groups, respectively (P < 0.01). In summary, the use of a semen

  6. Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and eland antelope.

    PubMed

    Nel-Themaat, L; Gómez, M C; Damiani, P; Wirtu, G; Dresser, B L; Bondioli, K R; Lyons, L A; Pope, C E; Godke, R A

    2007-01-01

    Semen and milk are potential sources of somatic cells for genome banks. In the present study, we cultured and characterised cells from: (1) cooled sheep milk; (2) fresh, cooled and frozen-thawed semen from Gulf Coast native (GCN) sheep (Ovis aries); and (3) fresh eland (Taurotragus oryx) semen. Cells attached to the culture surface from fresh (29%), cooled (43%) and slow-frozen (1 degrees C/min; 14%) ram semen, whereas no attachment occurred in the fast-frozen (10 degrees C/min) group. Proliferation occurred in fresh (50%) and cooled (100%) groups, but no cells proliferated after passage 1 (P1). Eland semen yielded cell lines (100%) that were cryopreserved at P1. In samples from GCN and cross-bred milk, cell attachment (83% and 95%, respectively) and proliferation (60% and 37%, respectively) were observed. Immunocytochemical detection of cytokeratin indicated an epithelial origin of semen-derived cells, whereas milk yielded either fibroblasts, epithelial or a mixture of cell types. Deoxyribonucleic acid microsatellite analysis using cattle-derived markers confirmed that eland cells were from the semen donor. Eland epithelial cells were transferred into eland oocytes and 12 (71%), six (35%) and two (12%) embryos cleaved and developed to morulae or blastocyst stages, respectively. In conclusion, we have developed a technique for obtaining somatic cells from semen. We have also demonstrated that semen-derived cells can serve as karyoplast donors for nuclear transfer.

  7. Frozen shoulder.

    PubMed Central

    Anton, H. A.

    1993-01-01

    The frozen shoulder is a common cause of shoulder pain and disability. Most patients slowly improve over 12 to 24 months. Some have prolonged loss of movement, pain, and associated disability. Treatments include physiotherapy, corticosteroid injections, and manipulation. Clinical trials of these treatments have produced conflicting results. PMID:8374364

  8. Visual Detection of Brucella spp. in Spiked Bovine Semen Using Loop-Mediated Isothermal Amplification (LAMP) Assay.

    PubMed

    Prusty, Bikash R; Chaudhuri, Pallab; Chaturvedi, V K; Saini, Mohini; Mishra, B P; Gupta, Praveen K

    2016-06-01

    Several pathogens including Brucella spp. are shed in semen of infected bulls and can be transmitted to cows through contaminated semen during artificial insemination. The present study reports omp2a and bcsp31 gene based loop-mediated isothermal amplification (LAMP) assays for detection of Brucella genomic DNA in semen from infected bulls. The positive results could be interpreted visually by change in colour of reaction mixture containing hydroxyl naphthol blue (HNB) dye from violet to sky blue. LAMP assays based on omp2a and bcsp31 could detect as little as 10 and 100 fg of B. abortus S19 genomic DNA, respectively. Sensitivity of omp2a and bcsp31 LAMP assays for direct detection of organisms in bovine semen was 2.28 × 10(1) CFU and 2.28 × 10(2) CFU of B. abortus S19 in spiked bovine semen, respectively. The omp2a LAMP assay was found equally sensitive to TaqMan probe based real-time PCR and 100 times more sensitive than conventional PCR in identifying Brucella in spiked semen. The diagnostic applicability of the omp2a LAMP assay was evaluated with seventy-nine bovine semen samples and results were re-evaluated through TaqMan probe based real-time PCR and conventional PCR. Taken together, the omp2a LAMP assay is easy to perform, rapid and sensitive in diagnosis of Brucella spp. in bovine semen.

  9. Enhanced early-life nutrition promotes hormone production and reproductive development in Holstein bulls.

    PubMed

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

    2015-02-01

    Holstein bull calves often reach artificial insemination centers in suboptimal body condition. Early-life nutrition is reported to increase reproductive performance in beef bulls. The objective was to determine whether early-life nutrition in Holstein bulls had effects similar to those reported in beef bulls. Twenty-six Holstein bull calves were randomly allocated into 3 groups at approximately 1 wk of age to receive a low-, medium-, or high-nutrition diet, based on levels of energy and protein, from 2 to 31 wk of age. Calves were on their respective diets until 31 wk of age, after which they were all fed a medium-nutrition diet. To evaluate secretion profiles and concentrations of blood hormones, a subset of bulls was subjected to intensive blood sampling every 4 wk from 11 to 31 wk of age. Testes of all bulls were measured once a month; once scrotal circumference reached 26cm, semen collection was attempted (by electroejaculation) every 2 wk to confirm puberty. Bulls were maintained until approximately 72 wk of age and then slaughtered at a local abattoir. Testes were recovered and weighed. Bulls fed the high-nutrition diet were younger at puberty (high=324.3 d, low=369.3 d) and had larger testes for the entire experimental period than bulls fed the low-nutrition diet. Bulls fed the high-nutrition diet also had an earlier and more substantial early rise in LH than those fed the low-nutrition diet and had increased concentrations of insulin-like growth factor-I (IGF-I) earlier than the bulls fed the low-nutrition diet. Furthermore, we detected a temporal association between increased IGF-I concentrations and an early LH rise in bulls fed the high-nutrition diet. Therefore, we inferred that IGF-I had a role in regulating the early gonadotropin rise (in particular, LH) and thus reproductive development of Holstein bulls. Overall, these results support our hypothesis that Holstein bull calves fed a high-nutrition diet reach puberty earlier and have larger testes than

  10. Effective freezing rate for semen cryopreservation in endangered Mediterranean brown trout (Salmo trutta macrostigma) inhabiting the Biferno river (South Italy).

    PubMed

    Iaffaldano, Nicolaia; Di Iorio, Michele; Manchisi, Angelo; Esposito, Stefano; Gibertoni, Pier Paolo

    2016-10-01

    This study was designed to determine: (i) the in vitro effects of different freezing rates on post-thaw semen quality of Mediterranean brown trout (Salmo trutta macrostigma) from the Biferno river; and (ii) the in vivo fertilization and hatching percentage of freezing rate giving rise to the best post-thaw semen quality. Pooled semen samples were diluted 1:3 (v:v) in a freezing extender composed of 300 mM glucose, 10% egg yolk and 10% dimethyl sulfoxide (DMSO). The extended semen was packaged in 0.25 ml plastic straws and frozen at different heights above the liquid nitrogen surface (1, 5 or 10 cm) for 10 min to give three different freezing rates. Semen samples were thawed at 30°C for 10 s. The variables assessed after thawing were sperm motility, duration of motility and viability. Our results clearly indicate a significant effect of freezing rate on post-thaw semen quality. Semen frozen 5 cm above the liquid nitrogen surface showed the best quality after freezing/thawing. Based on these in vitro data, 2 groups of 200 eggs were fertilized with fresh semen or semen frozen 5 cm above the liquid nitrogen surface. Fertilization and hatching rates recorded for eggs fertilized with frozen semen were significantly lower (25.4% and 22.5%, respectively) than the ones obtained using fresh semen (87.8% and 75.5%, respectively). An effective freezing protocol will allow for the creation of a sperm cryobank to recover the original population of Mediterranean brown trout in the Biferno river.

  11. Cryopreservation of Sambar deer semen in Thailand.

    PubMed

    Vongpralub, Thevin; Chinchiyanond, Wittaya; Hongkuntod, Pornchai; Sanchaisuriya, Pitcharat; Liangpaiboon, Sanan; Thongprayoon, Areeya; Somphol, Noppadon

    2015-01-01

    Little is known of the different freezing and thawing techniques for post-thaw survival of spermatozoa in Sambar deer. So, this study determined the effect of seminal plasma, egg yolk and glycerol extenders and their concentrations, plus cooling, freezing, and thawing protocols on the post-thaw quality of their semen. Semen samples were collected by electro-ejaculation from four Thai Sambar deer stags (Cervus unicolor equinus). As evaluated by post-thaw progressive motility and acrosome integrity removal of seminal plasma was beneficial; Tris-egg yolk was the most efficient extender; a 20% egg yolk concentration was better than the 0%, 10%, or 30%; and a 3% glycerol concentration was better than 5%, 7%, or 9%. Using the optimum dilution techniques, semen was loaded in 0.5 ml plastic straws. Cooling times from ambient temperature to 5°C in 3 hr resulted in higher post-thaw progressive motility and acrosome integrity than 1, 2, or 4 hr. Suspending the straws 4 cm above the surface for 15 min before plunging into liquid nitrogen was better than suspending at 2 or 6 cm. For thawing frozen semen, an intermediate thawing (50°C, 8 sec) protocol was more effective than the slower (37°C, 10 sec) or faster (70°C, 5 sec) thawing rates. Timed insemination following estrus synchronization of 10 hinds resulted in six confirmed pregnancies at 60 days. Five hinds delivered live fawns. This study provides an effective approach for semen cryopreservation and artificial insemination (AI), which should be valuable to scientists for genetics and reproductive management of Sambar deer in developing countries.

  12. Application of liquid semen technology improves conception rate of sex-sorted semen in lactating dairy cows.

    PubMed

    Xu, Z Z

    2014-11-01

    were not yet available. The percentage of heifer calves born to AI with SS semen was 87.0% for 2011 and 85.8% for 2012, both of which were lower than the expectation of 90% mainly due to misidentification of calf dams in seasonal dairy herds calving on pasture. In summary, results in this report showed that liquid SS semen only required half the dose rate of frozen SS semen to achieve a reproductive performance of over 94% of CON semen in lactating dairy cows. Careful planning and a robust distribution network are required to avoid semen wastage and to maximize the benefit of liquid SS semen.

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

    PubMed

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

    2017-02-01

    Crossbred cattle in some sectors of the world have a significant role in enhancing milk production thereby enhancing the per capita milk availability as a human food source. However, there are certain constraints associated with crossbred animals, such as disease susceptibility, increased reproductive problems, repeat breeding and poor seminal quality. The semen of crossbred bulls has a poor freezing capacity, increased cryo-damage, poor mass cell motility, greater percentages of dead/abnormal sperm and poor initial and post-freeze cell motility. The rejection rate of crossbred bulls for cryostorage of semen has been reported to be as great as 50% as a result of unacceptable semen quality. The identification of superior bulls using molecular technologies is needed which necessitates identification of the genes having a role in sperm function. The present study was, therefore, conducted to gain information on identification and expression of genes having a role in sperm motility in crossbred bulls. The gene transcripts in bulls with sperm of superior and inferior quality were profiled in Vrindavani crossbred cattle by microarray analyses and the results were verified by real time-quantitative PCR. Microarray analyses revealed 19,454 genes which were differentially expressed. At a two-fold cut off, 305 genes were differentially (P<0.01) expressed with 160 genes upregulated and 145 genes down regulated. Some of the upregulated candidate genes were further validated by RT-qPCR. These genes had a four to 16 fold upregulation in sperm with inferior motility as compared to sperm of crossbred bulls with superior motility.

  14. Ways to improve the biosecurity of bovine semen.

    PubMed

    de Ruigh, L; Bosch, J C; Brus, M C; Landman, B; Merton, J S

    2006-08-01

    Semen production and trade is a worldwide industry. A framework, based on international standards is awaiting international and national regulation. In the perspective of biosecurity of the final product, critical notes can be made according to the semen production regulation and product safety. Process description brings the obligatory health standards for the production bulls, collection and processing of semen, identification, registration, worldwide distribution and insemination into discussion. Test frequency, test quality and demands, way of sampling and test consistency can influence product safety. New scientific knowledge can influence the value of the regulation. Whether a country is free of notifiable disease should influence decisions regarding necessary tests for the production bulls. The biosecurity of the semen production process is influenced by several factors. The effectiveness of the antibiotics used is questionable. The extenders for cryopreservation added to the semen can affect product safety. The way materials and storage containers have to be disinfected must be clear. In modern industry, tracking and tracing is an important issue. Worldwide differences in ways of identification of straws do not benefit a proper identification and registration throughout the process. Regulation could help improve the transparency of production and trade. Before anything concerning biohazard is implemented in regulation, each rule should be thoroughly based on scientific research where possible and furthermore it must be possible to enforce the regulation. The effort it takes to enforce the regulation should be in balance with the benefit it provides. An approach to alter regulation quickly is advisable. To produce a safe product that is accepted for international trade is of vital interest for the survival of artificial insemination (AI) in cattle.

  15. Application and commercialization of flow cytometrically sex-sorted semen.

    PubMed

    Rath, D; Johnson, L A

    2008-07-01

    The current technology to sort X and Y chromosome bearing sperm population requires individual identification and selection of spermatozoa in a modified high-speed flow cytometer. For farm animal species, the technology is capable of producing sexed sperm at greater than 90% purity. However, only in the bovine, the technology has reached a developmental level that allows its commercial application. Meanwhile, the demand for female calves has grown rapidly, which encourages the demand for sex-sorted semen from high genetic value bulls. The success of the technology will depend mainly on the fertilizing capacity of the sorted spermatozoa, as this is the most affecting and economically relevant factor. To date, fertility is still variable and is quite dependent on post-sort processing. New processing techniques are under investigation and will likely be able to improve the fertility rates after AI with sex-sorted semen. It is of great importance to select the right bulls and to test the sorted samples on a routine basis. In addition to the demand for sex-sorted semen by the cattle industry, there is also a significant demand expressed by pig farmers. However, it is still unknown if the use of sex-sorted semen through commercial pig AI will be economically feasible. For the pig, the combination of in vitro fertilization with sexed semen and non-surgical embryo transfer is an alternative that merits further scientific attention. Recent developments in ovine AI and ET will make it very likely that commercial sheep industry will adopt the sexing technology in their breeding concepts.

  16. 29 CFR 1918.84 - Bulling cargo.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 7 2012-07-01 2012-07-01 false Bulling cargo. 1918.84 Section 1918.84 Labor Regulations...) SAFETY AND HEALTH REGULATIONS FOR LONGSHORING Handling Cargo § 1918.84 Bulling cargo. (a) Bulling cargo shall be done with the bull line led directly from the heel block. However, bulling may be done from...

  17. 29 CFR 1918.84 - Bulling cargo.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 7 2013-07-01 2013-07-01 false Bulling cargo. 1918.84 Section 1918.84 Labor Regulations...) SAFETY AND HEALTH REGULATIONS FOR LONGSHORING Handling Cargo § 1918.84 Bulling cargo. (a) Bulling cargo shall be done with the bull line led directly from the heel block. However, bulling may be done from...

  18. 29 CFR 1918.84 - Bulling cargo.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 7 2014-07-01 2014-07-01 false Bulling cargo. 1918.84 Section 1918.84 Labor Regulations...) SAFETY AND HEALTH REGULATIONS FOR LONGSHORING Handling Cargo § 1918.84 Bulling cargo. (a) Bulling cargo shall be done with the bull line led directly from the heel block. However, bulling may be done from...

  19. 29 CFR 1918.84 - Bulling cargo.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

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

  20. 29 CFR 1918.84 - Bulling cargo.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 7 2011-07-01 2011-07-01 false Bulling cargo. 1918.84 Section 1918.84 Labor Regulations...) SAFETY AND HEALTH REGULATIONS FOR LONGSHORING Handling Cargo § 1918.84 Bulling cargo. (a) Bulling cargo shall be done with the bull line led directly from the heel block. However, bulling may be done from...

  1. BILL E. KUNKLE INTERDISCIPLINARY BEEF SYMPOSIUM: Does tall fescue toxicosis negatively impact bull growth and breeding potential?

    PubMed

    Pratt, S L; Andrae, J G

    2015-12-01

    The predominant cool-season forage in the southeastern United States is the tall fescue cultivar Kentucky 31 (KY31). Kentucky 31 possesses an endophyte (), which produces a family of toxins called ergot alkaloids. These toxins negatively affect the physiology of animals on consumption and result in the syndrome known as fescue toxicosis. Currently, the United States annually produces approximately 11.4 billion kg of beef, of which 25% originates in the southeastern region of the United States where forage systems frequently are tall fescue based. Cattle within this forage system exhibit reduced gains and reproductive performance. The result is a reduction in the nation's beef supply with annual revenue losses recently estimated at approximately US$1 billion. Our hypothesis is that exposure to these ergot alkaloids in conjunction with limited availability of nutrients decreases bull semen quality and fertility. Although the literature is clear that these toxins affect BW, body temperature, blood flow, hair growth, and female reproduction in cattle, their effect on bull reproduction and the mechanisms through which the toxins act are not well defined. Six studies published from 2004 to 2015 assessed bull growth, body composition, and semen quality of young beef bulls exposed to ergot alkaloids. If semen quality or fertility is altered, the mechanisms involved may be either direct effects of ergot alkaloids through neurotransmitter receptors or indirect effects such as inhibiting the release of prolactin (PRL). The possible effects of ergot alkaloids or PRL require establishing the presence or absence of dopamine, adrenergic, serotonin, or PRL receptors in the testis, epididymis, and sperm cell of the bull. The objective of this review is to relate our findings to the few previous studies conducted that evaluated the impact of fescue toxicosis on bull reproduction and to propose possible mechanisms of action for lowered semen quality.

  2. Effects of dietary energy on sexual maturation and sperm production in Holstein bulls.

    PubMed

    Harstine, B R; Maquivar, M; Helser, L A; Utt, M D; Premanandan, C; DeJarnette, J M; Day, M L

    2015-06-01

    In prepubertal bulls and heifers of dairy and beef breeds, puberty can be induced to occur earlier than typical with targeted high-energy diets due to precocious activation of the endocrine mechanisms that regulate puberty. Precocious activation of puberty in bulls intended for use in the AI industry has the potential to hasten and perhaps increase sperm production. It was hypothesized that feeding bulls a high-energy diet beginning at 8 wk of age would advance the prepubertal rise in LH and lead to advanced testicular maturation and age at puberty. From 58 to 230 ± 0.3 d of age, Holstein bulls received either a high-energy diet (HE;n = 9; targeted ADG 1.5 kg/d) or a control diet (CONT;n = 10; targeted ADG 0.75 kg/d). Thereafter, all bulls were fed a similar diet. The HE treatment increased LH secretion at 125 d of age, testosterone concentrations from 181 to 210 d, and scrotal circumference (SC) from 146 to 360 d of age relative to the CONT treatment. Beginning at 241 ± 5 d of age, semen collection (artificial vagina) was attempted every 14 d in bulls from the HE (n = 8) and CONT (n = 7) treatment until each bull attained puberty (ejaculate containing 50 × 10 spermatozoa with 10% motility). To assess semen production as mature bulls, semen was collected thrice weekly beginning at 541 ± 5 d of age until slaughter at 569 ± 5 d of age. After slaughter, epididymal and testicular measurements were collected and testicular tissue was fixed to determine seminiferous tubule diameter. Age at puberty did not differ between treatments (310 ± 35 d). Although testis and epididymal weight and testis volume were greater (P < 0.05) in the HE than the CONT treatment, sperm production of mature bulls did not differ between treatments. Diameter of seminiferous tubules also did not differ between treatments. We conclude that the HE advanced aspects of sexual maturation and increased testes size, but this was not reflected in hastened puberty or sperm production in the present

  3. 9 CFR 78.14 - Rodeo bulls.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Interstate Movement of Cattle Because of Brucellosis § 78.14 Rodeo bulls. (a) A rodeo bull that is test... date of being tested, may be moved interstate only if the bull meets the requirements for cattle...

  4. 9 CFR 78.14 - Rodeo bulls.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Interstate Movement of Cattle Because of Brucellosis § 78.14 Rodeo bulls. (a) A rodeo bull that is test... date of being tested, may be moved interstate only if the bull meets the requirements for cattle...

  5. 14. BULL SHAFT, BULL RING AND PINION, AND DRUM. TOP ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    14. BULL SHAFT, BULL RING AND PINION, AND DRUM. TOP OF PIER III, GRANITE COPING, AND PLAIN CONCRETE PIER BELOW. DETAILS OF WEST PIER PROTECTION FRAMING AT PIER. WILLBRIDGE IN BACKGROUND. - Burlington Northern Railroad Bridge, Spanning Willamette River at River Mile 6.9, Portland, Multnomah County, OR

  6. Seasonal effects of semen collection and artificial insemination on dairy cow conception.

    PubMed

    Haugan, T; Reksen, O; Gröhn, Y T; Kommisrud, E; Ropstad, E; Sehested, E

    2005-11-01

    The effects of four seasons of semen collection and of artificial insemination on conception in dairy cows were studied. The solstices and equinoxes (December, March, June and September) defined the beginning and/or end of each season. Semen was collected from 973 progeny-test bulls over 8 years at the two Norwegian AI stations at 60.8 degrees N and 63.4 degrees N where artificial light was used to provide a minimum photoperiod of 10 h/day. The effect of using semen of elite bulls during progeny testing and after selection as elite sires also was investigated. Norwegian Red (NRF) cows were inseminated over a 7-year period using progeny test semen and over the last 4 years of the same period using the semen of the elite sires. The probability of conception to only first inseminations for cows up to, and including, the fifth lactation was assessed by 56-day non-return rate (56d NRR) and calving rate. Two data sets were analysed which excluded cows culled within 270 days of AI or included such cows as non-calving. The reasons for culling were categorised as those for fertility problems or all other reasons. Semen was used for AI irrespective of the season in which it had been collected. Season of semen collection did not affect 56d NRR but calving rate was significantly higher (by 0.5-0.8%, approximately; P < 0.01) for semen collected in the December-March period, when photoperiod was increasing, than at other times of the year. The season in which AI was performed showed a peak of 56d NRR in spring for heifers (P < 0.01) and in summer for parous animals (P < 0.01). For calving rate, however, no seasonal peak was found in heifers, whereas pluriparous cows had much higher calving rates in summer and autumn/early winter than late winter and spring (P < 0.01). Semen of elite sires resulted in higher calving rates by 0.5 (NS) to 1.9% (P < 0.01) when used after selection than when used during progeny testing. The difference between the calving rate achieved when the semen

  7. Freezing African Elephant Semen as a New Population Management Tool

    PubMed Central

    Hermes, Robert; Saragusty, Joseph; Göritz, Frank; Bartels, Paul; Potier, Romain; Baker, Barbara; Streich, W. Jürgen; Hildebrandt, Thomas B.

    2013-01-01

    Background The captive elephant population is not self-sustaining and with a limited number of breeding bulls, its genetic diversity is in decline. One way to overcome this is to import young and healthy animals from the wild. We introduce here a more sustainable alternative method - importation of semen from wild bulls without removing them from their natural habitat. Due to the logistics involved, the only practical option would be to transport cryopreserved sperm. Despite some early reports on African elephant semen cryopreservation, the utility of this new population management tool has not been evaluated. Methodology/Principal Findings Semen was collected by electroejaculation from 14 wild African savanna elephant (Loxodonta africana) bulls and cryopreserved using the directional freezing technique. Sperm treatments evaluated included the need for centrifugation, the use of hen or quail yolk, the concentration of glycerol (3%, 5% or 7%) in the extender, and maintenance of motility over time after thawing. Our results suggest that dilution in an extender containing hen yolk and 7% glycerol after centrifugation best preserved post-thaw sperm motility when compared to all other treatments (P≤0.012 for all). Using this approach we were able to achieve after thawing (mean ± SD) 54.6±3.9% motility, 85.3±2.4% acrosome integrity, and 86.8±4.6% normal morphology with no decrease in motility over 1 h incubation at 37°C. Sperm cryopreserved during this study has already lead to a pregnancy of a captive female elephant following artificial insemination. Conclusions/Significance With working techniques for artificial insemination and sperm cryopreservation of both African and Asian elephants in hand, population managers can now enrich captive or isolated wild elephant populations without removing valuable individuals from their natural habitat. PMID:23483917

  8. Effect of widespread and limited use of sexed semen on genetic progress and reproductive performance of dairy cows.

    PubMed

    Khalajzadeh, S; Nejati-Javaremi, A; Mehrbani Yeganeh, H

    2012-09-01

    Stochastic simulation was used for studying the impacts of sexed semen on genetic progress and reproductive performance of dairy cows. Three strategies were compared: WSS (use unsexed semen in cows and heifers), SSH (use sexed semen in heifers and unsexed semen in cows) and SSCH (use sexed semen in both cows and heifers). Conception rate (CR) of unsexed semen was considered to be 35% and 65% in cows and heifers, respectively. CR of sexed semen was considered to be 15 (20% in cows and 50% in heifers), 10, 5 and 0 percentage points lower than unsexed semen. Thus, four subschemes were compared under SSCH (SSCH15, SSCH10, SSCH5, SSCH0) and SSH (SSH15, SSH10, SSH5, SSH0). Moreover, the effect was studied in four distinct paths of selection: active sires (AS), young bulls (YB), bull dams (BD) and milking cows (CW). The average genetic superiority of CW was 12% and 9.5% in SSCH15 and SSH15 strategies relative to a base scheme, respectively. The average genetic superiority of CW was 19% and 10.5% in SSCH0 and SSH0, respectively. Regression analysis showed that genetic superiority of CW increased significantly, that is, 0.5% and 0.1% per every 1% increase in CR in SSCH and SSH, respectively. The result showed that there is a significant difference between genetic superiority of cows in SSCH and SSH schemes. Widespread and limited use of sexed semen in commercial dairy herds resulted in a large genetic advantage in CW. The genetic advantage of gender control was minimal in the selection paths of AS, YB and BD. Open days and services per conception reached to 153 v. 125 days and 5 v. 2.86 under SSCH15 compared with WSS. The age at first calving increased from 774 to 790 days in SSH15 and SSCH15 strategies. Mean of parities decreased to 2.26 v. 2.42 by using sexed semen. The widespread use of sexed semen increased the age average of cows in all parities. Sexed semen increased selection intensity in the CW path, and this contributed to the genetic merit of future cows. On the

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

    PubMed

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

    2009-06-01

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

  10. Short communication: Genetic analysis of dairy bull fertility from field data of Brown Swiss cattle.

    PubMed

    Tiezzi, F; Maltecca, C; Penasa, M; Cecchinato, A; Bittante, G

    2013-01-01

    The aim of this study was to estimate heritability and repeatability of dairy bull fertility in Italian Brown Swiss cattle. Bull fertility indicators were calving per service and nonreturn rate at 56 d after service. Data included 124,206 inseminations carried out by 86 technicians on 28,873 heifers and cows in 1,400 herds. Services were recorded from 1999 to 2008 and were performed with semen from 306 AI Brown Swiss bulls. Data were analyzed with a threshold animal model, which included the fixed effects of parity by class of days in milk of the inseminated cow (age at insemination for heifers), year-season of insemination, and status of the service bull at the time of insemination (progeny testing or proven), and the random effects of herd, technician, additive genetic, and permanent environment of inseminated heifer/cow and service bull, and residual. Also, genetic covariance between heifer/cow and service bull effects was considered in the model. Heritability and repeatability were 0.0079 and 0.0100 for nonreturn rate at 56 d after service, and 0.0153 and 0.0202 for calving per service, respectively. The low estimates obtained in the present study indicate that selection for male fertility using field data is hardly pursuable.

  11. Impact of using a fast-freezing technique and different thawing protocols on viability and fertility of frozen equine spermatozoa.

    PubMed

    Pugliesi, G; Fürst, R; Carvalho, G R

    2014-01-01

    The effects of freezing technique and thawing protocol on thawed semen viability and fertility were studied. Ejaculates from 5 stallions (n = 25) were frozen by conventional or a fast-freezing technique. Frozen semen was thawed by two thawing protocols (37 °C 30 s(-1) or 75 °C 7 s(-1) ). Thawed semen was evaluated by progressive motility, vigour, morphology and plasma membrane integrity. Mares (n = 25) were inseminated with 300 (n = 11) or 150 (n = 14) million spermatozoa. A greater (P < 0.05) vigour and progressively motile spermatozoa were detected, respectively, at thawing and after 20 min post-thawing in the fast-freezing technique than in the conventional one. Plasma membrane integrity was also greater (P < 0.05) in semen frozen with the fast-freezing technique. Semen viability was not affected by thawing protocol. Pregnancy rate using the fast-freezing technique was 76% (19/25), and did not differ (P > 0.05) between insemination doses. We concluded that the 150 million progressively motile spermatozoa per dose using a deep-horn insemination maximises the use of equine semen. The fast-freezing technique, as compared to the conventional one, efficiently preserves the viability and fertilising capacity of spermatozoa, indicating a new method to improve the fertility of frozen equine semen.

  12. Status of Oregon's Bull Trout.

    SciTech Connect

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

    1997-10-01

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

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

    PubMed

    Attia, S; Katila, T; Andersson, M

    2016-02-01

    Good-quality semen is a prerequisite for successful and profitable artificial insemination (AI) of modern dairy cattle. Fertility of the bulls is evaluated with andrological examinations and semen analyses, such as morphology. However, little attention has been paid to the inheritance of bull fertility. In this study, we correlated sperm morphology, birth year and station of 695 AI bulls with calving rate (CR). Sperm morphology was clearly associated with CR underlining the usefulness of morphological examination in the assessment of fertility. The correlation between the proportion of normal spermatozoa and CR was significant (p < 0.001). No significant differences were detected between stations or birth years. We also compared the CR of 695 AI bulls with the CR of their 27 sires to study the inheritance of fertility. Sire's CR did not correlate with the CR of the sons (p = 0.218). This result indicates that at least when sires of acceptable CR are used to produce sons for use in AI the inheritance of CR is not significantly correlated.

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

    PubMed Central

    Patel, Akhil; Saxena, Atul; Swain, Dilip Kumar; Yadav, Dushyant; Yadav, Sanjay Singh; Kumar, Abhishek; Kumar, Anuj

    2015-01-01

    Aim: This study was aimed to see the beneficial effect of butylated hydroxytoluene (BHT) as a semen additive of Hariana bull semen. Materials and Methods: The study was carried out in Hariana bulls. Twenty-four ejaculates from two bulls were used for this study. Each ejaculate was extended with standard glycerolated egg yolk tris extender and supplemented with BHT at two concentrations as 0.5 mM (T1) and 1.0 mM (T2). After dilution, equilibration and 24 h of cryopreservation, the samples were analyzed for progressive motility, sperm viability and membrane integrity. Results: Progressive motility, sperm viability and sperm membrane integrity were significantly (p<0.05) increased in the samples fortified with BHT as compared to the control during the process of cryopreservation and thawing. The BHT concentration of 1 mM revealed better results as compared to 0.5 mM. Conclusion: Addition of 1.0 mM BHT was found better in cryopreservation of Hariana bull semen compared to 0.5 mM BHT and control samples. The addition of BHT has improved the sperm quality by acting as an antioxidant thereby reducing the lipid peroxidation of the sperms. PMID:27065652

  15. Effects of bull elk demographics on age categories of harem bulls

    USGS Publications Warehouse

    Bender, L.C.

    2002-01-01

    Many management strategies for elk (Cervus elaphus) emphasize increasing numbers of mature bulls in the population. These strategies are usually assumed to enhance productivity via increased breeding by mature bulls. I compared age classes of harem bulls during the peak of the rut under 4 bull harvest strategies that resulted in different bull:cow ratios, mature bull:cow ratios, bull mortality rates, and proportions of mature bulls in the autumn (pre-hunting season) population. Proportions of harems held by differing age classes of bulls [mature (P84% of harems only in populations where mature bull:cow ratios exceeded 21:100 in the autumn population. Interaction of mature bull ratios in the autumn population, harem size, and bull selectivity in the harvest strategy must be considered if increased breeding by mature harem bulls is a management goal.

  16. Effect of growth rate from 6 to 16 months of age on sexual development and reproductive function in beef bulls.

    PubMed

    Brito, L F C; Barth, A D; Wilde, R E; Kastelic, J P

    2012-04-15

    Sexual development and reproductive function were studied in 22 Angus × Charolais and 17 Angus bulls from 6 to 16 mo of age. Associations of average daily gain (ADG) and body weight with ages at puberty and at maturity (satisfactory semen quality), scrotal circumference, paired-testes volume and weight, testicular vascular cone diameter and fat thickness, scrotal temperature, sperm production and morphology, and testicular histology, were determined. There were no significant correlations between cumulative average daily gain and any of the end points investigated. Body weight at various ages was negatively correlated with ages at puberty and maturity in Angus × Charolais bulls, positively correlated with paired-testes weight in Angus × Charolais and Angus bulls, and positively correlated with seminiferous tubule volume in Angus bulls (P < 0.05). Semen quality improved gradually with age and the interval between puberty and maturity (mean ± SD; 309.4 ± 29.7 and 357 ± 42 days of age) was approximately 50 days. Age, weight, scrotal circumference, and paired-testes volume were all good predictors of pubertal and mature status, with moderate to high sensitivity and specificity (71.6% to 92.4%). In summary, growth rate between 6 and 16 mo of age did not affect sexual development and reproductive function in beef bulls. However, greater body weight at various ages was associated with reduced age at puberty and maturity, and with larger testes at 16 mo of age, indicating that improved nutrition might be beneficial, but only when offered before 6 mo of age. Average daily gains of approximately 1 to 1.6 kg/day did not result in excessive fat accumulation in the scrotum, increased scrotal temperature, or reduction in sperm production and semen quality, and could be considered "safe" targets for growing beef bulls.

  17. Methylation patterns in fetal tissues generated from gilts inseminated with fresh or cryopreserved semen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Environmental influences, such as pollutants, climate, or diet, can alter the epigenetic configuration of gametes. The objective of our study was to evaluate differences in methylation activity of fetal placenta and liver from porcine pregnancies derived from fresh or frozen/thawed semen. Thirty cyc...

  18. Collection, analysis and cryopreservation of semen from Malayan gaur (Bos gaurus hubbacki): A preliminary study

    PubMed Central

    Iswadi, M.I.; Ann, Z.F.; Hafiz, M.M.; Hafiz, M.D.; Fahrul, F.J.; Hajarian, H.; Wahid, H.; Zawawi, I.; Khairiah, M.S.; Mazni, O.A.

    2012-01-01

    The Malayan gaur (Bos gaurus hubbacki) or Seladang is classified as vulnerable by the International Union for Conservation of Nature and Natural Resources (IUCN). The Malayan gaur is mainly distributed in the tropical woodlands of Peninsular Malaysia and Southern Thailand. The aim of this study was to collect, analyze and cryopreserve the semen of wild Malayan gaur. Transrectal massage (TM) and electroejaculation (EEJ) technique was applied in semen collection of the Malayan gaur. The semen was then cryopreserved in liquid nitrogen using slow freezing technique. Makler counting chamber was used to evaluate sperm concentration and motility, while the sperm viability and morphology of fresh and post-thaw sperm was determined using eosin-nigrosin staining protocol. As a result, we have successfully collected the Malayan gaur semen using EEJ technique. Sperm motility, viability and morphological changes of the post-thaw semen of Malayan gaur were found undesirable due to the complication of the cryopreservation process. On the basis of current study it can be concluded that Malayan gaur bulls semen can be obtain by EEJ with no evidence of rectal trauma. Optimization of the process of cryopreservation for Malayan gaur sperm is needed to maintain the cryoviability of the good sperm quality. The data generated in this study would be useful in conservation of genetic diversity program for Malayan gaur. PMID:26623302

  19. Growth hormone treatment of breeding bulls used for artificial insemination improves fertilization rates.

    PubMed

    Sauerwein, H; Breier, B H; Gallaher, B W; Götz, C; Küfner, G; Montag, T; Vickers, M; Schallenberger, E

    2000-01-01

    To evaluate new therapeutical concepts for male subfertility, we tested the effects of exogenous recombinant bovine growth hormone (rbGH) on various endocrine and metabolic parameters both in blood and in seminal plasma of bulls. Sperm quality was assessed morphometrically and by monitoring the number of successful artificial inseminations (AIs) defined as non-return rates (NRR). Aliquots of 450 semen samples were used from each bull and each experimental period (4 wk before, 14 weeks during and 6 wk after treatment). Six out of ten sires (average age 8.4 years) were treated every two weeks with 640-mg depot formulated rbGH (Eli Lilly). Four bulls received vehicle only. Blood plasma bGH, IGF-I, insulin and glucose concentrations were increased with rbGH treatment. In seminal plasma there was no effect of rbGH treatment on fructose and citrate or on testosterone concentrations. With one exception, rbGH-treated bulls had greater IGFBP-3 concentrations in seminal plasma. Motility of spermatozoa after freezing and thawing was increased compared with pretreatment rates. Most interestingly, the number of successful AIs was increased by an average of 6.0% NRR when ejaculates from rbGH-treated bulls were used.

  20. Cryopreserving turkey semen in straws and nitrogen vapour using DMSO or DMA: effects of cryoprotectant concentration, freezing rate and thawing rate on post-thaw semen quality.

    PubMed

    Iaffaldano, N; Di Iorio, M; Miranda, M; Zaniboni, L; Manchisi, A; Cerolini, S

    2016-04-01

    1. This study was designed to identify a suitable protocol for freezing turkey semen in straws exposed to nitrogen vapour by examining the effects of dimethylacetamide (DMA) or dimethylsulfoxide (DMSO) as cryoprotectant (CPA), CPA concentration, freezing rate and thawing rate on in vitro post-thaw semen quality. 2. Pooled semen samples were diluted 1:1 (v:v) with a freezing extender composed of Tselutin diluent containing DMA or DMSO to give final concentrations of 8% or 18% DMA and 4% or 10% DMSO. The semen was packaged in 0.25 ml plastic straws and frozen at different heights above the liquid nitrogen (LN2) surface (1, 5 and 10 cm) for 10 min. Semen samples were thawed at 4°C for 5 min or at 50°C for 10 s. After thawing, sperm motility, viability and osmotic tolerance were determined. 3. Cryosurvival of turkey sperm was affected by DMSO concentration. Freezing rate affected the motility of sperm cryopreserved using both CPAs, while thawing rates showed an effect on the motility of sperm cryopreserved using DMA and on the viability of sperm cryopreserved using DMSO. Significant interactions between freezing rate × thawing rate on sperm viability in the DMA protocol were found. 4. The most effective freezing protocol was the use of 18% DMA or 10% DMSO with freezing 10 cm above the LN2 surface and a thawing temperature of 50°C. An efficient protocol for turkey semen would improve prospects for sperm cryobanks and the commercial use of frozen turkey semen.

  1. History of commercializing sexed semen for cattle.

    PubMed

    Garner, D L; Seidel, G E

    2008-04-15

    Although the basic principles controlling the sex of mammalian offspring have been known for a relatively long time, recent application of certain modern cellular methodologies has led to development of a flow cytometric system capable of differentiating and separating living X- and Y-chromosome-bearing sperm in amounts suitable for AI and therefore, commercialization of this sexing technology. After a very long history of unsuccessful attempts to differentiate between mammalian sperm that produce males from those that produce females, a breakthrough came in 1981 when it was demonstrated that precise DNA content could be measured. Although these initial measurements of DNA content killed the sperm in the process, they led to the ultimate development of a sperm sorting system that was capable, not only of differentiating between live X- and Y-sperm, but of sorting them into relatively pure X- and Y-sperm populations without obvious cellular damage. Initial efforts to predetermine the sex of mammalian offspring in 1989 required surgical insemination, but later enhancements provided sex-sorted sperm in quantities suitable for use with IVF. Subsequent advances in flow sorting provided minimal numbers of sperm sufficient for use in AI. It was not until the flow cytometric sorting system was improved greatly and successful cryopreservation of sex-sorted bull sperm was developed that efficacious approaches to commercialization of sexed semen could be implemented worldwide in cattle. A number of companies now offer sex-sorted bovine sperm. Innovative approaches by a diverse group of scientists along with advances in computer science, biophysics, cell biology, instrumentation, and applied reproductive physiology provided the basis for commercializing sexed semen in cattle.

  2. Cholesterol concentration in seminal plasma as a predictive tool for quality semen evaluation.

    PubMed

    Beer-Ljubić, B; Aladrović, J; Marenjak, T S; Laskaj, R; Majić-Balić, I; Milinković-Tur, S

    2009-11-01

    The aim of this study was to investigate the relationship between lipid composition of bovine serum and seminal plasma, seasonality, and semen quality. The experiment was carried out in two groups of Simmental breeding bulls: Group I (ages 2 to 4 yr) and Group II (ages 5 to 10 yr). Blood samples were collected from jugular vein, and bovine semen was sampled with an artificial vagina once per season. Serum concentrations of total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triacylglycerols, nonesterified fatty acids (NEFAs), and lipoprotein electrophoretic patterns were determined. Seminal plasma concentrations of total cholesterol, HDL-C, and LDL-C were assayed. Serum concentration of triacylglycerols in young bulls was significantly higher in winter compared with that in autumn, whereas serum NEFA concentration was significantly higher in autumn compared with that in other seasons. Serum concentration of total cholesterol, LDL-C, and LDL lipoproteins in older bulls was significantly higher in winter than in spring. Seminal plasma concentration of total cholesterol in young bulls was significantly higher in spring compared with that in summer, whereas in older bulls it was significantly higher in winter compared with that in autumn samples. Sperm volume of both groups was significantly higher in summer compared with that in autumn and winter. Sperm motility in young bulls was lowest in summer and differed significantly from the values recorded in other seasons. The changes observed in seminal plasma cholesterol concentration were associated with extracellular lipid use and appeared to be applicable as a biochemical marker of sperm quality.

  3. Mathematical prediction of freezing times of bovine semen in straws placed in static vapor over liquid nitrogen.

    PubMed

    Santos, M V; Sansinena, M; Zaritzky, N; Chirife, J

    2013-02-01

    A widespread practice in cryopreservation is to freeze spermatozoa by suspending the straws in stagnant nitrogen vapor over liquid nitrogen (N(2)V/LN(2)) for variable periods of time before plunging into liquid nitrogen (-196°C) for indefinite storage. A mathematical heat transfer model was developed to predict freezing times (phase change was considered) required for bull semen and extender packaged in 0.5ml plastic straws and suspended in static liquid nitrogen vapor. Thermophysical properties (i.e. thermal conductivity, specific heat, density, initial freezing temperature) of bovine semen and extender as a function of temperature were determined considering the water change of phase. The non-stationary heat transfer partial differential equations with variable properties (nonlinear mathematical problem) were numerically solved considering in series thermal resistances (semen suspension-straw) and the temperature profiles were obtained for both semen suspension and plastic straw. It was observed both the external heat transfer coefficient in stagnant nitrogen vapor and its temperature (controlled by the distance from the surface of liquid nitrogen to the straw) affected freezing times. The accuracy of the model to estimate freezing times of the straws was further confirmed by comparing with experimental literature data. Results of this study will be useful to select "safe" holding times of bull semen in plastic straws placed N(2)V/LN(2) to ensure that complete freezing of the sample has occurred in the nitrogen vapor and avoid cryodamage when plunging in LN(2). Freezing times predicted by the numerical model can be applied to optimize freezing protocols of bull semen in straws.

  4. Critical sources of bacterial contamination and adoption of standard sanitary protocol during semen collection and processing in Semen Station

    PubMed Central

    Sannat, Chandrahas; Nair, Ajit; Sahu, S. B.; Sahasrabudhe, S. A.; Kumar, Ashish; Gupta, Amit Kumar; Shende, R. K.

    2015-01-01

    Aim: The present investigation was conducted to locate the critical sources of bacterial contamination and to evaluate the standard sanitation protocol so as to improve the hygienic conditions during collection, evaluation, and processing of bull semen in the Semen Station. Materials and Methods: The study compared two different hygienic procedures during the collection, evaluation and processing of semen in Central Semen Station, Anjora, Durg. Routinely used materials including artificial vagina (AV) inner liner, cone, semen collection tube, buffer, extender/diluter, straws; and the laboratory environment like processing lab, pass box and laminar air flow (LAF) cabinet of extender preparation lab, processing lab, sealing filling machine, and bacteriological lab were subjected to bacteriological examination in two phases of study using two different sanitary protocols. Bacterial load in above items/environment was measured using standard plate count method and expressed as colony forming unit (CFU). Results: Bacterial load in a laboratory environment and AV equipments during two different sanitary protocol in present investigation differed highly significantly (p<0.001). Potential sources of bacterial contamination during semen collection and processing included laboratory environment like processing lab, pass box, and LAF cabinets; AV equipments, including AV Liner and cone. Bacterial load was reduced highly significantly (p<0.001) in AV liner (from 2.33±0.67 to 0.50±0.52), cone (from 4.16±1.20 to 1.91±0.55), and extender (from 1.33±0.38 to 0) after application of improved practices of packaging, handling, and sterilization in Phase II of study. Glasswares, buffers, and straws showed nil bacterial contamination in both the phases of study. With slight modification in fumigation protocol (formalin @600 ml/1000 ft3), bacterial load was significantly decreased (p<0.001) up to 0-6 CFU in processing lab (from 6.43±1.34 to 2.86±0.59), pass box (from 12.13±2

  5. Seminal quality and sperm production in beef bulls with chronic dietary vitamin A deficiency and subsequent re-alimentation.

    PubMed

    Rode, L M; Coulter, G H; Kastelic, J P; Bailey, D R

    1995-05-01

    Sixteen Hereford bulls (16 mo of age, 462 kg average body weight) were used in each of 2 yr to evaluate the effects of hypovitaminosis A on seminal quality and sperm production. Bulls were fed a high-concentrate diet with (+VIT) or without (-VIT) supplemental Vitamin A until the apparent onset of hypovitaminosis A (28 and 32 wk in Year 1 and 2, respectively). Half of the bulls on each treatment were then slaughtered and those remaining were re-alimented with Vitamin A. Plasma retinol concentration in -VIT bulls reached a nadir at approximately 25 wk. In Year 1, the proportion of progressively motile spermatozoa was lower in -VIT bulls after 17 wk but returned to that of the +VIT group after re-alimentation. The proportion of spermatozoa with primary morphological defects appeared to be greater in -VIT bulls compared to +VIT bulls by 26 and 24 wk in Year 1 and 2, respectively. The incidence of these defects declined in -VIT bulls upon re-alimentation, and approached the incidence observed in +VIT bulls by 8 to 12 wk of re-alimentation. Hypovitaminosis A decreased paired testes weight, daily sperm production, and epididymal sperm reserves but did not affect daily gain. Prolonged dietary Vitamin A deficiency impaired semen quality and sperm production in the absence of other clinical symptoms. However, under practical feeding conditions, diets that result in long-term, marginal Vitamin A deficiency or a relatively short-term absence of Vitamin A intake probably would have minimal effects on spermatogenesis.

  6. Purification and properties of hyaluronidase from bull sperm.

    PubMed

    Yang, C H; Srivastava, P N

    1975-01-10

    Hyaluronidase from bull sperm was fractionated by ammonium sulfate and further purified by DEAE-cellulose and Sephadex chromatography. The highly purified hyaluronidase preparation showed 2,370 units per mg of protein (68,730 N.F. units per mg of protein), i.e. 182-fold purification. Disc gel electrophoresis showed one major component. The molecular weight of bull sperm hyaluronidase was 62,000 by sodium dodecyl sulfate gel electrophoresis. Hyaluronidase from bull sperm has optimum activity at pH 3.8 and an absolute requirement for cations. Kplus and Naplus have a greater effect than Ca2plus, Mg2plus, and Mn2plus, whereas Co2plus, Cu2plus, and Zn2plus do not affect the enzyme activity. Purified preparations are less stable than crude extracts stored frozen at minus 15 degrees. Km of hyaluronidase with hyaluronic acid as substrate is 3.7 mg per ml and Vmax is 2.4 mumol per min by Hofstee plot.

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

    PubMed

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

    2014-02-01

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

  8. Efficacy of caudal epidural injection of lidocaine, xylazine and xylazine plus hyaluronidase in reducing discomfort produced by electroejaculation in bulls.

    PubMed

    Pagliosa, Ronaldo C; Derossi, Rafael; Costa, Deiler S; Faria, Fabio J C

    2015-11-01

    To test the hypothesis that epidural administration of lidocaine, xylazine or xylazine plus hyaluronidase provides reduced pain and stress during electroejaculation in bulls, eight 30-month-old Nellore bulls received saline solution (control), 2% lidocaine, 2% xylazine or 2% xylazine plus hyaluronidase injected into the first intercoccygeal (Co1-Co2) epidural space in randomized order. Heart rate, respiratory rate, mean arterial pressure, analgesia, animal behavior and motor blockade were evaluated before treatment and at predetermined intervals during and after treatment. Pain and stress were scored subjectively, and semen quality was evaluated. The onset of anesthetic action was significantly faster with lidocaine (3.0 ± 1.2 min) than with xylazine or xylazine plus hyaluronidase (8.9 ± 1.5 and 5.5 ± 2.6 min, P=0.021 and P=0.012, respectively), and the onset of anesthesia with xylazine plus hyaluronidase was significantly faster than that with xylazine alone (P=0.032). Treatment with xylazine or xylazine plus hyaluronidase resulted in less discomfort than treatment with lidocaine, as indicated by animal behavior. Changes in heart rate, respiratory rate and arterial pressure were within acceptable limits. Penile protrusion and semen emission occurred in all animals during all four treatments. Our results suggest that xylazine plus hyaluronidase reduced discomfort during electroejaculation more effectively than xylazine or lidocaine alone. Further experiments are necessary to determine whether electroejaculation with xylazine plus hyaluronidase is feasible for obtaining semen from Nellore bulls unaccustomed to being handled or restrained.

  9. Assessment of sperm morphology in zebu bulls, under field conditions in the tropics.

    PubMed

    Chacón, J

    2001-04-01

    Sperm morphology was studied in 302 extensively managed Zebu bulls (aged 1.5-9 years), classified as sound (n=166) or unsound (n=136) for breeding, under field conditions in the dry tropics of Costa Rica. Single semen samples were collected by electro-ejaculation and fixed in formol-saline solution immediately after collection. Sperm morphology was determined in the field on wet smears using a microscope equipped with phase-contrast optics, and further determined in the laboratory on air-dried smears stained with carbol-fuchsin. The frequencies of sperm abnormalities (such as abnormal acrosome, head, neck, mid-piece, tail, and presence of cytoplasmic droplets) were recorded as a percentage of the total number of counted spermatozoa (400 cells). Zebu bulls considered unsound for breeding showed a higher mean prevalence (p < 0.05) of knobbed acrosomes (4.0 versus 0.9%), head defects [specifically, nuclear invaginations and heads with abnormal shapes and sizes (27.6 versus 4.0%)], abnormal tails (11.2 versus 4.7%), and proximal droplets (8.4 versus 1.6%), compared with bulls considered sound for breeding. In these latter bulls, the abnormality most commonly seen was the presence of single bent tails with an entrapped cytoplasmic droplet (3.0 +/- 3.7%). Young Zebu bulls (i.e. bulls under 2 years of age) showed a higher percentage of missing acrosomes, and proximal cytoplasmic droplets, than older sires (12.1 versus 2.4%, and 23.9 versus 3.6%, respectively; p < 0.05), interpreted as an indication of low ejaculation frequency and sexual immaturity, respectively. Bulls with a long scrotum and soft testicular consistency (TC) at palpation showed higher percentages of abnormal sperm heads in the ejaculate than bulls with a normal scrotal length (SL) and a normal TC (32.7 versus 12.8% and 30.7 versus 10.3%, respectively; p < 0.05). In addition, Zebu bulls with a scrotal circumference (SC) < or = 30 cm showed a higher prevalence of proximal cytoplasmic droplets than bulls

  10. Bulling among yearling feedlot steers.

    PubMed

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

    1976-09-01

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

  11. Studies on Freezing RAM Semen in Absence of Glycerol.

    NASA Astrophysics Data System (ADS)

    Abdelnaby, Abdelhady Abdelhakeam

    1988-12-01

    Glycerol is widely used as a major cryoprotective agent for freezing spermatozoa of almost all species. However, it reduces fertility of sheep inseminated cervically compared with intrauterine insemination. Studies were conducted to develop a method and procedure for freezing ram semen in the absence of glycerol. Post -thaw survival of ram spermatozoa frozen in the absence of glycerol was affected by time and temperature after collection and before dilution and time after dilution and before freezing. Increase in time at 5^ circC before or after dilution and before freezing increased both post-thaw motility and number of cells passing through Sephadex filter. A cold dilution method was developed. Slow cooling of fresh ram semen and diluting at 5^circ C 2-3 hr. after collection, then freezing 1 hr. after dilution improved both post-thaw motility and number of cells passing through Sephadex filter compared with immediate dilution at 30-37^circC after collection and freezing 3-4 hr. later (P < 0.05). An extender was developed to freeze ram semen in the absence of glycerol. An increase in post-thaw motility was obtained when semen was extended in TES titrated with Tris to pH 7.0 (TEST) and osmotic pressure of 375-400 mOsm/kg, containing 25-30% (v/v) egg yolk and 10% (v/v) maltose. A special device (boat) for freezing was constructed to insure the same height of the sample above LN _2 and thus the same freezing rate from freeze to freeze. Freezing of semen in 0.25cc straws at 5-10 cm above LN_2 (73.8 to 49.5 ^circC/min) yielded higher post-thaw motility than the rates resulted from freezing at 15 cm above LN_2 or 1 cm above LN _2. Faster Thawing in 37^ circC water for 30 sec. (7.8^ circC/sec.) increased post-thaw motility compared with slower thawing in 5 or 20^circ C water (P < 0.05). A lambing rate of 52.2% was obtained in one fertility trial conducted with ram semen frozen without glycerol and 17.1% in a second trial. One injection (IM) of 15 mg PGF_{2alpha}/ewe for

  12. Preservation of mithun (Bos frontalis) semen at refrigeration temperature.

    PubMed

    Karunakaran, M; Dhali, A; Mech, A; Khate, K; Rajkhowa, C; Mishra, D P

    2007-10-01

    The objective of the present study was to investigate the possibility of preserving mithun (Bos frontalis) spermatozoa at refrigeration temperature using tris-egg yolk diluent. Semen samples were collected from four adult mithun bulls through rectal massage method. Good quality semen samples (n=30) were preserved at 4 degrees C using tris-egg yolk diluent for 72 h. Progressive motility, live spermatozoa count and morphological abnormalities were evaluated every 12 h until 72 h of preservation. The colour, consistency and mass activity of fresh semen samples were found to be creamy white, medium and 3+ to 4+ (5+ scale), respectively. The average (mean+/-S.E.) volume (ml), pH and spermatozoa concentration (10(6) ml(-1)) of fresh semen samples were found to be 0.6+/-0.01, 6.8+/-0.03 and 425+/-48, respectively. Progressive motility and live spermatozoa count were found to be less than 30% (P<0.01) after 48 h of storage. Head (P<0.05), midpiece (P<0.05), tail (P<0.01) and total (P<0.01) abnormalities were found to be increased significantly over the time of storage. It was observed that progressive motility and live spermatozoa count remained above 30% and 40%, respectively, until 36 h of storage. Simultaneously the percentage of morphologically abnormal spermatozoa was found to be significantly low until 36 h of storage. The results indicate that it is possible to preserve mithun spermatozoa at refrigeration temperature in tris-egg yolk diluent, which can be further used for artificial insemination within 36 h of storage.

  13. Effect of manganese supplementation on the membrane integrity and the mitochondrial potential of the sperm of grazing Nelore bulls.

    PubMed

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

    2014-11-10

    The effect of dietary manganese (Mn(2+)) supplementation on the reproductive performance of Nelore bulls was evaluated by assessment of sperm membrane integrity. Sixty Nelore bulls (Bos taurus indicus) aged 18-20 mo were randomly divided into four groups (n=15) receiving dietary Mn(2+) supplementation at 540, 1300, 3800 and 6300mg/kg (treatments TC, T1300, T3800 and T6300, respectively). The diets were changed for the groups every 70d. Semen samples were obtained 15 and 56d after the diet change, which corresponded to the period of adjustment to the new diet and the time required for a complete spermatogenesis cycle, respectively. Sperm integrity was assessed by detection of: intact (IMe) or damaged (DMe) membranes, intact (IA) or damaged (DA) acrosomes, and high (HM) or low (LM) mitochondrial membrane potentials. Only bulls from the TC treatment showed a significant increase in the production of intact sperm [IMe/IA/LM] and decrease in the production of sperm with damaged acrosome [IMe/DA/LM] or completely damaged sperm [DMe/DA/LM] (P<0.05). The Mn(2+) concentrations in the semen were positively correlated with the incidence of sperm with IMe, DA, and LM and negatively correlated with number of sperm with DMe, IA, and LM. Therefore, dietary Mn(2+) supplementation for Nelore bulls must be limited to 540mg of Mn(2+)/kg given that higher doses are detrimental to the integrity of the plasma and acrosomal sperm membranes.

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

    PubMed

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

    2011-07-15

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

  15. Fertility of yearling beef bulls during mating.

    PubMed

    Ellis, R W; Rupp, G P; Chenoweth, P J; Cundiff, L V; Lunstra, D D

    2005-08-01

    Crossbred (Bos taurus) yearling beef bulls were assessed for breeding soundness and physical traits prior to multi-sire natural mating at pasture. Bulls (n = 60) were assigned to six groups of nine or 10 bulls and two bull-groups were rotated on 14-day intervals during a 63-day mating season in each breeding herd (n = 3) of 191-196 cows. The remaining bulls (n = 14) were maintained under similar environmental conditions without mating exposure. Bulls were observed during mating and assessed for breeding soundness and changes following mating. Bulls used for breeding (UFB) lost 77 kg of body weight and declined from body condition scores of 6 to 4.5, whereas bulls not used for breeding (NUB) maintained body condition scores of 6 and gained 27 kg. The UFB bulls incurred a 75% total injury rate with 63% incidence of lameness and 12% incidence of reproductive injuries, resulting in a 22% attrition rate. Only 45% were physically sound at the end of mating. Scrotal circumference declined in UFB bulls (-4.58%) and increased in NUB bulls (2.49%). From the 98% BSE-satisfactory rate (UFB) prior to breeding, only 61% were BSE-satisfactory post-breeding. The NUB bulls declined from 57 to 36% satisfactory. The BSE classification was influenced by significant increases in abnormal spermatozoa (primary and secondary), which was significantly associated with injuries incurred during mating. Group and breed differences in injury rates and BSE-status following mating were evident. Environmental conditions and mating activity influenced bull seminal quality and physical condition. Pregnancy rates in all three breeding herds (91-96%) were similar, with insignificant differences between bull-groups; the effects of physical and reproductive changes on individual bull fertility were immeasurable.

  16. Simple and Effective Methods of Freezing Capercaillie (Tetrao urogallus L.) Semen

    PubMed Central

    Kowalczyk, Artur; Łukaszewicz, Ewa

    2015-01-01

    A continuous decline in the number and range of capercaillie (Tetrao urogallus L.) in many European countries can be observed, mostly due to habitat destruction by human activity, unecological forestry management, and increased density of natural predators. Ex situ in vitro gene banks provide a unique opportunity to preserve the genetic material for future generations. Simple and effective cryopreservation methods for capercaillie semen are discussed. Semen was collected from seven males kept in the Capercaillie Breeding Centre at Forestry Wisła in Poland. Within five minutes after collection, ejaculates were diluted with EK diluent, then divided into two parts, and subjected to two freezing procedures: in pellets and in straws. In fresh semen, ejaculate clearness, viscosity, color and volume, as well as sperm concentration, motility and morphology, were evaluated, while in frozen-thawed semen only motility and morphology of sperm were determined. Fertilizing ability of thawed semen was examined for samples frozen in straws. Significant (P<0.05) differences between individual males were found in relation to the majority of fresh semen traits: ejaculate volume averaged 102.1 µL (varying from 49.0 to 205.0); average sperm concentration was 632.5 x106 mL-1 (178.8–1257.1); percentage of live normal cells varied from 39.2 to 70.3% (58.7% on an average); percentage of motile cells ranged from 76.0 to 85.7%) and motility parameters were male dependent, as well. Both cryopreservation methods had a negative effect on morphology and motility of frozen-thawed semen; however, the straw method yielded 60.7% and the pellet method 42.5% of live cells in total in thawed semen (P<0.05), while the number of live normal (intact) cells was similar (22.4 and 22.2%, respectively). Egg fertility varied between 77.8 and 91.7% (average 84.4%). Both freezing procedures seem to be effective in obtaining acceptable viability and high fertilizing potency of thawed sperm and can be used to

  17. The 1000 bull genome project

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To meet growing global demands for high value protein from milk and meat, rates of genetic gain in domestic cattle must be accelerated. At the same time, animal health and welfare must be considered. The 1000 bull genomes project supports these goals by providing annotated sequence variants and ge...

  18. Reproductive characteristics and thyroidal function in relation with season in Khuzestan buffalo (Bubalus bubalis) bulls

    PubMed Central

    Mayahi, Sadegh; Mamouei, Morteza; Tabatabaei, Saleh; Mirzadeh, Khalil

    2014-01-01

    High ambient temperature is the major constraint on Buffalo productivity. The aim of this study was to evaluate the reproductive performance and thyroid gland function in winter and summer seasons in Khuzestan buffalo bulls. Six male indigenous buffaloes of Khuzestan with nearly the same age (2-3 years old) and weight were used. Semen and blood samples through jugular vein were collected, every two weeks throughout the summer and winter seasons. The thyroid hormones and thyrotropin stimulating hormone (TSH) concentration in blood serum were measured by radioimmunoassay method. Semen quality was determined, using computer assisted sperm analyzer (CASA) and routine methods. The concentration of thyroxin (T4) was lower in winter than summer (p ≤ 0.05). The level of T3 uptake was higher in cold season than that of in hot season (p ≤ 0.05). The differences of tri-iodotyronine (T3) and TSH concentrations, as well as free thyroxin index were not significant between seasons. The semen volume and spermatozoa parameters including concentration, progressive motility, linear velocity, mean velocity, beat cross frequency, linear coefficient and straightness coefficient were higher in winter than summer (p ≤ 0.05). Semen pH and amplitude of lateral head displacement of spermatozoa were higher in summer than winter (p ≤ 0.05). In winter, there was positive correlation between spermatozoa concentration and T3 value of blood serum (p ≤ 0.05). There were positive correlations between values of semen volume and T4, progressive spermatozoa motility percent and TSH, as well as, total motility of spermatozoa and TSH in summer (p ≤ 0.05). In general, thyroid function and semen quality of Khuzestan buffaloes may be affected by seasons. PMID:25568719

  19. Swimming speed distributions of bull spermatozoa as determined by quasi-elastic light scattering.

    PubMed Central

    Hallett, F R; Craig, T; Marsh, J

    1978-01-01

    88 semen samples from 39 bulls have been investigated by the quasi-elastic light scattering technique. Normal, defective, and dead cells each yielded characteristic autocorrelation functions. The form of these functions indicates that the swimming speed distribution of normal cells is a gamma distribution with two degrees of freedom while that for defective or circular swimmers is a gamma distribution with one degree of freedom. The resulting analysis of the experimental autocorrelation functions yields the fraction of the sample that is normal, the fraction that is defective, and the average speed of each group. The average helical swimming speed of normal cells was found to be 384 micron/s, while the average trajectory speed of the circular swimmers was found to be 103 micron/s. The overall quality of the semen samples as determined by light scattering is compared to quality determination on the same samples by technicians from the artificial insemination industry. PMID:630041

  20. Effect of prolonged freezing of semen on exosome recovery and biologic activity.

    PubMed

    Welch, Jennifer L; Madison, Marisa N; Margolick, Joseph B; Galvin, Shannon; Gupta, Phalguni; Martínez-Maza, Otoniel; Dash, Chandravanu; Okeoma, Chioma M

    2017-03-24

    Exosomes are important vehicles of intercellular communication that shape host responses to physiologic, tumorigenic, and pathogenic conditions. The composition and function of exosomes are dynamic and depends on the state and condition of the cellular source. In prior work, we found that semen exosomes (SE) from healthy donors who do not use illicit drugs potently inhibit HIV-1. Following semen donation, specimens are either used immediately or frozen for use at a later time. It has been shown that short-term freezing of semen has no effect on SE-mediated HIV-1 inhibition. However, the effect of illicit drugs and prolonged freezing on SE bioactivity is unknown. Here, we show preservation of SE physical properties, (morphology, concentration, intensity/size) irrespective of illicit drug use or duration of semen freezing. Interestingly, illicit drugs and prolonged freezing decreased the levels of SE-bound CD63/CD9 and acetylcholinesterase activity respectively. Furthermore, we show differential effects of illicit drug use and prolonged freezing on SE-mediated HIV-1 inhibition. Our results highlight the importance of the source of SE and condition of semen storage on SE content and function. In-depth evaluation of donor drug-use and duration of semen storage on SE cargo and bioactivity will advance our understanding of SE composition and function.

  1. Effect of prolonged freezing of semen on exosome recovery and biologic activity

    PubMed Central

    Welch, Jennifer L.; Madison, Marisa N.; Margolick, Joseph B.; Galvin, Shannon; Gupta, Phalguni; Martínez-Maza, Otoniel; Dash, Chandravanu; Okeoma, Chioma M.

    2017-01-01

    Exosomes are important vehicles of intercellular communication that shape host responses to physiologic, tumorigenic, and pathogenic conditions. The composition and function of exosomes are dynamic and depends on the state and condition of the cellular source. In prior work, we found that semen exosomes (SE) from healthy donors who do not use illicit drugs potently inhibit HIV-1. Following semen donation, specimens are either used immediately or frozen for use at a later time. It has been shown that short-term freezing of semen has no effect on SE-mediated HIV-1 inhibition. However, the effect of illicit drugs and prolonged freezing on SE bioactivity is unknown. Here, we show preservation of SE physical properties, (morphology, concentration, intensity/size) irrespective of illicit drug use or duration of semen freezing. Interestingly, illicit drugs and prolonged freezing decreased the levels of SE-bound CD63/CD9 and acetylcholinesterase activity respectively. Furthermore, we show differential effects of illicit drug use and prolonged freezing on SE-mediated HIV-1 inhibition. Our results highlight the importance of the source of SE and condition of semen storage on SE content and function. In-depth evaluation of donor drug-use and duration of semen storage on SE cargo and bioactivity will advance our understanding of SE composition and function. PMID:28338013

  2. Cryopreservation of Indian red jungle fowl (Gallus gallus murghi) semen.

    PubMed

    Rakha, B A; Ansari, M S; Akhter, S; Hussain, I; Blesbois, E

    2016-11-01

    The population of red jungle fowl is declining and needs special attention for its conservation with suitable approaches. For ex situ in vitro conservation of Indian red jungle fowl, establishment of semen cryobank is an appropriate option, for which an extender with adequate retrieval capacity for functional spermatozoa is required. Therefore, studies were designed to evaluate a wide range of extenders for cryopreservation of Indian red jungle fowl (Gallus gallus murghi) sperm to achieve maximal post-thawed semen quality and fertility. For this purpose, semen from eight mature cocks were collected, initially evaluated (percent sperm motility, volume and concentration), pooled, assessed for motility, plasma membrane integrity, viability and acrosome integrity, and divided into six aliquots for dilution (1:5; 37°C) in Beltsville poultry, red fowl extender, Lake, EK, Tselutin poultry and chicken semen extenders. Diluted semen was cooled from 37°C to 4°C @ -0.275°C/min. Glycerol (20%) was added to chilled semen, equilibrated for 10min, filled in 0.5mL French straws, kept over LN2 vapours for 10min and plunged into LN2 and stored at -196°C. Percentages of motility, plasma membrane integrity, viability and acrosome integrity were higher (P<0.05) in red fowl extender at 0, 2 and 4h of incubation post-thaw. After cryopreservation and post-thawing at 37°C the highest (P<0.05) recovery rates and absolute livability index was also recorded in red fowl extender that was thus used for further artificial insemination of cooled-diluted (Liquid) and cryopreserved sperm. The no. of fertilized eggs (Liquid, 20.6±0.4; Cryopreserved, 12.6±0.5), percent fertility (86.7±2.2; 57.2±3.9), no. of hatched chicks (18.2±0.8; 10.0±0.3), percent hatch (76.5±2.7; 45.3±2.2) and hatchability of fertilized eggs (88.3±3.4; 79.6±3.4) were higher with sperm respectively freshly cooled-diluted or cryopreserved in red fowl extender. However, the rates obtained with frozen-thawed sperm

  3. TRIHALOMETHANE LEVELS AND SEMEN QUALITY

    EPA Science Inventory

    Trihalomethanes (THMs) are common byproducts of chlorinating drinking water. The effects of disinfection byproducts on semen quality have not yet been studied in humans, despite animal studies linking exposure to sperm abnormalities. We are currently analyzing the relationship of...

  4. Use of cholesterol-loaded cyclodextrin in donkey semen cryopreservation improves sperm viability but results in low fertility in mares.

    PubMed

    Oliveira, R R; Rates, D M; Pugliesi, G; Ker, P G; Arruda, R P; Moraes, E A; Carvalho, G R

    2014-10-01

    The use of cholesterol-loaded cyclodextrin (CLC) on semen cryopreservation has been related with better sperm viability in several species; however, the effect on fertility is not known in donkey semen. Ejaculates (n = 25) from five donkeys were diluted in S-MEDIUM with 0, 1, 2 or 3 mg of CLC/120 × 10(6) spermatozoa. Semen was frozen, and thawed samples were evaluated by computer-assisted sperm analyser system (CASA), supravital test, hyposmotic swelling test and fluorescent dyes to assess the integrity of sperm membranes. Mares (n = 60) were inseminated with frozen-thawed semen treated with the doses of 0 or 1 mg CLC. Percentages of sperm with progressive motility and with functional plasma membrane were greater (p < 0.05) in the CLC-treated groups than in the control. Percentages of intact plasma membrane and intact plasma membrane and acrosome detected by fluorescent dyes were also greater (p < 0.05) in CLC-treated groups. Although no difference (p > 0.05) in conception rates was detected between groups (control, 3/30, 10%; CLC-treated, 1/30, 3.3%), fertility was low for artificial insemination programs in mares. Therefore, we firstly demonstrated that frozen semen treated with CLC in S-MEDIA extender before freezing improves the in vitro sperm viability, but semen treated or not with CLC in S-MEDIUM extender results in a very low conception rate in mares inseminated with thawed donkey semen.

  5. Comparison of two dilution rates on canine semen quality after cryopreservation in a coconut water extender.

    PubMed

    de Cássia Soares Cardoso, Rita; Silva, Alexandre Rodrigues; da Silva, Lúcia Daniel Machado

    2006-05-01

    The objective of the present study was to evaluate the effect of sperm dilution (one part semen:one part extender or at 200 x 10(6) spermatozoa/mL) using a coconut water extender on the post-thaw sperm quality. Twelve ejaculates were collected from six dogs. Semen was divided into two aliquots, one for dilution one part semen:one part extender (group 1) and another for a concentration of 200 x 10(6) spermatozoa/mL (group 2). Semen was initially extended at 37 degrees C at a proportion of one part semen:half part extender (1:1/2) for group 1 (A-fraction). For group 2, the volume for a concentration of 200 x 10(6) spermatozoa/mL was calculated and a half of this volume was used for the initial dilution (A-fraction, 37 degrees C). Coconut water extender containing 20% egg yolk was used for this initial dilution in both groups. After dilution, the semen was cooled for 40 min in a thermal box (15 degrees C) and for 30 min in a refrigerator. The other half of the extender (B-fraction) containing egg yolk and glycerol (12%) was added to semen in both groups. Subsequently, the final concentration of glycerol in the extender was 6%. Ejaculates were frozen in 0.25 mL straws 5 cm above the surface of liquid nitrogen and stored at -196 degrees C. After 1 week, straws were thawed at 37 degrees C for 1 min and the microscopic criteria were evaluated. The dilution method had no influence on sperm motility, vigor and normal spermatozoa (71.4 compared with 67.7%). There was no effect of dog, ejaculate within male on post-thaw semen quality. Moreover, there was not a male x treatment interaction. Both treatments were efficient in preserving sperm quality.

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

    PubMed

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

    2012-05-01

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

  7. Development of a SYBR Green I based duplex real-time PCR for detection of bovine herpesvirus-1 in semen.

    PubMed

    Pawar, Sachin S; Meshram, Chetan D; Singh, Niraj K; Saini, Mohini; Mishra, B P; Gupta, Praveen K

    2014-11-01

    Bovine herpesvirus-1 (BoHV-1) is a viral pathogen found in infected bull semen, which is transmitted to inseminated cows by artificial insemination. BoHV-1 infection can cause reproductive disorders leading to significant economic loss to cattle industry. To detect BoHV-1 in semen, in this study, a SYBR Green I based duplex real-time PCR was developed. The assay included primers from BoHV-1 glycoprotein C (gC) and bovine growth hormone (bGH) genes for simultaneous detection in single tube. The result was interpreted by analysing melting temperature (Tm) peaks obtained after melt curve analysis of the amplified products at the end of reaction. The Tm peaks for BoHV-1-gC indicated presence of BoHV-1 while the bGH peak indicated reaction without inhibition. The sensitivity of the assay was to detect ten BoHV-1 genome copies per reaction. The analytical sensitivity was to detect 0.21 TCID50 infectious BoHV-1 in spiked semen. The assay was validated with 80 semen samples collected from breeding bulls. The diagnostic sensitivity and specificity of the assay was 100% with OIE recommended TaqMan probe based real-time PCR.

  8. Rhodiola sacra aqueous extract (RSAE) improves biochemical and sperm characteristics in cryopreserved boar semen.

    PubMed

    Zhao, H-W; Li, Q-W; Ning, G-Z; Han, Z-S; Jiang, Z-L; Duan, Y-F

    2009-03-15

    Although Rhodiola sacra aqueous extract (RSAE) has been used in many studies as an antioxidant, its effects on semen characteristics and its antioxidant properties during cryopreservation of boar sperm have never been evaluated. Semen was collected from five Duroc boars (2-4-year-old) twice weekly and frozen-thawed in extender with RSEA. Motion characteristics were assessed with a computer-aided semen analysis (CASA) system, whereas other sperm quality end points were assessed by routine methods. The effective concentration of RSEA in extender ranged from 4 to 8mg/L and the effect of RSEA on sperm quality was better in glycerol-free extender than extender containing glycerol (P<0.05). In frozen-thawed boar semen, there was a direct correlation (P<0.05) between RSEA concentration and glutathione (GSH) concentrations, mitochondrial activity, and hypoosmotic swelling test (HOST), and an inverse correlation (r=-0.982, P<0.05) between RSEA concentration and malondialdehyde (all end points were significantly higher at 6mg/L than in the control group). In summary: (i) the effective concentration of RSEA in extender ranged from 4 to 8mg/L; (ii) the effect of RSEA on sperm quality was better in extender without glycerol; and (iii) there was a significant correlation between RSEA concentrations and concentrations of GSH and MAD in frozen-thawed boar semen (antioxidant effects of RSEA were concentration-dependent). Further studies are needed to define the active ingredient in RSEA that protects boar sperm against ROS.

  9. Collection and preservation of pygmy hippopotamus (Choeropsis liberiensis) semen.

    PubMed

    Saragusty, J; Hildebrandt, T B; Bouts, T; Göritz, F; Hermes, R

    2010-09-01

    Knowledge about the reproduction of the endangered pygmy hippopotamus is almost non-existent. This study takes the first step toward changing this by devising a protocol for the collection, evaluation, and short-term preservation of semen of this endangered species. Semen was collected successfully from seven bulls by electroejaculation, using a specially designed rectal probe. Mean +/- SEM values of native sperm parameters from combined best fractions were: motility-80.0 +/- 4.1%, concentration-2421 +/- 1530 x 10(6) cells/mL, total collected cell number-759 +/- 261 x 10(6) cells, intact acrosome-87.8 +/- 1.2%, intact morphology-52.7 +/- 4.3%, and, for some, hypoosmotic swelling test-79.3 +/- 4.4% and seminal plasma osmolarity-297.5 +/- 3.3 mOsm. Seven different extenders were tested for sperm storage under chilling conditions: Berliner Cryomedium (BC), Biladyl, modification of Kenney modified Tyrode's medium (KMT), MES medium, Androhep((R)), boar M III() extender and Human Sperm Refrigeration Medium. While differences between males were apparent, the BC was consistently superior to all other extenders in sperm motility and facilitated storage for 7 d with up to 30% motility and some motility even after 3 weeks. With this knowledge in hand, the obvious two directions for future research are to conduct artificial insemination and to develop a technique for sperm cryopreservation.

  10. The development of beef breeding bulls.

    PubMed

    Engelken, T J

    2008-08-01

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

  11. Recent advances in cooled-semen technology.

    PubMed

    Aurich, Christine

    2008-09-01

    The majority of horse registries approve the use of artificial insemination, and horse breeding has widely taken benefit from the use of cooled-stored semen. New insights into cooled-semen technology open possibilities to reduce problems such as impaired semen quality after cooled-storage in individual stallions. The stallion itself has major impacts on quality and fertility of cooled-stored semen. Dietary supplementation of antioxidants and polyunsaturated fatty acids improves semen quality in a variety of species, but only few studies on this topic exist in the horse. Proper semen collection and handling is the main key to the maintenance of semen quality during cooled-storage. Semen collection should be achieved by minimal sexual stimulation with a single mount; this results in high sperm concentration, low content of seminal plasma and minimal contamination with bacteria. Milk-based semen extenders are most popular for semen processing and storage. The development of more defined extenders containing only the beneficial milk ingredients has made extender quality more constant and reliable. Semen is often centrifuged to decrease the seminal plasma content. Centrifugation results in a recovery rate of only 75% of spermatozoa in the semen pellet. Recovery rates after centrifugation may be improved with use of a "cushion technique" allowing higher centrifugation force and duration. However, this is not routinely used in cooled-semen technology. After slow-cooling, semen-storage and shipping is best performed at 5 degrees C, maintaining semen motility, membrane integrity and DNA integrity for up to 40 h after collection. Shipping containers created from Styrofoam boxes provide maintenance of semen quality at low cost.

  12. Frozen gene pools - A future for species otherwise destined for extinction

    USGS Publications Warehouse

    Gee, G.F.

    1986-01-01

    Conclusion: Semen banks and ova and embryo banks can be practical methods to maintain gene pools. Gene pool preservation is desperately needed today due to the rapid decline in number of species and their habitat, a matter that is of concern to.biologists, economists, and politicians worldwide. Techniques are available for the cryopreservation of semen from many animals (and embryos from a few mammals) and adaptations of these techniques to other animals should be possible. A frozen gene pool in conjunction with existing programs makes it possible to preserve gene pools at less cost or in.some cases where no other alternative to extinction existed.

  13. Effect of pre-freezing conditions on the progressive motility recovery rate of human frozen spermatozoa.

    PubMed

    Zhang, X; Zhou, Y; Xia, W; Wu, H; Yao, K; Liu, H; Xiong, C

    2012-10-01

    We evaluated the effects of sperm concentration, progressive motility, sperm morphology, duration of abstinence and collection season on the progressive motility recovery rate of human frozen spermatozoa to identify characteristics that predict the progressive motility recovery rate of human frozen spermatozoa and improve the protocol for sperm collecting in sperm banks. A total of 14 190 semen samples donated at Zhejiang human sperm bank of China between September 2006 and June 2011 were collected from 1624 donors. Semen was evaluated according to WHO standard procedures for sperm concentration. Progressive motility, sperm morphology, ejaculate collection season and abstinence time were recorded. After freezing and thawing, the progressive motility was assessed. Results showed that sperm concentration, progressive motility and normal morphology were significantly associated with the progressive motility recovery rate of human frozen spermatozoa. In addition, the abstinence time and collection season also significantly affected progressive motility recovery rate. Our results indicated that sperm concentration, progressive motility and normal morphology could be valuable in predicting the progressive motility recovery rate of human frozen spermatozoa. As such, progressive motility recovery may be improved by donating semen when abstinent for 3-5 days and during seasons other than summer.

  14. Successful ram semen cryopreservation with lyophilized egg yolk-based extender.

    PubMed

    Alcay, Selim; Berk Toker, M; Gokce, Elif; Ustuner, Burcu; Tekin Onder, N; Sagirkaya, Hakan; Nur, Zekariya; Kemal Soylu, M

    2015-10-01

    The aim of this study was to evaluate the effects of lyophilized egg yolk extender on ram semen cryopreservation. Ejaculates with a thick consistency, rapid wave motion (3-5 on a 0-5 scale) and >75% initial motility were pooled. Sperm were diluted to final concentration of 1/5 (semen/extender) in lyophilized egg yolk or fresh egg yolk extenders using two-step dilution method. The equilibrated semen was frozen in 0.25 mL straws. Semen samples were assessed for sperm motility, plasma membrane functional integrity using hypoosmotic swelling test (HOST), damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) and DNA integrity using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) at three time points: after dilution with extender A, equilibration and post-thaw. The results showed that freezing and thawing procedures (dilution, equilibration and thawing) had negative effects on motility (P<0.001), plasma membrane integrity (P<0.001), acrosome integrity (P<0.001) and DNA integrity (P<0.001). In the study, there were no significant differences between lyophilized and fresh egg yolk extenders when comparing motility, plasma membrane integrity, acrosome integrity and DNA integrity between groups. In conclusion, lyophilized egg yolk extender provided similar cryoprotective effects with fresh egg yolk extender to cryopreserve ram semen.

  15. Artificial insemination of captive European brown hares (Lepus europaeus PALLAS, 1778) with fresh and cryopreserved semen derived from free-ranging males.

    PubMed

    Hildebrandt, T B; Roellig, K; Goeritz, F; Fassbender, M; Krieg, R; Blottner, S; Behr, B; Hermes, R

    2009-11-01

    This study aimed to establish artificial insemination (AI) protocols to predictably initiate pregnancy during the breeding season in the European brown hare (EBH) (Lepus europaeus PALLAS, 1778). Semen was collected from seven captive and eight free-ranging males by means of electroejaculation. Semen from the free-ranging males was cryopreserved using directional freezing. Total motility/integrity of fresh and frozen-thawed semen was 91.6%/87.7% and 46.9%/53.8%, respectively. Ovulation was induced in ultrasonographically preselected females using a gonadotropin-releasing hormone analogue. Each female was inseminated with 1 mL fresh (Group A, n=16) or frozen-thawed semen (Group B, n=9) at a concentration of 100x10(6) spermatozoa/mL. The use of ultrasonography (10 to 22 MHz) confirmed the intracervical semen deposit, the success of artificial ovulation induction (formation of postovulatory corpus luteum), and permitted the monitoring of individual pregnancies. Although sperm motility/integrity was significantly different between groups, no significant difference was detected in conception rates (A, 87.50%; B, 77.78%). Because of embryonic resorption, there was a slight difference in fertility rate between groups (A, 62.5%; B, 77.78%). Overall, AI in captive EBH using fresh and frozen-thawed semen achieved successful fertility rates. Long-term cryopreserved semen was used to bring new genetic material from the wild into a genetically limited captive population without extensive animal transport. Therefore, AI has the potential to enhance breeding programs for EBH especially when cryopreserved semen from wild donors is used.

  16. Characteristics of frozen-thawed spermatozoa cryopreserved with different concentrations of glycerol in captive Japanese black bears (Ursus thibetanus japonicus).

    PubMed

    Okano, Tsukasa; Nakamura, Sachiko; Komatsu, Takeshi; Murase, Tetsuma; Miyazawa, Kiyoshi; Asano, Makoto; Tsubota, Toshio

    2006-10-01

    Seven mature Japanese black bears were used as semen donors, and a total of 7 semen samples collected from the animals by the electroejaculation method were cryopreserved in liquid nitrogen. Egg yolk-TRIS-citrate-glucose extender was used, and the effects of different final concentrations of glycerol, at 4-12% (v/v), on frozen-thawed spermatozoa were examined. No significant difference was observed in percent motility or percent abnormal morphology of frozen-thawed spermatozoa among the different glycerol concentrations. Percent viability and percent intact acrosomes of spermatozoa cryopreserved with 4 and 6% glycerol were significantly higher than those with 10 and 12% glycerol. These results suggest that a suitable glycerol concentration for freezing Japanese black bear semen within the range tested would be 4-6%.

  17. Tris-egg yolk-glycerol (TEY) extender developed for freezing dog semen is a good option to cryopreserve bovine epididymal sperm cells.

    PubMed

    Lopes, G; Soares, L; Ferreira, P; Rocha, A

    2015-02-01

    Cryopreservation of epididymal spermatozoa is often performed after shipping the excised testis-epididymis complexes, under refrigeration, to a specialized laboratory. However, epididymal spermatozoa can be collected immediately after excision of the epididymis and sent extended and refrigerated to a laboratory for cryopreservation. In this experiment, we evaluated the effect of both methods of cold storage bovine epididymal spermatozoa as well as of two different extenders on spermatozoa characteristics after freeze-thawing. For that, spermatozoa collected from the caudae epididymis of 19 bulls were extended and cryopreserved in either AndroMed(®) or a Tris-egg yolk (TEY)-based extender. Cryopreservation of sperm cells was performed immediately after castration (Group A, n = 9) or after cold storage for 24 h diluted in the two extenders and (Group B, n = 9) and also after cold storage for 24 h within the whole epididymis (Group C, n = 10). Sperm subjective progressive motility (light microscopy), plasma membrane integrity (hypoosmotic swelling test) and sperm viability (eosin-nigrosin) were evaluated. In vitro fertilization and culture (IVF) was performed to assess the blastocyst rate. No differences (p > 0.05) were observed on post-thaw sperm parameters between samples from Group A, B and C. TEY extended samples presented a higher (p < 0.01) percentage of progressive motile and live sperm, than those extended in AndroMed(®) . Blastocyst rate after IVF differed only (p < 0.05) between the reference group (IVF performed with frozen semen with known in vitro fertility) and Group A extended in AndroMed(®) . We conclude that when cryopreservation facilities are distant from the collection site, bovine epididymal sperm can be shipped chilled overnight either within the epididymal tail or after dilution without deleterious effect on post-thaw sperm quality. TEY extender was more suitable for cold storage and freezing bovine epididymal sperm, than the commercial

  18. 9 CFR 78.14 - Rodeo bulls.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... INTERSTATE TRANSPORTATION OF ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS BRUCELLOSIS Restrictions on Interstate Movement of Cattle Because of Brucellosis § 78.14 Rodeo bulls. (a) A rodeo bull that is test... classified as brucellosis negative based upon an official test conducted less than 365 days before the...

  19. Factors affecting spermatozoa morphology in beef bulls

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to evaluate factors affecting sperm morphology of bulls (n=908) collected at 320 days of age. Bulls were a composite breed (50% Red Angus, 25% Charolais, and 25% Tarentaise) born from 2002 to 2008 to dams fed levels of feed during mid and late gestation that were expe...

  20. Yearling Bull Breeding Soundness Examination: Special Considerations.

    PubMed

    Schrag, Nora; Larson, Robert L

    2016-07-01

    Accurate assessment of yearling bulls is important for the bottom line of all interested parties: the buyer, the seller, and the veterinarian performing the BSE. Special considerations and current research are highlighted and their application to the evaluation of yearling bulls is discussed.

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

    PubMed

    Vanha-Perttula, T; Remes, E

    1990-01-01

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

  2. Seminal plasma protein profiles of ejaculates obtained by internal artificial vagina and electroejaculation in Brahman bulls.

    PubMed

    Rego, J P A; Moura, A A; Nouwens, A S; McGowan, M R; Boe-Hansen, G B

    2015-09-01

    The present study was conducted to investigate if differences exist in the seminal plasma protein profile from mature Brahman bulls using two methods of semen collection: internal artificial vagina (IAV) and electroejaculation (EEJ). Semen was collected four times from three bulls on the same day and parameters were assessed immediately post-collection. Seminal plasma proteins were evaluated by 2-D fluorescence difference gel electrophoresis and identified by mass spectrometry. Semen volume was greater (P < 0.05) for EEJ (4.6 ± 0.35 mL) than for IAV (1.86 ± 0.24 mL) but sperm concentration was greater in IAV (1505 ± 189 × 10(6) sperm/mL) than in EEJ samples (344 ± 87 × 10(6) sperm/mL). Sperm motility and the percentage of normal sperm were not different between treatments. Total concentration of seminal plasma proteins was greater for samples collected by IAV as compared to EEJ (19.3 ± 0.9 compared with 13.0 ± 1.8 mg/mL, P < 0.05; respectively). Based on 2-D gels, 22 spots had a greater volume (P < 0.05) in gels derived from IAV samples, corresponding to 21 proteins identified as transferrin, albumin, epididymal secretory glutathione peroxidase, among others. Thirty-three spots, corresponding to 26 proteins, had a greater volume (P < 0.05) in gels derived from EEJ samples. These proteins were identified as spermadhesin-1, Bovine Sperm Protin 1, 3 and 5 isoforms, angiogenin-1, alpha-1B-glycoprotein, clusterin, nucleobindin-1, cathepsins, spermadhesin Z13, annexins, among others. Thus, proteins in greater amounts in samples obtained by IAV and EEJ were mainly of epididymal origin and accessory sex glands, respectively.

  3. Effect of inbreeding depression on bull sperm quality and field fertility.

    PubMed

    Dorado, Jesús; Cid, Rosa Morales; Molina, Antonio; Hidalgo, Manuel; Ariza, Julia; Moreno-Millán, Miguel; Demyda-Peyrás, Sebastián

    2015-12-18

    The present study investigated the effect of inbreeding depression on sperm quality using automated and objective methods and subsequent effects on beef bull field fertility. Individual inbreeding coefficient (F) values and field fertility data were determined using a dataset of AI bulls belonging to the Spanish Retinta Breeders Association (Asociación Nacional de Criadores de Ganado Vacuno Selecto de Raza Retinta (ANCRE)). Animals were clustered in two groups according to the F values as follows: (1) a high inbreeding group (HI; F ≥ 13.5%, mean 16.3); and (2) a non-inbreeding group (NI; F = 0%). In total, 17 different assessments were performed in both experimental groups, including evaluation of sperm morphology, acrosomal and DNA status, sperm plasma membrane integrity and function (hypo-osmotic swelling test), 10 kinetic parameters and the structure of sperm subpopulations. Sperm morphology, acrosomal and DNA status and osmotic tolerance were similar in both groups. Three velocity parameters (curvilinear velocity, straight line velocity and average path velocity) and the amplitude of lateral head displacement were higher in HI (P < 0.05). Cluster analysis of kinematic parameters revealed three different sperm subpopulations (sP1, sP2 and sP3), with the proportion of the sP1 population (highly active but non-progressive spermatozoa) being significantly (P < 0.05) higher in the HI group. Field fertility was assessed using two calving record datasets. In a smaller database including only bulls evaluated in the present study, there was a significant increase in the calving interval of cows sired with HI bulls. Conversely, in an extended genetic analysis of the ANCRE database, inbreeding only explained a small part of the variation in calving interval, and the results of regression analysis were not significant among bulls. The findings of the present study suggest that high inbreeding levels have a moderate effect on bull semen quality, with an increased

  4. A comparative study on the cryogenic preservation of semen from the sandhill crane and the domestic fowl

    USGS Publications Warehouse

    Sexton, T.J.; Gee, G.F.; Watson, P.F.

    1978-01-01

    SYNOPSIS: Recent findings on the cryogenic preservation of semen from the crane, Grus canadensis pratensis and the domestic fowl, Gallus domesticus, are compared. Highest levels of post-thaw motility for crane semen (55%) were obtained when semen was diluted 1:1 with the Beltsville Poultry Semen Extender (BPSE) and held for 30 min at 5 C before it was equilibrated with 4% dimethyl sulfoxide (DMSO) for 15 min. In contrast, post-thaw motility for fowl spermatozoa was highest (80%) when semen was diluted 1:3 with BPSE and held for 60 min at 5 C before it was equilibrated with 4% DMSO for 60 min. Post-thaw motility of spermatozoa of both species was highest when the following freezing rates were used: l C per min from +5 to -20 C, 50 C per min from -20 to -80 C, then plunging into liquid nitrogen which resulted in a rate of 160 C per min from -80 to -196 C. One of four crane eggs resulting from insemination with frozen-thawed semen was fertile, whereas 27 of 55 fowl eggs were fertile, but this difference may have been due largely to fewer spermatozoa being inseminated into the female crane than into the fowl.

  5. Cumene hydroperoxide induced changes in oxidation-reduction potential in fresh and frozen seminal ejaculates.

    PubMed

    Agarwal, A; Sharma, R; Henkel, R; Roychoudhury, S; Sikka, S C; du Plessis, S; Sarda, Y B; Speyer, C; Nouh, M; Douglas, C; Kayali, Z; Ahmed, E S; Sabanegh, E

    2017-03-15

    Oxidation-reduction potential (ORP) is a newer integrated measure of the balance between total oxidants (reactive oxygen species-ROS) and reductants (antioxidants) that reflects oxidative stress in a biological system. This study measures ORP and evaluates the effect of exogenous induction of oxidative stress by cumene hydroperoxide (CH) on ORP in fresh and frozen semen using the MiOXSYS Analyzer. Semen samples from healthy donors (n = 20) were collected and evaluated for sperm parameters. All samples were then flash-frozen at -80°C. Oxidative stress was induced by CH (5 and 50 μmoles/ml). Static ORP (sORP-(mV/10(6) sperm/ml) and capacity ORP (cORP-μC/10(6) sperm/ml) were measured in all samples before and after freezing. All values are reported as mean ± SEM. Both 5 and 50 μmoles/ml of CH resulted in a significant decline in per cent motility compared to control in pre-freeze semen samples. The increase in both pre-freeze and post-thaw semen samples for sORP was higher in the controls than with 50 μmoles/ml of CH. The change from pre-freeze to post-thaw cORP was comparable. The system is a simple, sensitive and portable tool to measure the seminal ORP and its dynamic impact on sperm parameters in both fresh and frozen semen specimens.

  6. Seasonal and genera-specific variations in semen availability and semen characteristics in large parrots.

    PubMed

    Bublat, A; Fischer, D; Bruslund, S; Schneider, H; Meinecke-Tillmann, S; Wehrend, A; Lierz, M

    2017-03-15

    In large parrots electro-stimulation is suitable for collecting semen, and therefore, to facilitate semen examination and artificial insemination. Previous studies have detected differences in the semen collection success rate and semen parameters between psittacine genera. It remained unclear whether these differences were genera-related, seasonal variations or depend on the males' relationship status. To answer these questions, semen collection and spermatological analysis were performed for four psittacine groups (macaws, amazons, eclectus parrots and cockatoos) over 13 months. In one breeding facility, semen collection was attempted in 82 males using electro-stimulation twice monthly. A complete spermatological evaluation was performed on 435 semen samples. Volume, color, consistency, contamination and pH of semen, as well as motility, progressive motility, sperm concentration, total sperm count, viability, and morphology of spermatozoa were evaluated. Seasonality affected the collection success rate in macaws and amazons. Thereby, in amazons a distinct peak was observed several days before and around oviposition, whereas eclectus parrots and cockatoos produced semen all year round. The average sperm concentration was highest in eclectus parrots (2.7 × 10(6) sperm/μl) and lowest in macaws (35.6 × 10(3) sperm/μl). The differences in the semen collection success rate and semen parameters seem to coincide with the bird's breeding biology. The collected data allows a prognostic estimation when semen collection seems favorable, and may be taken as orientation values for semen analysis in these species.

  7. Polymorphisms in luteinizing hormone receptor and hypothalamic gonadotropin-releasing hormone genes and their effects on sperm quality traits in Chinese Holstein bulls.

    PubMed

    Sun, Li-Ping; Du, Qing-Zhi; Song, Ya-Pan; Yu, Jun-Na; Wang, Shu-Juan; Sang, Lei; Song, Luo-Wen; Yue, Yao-Min; Lian, Yu-Ze; Zhang, Sheng-Li; Hua, Guo-Hua; Zhang, Shu-Jun; Yang, Li-Guo

    2012-06-01

    Genes of hypothalamic-pituitary-gonadal axis play a key role in male reproductive performance. This study evaluated the polymorphisms of luteinizing hormone receptor (LHR) and hypothalamic gonadotropin-releasing hormone (GnRH) genes and their effects on sperm quality traits including semen volume per ejaculate (VOL), sperm density (SD), fresh sperm motility (FSM), thawed sperm motility (TSM), acrosome integrity rate (AIR), and abnormal sperm rate (ASR) collected from 205 Chinese Hostein bulls. The study bulls consisted of 205 mature Chinese Holstein, 27 Simmental, 28 Charolais, and 14 German yellow cattle. One single nucleotide polymorphism (SNP) (A883G) in exon 2 of GnRH and two SNPs (A51703G and G51656T) in intron 9 of LHR were identified in 274 bulls. Analysis of variance in 205 Chinese Holstein bulls showed that age had significant effect on both SD and FSM (P < 0.01), and ASR (P < 0.05). With regards to genotype and its interaction with age, only the SNP of G51656T in LHR gene had significant effect on SD (P < 0.05, P < 0.01; respectively). The association result showed that bulls with AG genotype had higher FSM than bulls with AA and GG genotype in LHR at 51,703 locus (P < 0.10), and bulls with GG genotype had higher SD than bulls with TT genotype in LHR at G51656T locus (P < 0.10). Phenotypic correlation among the traits revealed that significant negative correlations were observed between ASR and AIR (r = -0.736, P < 0.01), ASR and AIR (r = -0.500, P < 0.01). There were moderate positive correlations between VOL and SD (r = 0.422, P < 0.01), as well as FSM (r = 0.411, P < 0.01). In conclusion, LHR may be a potential marker for sperm quality of SD and FSM.

  8. Management and Breeding Soundness of Mature Bulls.

    PubMed

    Palmer, Colin W

    2016-07-01

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

  9. Cryoprotectant redistribution along the frozen straw probed by Raman spectroscopy.

    PubMed

    Karpegina, Yu A; Okotrub, K A; Brusentsev, E Yu; Amstislavsky, S Ya; Surovtsev, N V

    2016-04-01

    The distribution of cryoprotectant (10% glycerol) and ice along the frozen plastic straw (the most useful container for freezing mammalian semen, oocytes and embryos) was studied by Raman scattering technique. Raman spectroscopy being a contactless, non-invasive tool was applied for the straws filled with the cryoprotectant solution and frozen by controlled rate programs commonly used for mammalian embryos freezing. Analysis of Raman spectra measured at different points along the straw reveals a non-uniform distribution of the cryoprotectant. The ratio between non-crystalline solution and ice was found to be increased by several times at the bottom side of the solution column frozen by the standard freezing program. The increase of the cryoprotectant fraction occurs in the area where embryos or oocytes are normally placed during their freezing. Possible effects of the cooling rate and the ice nucleation temperature on the cryoprotectant fraction at the bottom side of the solution column were considered. Our findings highlight that the ice fraction around cryopreserved embryos or oocytes can differ significantly from the averaged one in the frozen plastic straws.

  10. Effects of open-entry spike-bull, limited-entry branched-bull harvesting on elk composition in Washington

    USGS Publications Warehouse

    Bender, L.C.; Fowler, P.E.; Bernatowicz, J.A.; Musser, J.L.; Stream, L.E.

    2002-01-01

    The Washington Department of Fish and Wildlife implemented an open-entry spike-bull, limited-entry branched-bull elk (Cervus elaphus) harvest strategy in the Blue Mountains (1989), Yakima (1994), and Colockum (1994) herd areas of Washington state with goals of increasing numbers of adult bulls to increase breeding efficiency and possibly calf recruitment. Numbers of total bulls/100 cows (x??=5.4) and branched bulls/100 cows (x??=5.3) increased with the change in harvest strategy, while yearling bulls/100 cows remained unchanged; calves/100 cows declined (x??=-8.6). Calves/100 cows were always negatively correlated with both total bulls/100 cows and branched bulls/100 cows in each area; correlations were significant in 5 of 9 comparisons with total-bull ratios and 5 of 9 comparisons with branched-bull ratios. Open-entry spike-bull, limited-entry branched-bull harvesting can be used to increase total-bull and branched-bulls ratios in hunted elk populations. However, the increased ratios of bulls and branched bulls were unimportant in influencing calf recruitment, likely because of the importance of female condition on production and survival of young.

  11. Ejaculate collection efficiency and post-thaw semen quality in wild-caught Griffon vultures from the Sardinian population.

    PubMed

    Madeddu, Manuela; Berlinguer, Fiammetta; Ledda, Massimo; Leoni, Giovanni G; Satta, Valentina; Succu, Sara; Rotta, Andrea; Pasciu, Valeria; Zinellu, Angelo; Muzzeddu, Marco; Carru, Ciriaco; Naitana, Salvatore

    2009-02-19

    This study aimed to test the feasibility of a programme of semen collection and cryopreservation in Griffon vultures. Four wild-caught individuals kept in captivity because of unrecoverable traumas were used. Semen collection attempts were made twice a week during three consecutive reproductive seasons (December - March) using the abdominal massage method. Ejaculation was successfully induced between late January and late February. Semen collection efficiency was rather low (27.9%) and it did not vary among individuals (p > 0.05). No differences were found in ejaculate volumes (12.5 +/- 9.1 microl), spermatozoa concentration (28.4 +/- 30.9 million cells/ml) and viability (61.3 +/- 13.9%) among the 4 vultures. ATP values differed among the four vultures (p < 0.001); B showed higher nucleotide concentration than both C and D, while it did not differ form A, whose values were higher compared with D. After freezing and thawing, semen in vitro viability, DNA integrity and ATP intracellular concentration were determined. Spermatozoa viability after thawing did not differ among the four individuals (52.6 +/- 5.8 in A, 53.4 +/- 4.6 in B, 50.4 +/- 3.2 in C, 42.5 +/- 2.7 in D), but it decreased significantly compared to fresh semen (p < 0.05). During 4 hrs in vitro culture, spermatozoa collected from B maintained over time a higher viability in vitro when compared to A, C and D. As evaluated by the comet assay method, DNA fragmentation after freezing and thawing did not differ in the 4 vultures. ATP concentration in frozen/thawed semen was significantly lower than in fresh semen (p < 0.0001). This study indicates that semen cryopreservation can be considered as a useful tool in the conservation of Griffon vulture genetic resources, but further studies are needed to optimize this technique.

  12. Semen molecular and cellular features: these parameters can reliably predict subsequent ART outcome in a goat model

    PubMed Central

    Berlinguer, Fiammetta; Madeddu, Manuela; Pasciu, Valeria; Succu, Sara; Spezzigu, Antonio; Satta, Valentina; Mereu, Paolo; Leoni, Giovanni G; Naitana, Salvatore

    2009-01-01

    Currently, the assessment of sperm function in a raw or processed semen sample is not able to reliably predict sperm ability to withstand freezing and thawing procedures and in vivo fertility and/or assisted reproductive biotechnologies (ART) outcome. The aim of the present study was to investigate which parameters among a battery of analyses could predict subsequent spermatozoa in vitro fertilization ability and hence blastocyst output in a goat model. Ejaculates were obtained by artificial vagina from 3 adult goats (Capra hircus) aged 2 years (A, B and C). In order to assess the predictive value of viability, computer assisted sperm analyzer (CASA) motility parameters and ATP intracellular concentration before and after thawing and of DNA integrity after thawing on subsequent embryo output after an in vitro fertility test, a logistic regression analysis was used. Individual differences in semen parameters were evident for semen viability after thawing and DNA integrity. Results of IVF test showed that spermatozoa collected from A and B lead to higher cleavage rates (0 < 0.01) and blastocysts output (p < 0.05) compared with C. Logistic regression analysis model explained a deviance of 72% (p < 0.0001), directly related with the mean percentage of rapid spermatozoa in fresh semen (p < 0.01), semen viability after thawing (p < 0.01), and with two of the three comet parameters considered, i.e tail DNA percentage and comet length (p < 0.0001). DNA integrity alone had a high predictive value on IVF outcome with frozen/thawed semen (deviance explained: 57%). The model proposed here represents one of the many possible ways to explain differences found in embryo output following IVF with different semen donors and may represent a useful tool to select the most suitable donors for semen cryopreservation. PMID:19900288

  13. Ejaculate collection efficiency and post-thaw semen quality in wild-caught Griffon vultures from the Sardinian population

    PubMed Central

    Madeddu, Manuela; Berlinguer, Fiammetta; Ledda, Massimo; Leoni, Giovanni G; Satta, Valentina; Succu, Sara; Rotta, Andrea; Pasciu, Valeria; Zinellu, Angelo; Muzzeddu, Marco; Carru, Ciriaco; Naitana, Salvatore

    2009-01-01

    This study aimed to test the feasibility of a programme of semen collection and cryopreservation in Griffon vultures. Four wild-caught individuals kept in captivity because of unrecoverable traumas were used. Semen collection attempts were made twice a week during three consecutive reproductive seasons (December – March) using the abdominal massage method. Ejaculation was successfully induced between late January and late February. Semen collection efficiency was rather low (27.9%) and it did not vary among individuals (p > 0.05). No differences were found in ejaculate volumes (12.5 +/- 9.1 μl), spermatozoa concentration (28.4 +/- 30.9 million cells/ml) and viability (61.3 +/- 13.9%) among the 4 vultures. ATP values differed among the four vultures (p < 0.001); B showed higher nucleotide concentration than both C and D, while it did not differ form A, whose values were higher compared with D. After freezing and thawing, semen in vitro viability, DNA integrity and ATP intracellular concentration were determined. Spermatozoa viability after thawing did not differ among the four individuals (52.6 +/- 5.8 in A, 53.4 +/- 4.6 in B, 50.4 +/- 3.2 in C, 42.5 +/- 2.7 in D), but it decreased significantly compared to fresh semen (p < 0.05). During 4 hrs in vitro culture, spermatozoa collected from B maintained over time a higher viability in vitro when compared to A, C and D. As evaluated by the comet assay method, DNA fragmentation after freezing and thawing did not differ in the 4 vultures. ATP concentration in frozen/thawed semen was significantly lower than in fresh semen (p < 0.0001). This study indicates that semen cryopreservation can be considered as a useful tool in the conservation of Griffon vulture genetic resources, but further studies are needed to optimize this technique. PMID:19228408

  14. Fresh Frozen Plasma

    DTIC Science & Technology

    2009-03-01

    therapeutic means). FFP can be prepared either by separation from whole blood or collection via plasmapheresis . Fresh frozen plasma contains the...FFP can be further separated into cryoprecipitate and what is known as “cryo-poor plasma,” a product rarely used for therapeutic means. Plasma is the

  15. The Frozen Price Game

    ERIC Educational Resources Information Center

    Alden, Lori

    2003-01-01

    In this article, the author discusses the educational frozen price game she developed to teach the basic economic principle of price allocation. In addition to demonstrating the advantages of price allocation, the game also illustrates such concepts as opportunity costs, cost benefit comparisons, and the trade-off between efficiency and equity.…

  16. Effect of pasteurized egg and Rosmarinus officinalis supplementation on quality of cryopreserved ram semen.

    PubMed

    Mascaro, F; Gil, L; Malo, C; Gonzales, N; Martinez, F; de Blas, I

    2013-01-01

    The aim was to assess the in vitro effect of pasteurized egg (PE) and rosemary (Rosmarinus officinalis) on frozen-thawed ram semen. Ejaculates from three mature rams of the Rasa Aragonesa breed were cryopreserved using a 2-step dilution method (Fraction 1: F1; Fraction 2: F2). In Experiment 1, semen was frozen in egg yolk (EY) or PE extenders. After thawing, similar results were obtained in terms of total and progressive motility, viability, hypo-osmotic swelling test (HOST) and acrosome integrity after 2 h incubation. In Experiment 2, addition of rosemary to F1, F2 or both fractions to EY extenders was evaluated. Rosemary in F1 decreased progressive motility (p = 0.013) after 2 h incubation. Finally, PE can be used as a substitute for EY to reduce hygienic risks in extenders and is easier to standardize. Supplementation of EY extender with rosemary in F1 reduced progressive motility. Rosemary supplementation in F2 does not affect semen quality.

  17. Evaluation of semen from nondomestic birds

    USGS Publications Warehouse

    Gee, G.F.; Bakst, M.R.; Cecil, H.C.

    1997-01-01

    Aspects of poultry Al technology are applicable to nondomestic birds. However, modifications in the methods of semen collection, evaluation, and insemination are often necessary to accomodate either the bird's size, sperm numbers, or. female anatomy. This section provides a brief overview of procedures used to evaluate semen from nondomestic birds. Unless specified, materials, reagents, etc., are identical to those used in evaluating poultry semen (see appropriate chapters).

  18. Foods - fresh vs. frozen or canned

    MedlinePlus

    Frozen foods vs. fresh or canned; Fresh foods vs. frozen or canned; Frozen vegetables versus fresh ... a well-balanced diet. Many people wonder if frozen and canned vegetables are as healthy for you ...

  19. Cryopreservation of rabbit semen using non-permeable cryoprotectants: effectiveness of different concentrations of low-density lipoproteins (LDL) from egg yolk versus egg yolk or sucrose.

    PubMed

    Iaffaldano, N; Di Iorio, M; Rosato, M P; Manchisi, A

    2014-12-30

    This study was designed to identify the most effective non-permeable cryoprotectant (CPA) for the cryopreservation of rabbit semen by comparing the effects of different concentrations of low-density lipoproteins (LDL) on post-thaw sperm quality with those of whole egg yolk or sucrose. In a second experiment, the performance of the non-permeable CPAs identified as most effective was assessed in vivo by determining reproductive performances. Pooled semen samples were diluted to a ratio of 1:1 (v:v) in freezing extender (Tris-citrate-glucose and 16% dimethylsulfoxide as permeable CPA) containing as non-permeable CPAs 6, 8, 10 or 15% LDL from egg yolk, 0.1M sucrose, or 15% egg yolk. The semen was loaded in 0.25mL straws and frozen in liquid nitrogen vapor. After thawing, we determined sperm motility, viability, osmotic resistance, and acrosome and DNA integrity. Our results clearly revealed a significant effect of LDL concentration on semen quality. Also, at an optimal concentration of 10%, motility and acrosome integrity were improved over the values recorded for egg yolk (P<0.05). Based on the in vitro data, 3 groups of does (n=30 each) were inseminated with fresh semen or semen frozen using sucrose or 10% LDL. Sucrose led to a significantly higher conception rate than LDL and reproductive performance was similar to that observed for fresh semen. Our findings indicate the markedly better performance of sucrose in vivo as a non-permeable CPA for the cryopreservation of rabbit semen.

  20. Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4 °C.

    PubMed

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

    2011-03-01

    An extender has been developed with low-density lipoproteins (LDLs) that eliminates the microbial risks associated with the use of whole egg yolk. The objective of this study was to assess the effects of substituting egg yolk with LDLs for use as an extender in sperm preservation at 4 °C, as well as on spermatozoa motility, plasma membrane and acrosome integrity, at two different concentrations (80×10(6) and 240×10(6) sperm per ml) for 8 days and to evaluate glycerol toxicity in both extenders. A total of 12 ejaculates were collected from three bulls. Spermatozoa motility was examined using computer-assisted semen analysis. Plasma membrane integrity was determined using the hypo-osmotic swelling test and acrosome integrity with the fluorescein isothiocyanate-Pisum sativum agglutinin test. The semen was subsequently divided into four aliquots and diluted with Tris-egg yolk-glycerol (TEG), Tris-egg yolk without glycerol (TE), LDL with glycerol (LDL(+)) and LDL without glycerol (LDL(-)), at 80×10(6) and 240×10(6) sperm per ml. This study showed that the LDL(+) and LDL(-) extenders were more effective at preserving spermatozoa motility, plasma membrane integrity and acrosome integrity than TEG and TE (P<0.05) during 8 days of incubation. After 3 days of incubation, a toxicity of glycerol was observed in TEG, whereas no significant difference was observed between LDL(+) and LDL(-). We can therefore conclude that the LDL extender can be used to refrigerate semen at 4 °C instead of TEG and TE at 80×10(6) and 240×10(6) sperm per ml for elite bulls. This finding can be used to define a policy for the storage of high-quality bull semen.

  1. Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4 °C

    PubMed Central

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

    2011-01-01

    An extender has been developed with low-density lipoproteins (LDLs) that eliminates the microbial risks associated with the use of whole egg yolk. The objective of this study was to assess the effects of substituting egg yolk with LDLs for use as an extender in sperm preservation at 4 °C, as well as on spermatozoa motility, plasma membrane and acrosome integrity, at two different concentrations (80×106 and 240×106 sperm per ml) for 8 days and to evaluate glycerol toxicity in both extenders. A total of 12 ejaculates were collected from three bulls. Spermatozoa motility was examined using computer-assisted semen analysis. Plasma membrane integrity was determined using the hypo-osmotic swelling test and acrosome integrity with the fluorescein isothiocyanate–Pisum sativum agglutinin test. The semen was subsequently divided into four aliquots and diluted with Tris–egg yolk–glycerol (TEG), Tris–egg yolk without glycerol (TE), LDL with glycerol (LDL+) and LDL without glycerol (LDL−), at 80×106 and 240×106 sperm per ml. This study showed that the LDL+ and LDL− extenders were more effective at preserving spermatozoa motility, plasma membrane integrity and acrosome integrity than TEG and TE (P<0.05) during 8 days of incubation. After 3 days of incubation, a toxicity of glycerol was observed in TEG, whereas no significant difference was observed between LDL+ and LDL−. We can therefore conclude that the LDL extender can be used to refrigerate semen at 4 °C instead of TEG and TE at 80×106 and 240×106 sperm per ml for elite bulls. This finding can be used to define a policy for the storage of high-quality bull semen. PMID:21113174

  2. Efficacious long-term cooling and freezing of Sapajus apella semen in ACP-118(®).

    PubMed

    Leão, D L; Miranda, S A; Brito, A B; Lima, J S; Santos, R R; Domingues, S F S

    2015-08-01

    The objectives of the present study were to test the effect of coconut water solution (CWS), TES-TRIS and ACP-118(®) on the seminal cooling and cryopreservation of semen from capuchin monkeys (Sapajus apella). Semen was collected from six males by electro-ejaculation, diluted in TES-TRIS, CWS or ACP-118(®), and maintained at 4°C for 28h. Semen was subsequently evaluated (Experiment I) or cryopreserved in the presence of different glycerol concentrations (3%, 5% or 7%) (Experiment II). ACP-118(®) was the preferred extender to preserve sperm motility and viability after 28h incubation at 4°C. Cooled sperm were successfully frozen-thawed in a medium containing 3% glycerol. After thawing, sperm retained the capacity to fertilize oocytes and zygotes were obtained. In conclusion, ACP-118(®) can be effectively and efficiently used as extender for the cooling of S. apella semen. Furthermore, cryopreservation using ACP-118(®) by adding 3% glycerol is suitable to maintain sperm morphology and the capacity of these cells to fertilize in vitro.

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

    PubMed

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

    2015-08-01

    This study was undertaken to examine the effect of cholesterol-loaded-cyclodextrin (CLC) on boar sperm viability and spermatozoa cryosurvival during boar semen cryopreservation, and methyl-β-cyclodextrin (MBCD) was treated for comparing with CLC. Boar semen treated with CLC and MBCD before freezing process to monitor the effect on survival and capacitation status by flow cytometry with appropriate fluorescent probes. Sperm viability was higher in 1.5mg CLC-treated sperm (76.9±1.01%, P<0.05) than un-treated and MBCD-treated sperm before cryopreservation (58.7±1.31% and 60.3±0.31%, respectively). For CTC patterns, F-pattern was higher in CLC treated sperm than MBCD-treated sperm, for B-pattern was higher in CLC-treated sperm than fresh sperm (P<0.05). For AR pattern (an acrosome-reacted sperm) was lower in CLC-treated sperm than MBCD-treated sperm (P<0.05). Moreover, we examined in vitro development of porcine oocytes after in vitro fertilization using CLC-treated frozen-thawed semen, in which CLC treatment prior to freezing and thawing increased the development of oocytes to blastocyst stage in vitro. In conclusion, CLC could protect the viability of spermatozoa from cryodamage prior to cryopreservation in boar semen.

  4. Use of Aloe vera-based extender for chilling and freezing collared peccary (Pecari tajacu) semen.

    PubMed

    Souza, A L P; Lima, G L; Peixoto, G C X; Silva, A M; Oliveira, M F; Silva, A R

    2016-05-01

    As an alternative for the conservation of collared peccary semen, this research aims at evaluating the use of Aloe vera (AV) extract as a cryoprotectant for semen chilling and freezing. Five ejaculates were divided in two aliquots that were diluted in Tris plus egg yolk (EY; 20%) or AV extract (20%) and chilled at 5 °C. In both treatments, an adequate semen conservation was achieved and values closer to 40% motile sperm with viability and osmotic response ranging from 20% to 40%, and normal morphology of 80% were found after 36 hours of storage. Moreover, 12 other ejaculates were diluted in Tris plus EY (20%) or AV extract (5, 10, or 20%) and glycerol (3%). Samples were frozen in liquid nitrogen and thawed after 1 week. After thawing, all the treatments containing EY or AV provided similar values for sperm morphology, viability, osmotic response, membrane integrity, sperm motility, amplitude of lateral head, beat cross frequency, and rapid, low, and static subpopulations, but the highest values for straightness and the lowest values for curvilinear velocity were found using 20% AV (P < 0.05). In conclusion, we found that AV extract at a 20% concentration could be used as an alternative substitute to EY in the formulation of Tris extenders for collared peccaries' semen chilling or freezing.

  5. Enrichment of high-quality spermatozoa in bovine semen: relative effectiveness of three filtration matrixes.

    PubMed

    Cisale, H O; Fischman, M L; Blasi, C D; Fernandez, H A; Gledhill, B L

    2001-05-01

    To be practical, any method for improving bull semen must yield a large quantity of motile spermatozoa. Some separation methods based on physical properties, e.g. filtration, chromatography, centrifugation, washing and pooling, have been reported as satisfactory, but generally are not repeatable. Nevertheless, filtration methods appear to allow the attainment of an acceptable number of spermatozoa, thus allowing such a technique to be introduced in the production of standard bovine semen doses for artificial insemination. The aim of this work was to evaluate systematically the relative effects of three filtration matrixes (silica oxide, glass beads or Sephadexätrade mark) on the improvement of whole ejaculate quality. Analysis of the type of matrix and the volume and height of the filtration column was performed. The only characteristic of the columns that appears to influence ejaculate quality after filtering is the matrix volume. While all matrixes produced improvement of semen quality, SephadexTM was better than the other matrixes tested. An explanation for the mechanism of column filtration is proposed.

  6. Texture of Frozen Food

    NASA Astrophysics Data System (ADS)

    Wani, Kohmei

    Quantitative determination of textural quality of frozen food due to freezing and storage conditions is complicated,since the texture is consisted of multi-dimensiona1 factors. The author reviewed the importance of texture in food quality and the factors which is proposed by a priori estimation. New classification of expression words of textural properties by subjective evaluation and an application of four elements mechanical model for analysis of physical characteristics was studied on frozen meat patties. Combination of freezing-thawing condition on the subjective properties and physiochemical characteristics of beef lean meat and hamachi fish (Yellow-tail) meat was studied. Change of the plasticity and the deformability of these samples differed by freezing-thawing rate and cooking procedure. Also optimum freezing-thawing condition was differed from specimens.

  7. Curation of Frozen Samples

    NASA Technical Reports Server (NTRS)

    Fletcher, L. A.; Allen, C. C.; Bastien, R.

    2008-01-01

    NASA's Johnson Space Center (JSC) and the Astromaterials Curator are charged by NPD 7100.10D with the curation of all of NASA s extraterrestrial samples, including those from future missions. This responsibility includes the development of new sample handling and preparation techniques; therefore, the Astromaterials Curator must begin developing procedures to preserve, prepare and ship samples at sub-freezing temperatures in order to enable future sample return missions. Such missions might include the return of future frozen samples from permanently-shadowed lunar craters, the nuclei of comets, the surface of Mars, etc. We are demonstrating the ability to curate samples under cold conditions by designing, installing and testing a cold curation glovebox. This glovebox will allow us to store, document, manipulate and subdivide frozen samples while quantifying and minimizing contamination throughout the curation process.

  8. Effects of toxicosis on bull growth, semen characteristics and breeding soundness evaluation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tall fescue (Lolium arundinaceum) possesses heat, drought, and pest resistance conferred to the plant by its mutualistic relationship with the ergot alkaloid producing fungal endophyte, Neotyphodium coenophialum. The objective of this study was to evaluate the impact of ergot alkaloid consumption on...

  9. Ebola Virus Persistence in Semen Ex Vivo.

    PubMed

    Fischer, Robert J; Judson, Seth; Miazgowicz, Kerri; Bushmaker, Trent; Munster, Vincent J

    2016-02-01

    On March 20, 2015, a case of Ebola virus disease was identified in Liberia that most likely was transmitted through sexual contact. We assessed the efficiency of detecting Ebola virus in semen samples by molecular diagnostics and the stability of Ebola virus in ex vivo semen under simulated tropical conditions.

  10. Ebola Virus Persistence in Semen Ex Vivo

    PubMed Central

    Fischer, Robert J.; Judson, Seth; Miazgowicz, Kerri; Bushmaker, Trent

    2016-01-01

    On March 20, 2015, a case of Ebola virus disease was identified in Liberia that most likely was transmitted through sexual contact. We assessed the efficiency of detecting Ebola virus in semen samples by molecular diagnostics and the stability of Ebola virus in ex vivo semen under simulated tropical conditions. PMID:26811984

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

    SciTech Connect

    Du, J.; Kleinhans, F.W.; Spitzer, V.J.; Critser, J.K. . Dept. of Medical Research); Horstman, L. . School of Veterinary Medicine); Mazur, P. )

    1990-01-01

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

  12. Effect of Addition of Taurine on the Liquid Storage (5°C) of Mithun (Bos frontalis) Semen

    PubMed Central

    Perumal, P.; Vupru, Kezhavituo; Rajkhowa, C.

    2013-01-01

    The present study was undertaken to assess the effect of taurine on sperm motility, viability, total sperm abnormalities, acrosomal and plasma membrane integrity, enzymatic profiles such as reduced glutathione (GSH), glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT), and biochemical profiles such as cholesterol efflux and malondialdehyde (MDA) production. A total of 50 ejaculates were collected twice a week from 8 mithun bulls, and semen was split into 4 equal aliquots and diluted with the TEYC extender. Group 1: semen was without additives (control); groups 2 to 4: semen was diluted with 25 mM, 50 mM, and 100 mM of taurine, respectively. Seminal parameters and enzymatic and biochemical profiles were assessed at 5°C. Inclusion of taurine into diluent resulted in significant (P < 0.05) decreases in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities after liquid storage compared with the control group. Additionally, taurine at 50 mM has significant improvement in quality of mithun semen than taurine at 25 or 100 mM stored in in vitro at 5°C. It was concluded that the possible protective effects of taurine on sperm parameters are from enhancing the function of antioxidant enzymes, preventing efflux of cholesterol from cell membranes and decreased MDA production. PMID:23853737

  13. Sperm DNA fragmentation and morphological degeneration in chilled elephant (Elephas maximus and Loxodonta Africana) semen collected by transrectal massage.

    PubMed

    O'Brien, J K; Steinman, K J; Montano, G A; Love, C C; Robeck, T R

    2013-05-01

    Ejaculates from nine Asian and two African elephants were analysed to gain a further understanding of mechanisms underlying variable semen quality after transrectal massage. Semen analysis was performed after collection (0 h; subjective motility parameters only) and after 24 h of chilled storage at 10 °C (24 h; all ejaculate and sperm characteristics). Ejaculates with ≤50% total motility (TM) at 24 h, which represented >90% of collection attempts, contained a sperm population with a high degree of DNA damage (64.2 ± 19.2% fragmented DNA) and an elevated incidence of detached heads (43.3 ± 22.5%). In contrast, good quality ejaculates designated as those with >50% TM at 24 h displayed higher (p < 0.05) values of sperm kinetic parameters, DNA integrity and normal morphology. Fertility potential was high for good quality ejaculates from two males (one Asian and one African bull) based on in vitro characteristics after chilled storage for up to 48 h post-collection. Urine contamination of semen, as assessed quantitatively by creatinine concentration, was confirmed as a significant factor in reduced elephant ejaculate quality. However, the identification of considerable DNA damage and morphological degeneration in the majority of ejaculates after only 24 h of chilled storage indicates that sperm ageing could be a primary contributor to inconsistent semen quality in the elephant.

  14. Assessment of the value of international genetic evaluations for yield in predicting domestic breeding values for foreign Holstein bulls.

    PubMed

    Nicolazzi, E L; Forabosco, F; Fikse, W F

    2011-05-01

    International genetic evaluations are a valuable source of information for decisions about the importation of (the semen of) foreign bulls. This study analyzed data from 6 countries (Australia, Canada, Italy, France, the Netherlands, and the United States) and compared international evaluations for production traits of foreign bulls (i.e., when no national daughter information was available) to their national breeding values in August 2009, which were based only on domestic daughters' data. A total of 821 bulls with highly reliable estimated breeding values (EBV) for milk, fat, and protein yield were analyzed. No evidence of systematic over- or underestimation was found in most of the countries analyzed. Observed correlations between national and international evaluations were close to 0.9 and, for most countries, generally close to their expected values (calculated from national and international EBV reliabilities). In Italy, however, higher differences between observed and expected correlations and significant mean differences between EBV for more than one trait were observed in bulls progeny-tested in the United States and in other European countries (with differences up to 33.1% of the genetic standard deviation). These results were probably induced by a relatively recent change in the model for national evaluation. The findings in this study reflect a conservative estimate of the real value of international evaluations, as changes in methodologies in either the national or the international evaluations decreased the ability of past international evaluations to predict current national evaluations. Nevertheless, our results indicate that international evaluations based on foreign information for Holstein bulls were reasonably accurate predictors of the future national breeding values based only upon domestic daughters.

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

    PubMed

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

    2015-01-01

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

  16. Critical swimming speeds of wild bull trout

    USGS Publications Warehouse

    Mesa, M.G.; Weiland, L.K.; Zydlewski, G.B.

    2004-01-01

    We estimated the critical swimming speeds (Ucrit) of wild bull trout at 6??, 11??, and 15??C in laboratory experiments. At 11??C, 5 fish ranging from 11 to 19 cm in length had a mean Ucrit of 48.24 cm/s or 3.22 body lengths per second (BL/s). Also at 11??C , 6 fish from 32 to 42 cm had a mean Ucrit of 73.99 cm/s or 2.05 BL/s. At 15??C, 5 fish from 14 to 23 cm had a mean Ucrit of 54.66 cm/s or 2.88 BL/s. No fish successfully swam at 6??C. Swim speed was significantly influenced by fish length. Many bull trout performed poorly in our enclosed respirometers: of 71 Ucrit tests we attempted, only the 16 described above were successful. Bull trout that refused to swim held station within tunnels by using their pectoral fins as depressors, or they rested and later became impinged against a downstream screen. Several common techniques did not stimulate consistent swimming activity in these fish. Our estimates of U crit for bull trout provide an understanding of their performance capacity and will be useful in modeling efforts aimed at improving fish passage structures. We recommend that fishway or culvert designers concerned with bull trout passage maintain velocities within their structures at or below our estimates of Ucrit, thus taking a conservative approach to ensuring that these fish can ascend migratory obstacles safely.

  17. Genetic parameters for scrotal circumference, breeding soundness examination and sperm defects in young Nellore bulls.

    PubMed

    Silva, M R; Pedrosa, V B; Borges-Silva, J C; Eler, J P; Guimarães, J D; Albuquerque, L G

    2013-10-01

    A total of 51,161 records of scrotal circumference measurements at 18 mo of age (SC18) and 17,648 records of sperm defects and breeding soundness of Nellore bulls (mean age of 22.5 mo), raised under extensive conditions, were analyzed to estimate coefficients of heritability and genetic correlations of morphological semen traits by Bayesian inference. The observed semen traits were classified as minor (MID), major (MAD), and total sperm defects (TD). The animals were classified according to breeding soundness as satisfactory and unsatisfactory potential breeders. The (co)variance components and breeding values were estimated by Gibbs sampling using the GIBBS2F90 program under an animal model that included contemporary group as fixed effect, age of animal as linear covariate, and direct additive genetic effects as random effects. Heritabilities of 0.40 ± 0.02, 0.16 ± 0.02, 0.04 ± 0.01, 0.15 ± 0.01, and 0.10 ± 0.01 were obtained for SC18, MID, MAD, TD, and breeding soundness, respectively. The SC18 showed a positive and moderate correlation with breeding soundness (0.56 ± 0.04) and a negative and low correlation with MID (-0.23 ± 0.03), MAD (-0.16 ± 0.02), and TD (-0.24 ± 0.02). In conclusion, scrotal circumference showed the best response to selection among the traits studied and was favorably correlated with breeding soundness and sperm morphology in young Nellore bulls.

  18. Sequestration of PDC-109 protein by specific antibodies and egg yolk cryoprotects bull spermatozoa.

    PubMed

    Srivastava, N; Srivastava, S K; Ghosh, S K; Jerome, A; Das, G K; Mehrotra, S

    2013-10-01

    PDC-109, one of the most abundant proteins in bovine seminal plasma, has detrimental effect on spermatozoa in a time- and concentration-dependent manner. Therefore, we hypothesized that sequestration of detrimental protein from ejaculates would be beneficial following cryopreservation of sperm cells. To this aim, we evaluated the effect of sequestration of PDC-109 either by anti-PDC-109 antibodies (Ab) or egg yolk (EY) alone or by the synergistic action of EY + Ab in minimizing cryoinjury to bull spermatozoa. PDC-109 protein was purified by applying two-step chromatography procedures. The purified protein was injected in rabbits to raise antibodies which were isolated using ion-exchange chromatography. After checking the Ab cross-reactivity, they were quantitated and added to ejaculates, either alone or in addition to EY in Tris-glycerol (TG) extender. Thus, ejaculates were processed in extender containing EY + TG (group I), Ab + TG (group II) or EY + Ab + TG (group III). Semen quality parameters (SQPs) viz. viability and acrosome integrity (FITC-PSA), cryoinjury to spermatozoa (chlortetracycline, CTC assay) and in vitro fertility of protein-sequestered-semen (zona-penetration assay) were evaluated. A significant (p < 0.05) improvement in post-thaw SQPs as well as in non-capacitated spermatozoa observed at pre-freeze and post-thaw stages of cryopreservation in group III compared with other groups indicated reduction in protein-mediated cryoinjury. From this study, it can be concluded that sequestration of PDC-109 by synergistic action of EY+Ab as compared to either of them alone significantly improve sperm quality and minimize cryoinjury to bull spermatozoa upon storage at ultra-low temperatures.

  19. Cryopreservation of European Mouflon (Ovis Gmelini Musimon) semen during the non-breeding season is enhanced by the use of trehalose.

    PubMed

    Berlinguer, F; Leoni, G G; Succu, S; Mossa, F; Galioto, M; Madeddu, M; Naitana, S

    2007-04-01

    The influence of trehalose on European mouflon spermatozoa cryopreservation during the non-breeding season was tested. Semen was frozen in two different extenders: (a) recommended Tris-based ram extender (CTR); (b) CTR extender supplemented with trehalose 0.147 mm (TRH). Sperm viability and acrosome integrity were assessed using propidium iodide and fluorescein isothiocynate labelled Pisum Sativum agglutinin. Trehalose significantly enhanced sperm viability after thawing compared with CTR extender (62.7% vs 51.8%; p < 0.05), whereas no differences were observed on acrosome integrity (42.9% vs 42.1%). Trehalose influence was also evidenced in the in vitro fertility test performed with sheep oocytes matured in vitro. Both fertilization rates (60.9% TRH vs 43.6% CTR; p < 0.05) and cleavage rates (58% TRH vs 39.8% CTR; p < 0.001) were higher for trehalose frozen semen compared with control extender frozen semen. A higher percentage of zygotes resulting from fertilization with trehalose cryopreserved semen presented the first cleavage earlier if compared with the group fertilized with control semen (48.7% vs 31.5%, respectively; p < 0.01). This result was confirmed by embryo kinetic development. Fertilization with trehalose cryopreserved semen leaded to an higher percentage of blastocysts (40.2% vs 27.8% CTR; p < 0.05), and enhanced in particular the number of blastocysts that developed on the day 6th of culture (28.6% vs 17% CTR; p < 0.05). Our data demonstrated that, during mouflon non-breeding season, trehalose extender enhances spermatozoa viability and its in vitro fertilizing capacity, allowing the production of an higher number of blastocysts.

  20. Effect of genistein supplementation of thawing medium on characteristics of frozen human spermatozoa.

    PubMed

    Martinez-Soto, Juan Carlos; de DiosHourcade, Juan; Gutiérrez-Adán, Alfonso; Landeras, José Lorenzo; Gadea, Joaquín

    2010-05-01

    In this study, we evaluated the effects of genistein supplementation of the thawing extender on frozen-thawed human semen parameters. We analyzed the effect of supplementation on sperm motility, capacitation (membrane lipid disorder), reactive oxygen species (ROS) generation, chromatin condensation and DNA damage. Using this preliminary information, it maybe possible to improve the cryopreservation process and reduce the cellular damage. We have confirmed that the isoflavone genistein (10 micromol L(-1)) has antioxidant properties on the frozen-thawed spermatozoa. This results in a decreased ROS production that shows a slight improvement in the sperm motility, and decreases the membrane lipid disorder and DNA damage caused by cryopreservation. These results suggest an effect of genistein on sperm functionality that could be of interest for assisted reproduction treatments using frozen-thawed human spermatozoa, but further studies will be necessary to confirm our findings and to evaluate the possible clinical applications.

  1. 4. Photocopy from James H. Bull, Records of the Descendants ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    4. Photocopy from James H. Bull, Records of the Descendants of John and Elizabeth Bull, 1919 WEST AND NORTH ELEVATIONS - Mount Pleasant, Bulltown Road (East Nantmeal Township), East Nantmeal, Chester County, PA

  2. 58. DETAIL OF PINION AND BULL GEARS: Detail view towards ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    58. DETAIL OF PINION AND BULL GEARS: Detail view towards northeast of the pinion and bull gears of the winding machinery. - San Francisco Cable Railway, Washington & Mason Streets, San Francisco, San Francisco County, CA

  3. Electroejaculation and semen analysis and freezing in the giant panda (Ailuropoda melanoleuca).

    PubMed

    Platz, C C; Wildt, D E; Howard, J G; Bush, M

    1983-01-01

    Semen was collected by a standardized electroejaculation procedure from a giant panda on 4 occasions. Ejaculate volume, sperm count and % sperm motility were 2.3-3.6 ml, 62-562 x 10(6) spermatozoa/ml and 45-85%, respectively. The results, although limited to a single male, suggested a seasonal influence on ejaculate and gonadal parameters with improved ejaculate volume, sperm motility and increased testicular size in the season proximate to the female's oestrous period. Frozen-thawed spermatozoa were motile with no apparent abnormalities induced by the freezing procedure.

  4. Effect of freezing extender composition and male line on semen traits and reproductive performance in rabbits.

    PubMed

    Viudes-de-Castro, M P; Lavara, R; Safaa, H M; Marco-Jiménez, F; Mehaisen, G M K; Vicente, J S

    2014-05-01

    This study was conducted to elucidate the effect of different freezing extenders on two lines selected for hyperprolificacy and longevity (H and LP, respectively). In extender A, dimethyl sulphoxide (Me2SO) and sucrose were used as cryoprotectants. In extenders B and C, the sucrose was replaced by 20% egg yolk, and in extender C the Me2SO was substituted by acetamide. Semen was packaged in 0.25 ml plastic straws and cooled at 5°C for 45 min, and then was frozen in liquid nitrogen vapour for 10 min before being plunged into the liquid nitrogen. Thawing was carried out by immersing the straws in a water bath at 50°C for 10 s. Frozen-thawed semen characteristics and reproductive parameters were affected by freezing. Extender C showed significantly lower post-thawing quality traits than any of the three extenders. Acrosome integrity was significantly improved when Me2SO was used as cryoprotectant. Sucrose replacement by 20% egg yolk had no effect on acrosome integrity but provided significantly lower sperm motility and viability. Freezing extender affected fertility rate, total born, number of implantation sites and gestational losses, obtaining better results when extender A was used. The acrosomal integrity after frozen-thawed process showed a significant correlation with fertility at 12th day and also at birth, indicating that an increase in acrosomal integrity leads to an increase in both fertilities (12th day and at birth). A positive correlation between motility of semen and implantation sites was found. The post-thawing quality traits of semen were not affected by the genetic line, although LP line showed higher total born and lower foetal and gestational losses. The findings of this study suggest that freezing extender composition has a significant effect on the success of rabbit sperm for preservation, and when Me2SO was used as permeable cryoprotectant sucrose provided better protection compared with egg yolk and improved reproductive traits, and, on the

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

    PubMed

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

    2015-10-01

    The possibility of including amino acids for cryopreservation of ram semen to improve the quality of frozen semen was explored in this study in sheep model. 24 samples were collected in triplicate from 8 rams of 2-3 year old Bannur cross bred rams maintained at the Institute Experimental Livestock Unit. Semen was diluted in tris-egg yolk glycerol diluent and made into 7 aliquots as follows: aliquot 1 served as control, "l-alanine" was added at 100 and 135mM in the aliquots 2 and 3, "l-glutamine" was added at 20 and 25mM in the aliquots 4 and 5 and "l-proline" was added at 25 and 50mM in the aliquots 6 and 7, respectively. Diluted semen was filled in 0.25ml French straws and frozen in LN2. Inclusion of "l-proline" and "l-glutamine" in the diluent increased the percent live sperm (P<0.001), total motility (P<0.05) and maintained higher functional membrane and acrosomal integrity (P<0.001) by decreasing lipid peroxidation (P<0.001) compared to the control group. In contrast, "l-alanine" decreased the percentage of total motility, fast progressive spermatozoa and increased (P<0.01) the percentage of immotile spermatozoa. It can be concluded that 20mM "l-glutamine" and 25mM "l-proline" can be used as semen additive to freeze ram semen as they prevented cryoinjuries to sperm and improved the pre-freeze and post-thaw semen characteristics.

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

    PubMed

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

    2014-05-01

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

  7. Frozen Carbon Dioxide

    NASA Technical Reports Server (NTRS)

    2005-01-01

    1 August 2005 This Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) image shows a south polar residual cap landscape, formed in frozen carbon dioxide. There is no place on Earth that one can go to visit a landscape covering thousands of square kilometers with frozen carbon dioxide, so mesas, pits, and other landforms of the martian south polar region are as alien as they are beautiful. The scarps of the south polar region are known from thousands of other MGS MOC images to retreat at a rate of about 3 meters (3 yards) per martian year, indiating that slowly, over the course of the MGS mission, the amount of carbon dioxide in the martian atmosphere has probably been increasing.

    Location near: 86.9oS, 25.5oW Image width: width: 3 km (1.9 mi) Illumination from: upper left Season: Southern Spring

  8. Semen characteristics in pubertal boys. I. Semen quality after first ejaculation.

    PubMed

    Janczewski, Z; Bablok, L

    1985-01-01

    Semen specimens from 134 pubertal boys were examined, and some 274 assays were made. An analysis of the biological quality of semen in relation to the period of time after first ejaculation brings high values of statistical dependence of the volume of semen, its liquefaction, spermatozoal concentration, percentage of morphologically normal forms of spermatozoa, and normal spermatozoal motility on the period of time after first ejaculation. Normal figures for semen volume, semen liquefaction, spermatozoal concentration, and morphology are observed 12-14 months after first ejaculation. The percentage of normally motile spermatozoa becomes standard 21-23 months after first ejaculation. There were changes in semen characteristics from azoospermia through cryptozoospermia, oligozoospermia, and asthenozoospermia to normospermia. Azoospermia dominates until the fifth month after the first ejaculation, oligozoospermia from the sixth to the eleventh month, asthenozoospermia from the twelfth to the twentieth month, and normospermia from the twenty-first month.

  9. 21 CFR 158.170 - Frozen peas.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Frozen peas. 158.170 Section 158.170 Food and... CONSUMPTION FROZEN VEGETABLES Requirements for Specific Standardized Frozen Vegetables § 158.170 Frozen peas. (a) Identity—(1) Product definition. Frozen peas is the food in “package” form as that term...

  10. 21 CFR 160.110 - Frozen eggs.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Frozen eggs. 160.110 Section 160.110 Food and... CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.110 Frozen eggs. (a) Frozen eggs, frozen whole eggs, frozen mixed eggs is the food prepared by...

  11. 21 CFR 160.110 - Frozen eggs.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Frozen eggs. 160.110 Section 160.110 Food and... CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.110 Frozen eggs. (a) Frozen eggs, frozen whole eggs, frozen mixed eggs is the food prepared by...

  12. 21 CFR 160.110 - Frozen eggs.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Frozen eggs. 160.110 Section 160.110 Food and... CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.110 Frozen eggs. (a) Frozen eggs, frozen whole eggs, frozen mixed eggs is the food prepared by...

  13. 21 CFR 160.110 - Frozen eggs.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Frozen eggs. 160.110 Section 160.110 Food and... CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.110 Frozen eggs. (a) Frozen eggs, frozen whole eggs, frozen mixed eggs is the food prepared by...

  14. 21 CFR 160.110 - Frozen eggs.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Frozen eggs. 160.110 Section 160.110 Food and... CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.110 Frozen eggs. (a) Frozen eggs, frozen whole eggs, frozen mixed eggs is the food prepared by...

  15. Comparison between primary sex ratio in spermatozoa of bulls and secondary sex ratio in the deriving offspring.

    PubMed

    Amadesi, A; Frana, A; Gandini, L M; Bornaghi, V; Parati, K; Bongioni, G; Puglisi, R; Galli, A

    2015-01-15

    The objectives of the present work were to compare the primary sex ratio in sperm with the secondary sex ratio recorded in the offspring produced by artificial insemination (AI) with the same sperm and assess whether the primary sex ratio is influenced by sperm survival and motility after thawing. Calving data of 98 Holstein Friesian bulls used in AI were collected during 4 years, and commercial semen of the same bulls was analyzed immediately after thawing and after swim-up using a real-time polymerase chain reaction method developed and validated in our laboratory. Calving data relative to single bulls did not reveal any significant deviation between genders from the theoretical 1:1 for none of the bulls, being the mean values of male and female calves born 52.1 ± 2.80% and 47.9 ± 2.71%, respectively. Thereafter, calving events of bulls were classified and analyzed according to four classes of years: 2009 (n = 13,261), 2010 (n = 21,551), 2011 (n = 24,218), and 2012 (n = 41,726), and seasons categorized as winter, spring, summer, and fall. When data aggregated per years were analyzed, the difference between the two sexes was significant (P < 0.005) in favor of the male gender, whereas no influence of the season was evidenced. Real-time polymerase chain reaction did not evidence any difference between the mean values of frequency of Y chromosome-bearing sperm detected in three sperm batches of the same bulls analyzed immediately after thawing (51.1 ± 2.1), nor a difference with respect to the theoretical 1:1 ratio was reported after sperm analysis of one batch of sperm of the bulls analyzed after swim-up and immediately after thawing (50.1 ± 2.1 and 49.8 ± 1.8, respectively). The results are consistent with the observation of the farmers who often report a skewed sex ratio of the calves being born with AI in favor of the male gender. However, we have not evidenced differences in the primary sex ratio with respect to the theoretical 1:1 ratio both at thawing

  16. Associations between feed efficiency, sexual maturity and fertility-related measures in young beef bulls.

    PubMed

    Fontoura, A B P; Montanholi, Y R; Diel de Amorim, M; Foster, R A; Chenier, T; Miller, S P

    2016-01-01

    The beef industry has emphasized the improvement of feed utilization, as measured by modeling feed intake through performance traits to calculate residual feed intake (RFI). Evidence supports an inverse relationship between feed efficiency and reproductive function. The objective of this study was to determine the relationship of reproductive assessments and RFI unadjusted (RFI(Koch)) or adjusted for body composition (RFI(us)) and the relationship among fertility-related parameters. In total, 34 crossbred bulls were housed together for 112 days of performance evaluation, followed by assessment of scrotum IR imaging, scrotal circumference, testes ultrasonography and semen quality parameters at 377±33.4 days of age. Bulls were slaughtered at 389±34.0 days of age, and analyses of carcass composition, biometrics and histomorphometry of the testis and epididymis were conducted. Bulls were grouped into two subpopulations based on divergence of RFI, and within each RFI model either by including 50% of the population (Halves, high and low RFI, n=17) or 20.6% extremes of the population (Tails, high and low RFI, n=7). The means of productive performance and fertility-related measures were compared through these categories. Pearson's correlation was calculated among fertility-related measures. In the Halves subpopulation of the RFI(us), sperm of low-RFI bulls had decreased progressive motility (47.30% v. 59.90%) and higher abundance of tail abnormalities (4.30% v. 1.80%) than that of high-RFI bulls. In the Tails subpopulation of the RFI(Koch), low RFI displayed less variation in the scrotum surface temperature (0.62°C v. 1.16°C), decreased testis echogenicity (175.50 v 198.00 pixels) and larger (60.90 v. 56.80 mm(2)) but less-developed seminiferous tubules than high-RFI bulls. The evaluation of fertility-related parameters indicated that a higher percentage of immature seminiferous tubules was correlated with occurrence of sperm with distal droplets (r=0.59), a larger

  17. [Identification of semen Cuscutae by HPCE].

    PubMed

    Hu, P; Luo, G A; Wang, R J; Guo, H Z; Li, J S

    1997-07-01

    A method for identification of semen Cuscutae, a traditional Chinese herb, was developed. The proteins in semen Cuscutae were extracted under acid or basic conditions and separated by high performance capillary electrophoresis. The electrophoretograms of acidic or basic extractants from Cuscuta chinensis Lam., Cuscuta australis R. Br. and Cuscuta japonica Choisy showed significant differences, which can be used to identify the three different semen Cuscutae. The results of the identification for 13 pharmacognosical samples agreed well with those of scanning electronic microscopy and tissue microanalysis.

  18. Quality parameters for alpaca (Vicugna pacos) semen are affected by semen collection procedure.

    PubMed

    Morton, K M; Thomson, P C; Bailey, K; Evans, G; Maxwell, W M C

    2010-08-01

    Artificial insemination (AI) is poorly developed in camelids owing to the difficulty in collecting high quality semen and the highly viscous nature of the semen. Semen collected by artificial vagina (AV) is often of low quality and must be improved before any further development of AI technology can occur. The present study investigated the effects of adding a cervix-like stricture to the AV, presence of females, collecting semen into Androhep, skim-milk or Tris diluents, and catalase supplementation (0, 100, 200 or 600 units/ml) of Tris diluent on alpaca semen quality parameters. The addition of a cervix-like stricture increased mating length (p < 0.05), whilst the presence of females during semen collection did not improve semen quality parameters (p > 0.05). Collection of semen into Tris diluent improved sperm motility (58.0 +/- 11.9%) compared with the control (34.0 +/- 10.8%; p < 0.05), Androhep (33.5 +/- 10.7%) and skim-milk diluents (28.2 +/- 10.4%). Semen viscosity was reduced by collection into Androhep (4.6 +/- 1.7 mm) and skim-milk diluents (3.6 +/- 1.3 mm) compared with Tris diluent (5.7 +/- 2.1 mm) and no collection medium (9.3 +/- 3.5 mm; p < 0.05). Tris diluent supplemented with 100, 200 or 600 units/ml catalase increased semen viscosity (5.0 +/- 3.2 and 4.9 +/- 3.2 mm). Collection of alpaca semen by AV into Tris diluent increased semen quality facilitating further development of AI technology in alpacas.

  19. Association of soybean-based extenders with field fertility of stored ram (Ovis aries) semen: a randomized double-blind parallel group design.

    PubMed

    Khalifa, Tarek; Lymberopoulos, Aristotelis; Theodosiadou, Ekaterini

    2013-02-01

    Two consecutive randomized double-blind field fertility experiments were conducted over a 4-month period and aimed at evaluating the association of two commercial soybean lecithin-based extenders (AndroMed [Minitub, Tiefenbach, Germany] and BioXcell [IMV Technologies, L'Aigle, France]) with pregnancy rates of chilled-stored (CS) and frozen-thawed (FT) ram semen. Semen samples with more than 2 × 10(9) sperm per mL and 70% progressive motile spermatozoa were collected via an artificial vagina from twelve proven fertile Chios rams, split-diluted with the above mentioned extenders, packaged in 0.25 mL straws and either stored at 5 ± 1 °C for 30 to 36 hours or frozen and thawed. Non-lactating multiparous ewes were inseminated in progestagen-synchronized estrus either with CS (AndroMed: N = 212 and BioXcell: N = 206; intracervical AI) or with FT (AndroMed: N = 114 and BioXcell: N = 92; laparoscopic intrauterine AI) semen. Ovulation was confirmed in all ewes based on determination of blood plasma progesterone (>1 ng/mL) 8 days post AI. Ewes were screened for pregnancy diagnosis by transabdominal ultrasonography 65 days post AI. BioXcell was superior to AndroMed in preserving the fertilizing potential of CS (P < 0.05) and FT (P < 0.005) semen. In AndroMed-stored semen, young rams (1.5-2.5 years old, N = 8) had a pregnancy rate (59.1%; 124/210) lower than that (72.4%; 84/116) of mature rams (4.5 to 5.5 years, N = 4; P < 0.025). Compared with AndroMed extender, processing of young ram semen in BioXcell extender improved pregnancy rates of CS (66.7%; 88/132 vs. 83.9%; 94/112; P < 0.005) and FT (46.2%; 36/78 vs. 71.0%; 44/62; P < 0.01) spermatozoa. Both extenders were similarly effective in preserving pregnancy rates of mature ram semen (P > 0.05). Ram-by-extender interactions were significant for pregnancy rates of CS and FT semen. Irrespective of extenders, overall pregnancy rates after intracervical and intrauterine AI were 75.1% and 62.2%, respectively (P < 0.001). In

  20. Alternatives to Antibiotics in Semen Extenders: A Review

    PubMed Central

    Morrell, Jane M.; Wallgren, Margareta

    2014-01-01

    Antibiotics are added to semen extenders to be used for artificial insemination (AI) in livestock breeding to control bacterial contamination in semen arising during collection and processing. The antibiotics to be added and their concentrations for semen for international trade are specified by government directives. Since the animal production industry uses large quantities of semen for artificial insemination, large amounts of antibiotics are currently used in semen extenders. Possible alternatives to antibiotics are discussed, including physical removal of the bacteria during semen processing, as well as the development of novel antimicrobials. Colloid centrifugation, particularly Single Layer Centrifugation, when carried out with a strict aseptic technique, offers a feasible method for reducing bacterial contamination in semen and is a practical method for semen processing laboratories to adopt. However, none of these alternatives to antibiotics should replace strict attention to hygiene during semen collection and handling. PMID:25517429

  1. Ram seminal plasma improves pregnancy rates in ewes cervically inseminated with ram semen stored at 5 °C for 24 hours.

    PubMed

    López-Pérez, A; Pérez-Clariget, R

    2012-01-15

    In this study, we compared pregnancy rates obtained using ram semen stored at 5 °C for 24 h, with ram or bull seminal plasma (SP) added to TRIS-egg yolk extender. During the breeding period, 670 adult Corriedale ewes were cervically inseminated with semen (2 × 10(8) sperm in a volume of 0.2 mL) from eight adult Corriedale rams. Ejaculates, obtained using an artificial vagina, were split into three aliquots and diluted with the following: TRIS-egg yolk based extender (T), T + 30% ram SP (R), or T + 30% bull SP (B). Samples were refrigerated and stored at 5 °C for 24 h until used for AI. Pregnancy was assessed by ultrasonography 35 to 40 d after AI. Pregnancy rate was not affected by ram (P = 0.77) or breeding period (P = 0.43), and there were no interactions between extender and ram (P = 0.94), or extender and breeding period (P = 0.24). However, there was an effect of extender (P = 0.0009) on pregnancy rates; ram SP, but not bull SP, increased pregnancy rates compared with extender without SP (49.7, 38.1, and 31.1%, for R, B, and T respectively). In conclusion, ram SP added to TRIS-egg yolk extender had a beneficial effect on the pregnancy rate of ram sperm stored at 5 °C for 24 h and used for cervical insemination of ewes.

  2. [Morphological semen changes in Chlamydia trachomatis infection].

    PubMed

    Maciejewski, Z; Dziecielski, H; Swierczyński, W; Semmler, G

    1989-06-01

    Semen was examined in 150 men patients of the Andrology Clinic for demonstration of Chlamydia trachomatis and for analysis of the effect of this infection on semen quality depression. A correlation was noted between the degree of infection (large number of organisms per field of vision) and such changes as cryptozoospermia, azoospermia, asthenozoospermia, teratozoospermia, oligoasthenozoospermia, asthenoteratozoospermia. Of interest was a high proportion of infection (56%) with Ch. trachomatis in this group.

  3. Microbial contamination of embryos and semen during long term banking in liquid nitrogen.

    PubMed

    Bielanski, A; Bergeron, H; Lau, P C K; Devenish, J

    2003-04-01

    We report on microbial contamination of embryos and semen cryopreserved in sealed plastic straws and stored for 6-35 years in liquid nitrogen. There were 32 bacterial and 1 fungal species identified from randomly drawn liquid nitrogen, frozen semen, and embryos samples stored in 8 commercial and 8 research facility liquid nitrogen (LN) tanks. The identified bacteria represented commensal or environmental microorganisms and some, such as Escherichia coli, were potential or opportunistic pathogens for humans and animals. Stenotrophomonas maltophilia was the most common contaminant identified from the samples and was further shown to significantly suppress fertilization and embryonic development in vitro. Analysis of the strains by pulsed field gel electrophoresis revealed restriction patterns with no relatedness indicating that there was no apparent cross-contamination of S. maltophilia strains between the germplasm and liquid nitrogen samples. In addition, no transmission of bovine viral diarrhea virus (BVDV) and bovine herpesvirus-1 (BHV-1) from infected semen and embryos straws to clean germplasm stored in the same LN tanks or LN was detected.

  4. Objective assessment of the cryoprotective effects of dimethylformamide for freezing goat semen.

    PubMed

    Bezerra, Francisco Silvestre B; Castelo, Thibério S; Alves, Heron M; Oliveira, Isabelle R S; Lima, Gabriela L; Peixoto, Gislayne C X; Bezerra, Ana Carla S D; Silva, Alexandre R

    2011-12-01

    The aim of this work was to assess the cryoprotective effects of dimethylformamide (DMF) for freezing goat semen, using an objective analysis by computer-assisted sperm analysis (CASA). Twenty-one ejaculates (seven per animal) were collected from three stud bucks with the aid of an artificial vagina and immediately evaluated for gross and microscopic characteristics. The semen was diluted in two steps with a Tris-egg yolk extender containing 6% glycerol or 6% DMF, frozen in 0.50-mL straws, and stored in liquid nitrogen. Samples were accessed for sperm morphology, sperm membrane structural and functional integrity, and by CASA, immediately after thawing. There were differences (P<0.05) between glycerol and DMF with regard to subjective progressive motility (23.9±2.2% vs. 16.6±2.0%), objective progressive motility (3.5±0.4% vs. 1.8±0.3%), linearity (53.9±1.6% vs. 48.1±1.4%) and amplitude of lateral head (2.3±0.1 vs. 2.9±0.1 mm), which confirmed the efficiency of glycerol. In conclusion, dimethylformamide could be used as an alternative cryoprotectant for goat semen freezing. However it was showed that no benefits were derived by using dimethylformamide to replace glycerol at an equal 6% concentration.

  5. Effects of hypotaurine, cysteamine and aminoacids solution on post-thaw microscopic and oxidative stress parameters of Angora goat semen.

    PubMed

    Bucak, Mustafa Numan; Tuncer, Pürhan Barbaros; Sariözkan, Serpil; Ulutaş, Pinar Alkim; Coyan, Kenan; Başpinar, Nuri; Ozkalp, Birol

    2009-12-01

    This study was conducted to determine the effects of cysteamine, hypotaurine and aminoacids solution (BME) on standard semen parameters, lipid peroxidation and antioxidant activities of Angora goat semen after the freeze-thawing process. Ejaculates collected from four Angora goats were evaluated and pooled at 37 degrees C. Semen samples, which were diluted with a Tris-based extender containing the antioxidants hypotaurine (5 mM) and cysteamine (5 mM), and an aminoacid solution (13%), and an extender containing no antioxidants (control), were cooled to 5 degrees C and frozen in 0.25-ml French straws in liquid nitrogen. Frozen straws were thawed individually at 37 degrees C for 20s in a water bath for evaluation. Supplementation with cysteamine, hypotaurine and BME caused significant (P<0.05) increases in sperm motility, and significant (P<0.05) decreases in total abnormality rates in comparison to the control group. While all in vitro treatments did not affect the acrosomal abnormality rates, hypotaurine and BME but not cysteamine significantly (P<0.05) increased the HOST results as compared to the control group. Supplementation with antioxidants and BME did not significantly affect MDA levels and CAT activity in comparison to the control group (P>0.05). The antioxidants hypotaurine and cysteamine decreased SOD activity when compared to the BME group and controls (P<0.001).

  6. Diagnosis of lumbosacral diskospondylosis in a bucking bull assisted by high-definition thermal and nuclear scintigraphic imaging.

    PubMed

    Caldwell, Marc; Passler, Thomas; Purohit, Ram C; Pascoe, David; Wolfe, Dwight F

    2017-03-01

    CASE DESCRIPTION An 8-year-old Brahman-cross bull was evaluated for left hind limb lameness of 2 months' duration. The lameness was first noticed during a rodeo bucking performance, immediately after the bull appeared to land inappropriately on the affected limb. CLINICAL FINDINGS Physical examination findings revealed left hind limb lameness, ataxia, and left-sided epaxial muscle atrophy. Palpation per rectum along the lumbar portion of the vertebral column revealed evidence of exostosis of the ventral aspect. High-definition infrared thermal imaging revealed a pattern of reduced skin temperature in the area of the left lumbar and gluteal regions suggestive of a disruption in the sympathetic control of peripheral blood flow. Nuclear scintigraphy revealed a focal area of increased radioisotope uptake on the left ventrolateral aspect of the L2-3 intervertebral joint. A presumptive diagnosis of ventrolateral vertebral spondylosis resulting in spinal nerve impingement was made. TREATMENT AND OUTCOME 200 mg of methylprednisolone was epidurally injected at the site of the lesion, and treatment with polysulfated glycosaminoglycans was initiated (500 mg, IM, every 4 days for 7 treatments, then monthly thereafter). The lameness and ataxia observed in the left hind limb resolved within 1 week after treatment began. Subsequently, the bull was discharged from the hospital and was used successfully for semen collection and live-cover breeding. CLINICAL RELEVANCE Use of thermography for the bull of this report provided additional insight into neurovascular physiologic function that classical imaging modalities are unable to provide and, when combined with nuclear scintigraphy, aided in identifying the most critical lesion in a complex clinical case.

  7. Fertility-associated metabolites in bull seminal plasma and blood serum: 1H nuclear magnetic resonance analysis.

    PubMed

    Kumar, Ajeet; Kroetsch, Tom; Blondin, Patrick; Anzar, Muhammad

    2015-02-01

    Early estimation of bull fertility is highly desirable for the conservation of male genetics of endangered species and for the exploitation of genetically superior sires in artificial insemination programs. The present work was conducted as a proof-of-principle study to identify fertility-associated metabolites in dairy bull seminal plasma and blood serum using proton nuclear magnetic resonance ((1)H NMR). Semen and blood samples were collected from high- and low-fertility breeding bulls (n = 5 each), stationed at Semex, Guelph, Canada. NMR spectra of serum and seminal plasma were recorded at a resonance frequency of 500.13 MHz on a Bruker Avance-500 spectrometer equipped with an inverse triple resonance probe (TXI, 5 mm). Spectra were phased manually, baseline corrected, and calibrated against 3-(trimethylsilyl) propionic-2,2,3,3-d4 acid at 0.0 parts per million (ppm). Spectra were converted to an appropriate format for analysis using Prometab software running within MATLAB. Principal component analysis was used to examine intrinsic variation in the NMR data set, and to identify trends and to exclude outliers. Partial least square-discriminant analysis was performed to identify the significant features between fertility groups. The fertility-associated metabolites with variable importance in projections (VIP) scores >2 were citrate (2.50 ppm), tryptamine/taurine (3.34-3.38 ppm), isoleucine (0.74 ppm), and leucine (0.78 ppm) in the seminal plasma; and isoleucine (1.14 ppm), asparagine (2.90-2.94 ppm), glycogen (3.98 ppm), and citrulline (1.54 ppm) in the serum. These metabolites showed identifiable peaks, and thus can be used as biomarkers of fertility in breeding bulls.

  8. Performance, growth, and maturity of Nellore bulls.

    PubMed

    Costa e Silva, Luiz Fernando; Valadares Filho, Sebastião de Campos; Detmann, Edenio; Rotta, Polyana Pizzi; Zanetti, Diego; Villadiego, Faider Alberto Castaño; Pellizzoni, Samantha Gusmão; Pereira, Rafael Moura Guimarães

    2013-03-01

    The objectives of this study were to evaluate the dry matter intake (DMI), digestibility, average daily gain (ADG), microbial efficiency, empty body weight (EBW) gain, and body composition of Nellore bulls. Additionally, Nellore bull maturity was estimated, and the prediction equation for DMI, suggested by the Brazilian nutrient requirements system (BR CORTE; Azevêdo et al. 2010), was evaluated. Thirty-three Nellore bulls, with a mean initial weight of 259 ± 25 kg and age of 14 ± 1 months, were used in this study. Five animals were slaughtered at the beginning of the experiment (control group), and the remaining 28 were divided into 4 groups, each slaughtered at 42-day intervals. Their diet was composed of corn silage and concentrate (55:45). The power model was used to estimate muscle tissue, bone tissue, crude protein (CP), mineral matter (MM), and water present in the empty body, while the exponential model was used to estimate adipose tissue and ether extract (EE) present in the empty body. When expressed in kilograms per day, differences were observed (P < 0.05) only for the intake of EE and neutral detergent fiber as a function of feedlot time periods. Although there was a difference in relation to nutrient intake, it did not affect (P > 0.05) digestibility, with the exception of EE digestibility. The equation suggested by BR CORTE correctly estimates the DMI of Nellore bulls. ADG was not affected (P > 0.05) by time spent in the feedlot. No differences were observed (P > 0.05) for microbial efficiency; a mean value of 142 g microbial crude protein/kg total digestible nutrients was achieved. The muscle and bone tissues, CP, MM, and water present in the empty body increased as the animal grew, although at a lower rate. The adipose tissue and EE present in the empty body increased their deposition rate when the animal reached its mature weight. Maturity is defined as when an animal reaches 22 % EE in the empty body, which

  9. Effect of dilution in sperm maturation media and time of storage on sperm motility and fertilizing capacity of cryopreserved semen of sex-reversed female rainbow trout.

    PubMed

    Judycka, Sylwia; Ciereszko, Andrzej; Dobosz, Stefan; Zalewski, Tomasz; Dietrich, Grzegorz J

    2017-05-01

    Masculinized females, also called neomales or sex-reversed females have a male phenotype but retain the female genotype (XX). Therefore, all spermatozoa produced in their functional testes carry an X chromosome, which is desired for the production of all-female rainbow trout populations. Semen of sex-reversed female rainbow trout is of low quality and in vitro maturation is required, which includes dilution of sperm suspensions with specially formulated maturation solutions. The aim of this study was to determine the effect of dilution in different maturation media on sperm quality (sperm motility characteristics and fertilizing capacity) of frozen/thawed sperm of sex-reversed female rainbow trout. The effect of time of post-thaw storage (0, 15, 60 and 120min) on semen quality was also tested. Sperm motility parameters and fertilization rate at the eyed and hatching stages were assessed for post-thaw semen diluted in different media. The cryopreservation procedure resulted in high post-thaw sperm motility of about 57% and did not differ from fresh semen. Unexpectedly, maturation media decreased sperm activation capacity immediately after dilution; however, sperm motility increased over time. Fertilization rates of frozen/thawed semen were high (71-87%) and did not differ significantly between experimental variants at any of tested periods of storage. Our results demonstrated that the effect of the maturation media on frozen/thawed sperm is different from that of fresh sperm. The progressive increase in post-thaw sperm motility in maturation media can potentially be applied to routine hatchery practice.

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

    PubMed

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

    2014-03-01

    Cryodamage is a major problem in semen cryopreservation, causing changes in the levels of proteins that influence the function and motility of spermatozoa. In this study, protein samples prepared from fresh and frozen-thawed boar spermatozoa were compared using the isobaric tags for relative and absolute quantification (iTRAQ) labeling technique coupled to 2D LC-MS/MS analysis. A total of 41 differentially expressed proteins were identified and quantified, including 35 proteins that were present at higher levels and six proteins that were present at lower levels in frozen-thawed spermatozoa by at least a mean of 1.79-fold (P<0.05). On classifying into ten distinct categories using bioinformatic analysis, most of the 41 differentially expressed proteins were found to be closely relevant to sperm premature capacitation, adhesions, energy supply, and sperm-oocyte binding and fusion. The expression of four of these proteins, SOD1, TPI1, ODF2, and AKAP3, was verified by western blot analysis. We propose that alterations in these identified proteins affect the quality of cryopreserved semen and ultimately lower its fertilizing capacity. This is the first study to compare protein levels in fresh and frozen-thawed spermatozoa using the iTRAQ technology. Our preliminary results provide an overview of the molecular mechanisms of cryodamage in frozen-thawed spermatozoa and theoretical guidance to improve the cryopreservation of boar semen.

  11. Verminous myelitis in a pit bull puppy.

    PubMed

    Snook, Eric R; Baker, David G; Bauer, Rudy W

    2009-05-01

    A 10-week-old, male pit bull dog presented to the referring veterinarian with hind limb paresis and epaxial muscle atrophy. No spinal lesions were identified at gross necropsy; however, histologically there was marked granulomatous myelitis in the spinal cord between T13 and L2 with occasional, intralesional nematode larvae. Based on morphologic characteristics, the nematode larvae were identified as Strongyloides spp., possibly Strongyloides stercoralis.

  12. Mauling by pit bull terriers: case report.

    PubMed

    Baack, B R; Kucan, J O; Demarest, G; Smoot, E C

    1989-04-01

    A child with extensive soft-tissue defects following an attack by four pit bull terriers is presented. Some future procedures are required and she will have a permanent gait disability. The multidisciplinary management of this patient is described. The escalating problem of dog attacks in the United States is discussed. It is hoped that increased physician and public awareness will expedite the enactment and enforcement of effective vicious-dog legislation.

  13. Bull heading to kill live gas wells

    SciTech Connect

    Oudeman, P.; Avest, D. ter; Grodal, E.O.; Asheim, H.A.; Meissner, R.J.H.

    1994-12-31

    To kill a live closed-in gas well by bull heading down the tubing, the selected pump rate should be high enough to ensure efficient displacement of the gas into the formation (i.e., to avoid the kill fluid bypassing the gas). On the other hand, the pressures that develop during bull heading at high rate must not exceed wellhead pressure rating, tubing or casing burst pressures or the formation breakdown gradient, since this will lead, at best, to a very inefficient kill job. Given these constraints, the optimum kill rate, requited hydraulic horsepower, density and type of kill fluids have to be selected. For this purpose a numerical simulator has been developed, which predicts the sequence of events during bull heading. Pressures and flow rates in the well during the kill job are calculated, taking to account slip between the gas and kill fluid, hydrostatic and friction pressure drop, wellbore gas compression and leak-off to the formation. Comparison with the results of a dedicated field test demonstrates that these parameters can be estimated accurately. Example calculations will be presented to show how the simulator can be used to identify an optimum kill scenario.

  14. Novel and traditional traits of frozen-thawed porcine sperm related to in vitro fertilization success.

    PubMed

    Daigneault, Bradford W; McNamara, Kelli A; Purdy, Phillip H; Krisher, Rebecca L; Knox, Robert V; Miller, David J

    2014-07-15

    Cryopreserved semen allows the use of single ejaculates for repeated analyses, potentially improving IVF consistency by eliminating interejaculate variability observed with fresh semen. However, the freezing and thawing processes result in compromised sperm function and IVF success. Semen samples are often screened for motility before use for IVF. Samples that are below a designated motility threshold may be discarded. Our objectives were to determine if post-thaw sperm motility, other traits that may be indicative of sperm function, or a novel assay of oviduct binding were related to IVF success. Semen from 16 boars was cooled to 15 °C for overnight shipment before cryopreservation. Semen was thawed and motility was recorded microscopically and confirmed using computer-automated sperm assessment. Each sample was tested by IVF in two to three independent replicates. Regression and correlation analyses were employed to determine the interrelationships between sperm traits and the relationships between post-thaw motility, sperm-oviduct binding and IVF outcomes. Among the sperm traits examined, sperm acrosome integrity was negatively correlated with post-thaw motility (r(2) = 0.64) but not with IVF results. The number of sperm bound to oviduct aggregates was correlated with IVF polyspermy rates (r(2) = 0.62, P < 0.05) but less with overall IVF rates (r(2) = 0.31, P > 0.10). There was some relationship of post-thaw motility with IVF monospermic fertilization (P = 0.06, r(2) = 0.08) but not to other IVF outcomes. Our results indicate that post-thaw motility of frozen-thawed boar sperm is strongly related to acrosome integrity but has limited use for predicting IVF success. The number of sperm bound to oviduct cells was related to IVF polyspermy rates and may be more indicative of in vitro sperm function than traditional sperm motility and acrosome status evaluation.

  15. Semen quality in Peruvian pesticide applicators: association between urinary organophosphate metabolites and semen parameters

    PubMed Central

    Yucra, Sandra; Gasco, Manuel; Rubio, Julio; Gonzales, Gustavo F

    2008-01-01

    Background Organophosphates are broad class of chemicals widely used as pesticides throughout the world. We performed a cross-sectional study of associations between dialkylphosphate metabolites of organophosphates and semen quality among pesticide applicators in Majes (Arequipa), Peru. Methods Thirty-one men exposed to organophosphate (OP) pesticides and 31 non-exposed were recruited (age, 20–60 years). In exposed subjects, semen and a blood sample were obtained one day after the last pesticide application. Subjects were grouped according to levels of OP metabolites in urine. Semen samples were analyzed for sperm concentration, percentage of sperm motility, percentage of normal morphology, semen leucocytes and concentrations of fructose and zinc. Exposure to OP was assessed by measuring six urinary OP metabolites (dimethyl and diethyl phosphates and thiophosphates) by gas chromatography using a single flame photometric detector. Results Diethyldithiophosphate (p = 0.04) and diethylthiophosphate (p = 0.02) better reflected occupational pesticide exposure than other OP metabolites. Semen analysis revealed a significant reduction of semen volume and an increase in semen pH in men with OP metabolites. Multiple regression analysis showed that both occupational exposure to pesticides and the time of exposure to pesticides were more closely related to alterations in semen quality parameters than the single measurement of OP metabolites in urine. Conclusion The study demonstrated that occupational exposure to OP pesticides was more closely related to alterations in semen quality than a single measurement of urine OP metabolites. Current measurement of OP metabolites in urine may not reflect the full risk. PMID:19014632

  16. Growth and pubertal development of F1 bulls from Hereford, Angus, Norwegian Red, Swedish Red and White, Friesian, and Wagyu sires.

    PubMed

    Casas, E; Lunstra, D D; Cundiff, L V; Ford, J J

    2007-11-01

    The objective of the study was to characterize body growth, testicular development, and puberty from 8 to 14 mo of age in bulls (n = 120) produced by mating sires from Hereford, Angus, Norwegian Red, Swedish Red and White, Friesian, and Wagyu breeds to MARC III ((1/4) Hereford, (1/4) Angus, (1/4) Red Poll, and (1/4) Pinzgauer) cows. Traits evaluated were birth weight, weaning weight (at 215 d), yearling weight, ADG from 8 to 14 mo of age, paired testicular volume growth from 8 to 14 mo of age, age at puberty (determined by production of 50 x 10(6) sperm with 10% motility), age at freezable semen (determined by production of 500 x 10(6) sperm with 50% motility), and, at 15 mo of age, paired testicular weight and daily sperm production per testis pair. There was an effect of sire breed (P = 0.03) for age at puberty; animals with Wagyu and Swedish Red and White inheritance reached puberty at a later date (302 and 302 d of age, respectively) compared with Angus-sired bulls (268 d). Age at puberty for Hereford-, Norwegian Red-, and Friesian-sired bulls was 270, 271, and 278 d, respectively. Differences in BW were observed (P = 0.03) at birth; bulls with Hereford and Friesian were heavier at birth (43 and 41 kg, respectively) compared with those with Norwegian Red, Swedish Red and White, and Wagyu inheritance (39, 38, and 38 kg, respectively). Differences in BW were also observed at 1 yr of age (P = 0.001), where the heaviest animals were those sired by Angus (450 kg), whereas the lightest animals were those sired by Wagyu (403 kg). Bulls with Wagyu inheritance had the lowest (P = 0.04) ADG (1.12 kg/d) compared with bulls with inheritance from Hereford (1.22 kg/d), Angus (1.28 kg/d), Norwegian Red (1.24 kg/d), Swedish Red and White (1.25 kg/d), and Friesian (1.27 kg/d). Differences in scrotal growth rate were not significant (P = 0.99). They ranged from 1.95 in Angus-sired to 1.66 cm3/d in Wagyu-sired bulls. There were no differences (P = 0.80) for age at freezable semen

  17. Urethral anatomy and semen flow during ejaculation

    NASA Astrophysics Data System (ADS)

    Kelly, Diane

    2016-11-01

    Ejaculation is critical for reproductive success in many animals, but little is known about its hydrodynamics. In mammals, ejaculation pushes semen along the length of the penis through the urethra. Although the urethra also carries urine during micturition, the flow dynamics of micturition and ejaculation differ: semen is more viscous than urine, and the pressure that drives its flow is derived primarily from the rhythmic contractions of muscles at the base of the penis, which produce pulsatile rather than steady flow. In contrast, Johnston et al. (2014) describe a steady flow of semen through the crocodilian urethral groove during ejaculation. Anatomical differences of tissues associated with mammalian and crocodilian urethral structures may underlie these differences in flow behavior.

  18. Ejaculatory process and related semen characteristics.

    PubMed

    Bravo, P W; Moscoso, R; Alarcon, V; Ordoñez, C

    2002-01-01

    South American camelids are dribble ejaculators, and urethral contractions occur throughout copulation, which may last 25 min. The urethral contractions and their association with semen characteristics during copulation were determined in llamas and alpacas. A transrectal probe was held in the rectum of the male while copulating an artificial vagina, which was accessed underneath the dummy through a hole. The semen-collecting tube was changed every 5 min. Semen characteristics, color, volume, consistency, motility, concentration, and percentage of live sperm were determined at 5-min intervals. Urethral contractions were evenly distributed during copulation: 40 in alpacas and 63 in llamas (p < .05), with a general range of 11 to 132. Semen color was milky in 63%, and translucent in 36.5% for alpacas; and creamy (9.9%), milky (47%), and translucent (42%) for llamas. The mean volume of ejaculate was 0.3, 0.4, 0.6, 0.7, 0.6, 0.8, 0.3, and 3.0 mL for 5, 10, 15, 20, 25, and 30 min, respectively. Semen consistency was variable: viscous (65%) and semiviscous (34%) in alpacas; and viscous (57%) and semiviscous (42%) in llamas. Spermatic motility varied between 60 and 80% for the llama, and 40 and 80% for the alpaca. Spermatic concentration varied between 60 and 188 x 10(3)/mm3 in llamas, and 30 and 170 x 10(3)/mm3 in alpacas. Percentage of live sperm varied the least: 81 to 90% in llamas and 65 to 90% in alpacas. The ejaculate of llamas and alpacas is not fractionated, urethral contractions are evenly distributed, during copulation, and semen characteristics are present throughout the copulatory period.

  19. Developmental competence of Dromedary camel oocytes fertilized in vitro by frozen-thawed ejaculated and epididymal spermatozoa

    PubMed Central

    Scholkamy, T. H.; El-Badry, D. A.; Mahmoud, K. Gh. M.

    2016-01-01

    The present study aimed to compare the in vitro fertilizing capacity of frozen-thawed ejaculated and epididymal spermatozoa in order to standardize the semen preparation protocol for camel in vitro fertilization (IVF). Semen samples were collected from 7 Dromedary camels by means of artificial vagina (AV). Ten cauda epididymes were obtained from slaughtered adult camels, isolated, incised and rinsed for obtaining the sperm rich fluid. Ejaculated and epididymal spermatozoa were processed for cryopreservation. Fresh and frozen-thawed ejaculated and epididymal spermatozoa were evaluated for motility, livability, membrane and acrosomal integrities. Frozen-thawed ejaculated and epididymal spermatozoa were used to fertilize camel mature oocytes in vitro. The results showed that, the progressive motility of freshly collected epididymal spermatozoa was significantly (P<0.05) higher than ejaculated spermatozoa (49.25 ± 1.75 vs. 38.50 ± 1.50%, respectively). The viability index of epididymal spermatozoa was significantly (P<0.05) higher than that of ejaculated spermatozoa (96.63 ± 2.45 vs. 84.00 ± 4.08, respectively). The post-thaw acrosome and membrane integrities of epididymal spermatozoa were significantly (P<0.05) higher than those of ejaculated spermatozoa. Morula and blastocyst rates of camel oocytes in vitro fertilized by frozen-thawed epididymal spermatozoa (59.4 ± 0.8, 19.12 ± 0.7 and 10.29 ± 0.7%, respectively) were significantly (P<0.05) higher than those fertilized by frozen-thawed ejaculated spermatozoa (48.27 ± 3.1, 11.63 ± 1.1 and 5.43 ± 0.8%, respectively). In conclusion, the Dromedary camel frozen epididymal spermatozoa have the capacity to endure cryopreservation, fertilize oocytes and produce embryos in vitro better than ejaculated sperm. PMID:28224009

  20. Strategies for Processing Semen from Subfertile Stallions for Cooled Transport.

    PubMed

    Varner, Dickson D

    2016-12-01

    Subfertility can be a confusing term because some semen of good quality can have reduced fertility following cooled transport if the semen is processed in an improper manner. General procedures aimed at processing stallion semen for cooled transport are well described. An array of factors could exist in reduced fertility of cool-transported semen. This article focuses on centrifugation techniques that can be used to maximize sperm quality of stallions whose semen is intended for cooled transport. Clinical cases are also provided for practical application of techniques.

  1. Weekly bull exchange shortens postpartum anestrus in suckled beef cows.

    PubMed

    Miller, V; Ungerfeld, R

    2008-05-01

    The duration of anestrus in cattle was usually shortened when cows were exposed to bulls. The objective of the present experiment was to determine if weekly bull exchange accelerated the resumption of cyclicity in postpartum suckled beef cows. We tested the hypothesis that exposure of postpartum, anestrous, suckled beef cows (extensively managed) to weekly exchange of bulls, accelerates the resumption of cyclicity, compared to continuous exposure to the same bulls. Ninety-one multiparous suckled Hereford and Hereford x Angus cows, <60d postpartum, were assigned to two homogeneous groups. Beginning on December 1st (late spring), the control group (C, n=45) remained with one pair of bulls throughout the breeding period (7 weeks), whereas in the "exchanged" (E, n=46) group two pairs of bulls were exchanged weekly. Based on weekly ultrasonographic examinations of all cows, none had a CL at the start of the experiment and for 2 weeks after the start of bull exposure. However, the accumulated frequency of cows with a CL was greater in group E than in group C cows on week 4 (P=0.024), as well as on weeks 5-7 (P<0.001) after the start of bull exposure. Furthermore, in group E versus group C, there was a higher pregnancy rate 30d after the end of bull exposure (26 of 46, 56.2% vs. 16 of 45, 35.6%; P=0.045). In conclusion, weekly exchange of two pairs of bulls shortened postpartum anestrus in suckled multiparous cows, compared to continuous exposure to a single pair of bulls.

  2. A field study on artificial insemination of swamp and crossbred buffaloes with sexed semen from river buffaloes.

    PubMed

    Lu, Yangqing; Liao, Yanqiong; Zhang, Ming; Yang, Bingzhuang; Liang, Xianwei; Yang, Xiaogan; Lu, Shengsheng; Wu, Zhuyue; Xu, Huiyan; Liang, Yunbin; Lu, Kehuan

    2015-10-01

    Sex preselection by flow sorting of X- and Y-sperm has been proven to be an efficient and economically feasible strategy for use in Holstein dairy cow breeding, and previous reports have demonstrated the feasibility of altering the sex ratio in buffalo species by using sexed semen in either artificial insemination or IVF. However, because buffalo reproductive physiology and farm management are different from Holsteins, factors involved in artificial insemination by sexed semen need to be further addressed before being applied in buffalo breeding at village-level husbandry. In this study, a total of 4521 swamp or crossbred (F1 or F2) buffaloes with natural estrus were inseminated with X-sorted sperm from river buffaloes, resulting in a 48.5% (2194 of 4521) pregnancy rate and 87.6% (1895 of 2163) sex accuracy in the derived calves. The pregnancy rate obtained with sexed semen from Murrah bulls was higher than that of Nili-Ravi, 52.5% (895 of 1706) versus 46.1% (1299 of 2815; P < 0.01), respectively. Also, significant variations were seen in pregnancy rates from inseminations performed in different seasons (P < 0.01) and by different technicians (P < 0.01). In contrast to Holsteins, no difference was seen in the pregnancy rate between heifers and parous buffalo cows, and buffalo cows with different genetic backgrounds (swamp type, crossbred F1 and F2) showed similar fertility after insemination with sexed semen. The findings in the present study under field conditions pave the way for application of sexing technology to buffalo breeding under village-level husbandry and diverse genetic backgrounds.

  3. Scrotal infrared digital thermography as a predictor of seasonal effects on sperm traits in Braford bulls.

    PubMed

    Menegassi, Silvio Renato Oliveira; Barcellos, Júlio Otavio Jardim; Dias, Eduardo Antunes; Koetz, Celso; Pereira, Gabriel Ribas; Peripolli, Vanessa; McManus, Concepta; Canozzi, Maria Eugênia Andrighetto; Lopes, Flávio Guiselli

    2015-03-01

    The aim of this study was to assess the seasonal effects of the environment on semen quality in bulls, using infrared thermography. Sperm motility (M), mass motion (MM), and vigor (VIG) were evaluated in sperm samples from 17 Bradford bulls aged approximately 24 months at the beginning of the study. Infrared thermography images and data were collected using an infrared FLIR T 300 camera and Quick Report 1.2 SP2 software to determine the temperature of the proximal and distal poles of the testis and to assess the testicular temperature gradient. The seasonal effects on physiological, seminal, and climatic variables were analyzed by the GLM ANOVA and CORR procedures using SAS®. The microclimatic factors were recorded in hourly intervals, and the daily mean temperature and mean relative humidity were calculated to determine the daily temperature-humidity index (THI) every day for 1 year. The temperature gradient (TG) variations of the testes were significantly higher in the autumn (4.5 °C), winter (4.0 °C), and spring (2.9 °C) compared to summer (0.9 °C) (P < 0.05). Ocular globe temperatures were lower in the winter (27.6 °C) and autumn (26.8 °C) compared to summer (33.9 °C) and spring (31.1 °C) (P < 0.05). The average MM (2.58), M (52.64), and VIG (2.70) of the semen decreased in the summer compared to other seasons (P < 0.01). The TG was negatively correlated with THI (-0.44; P < 0.05). For the seminal variables, MaD (-0.45; P < 0.05) and TD (-0.50; P < 0.01) presented a negative correlation with TG. The TG had a positive correlation between M and VIG, which had values of 0.36 and 0.35, respectively (P < 0.05). We have concluded that infrared thermography can be used to assess the testicular temperature gradient and its consequences on physical and quantitative aspects of sperm.

  4. Scrotal infrared digital thermography as a predictor of seasonal effects on sperm traits in Braford bulls

    NASA Astrophysics Data System (ADS)

    Menegassi, Silvio Renato Oliveira; Barcellos, Júlio Otavio Jardim; Dias, Eduardo Antunes; Koetz, Celso; Pereira, Gabriel Ribas; Peripolli, Vanessa; McManus, Concepta; Canozzi, Maria Eugênia Andrighetto; Lopes, Flávio Guiselli

    2015-03-01

    The aim of this study was to assess the seasonal effects of the environment on semen quality in bulls, using infrared thermography. Sperm motility (M), mass motion (MM), and vigor (VIG) were evaluated in sperm samples from 17 Bradford bulls aged approximately 24 months at the beginning of the study. Infrared thermography images and data were collected using an infrared FLIR T 300 camera and Quick Report 1.2 SP2 software to determine the temperature of the proximal and distal poles of the testis and to assess the testicular temperature gradient. The seasonal effects on physiological, seminal, and climatic variables were analyzed by the GLM ANOVA and CORR procedures using SAS®. The microclimatic factors were recorded in hourly intervals, and the daily mean temperature and mean relative humidity were calculated to determine the daily temperature-humidity index (THI) every day for 1 year. The temperature gradient (TG) variations of the testes were significantly higher in the autumn (4.5 °C), winter (4.0 °C), and spring (2.9 °C) compared to summer (0.9 °C) ( P < 0.05). Ocular globe temperatures were lower in the winter (27.6 °C) and autumn (26.8 °C) compared to summer (33.9 °C) and spring (31.1 °C) ( P < 0.05). The average MM (2.58), M (52.64), and VIG (2.70) of the semen decreased in the summer compared to other seasons ( P < 0.01). The TG was negatively correlated with THI (-0.44; P < 0.05). For the seminal variables, MaD (-0.45; P < 0.05) and TD (-0.50; P < 0.01) presented a negative correlation with TG. The TG had a positive correlation between M and VIG, which had values of 0.36 and 0.35, respectively ( P < 0.05). We have concluded that infrared thermography can be used to assess the testicular temperature gradient and its consequences on physical and quantitative aspects of sperm.

  5. Effects of storage temperature and semen extender on stored canine semen.

    PubMed

    Hori, Tatsuya; Yoshikuni, Ryuta; Kobayashi, Masanori; Kawakami, Eiichi

    2014-03-01

    The objective of the present study was to determine an optimum temperature and extender for short-term transport of canine ejaculated semen. There was no significant difference in the qualities of semen diluted with two kinds of extender, egg yolk Tris-citrate fructose (EYT-FC) or glucose (EYT-GC) extender, between the 2, 8 or 12 and the 4°C control groups during storage for up to 48 hr, while the 16-24°C groups showed decreased sperm motility during storage for 48 hr. However, the 2°C group showed slightly lower sperm motility and slightly higher sperm abnormality than the 4°C group. Therefore, we concluded that semen qualities can be maintained for up to 48 hr when canine semen samples are extended with EYT-FC or EYT-GC and stored at a temperature in the range of 4-12°C.

  6. Protocol for determining bull trout presence

    USGS Publications Warehouse

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

    2002-01-01

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

  7. The uses of infrared thermography to evaluate the effects of climatic variables in bull's reproduction

    NASA Astrophysics Data System (ADS)

    Menegassi, Silvio Renato Oliveira; Pereira, Gabriel Ribas; Dias, Eduardo Antunes; Koetz, Celso; Lopes, Flávio Guiselli; Bremm, Carolina; Pimentel, Concepta; Lopes, Rubia Branco; da Rocha, Marcela Kuczynski; Carvalho, Helena Robattini; Barcellos, Júlio Otavio Jardim

    2016-01-01

    The objective of this study was to evaluate the seasonal effects of the environment on sperm quality in subtropical region determined by temperature and humidity index (THI). We used 20 Brangus bulls (5/8 Angus × 3/8 Nellore) aged approximately 24 months at the beginning of the study. Semen evaluations were performed twice per season during 1 year. Climate THI data were collected from an automatic weather station from the National Institute of Meteorology. Infrared thermography images were used to determine the temperature of the proximal and distal poles of the testis to assess the testicular temperature gradient (TG). The seasonal effects on seminal and climatic variables were analyzed with ANOVA using MIXED procedure of SAS. Sperm motility in spring (60.1 %), summer (57.6 %), and autumn (64.5 %) showed difference compared to winter (73.0 %; P < 0.01). TG was negatively correlated with THI at 18 days (spermiogenesis) (-0.76; P < 0.05) and at 12 days (epididymal transit) (-0.85; P < 0.01). Ocular temperature (OcT) had a positive correlation with THI at 18 days (0.78; P < 0.05) and at 12 days (0.84; P < 0.01). Motility showed a negative correlation with THI only at 18 days (-0.79; P < 0.05). During spermiogenesis, the TG had higher negative correlation compared to OcT (-0.97; P < 0.01) and rectal temperature (-0.72; P < 0.05). Spermatozoa with distal midpiece reflex were correlated with THI during transit epididymis (0.72; P < 0.05). Seminal parameters are not affected when THI reaches 93.0 (spermiogenesis) and 88.0 (epididymal transit). We concluded that infrared thermography can be adopted as an indirect method in order to assess the effect of environmental changes in TG and OcT of Brangus bulls.

  8. Thallium toxicosis in a Pit Bull Terrier.

    PubMed

    Volmer, Petra A; Merola, Valentina; Osborne, Tanasa; Bailey, Keith L; Meerdink, Gavin

    2006-01-01

    Thallotoxicosis is described in an adult Pit Bull Terrier. The dog exhibited anorexia, emesis, weakness, conscious proprioceptive deficits, and a hemorrhagic diarrhea before death. A severe, acute necrotizing enterocolitis was evident upon histological examination, as was a multifocal to coalescing pulmonary edema. Liver and kidney thallium concentrations were 18 and 26 ppm, respectively. The source of the thallium was determined to be thallium sulfate obtained by a person with the intent to harm family members. Although thallium has not been produced in the United States for 20 years, this report demonstrates the need to consider thallium toxicosis as a differential diagnosis for animals presenting with vague and mixed gastrointestinal and neurological signs.

  9. Microheterogeneity in Frozen Protein Solutions

    PubMed Central

    Twomey, Alan; Kurata, Kosaku; Nagare, Yutaka; Takamatsu, Hiroshi; Aksan, Alptekin

    2015-01-01

    In frozen and lyophilized systems, the biological to be stabilized (e.g. therapeutic protein, biomarker, drug-delivery vesicle) and the cryo-/lyoprotectant should be co-localized for successful stabilization. During freezing and drying, many factors cause physical separation of the biological from the cryo-/lyoprotectant, called microheterogeneity (MH), which may result in poor stabilization efficiency. We have developed a novel technique that utilized confocal Raman microspectroscopy in combination with counter-gradient freezing to evaluate the effect of a wide range of freezing temperatures (−20 < TF < 0°C) on the MH generated within a frozen formulation in only a few experiments. The freezing experiments conducted with a model system (albumin and trehalose) showed the presence of different degrees of MH in the freeze-concentrated liquid (FCL) in all solutions tested. Mainly, albumin tended to accumulate near the ice interface, where it was physically separated from the cryoprotectant. In frozen 10 wt% trehalose solutions, heterogeneity in FCL was relatively low at any TF. In frozen 20 wt% trehalose solutions, the optimum albumin to trehalose ratio in the FCL can only be ensured if the solution was frozen within a narrow range of temperatures (−16 < TF < −10°C). In the 30 wt% trehalose solutions, freezing within a much more narrow range (−12 < TF < −10°C) was needed to ensure a fairly homogeneous FCL. The method developed here will be helpful for the development of uniformly frozen and stable formulations and freezing protocols for biological as MH is presumed to directly impact stability. PMID:25888798

  10. Microheterogeneity in frozen protein solutions.

    PubMed

    Twomey, Alan; Kurata, Kosaku; Nagare, Yutaka; Takamatsu, Hiroshi; Aksan, Alptekin

    2015-06-20

    In frozen and lyophilized systems, the biological to be stabilized (e.g. therapeutic protein, biomarker, drug-delivery vesicle) and the cryo-/lyo-protectant should be co-localized for successful stabilization. During freezing and drying, many factors cause physical separation of the biological from the cryo-/lyo-protectant, called microheterogeneity (MH), which may result in poor stabilization efficiency. We have developed a novel technique that utilized confocal Raman microspectroscopy in combination with counter-gradient freezing to evaluate the effect of a wide range of freezing temperatures (-20frozen formulation in only a few experiments. The freezing experiments conducted with a model system (albumin and trehalose) showed the presence of different degrees of MH in the freeze-concentrated liquid (FCL) in all solutions tested. Mainly, albumin tended to accumulate near the ice interface, where it was physically separated from the cryoprotectant. In frozen 10wt% trehalose solutions, heterogeneity in FCL was relatively low at any TF. In frozen 20wt% trehalose solutions, the optimum albumin to trehalose ratio in the FCL can only be ensured if the solution was frozen within a narrow range of temperatures (-16frozen and stable formulations and freezing protocols for biological as MH is presumed to directly impact stability.

  11. Effects of seasons on some semen parameters and bacterial contamination of Awassi ram semen.

    PubMed

    Azawi, O I; Ismaeel, M A

    2012-06-01

    The objectives of the present study were to determine the effects of season on some semen parameters and bacterial contamination of Awassi ram semen. Semen samples from six mature Awassi rams were used in this study. Semen collection was performed with artificial vagina every week, from September 2009 to October 2010. Volume, sperm concentration, mass motility, individual motility, percentage live sperm and sperm abnormalities were evaluated. Moreover, determination of viable bacterial count of the rams was also recorded weekly. Higher (p < 0.05) semen volume in the hot summer and spring months was observed of August (1.55 ± 0.08 ml) and March (1.27 ± 0.15 ml). Sperm concentration was highest (p < 0.05) in the breeding season (late summer to early autumn) of September (4.21 ± 0.86 × 10(9) sperm/ml). Sperm individual motility and percent of live sperm observed in August (summer) and May (end of spring) when the environmental temperature started to increase were recorded highest values and differed significantly (p < 0.05) from December and January (winter). The highest value of the mean sperm acrosomal defects (13.33 ± 0.63%) was recorded in December. The highest value of the mean viable bacterial count (138.3 ± 21.6) was recorded in July (summer). A significant decrease (p < 0.01) in the mean viable bacterial count was observed from the middle of winter towards the end of spring. The lowest bacterial count was noted in January (60.5 ± 2.98). It could be concluded from the results of the present study that there is an effect of season on ram semen quality, and summer high temperature in northern Iraq has no effect on Awassi ram semen. There is a significant effect of season on bacterial count on Awassi ram semen.

  12. [Rapid frozen sections in neuropathology].

    PubMed

    Haybaeck, J; von Campe, G; Hainfellner, J A

    2012-09-01

    Neuropathological evaluation of frozen sections requires a) special expertise in neuropathological specimen assessment and neurooncology as well as b) a trustful and open communication culture with the neurosurgeons. In addition to frozen sections, cytological examinations of smear and touch preparations as supporting methods are available to reach a correct diagnosis: these additional methods should therefore be performed whenever possible. Besides evaluation of biopsy specimens, appraisal of resection specimens and resection margin controls are of high clinical relevance. In the case of diffusely infiltrating central nervous system (CNS) neoplasms, in particular gliomas, resection margin control is often not feasible in contrast to other types of solid tumor.

  13. MALLEABLE IRON BULL LADLE, HOLDS IRON AFTER IT IS TAPPED ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    MALLEABLE IRON BULL LADLE, HOLDS IRON AFTER IT IS TAPPED OUT OF THE CUPOLA UNTIL IT NEEDED BY POURERS ON THE CONVEYOR LINES WHO FILL MOBILE LADLES ATTACHED TO OVERHEAD RAIL SYSTEMS AS THE BULL LADLE TIPS. - Stockham Pipe & Fittings Company, Malleable Foundry, 4000 Tenth Avenue North, Birmingham, Jefferson County, AL

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  15. Sire conception rate: New national AI bull fertility evaluation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bull fertility evaluations called Estimated Relative Conception Rate (ERCR) were provided to the industry by Dairy Records Management Systems (DRMS) from 1986 to November 2005. In May 2006, AIPL assumed responsibility for evaluation of U.S. bull fertility. As an initial step, AIPL implemented the ER...

  16. Similarities and differences between the sperm quality index and sperm mobility index of broiler breeder semen.

    PubMed

    Dumpala, P R; Parker, H M; McDaniel, C D

    2006-12-01

    The sperm quality index (SQI) and sperm mobility index (SMI) both predict sperm motility. Previous research was conducted comparing the SMI with the SQI; however, semen was diluted improperly for the SQI (40-fold). For an accurate comparison, semen must be diluted 10-fold for the SQI. Therefore, the overall objective of this study, utilizing 4 experiments, was to examine the relationship of the SQI with the SMI when: 1) active, heat-inactivated, and boiled sperm were incubated and analyzed over 60 min, 2) motile and immotile sperm were combined, 3) dead sperm (boiled or frozen) and live sperm were combined, and 4) sperm were rendered immotile by adding Zn. In experiment 1, the SQI was stable throughout incubation for active sperm and zero for heat-inactivated and boiled sperm. The SMI from active sperm steadily increased over incubation. However, the SMI from heat-inactivated sperm increased drastically resulting in values higher than active sperm after 35 min of incubation. The SMI from active sperm was higher than boiled sperm throughout incubation. For experiment 2, a cubic increase in the SQI occurred as active sperm increased from 0 to 100%. However, there was no difference in SMI readings until samples contained 80% active sperm. In experiment 3, for both boiled and frozen sperm, as the percentage of viable sperm increased, there was a logarithmic increase in the SQI. The SMI values were similar for 0 and 100% viable sperm when using boiled sperm, and all SMI readings were similar to 0% viable for frozen sperm. For experiment 4, both the SQI and SMI values from semen incubated with Zn were lower than that of saline. The SQI from sperm incubated in saline was higher than that of Zn throughout incubation, yet there were no differences in SMI values after 90 min of incubation. In conclusion, both the SQI and SMI of sperm decrease in response to Zn. However, it appears that immotile and dead sperm are capable of increasing SMI values but not SQI values.

  17. Correlation of phthalate exposures with semen quality

    SciTech Connect

    Pant, Niraj Shukla, Manju; Kumar Patel, Devendra; Shukla, Yogeshwar; Mathur, Neeraj; Kumar Gupta, Yogendra; Saxena, Daya Krishna

    2008-08-15

    Phthalates are widely used man-made chemical released in the environment and human exposure is mainly through diet. As the phthalate plasticizers are not covalently bound to PVC, they can leach, migrate or evaporate into the environment and as a result have become ubiquitously contaminants. The present study investigates the correlation, if any, between the phthalate esters (DEP, DEHP, DBP, DMP, DOP) and sperm mitochondrial status, ROS, LPO, SCSA, and sperm quality. The study was conducted in the urban/rural population of Lucknow visiting Obstetrics and Gynecology Department, CSMMU, Lucknow. Semen analysis was performed according to the WHO guidelines while phthalate analysis by HPLC and LPO by spectrophotometer and the sperm mitochondrial status, ROS, SCSA using flow cytometry. The questionnaire data showed no significant difference in the demographic characteristics among the groups. In general, urban population was found to have statistically significant higher levels of phthalate esters than the rural. Further, infertile men showed statistically significant (p < 0.05) higher levels of pollutants in the semen than fertile men. A negative correlation between semen phthalate level viz DEHP and sperm quality and positive association with depolarized mitochondria, elevation in ROS production and LPO, DNA fragmentation was established. The findings are suggestive that phthalates might be one among the contributing factors associated with the deterioration in semen quality and these adverse effects might be ROS, LPO and mitochondrial dysfunction mediated.

  18. AIR POLLUTION EFFECTS ON SEMEN QUALITY

    EPA Science Inventory

    The potential impact of exposure to periods of high air pollution on male reproductive health was examined within the framework of an international project conducted in the Czech Republic. Semen quality was evaluated in young men (age 18) living in the Teplice District who are ex...

  19. Evaluation of thermal comfort, physiological, hematological, and seminal features of buffalo bulls in an artificial insemination station in a tropical environment.

    PubMed

    Barros, Daniel Vale; Silva, Lilian Kátia Ximenes; de Brito Lourenço, José; da Silva, Aluizio Otávio Almeida; E Silva, André Guimarães Maciel; Franco, Irving Montanar; Oliveira, Carlos Magno Chaves; Tholon, Patrícia; Martorano, Lucieta Guerreiro; Garcia, Alexandre Rossetto

    2015-06-01

    This study aimed to assess the variation over time in thermal comfort indices and the behavior of physiological parameters related to thermolysis, blood parameters, and semen in natura of buffalo bulls reared in tropical climate. The study was carried out in an artificial insemination station under a humid tropical climate (Afi according to Köppen). Ten water buffalo bulls (Bubalus bubalis) were used during the 5 months (April to August) of study. The environmental Temperature Humidity Index (THId) and the pen microclimate Temperature Humidity Index (THIp) were calculated. Every 25 days, respiratory rate (RR), heart rate (HR), rectal temperature (RT), and Benezra's thermal comfort index (BTCI) were assessed in the morning and in the afternoon. A blood assay was performed every month, while semen was collected weekly. THIp did not vary over the months (P > 0.05) and was higher in the afternoon than in the morning (77.7 ± 2.6 versus 81.8 ± 2.1, P < 0.05). RR, HR, and BTCI significantly increased over the months and were different between the periods of the day (P > 0.05) but within the physiological limits. RT varied between the periods of the day and decreased over the months, being the lowest in August (37.8 ± 0.7 °C), time-impacted hematocrit, mean corpuscular volume, hemoglobin levels, and spermatic gross motility and vigor (P < 0.05). Thus, buffalo bulls reared under a humid tropical climate may have variations in thermal comfort during the hotter periods but are able to efficiently activate thermoregulatory mechanisms and maintain homeothermy, hence preserving their physiological and seminal parameters at normal levels.

  20. Resolving disputes over frozen embryos.

    PubMed

    Robertson, J A

    1989-01-01

    The relation between respect for family and reproductive choice and use of IVF technology is in dispute in recent legal cases on the disposition of frozen embryos. Couples in IVF programs should be encouraged to stipulate in advance binding instructions regarding the disposition of such embryos.

  1. Frozen Scope and Grammatical Optimization

    ERIC Educational Resources Information Center

    Freedman, Michael

    2014-01-01

    The literature on quantifier scope has repeatedly observed that some otherwise expected permutations of scope taking elements are unavailable. Various methods have been proffered explaining these facts. This thesis aims to unify three disparate areas where the scope of operators seems to be frozen: the interaction of universal quantifiers with…

  2. 7 CFR 58.327 - Frozen cream.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Specifications for Dairy Plants Approved for USDA Inspection and Grading Service 1 Quality Specifications for Raw Material § 58.327 Frozen cream. To produce frozen cream eligible for official certification, the quality...

  3. 7 CFR 58.327 - Frozen cream.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Specifications for Dairy Plants Approved for USDA Inspection and Grading Service 1 Quality Specifications for Raw Material § 58.327 Frozen cream. To produce frozen cream eligible for official certification, the quality...

  4. 7 CFR 58.327 - Frozen cream.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Specifications for Dairy Plants Approved for USDA Inspection and Grading Service 1 Quality Specifications for Raw Material § 58.327 Frozen cream. To produce frozen cream eligible for official certification, the quality...

  5. 7 CFR 58.327 - Frozen cream.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Specifications for Dairy Plants Approved for USDA Inspection and Grading Service 1 Quality Specifications for Raw Material § 58.327 Frozen cream. To produce frozen cream eligible for official certification, the quality...

  6. The Eccentric Behavior of Nearly Frozen Orbits

    NASA Technical Reports Server (NTRS)

    Sweetser, Theodore H.; Vincent, Mark A.

    2013-01-01

    Frozen orbits are orbits which have only short-period changes in their mean eccentricity and argument of periapse, so that they basically keep a fixed orientation within their plane of motion. Nearly frozen orbits are those whose eccentricity and argument of periapse have values close to those of a frozen orbit. We call them "nearly" frozen because their eccentricity vector (a vector whose polar coordinates are eccentricity and argument of periapse) will stay within a bounded distance from the frozen orbit eccentricity vector, circulating around it over time. For highly inclined orbits around the Earth, this distance is effectively constant over time. Furthermore, frozen orbit eccentricity values are low enough that these orbits are essentially eccentric (i.e., off center) circles, so that nearly frozen orbits around Earth are bounded above and below by frozen orbits.

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

    PubMed

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

    2012-04-01

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

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

    PubMed

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

    2012-05-01

    The present study aimed to evaluate the ability of spermatozoa from individual boar ejaculates to withstand different semen-processing techniques. Eighteen sperm-rich ejaculate samples from six boars (three per boar) were diluted in Beltsville Thawing Solution and split into three aliquots. The aliquots were (1) further diluted to 3×10(7) sperm/mL and stored as a liquid at 17°C for 72 h, (2) frozen-thawed (FT) at 1×10(9) sperm/mL using standard 0.5-mL straw protocols, or (3) sex-sorted with subsequent liquid storage (at 17°C for 6 h) or FT (2×10(7) sperm/mL using a standard 0.25-mL straw protocol). The sperm quality was evaluated based on total sperm motility (the CASA system), viability (plasma membrane integrity assessed using flow cytometry and the LIVE/DEAD Sperm Viability Kit), lipid peroxidation (assessed via indirect measurement of the generation of malondialdehyde (MDA) using the BIOXYTECH MDA-586 Assay Kit) and DNA fragmentation (sperm chromatin dispersion assessed using the Sperm-Sus-Halomax(®) test). Data were normalized to the values assessed for the fresh (for liquid-stored and FT samples) or the sorted semen samples (for liquid stored and the FT sorted spermatozoa). All of the four sperm-processing techniques affected sperm quality (P<0.01), regardless of the semen donor, with reduced percentages of motile and viable sperm and increased MDA generation and percentages of sperm with fragmented DNA. Significant (P<0.05) inter-boar (effect of boars within each semen-processing technique) and intra-boar (effect of semen-processing techniques within each boar) differences were evident for all of the sperm quality parameters assessed, indicating differences in the ability of spermatozoa from individual boars to withstand the semen-processing techniques. These results are the first evidence that ejaculate spermatozoa from individual boars can respond in a boar-dependent manner to different semen-processing techniques.

  9. Semen collection and fertility in naturally fertile sandhill cranes

    USGS Publications Warehouse

    Chen, G.; Gee, G.F.; Nicolich, Jane M.; Taylor, J.A.; Urbanek, R.P.; Stahlecker, D.W.

    1997-01-01

    Aviculturists often ask if semen collection will interfere with fertility in naturally fertile pairs of cranes. We used 12 naturally fertile Florida sandhill crane (Grus canadensis pratensis) pairs for this study, 6 control and 6 experimental. All pairs had produced fertile eggs in previous years and were in out-of-doors pens scattered throughout different pen complexes, within auditory range but physically isolated. Semen was collected on Tuesday mornings and Friday afternoons from 26 February 1993 to 4 June 1993. We used standard artificial insemination methods to collect and to evaluate the semen and spermatozoa. Semen collection did not affect semen quality or quantity. Semen volume, sperm density, sperm motility, sperm morphology, sperm live, sperm number per collection, and male response to semen collection exhibited significant daily variation (P < 0.05). Although semen collection began 13 days before the first egg in the experimental group, we observed no differences in the date of first egg laid or in fertility between experimental and control groups. Also, we observed no differences in the interval between clutches or in the percentage of broken eggs between experimental and control groups. Sires consistently producing better semen samples produced fewer fertile eggs than sires producing poorer semen samples (r = 0.60).

  10. Associations of adiponectin and fertility estimates in Holstein bulls.

    PubMed

    Kasimanickam, Vanmathy R; Kasimanickam, Ramanathan K; Kastelic, John P; Stevenson, Jeffrey S

    2013-03-15

    Adiponectin is a pleiotropic regulator of numerous biological functions, including gonadal steroidogenesis and might play a role in sperm structures and functions. The objectives were to: (1) determine associations among serum concentrations of adiponectin, testosterone, and prolactin, and the sperm DNA fragmentation index; (2) associate sperm adiponectin mRNA abundance with estimates of fertility (sire conception rate); and (3) determine sperm protein expression of adiponectin and its receptor in pre- and postcapacitated sperm from Holstein bulls. In experiment 1, biweekly serum concentrations of adiponectin, prolactin, and testosterone were greater (P < 0.05) for high fertility bulls compared with average and low fertility bulls. Furthermore, sperm DNA fragmentation index was greater (P < 0.05) for low fertility compared with both average and high fertility bulls. In experiment 2, samples of sperm from a single collection from commercial Holstein bulls (N = 34) were used to evaluate relative sperm mRNA expression of adiponectin and its receptors, AdipoR1 and AdipoR2, and protein levels of adiponectin and its receptors, AdipoR1 and AdipoR2, in pre- and postcapacitation sperm. The mRNA abundance of adiponectin and its receptors, AdipoR1 and AdipoR2, were greater for high fertility bulls (>2 to ≤4 sire conception rate) compared with average (≥2 to ≤2) and low (>-2 to ≤-4) fertility bulls. Based on the sperm capacitation assay, average fertility bulls had a greater percentage of acrosome-reacted sperm at 5 hours than high and low fertility bulls, whereas high fertility bulls had a greater percentage of acrosome-reacted sperm than low fertility bulls. After capacitation, levels of adiponectin protein were lower in average fertility bulls, AdipoR1 was lower in all fertility groups, and AdipoR2 was lower in average and high fertility bulls. In conclusion, adiponectin and its receptors had vital roles in sperm structural and functional traits and consequently

  11. 7 CFR 58.327 - Frozen cream.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Frozen cream. 58.327 Section 58.327 Agriculture... Material § 58.327 Frozen cream. To produce frozen cream eligible for official certification, the quality of the cream used shall meet the requirements of cream acceptable for the manufacture of U.S. Grade AA...

  12. 5 CFR 1650.3 - Frozen accounts.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Frozen accounts. 1650.3 Section 1650.3... SAVINGS PLAN General § 1650.3 Frozen accounts. (a) All withdrawals from the TSP are subject to the rules... participant may not withdraw any portion of his or her account balance if the account is frozen due to...

  13. Extracellular calcium is involved in egg yolk-induced head-to-head agglutination of bull sperm.

    PubMed

    Yang, D H; McMillan, A G; Standley, N T; Shannon, P; Xu, Z Z

    2012-10-15

    Head-to-head agglutination of bull sperm occurs when semen is highly diluted in an egg yolk-citrate diluent without streptomycin. The objectives were to investigate causes of sperm agglutination and the underlying mechanism. Aliquots of bull semen were diluted in a base diluent (BD) supplemented with various test components and the percentage of agglutinated sperm (% AggSp) was quantified at 1, 5, 24, 48, and 72 h of incubation. When sperm were incubated at 22 °C, no agglutination was observed in BD for up to 72 h, whereas the % AggSp was 5.0, 41.7, 72.2, 91.1, and 92.8% in BD + 5% egg yolk (BD + EY) at 1, 5, 24, 48 and 72 h, respectively. However, no sperm agglutination was observed in BD + EY if incubation temperature was 37 °C. Addition of 5 or 10 mM ethylenebis (oxyethyleneni-trilo) tetra-acetic acid to BD + EY reduced the % AggSp from 95% to <5% at 72 h (P < 0.001), but addition of 5 mM CaCl(2) to BD failed to induce sperm agglutination in the absence of egg yolk, implicating calcium and other factors in egg yolk. Addition of the citrate-soluble fraction (CSF) of egg yolk to BD induced sperm agglutination similar to whole egg yolk, whereas water- and saline-soluble fractions of egg yolk were ineffective. The sperm-agglutinating efficacy of CSF (the % AggSp = 95% at 72 h) was reduced by dialysis (20%; P < 0.05), partially restored by addition of 5 mM CaCl2 (70%; P < 0.05), but the calcium effect was neutralized by addition of 5 mM ethylenebis (oxyethyleneni-trilo) tetra-acetic acid (1.7%; P < 0.05), again implicating calcium. Addition of 30 μM of a protein kinase A inhibitor (H-89) to an agglutinating diluent failed to inhibit sperm agglutination, whereas addition of 2 mM of a cAMP analogue, dbcAMP, to a nonagglutinating diluent failed to induce sperm agglutination. Agglutination status had no effect on sperm plasma membrane/acrosome status and mitochondrial membrane potential. In conclusion, calcium and other component(s) in the CSF of egg yolk induced head

  14. Recognition of FT-IR Data Cuscutae Semen, Japanese Dodder, and Sinapis Semen Using Discrete Wavelet Transformation and RBF Networks

    PubMed Central

    Hu, Tao; Weng, Xuexiang; Xu, Lishan; Cheng, Cungui; Yu, Peng

    2013-01-01

    Horizontal attenuation total reflection Fourier transformation infrared spectroscopy (HATR-FT-IR) studies on cuscutae semen and its confusable varieties Japanese dodder and sinapis semen combined with discrete wavelet transformation (DWT) and radial basis function (RBF) neural networks have been conducted in order to classify them. DWT is used to decompose the FT-IRs of cuscutae semen, Japanese dodder, and sinapis semen. Two main scales are selected as the feature extracting space in the DWT domain. According to the distribution of cuscutae semen, Japanese dodder, and sinapis semen's FT-IRs, three feature regions are determined at detail 3, and two feature regions are determined at detail 4 by selecting two scales in the DWT domain. Thus five feature parameters form the feature vector. The feature vector is input to the RBF neural networks to train so as to accurately classify the cuscutae semen, Japanese dodder, and sinapis semen. 120 sets of FT-IR data are used to train and test the proposed method, where 60 sets of data are used to train samples, and another 60 sets of FT-IR data are used to test samples. Experimental results show that the accurate recognition rate of cuscutae semen, Japanese dodder, and sinapis semen is average of 100.00%, 98.33%, and 100.00%, respectively, following the proposed method. PMID:24282653

  15. Recognition of FT-IR Data Cuscutae Semen, Japanese Dodder, and Sinapis Semen Using Discrete Wavelet Transformation and RBF Networks.

    PubMed

    Hu, Tao; Weng, Xuexiang; Xu, Lishan; Cheng, Cungui; Yu, Peng

    2013-01-01

    Horizontal attenuation total reflection Fourier transformation infrared spectroscopy (HATR-FT-IR) studies on cuscutae semen and its confusable varieties Japanese dodder and sinapis semen combined with discrete wavelet transformation (DWT) and radial basis function (RBF) neural networks have been conducted in order to classify them. DWT is used to decompose the FT-IRs of cuscutae semen, Japanese dodder, and sinapis semen. Two main scales are selected as the feature extracting space in the DWT domain. According to the distribution of cuscutae semen, Japanese dodder, and sinapis semen's FT-IRs, three feature regions are determined at detail 3, and two feature regions are determined at detail 4 by selecting two scales in the DWT domain. Thus five feature parameters form the feature vector. The feature vector is input to the RBF neural networks to train so as to accurately classify the cuscutae semen, Japanese dodder, and sinapis semen. 120 sets of FT-IR data are used to train and test the proposed method, where 60 sets of data are used to train samples, and another 60 sets of FT-IR data are used to test samples. Experimental results show that the accurate recognition rate of cuscutae semen, Japanese dodder, and sinapis semen is average of 100.00%, 98.33%, and 100.00%, respectively, following the proposed method.

  16. Increase in average testis size of Canadian beef bulls.

    PubMed

    García Guerra, Alvaro; Hendrick, Steve; Barth, Albert D

    2013-05-01

    Selection for adequate testis size in beef bulls is an important part of bull breeding soundness evaluation. Scrotal circumference (SC) is highly correlated with paired testis weight and is a practical method for estimating testis weight in the live animal. Most bulls presented for sale in Canada have SC included in the presale information. Scrotal circumference varies by age and breed, and may change over time due to selection for larger testis size. Therefore, it is important to periodically review the mean SC of various cattle breeds to provide valid bull selection criteria. Scrotal circumference data were obtained from bulls sold in western Canada from 2008 to 2011 and in Quebec from 2006 to 2010. Average scrotal circumferences for the most common beef breeds in Canada have increased significantly in the last 25 years. Differences between breeds have remained unchanged and Simmental bulls still have the largest SC at 1 year of age. Data provided here could aid in the establishment of new suggested minimum SC measurements for beef bulls.

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

    PubMed

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

    2007-06-01

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

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

    PubMed

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

    2013-01-01

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

  19. 21 CFR 101.95 - “Fresh,” “freshly frozen,” “fresh frozen,” “frozen fresh.”

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false âFresh,â âfreshly frozen,â âfresh frozen,â âfrozen... frozen,” “fresh frozen,” “frozen fresh.” The terms defined in this section may be used on the label or in... state and has not been frozen or subjected to any form of thermal processing or any other form...

  20. The effect of superoxide dismutase mimetic and catalase on the quality of postthawed goat semen.

    PubMed

    Shafiei, Mojtaba; Forouzanfar, Mohsen; Hosseini, Sayyed Morteza; Esfahani, Mohammad Hossein Nasr

    2015-05-01

    Manganese(III) meso-tetrakis(N-ethylpyridinium-2-yl)porphyrin chloride (MnTE) is a cell-permeable superoxide dismutase mimetic agent which can convert superoxide to hydrogen peroxide (H2O2). Supplementation of MnTE to a commercial semen extender can protect sperm from superoxide but not H2O2. Therefore, we proposed that addition of catalase (0.0, 200, or 400 IU/mL) in combination with MnTE (0.1 μM) may further improve the cryopreservation efficiency of goat semen in commercially optimized freezing media such as Andromed. Therefore, ejaculates were obtained from three adult bucks twice a week during the breeding season and diluted with Andromed supplemented with or without MnTE and catalase and were frozen in liquid nitrogen. Sperm parameters and reactive oxygen species contents were evaluated 2 hours after dilution (before freezing) and after freezing/thawing. The results revealed that all the treatments significantly (P ≤ 0.05) improved sperm motility, viability, and membrane integrity after freezing and reduced reactive oxygen species content compared with the control group, but maximum improvement was obtained in MnTE + 400 IU/mL catalase. In addition, supplementation with these antioxidants significantly (P ≤ 0.05) increases the cleavage rate after IVF. In conclusion, the results of present study suggest that addition of antioxidant MnTE or catalase to commercial optimized media, such as Andromed, improves total motility, membrane integrity, and viability of goat semen samples after thawing. But the degree of improvement for these parameters significantly (P ≤ 0.05) higher when MnTE and catalase were simultaneously added to the cryopreservation media.

  1. 9 CFR 98.35 - Declaration, health certificate, and other documents for animal semen.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... POULTRY) AND ANIMAL PRODUCTS IMPORTATION OF CERTAIN ANIMAL EMBRYOS AND ANIMAL SEMEN Certain Animal Semen... clinical signs of scrapie at the time of semen collection nor developed scrapie between the time of...

  2. 9 CFR 98.35 - Declaration, health certificate, and other documents for animal semen.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... POULTRY) AND ANIMAL PRODUCTS IMPORTATION OF CERTAIN ANIMAL EMBRYOS AND ANIMAL SEMEN Certain Animal Semen... clinical signs of scrapie at the time of semen collection nor developed scrapie between the time of...

  3. Antioxidant effect of quercetin in an extender containing DMA or glycerol on freezing capacity of goat semen.

    PubMed

    Seifi-Jamadi, Afshin; Ahmad, Ejaz; Ansari, Mahdi; Kohram, Hamid

    2017-04-01

    This study was performed to evaluate the effectiveness of quercetin as a non-enzymatic antioxidant in combination with glycerol or Dimethylacetamide (DMA), on freezability of goat semen. Ejaculates from four healthy mature Mahabadi goats were collected using an artificial vagina. After primary processing, semen was pooled and extended by egg yolk based extender supplemented with different concentrations of quercetin (10 or 20 μM) along with 5% glycerol or DMA. The extended semen was frozen and sperm motility parameters, viability, abnormality, membrane integrity and lipid peroxidation were assessed after thawing. Results showed that sperm viability, total motility, progressive motility, straightness (STR) and linearity (LIN) were higher (P < 0.05), and abnormality percentage and MDA concentration were lower (P < 0.05) in extender containing DMA. Similarly, higher (P < 0.05) total motility, progressive motility, viability and membrane integrity along with lower (P < 0.05) MDA level were noted in Q10 group. The lowest (P < 0.05) MDA level was observed in DMA extender containing moderate level of quercetin (Q10D). Also the STR was higher (P < 0.05) in Q10D compared to Q10G and Q20G groups. In conclusion, supplementation of extender with 10 μM quercetin in combination with DMA improves the goat sperm motion kinetics and suppresses lipid peroxidation after freezing and thawing. Furthermore, DMA is more effective cryoprotectant for the freezing of goat sperm.

  4. Cryopreservation of turkey semen: effect of breeding line and freezing method on post-thaw sperm quality, fertilization, and hatching.

    PubMed

    Long, Julie A; Purdy, Phillip H; Zuidberg, Kees; Hiemstra, Sipke-Joost; Velleman, Sandra G; Woelders, Henri

    2014-06-01

    Cryopreservation methods for poultry semen are not reliable for germplasm preservation, especially for turkeys, where fertility rates from frozen/thawed semen are particularly low. The objective was to evaluate cryopreservation methods for effectiveness in promoting cryosurvival and post-thaw function of sperm from five turkey lines: one commercial line and four research (RBC1; E; RBC2; F) lines from Ohio State University (OSU). The model for cryopreservation was set up as a 2×2×2×5 design for cryoprotectant (glycerol or dimethylacetamide (DMA)), cryopreservation medium (Lake or ASG), method of dilution (fixed dilution volume versus fixed sperm concentration) and turkey line, respectively. The final cryoprotectant concentrations were 11% glycerol or 6% DMA. Thawed sperm were evaluated for plasma membrane integrity and quality, motility, acrosome integrity and, after artificial insemination, for egg fertility and hatchability. Commercial turkey hens were used for all fertility trials, regardless of semen source. Turkey sperm frozen with glycerol exhibited higher membrane integrity and membrane quality upon thawing than turkey sperm frozen with DMA although no differences in total motility, and only minimal differences in progressive motility, were detected among the eight cryopreservation treatments. Within line, fertility was affected by cryoprotectant, medium and dilution method, where the overall highest percentages of fertile, viable embryos (Day 7) occurred for the DMA/ASG/fixed sperm concentration method, while high percentages (15.8-31.5%) of fertile, non-viable embryos (Day 1-6) were observed for multiple cryopreservation methods, including two glycerol treatments. From a single insemination, the duration of true and viable fertility in all lines was 10-13 weeks and 9-10 weeks, respectively. The duration of hatchability was 4-6 weeks after insemination for four of the turkey lines. The highest percentage of viable embryos was observed for the commercial

  5. Involuntary reduction in vigour of calves born from sexed semen.

    PubMed

    Djedović, Radica; Bogdanović, Vladan; Stanojević, Dragan; Nemes, Zsolt; Gáspárdy, András; Cseh, Sándor

    2016-06-01

    The objective of this study was to compare the reproductive traits of heifers and the development characteristics of their calves following artificial insemination (AI) with sexed and non-sexed semen. The analysed characteristics included conception rate, gestation length, calf birth weight, calf vigour, stillbirth rate, and twinning rate. Data of 530 calves produced with sexed and 1,163 calves produced with non-sexed semen were analysed. The General Linear Model (GLM) was applied to assess the influence of semen type, farm, season of insemination, the calf's sex and the inseminating sire on gestation length and calf birth weight. With the exception of gestation length (P > 0.05), all other traits studied were significantly (P < 0.01) influenced by the type of semen. The conception rate was 55% for conventional and 44% for sexed semen, and the average gestation length was 274.6 and 274.9 days, respectively. The mean calf birth weight was 37.47 kg for non-sexed and 36.75 kg for sexed semen. The stillbirth rate was 6.19% for conventional and 7.54% for sexed semen, while the twinning rate was 3.78% for conventional and 1.13% for sexed semen. The calves produced with non-sexed and sexed semen differed significantly in viability (P < 0.001), the latter having a lower calf vigour score. The use of conventional semen did not affect the ratio of female and male calves (52.7:47.3%; P > 0.05); however, artificial insemination with X-sorted sexed semen significantly altered the sex ratio of calves (85.1:14.9%, P < 0.01). The results obtained in this investigation are in agreement with the majority of studies which compared the fertility traits, sex ratio and calf characteristics depending on the application of artificial insemination with sexed or conventional semen.

  6. Sperm ubiquitination in epididymal feline semen.

    PubMed

    Vernocchi, Valentina; Morselli, Maria Giorgia; Varesi, Sara; Nonnis, Simona; Maffioli, Elisa; Negri, Armando; Tedeschi, Gabriella; Luvoni, Gaia Cecilia

    2014-09-01

    Ubiquitin is a 8.5-kDa peptide that tags other proteins for proteasomal degradation. It has been proposed that ubiquitination might be responsible for the elimination of defective spermatozoa during transit through the epididymis in humans and cattle, but its exact biological function in seminal plasma has not yet been clarified. In the domestic cat (Felis catus), the percentage of immature, unviable, and abnormal spermatozoa decreases during the epididymal transit, indicating the existence of a mechanism that removes defective spermatozoa. Magnetic cell separation techniques, based on the use of magnetic beads coated with anti-ubiquitin antibodies, may allow the selective capture of ubiquitinated spermatozoa from semen, thus contributing to the identification of a potential correlation between semen quality and ubiquitination process. Moreover, the selective identification of all the ubiquitinated proteins in different epididymal regions could give a better understanding of the ubiquitin role in feline sperm maturation. The aims of this study were as follows: (1) to verify the possibility of separating ubiquitinated spermatozoa with magnetic ubiquitin beads and identify the morphological and acrosomal differences between whole sample and unbound gametes, (2) to characterize all the ubiquitinated proteins in spermatozoa retrieved in the three epididymal regions by a proteomic approach. The data indicated the presence of ubiquitinated proteins in cat epididymal semen. However, a correlation between abnormal and ubiquitinated spermatozoa has not been found, and ubiquitin cannot be considered as a biomarker of quality of epididymal feline spermatozoa. To the author's knowledge, this is the first identification of all the ubiquitinated proteins of cat spermatozoa collected from different epididymal regions. The proteomic pattern allows a further characterization of cat epididymal semen and represents a contribute to a better understanding of the ubiquitin role in

  7. The JLab Frozen Spin Target

    SciTech Connect

    Keith, C. D.

    2009-08-04

    A polarized, frozen spin target has been designed and constructed at Jefferson Lab for use inside the CEBAF Large Acceptance Spectrometer. Protons in TEMPO-doped butanol are polarized via dynamic nuclear polarization (DNP) to approximately 90% using microwaves and an external, 5 T solenoid magnet. The target sample is then cooled to approximately 30 mK while an internal 0.56 T superconducting magnet is used to maintain the polarization. Relaxation times in excess of 3500 hours have been observed.

  8. Milk quality, coagulation properties, and curd firmness modeling of purebred Holsteins and first- and second-generation crossbred cows from Swedish Red, Montbéliarde, and Brown Swiss bulls.

    PubMed

    Malchiodi, F; Cecchinato, A; Penasa, M; Cipolat-Gotet, C; Bittante, G

    2014-07-01

    The objective of the present study was to investigate how the crossbreeding of Holstein (HO) cows with bulls from Nordic and Alpine European breeds affect milk quality traits, traditional milk coagulation properties (MCP), and curd firmness modeling obtained from individual milk samples. A total of 506 individual milk samples were collected from evening milking at 3 commercial farms located in Northern Italy. Over the past decade, the 3 farms have followed crossbreeding programs in part of their herds, whereas the remainder of the animals consisted of purebred HO. The basic scheme was a 3-breed rotation based on the use of Swedish Red (SR) semen on HO cows (SR × HO), the use of Montbéliarde (MO) semen on first-cross cows [MO × (SR × HO)], and the use of HO semen in the third cross. In all herds, a smaller proportion of purebred HO were mated to M and Brown Swiss (BS) bulls, and these first crosses were mated to SR and MO bulls, respectively. Milk samples were analyzed for milk composition and MCP, and parameters for curd firmness were modeled. Compared with purebred HO, crossbred cows produced less milk with lower lactose content, higher fat and protein content, and a tendency for higher casein content. Crossbred cows generally produced milk with a more favorable curd-firming rate (k₂₀) and curd firmness 30 min after rennet addition, among traditional MCP, and better trends of curd firmness measures as shown by model parameters: estimated rennet coagulation time, asymptotical potential value of curd firmness, and curd-firming instant rate constant. Among crossbred cows, SR × HO presented longer rennet coagulation time compared with MO × HO and BS × HO cows, and MO × HO showed shorter k₂₀ compared with BS × HO cows. Among second-generation cows, those sired by SR bulls showed a lower incidence of noncoagulated samples, higher curd firmness 30 min after rennet addition and asymptotical potential value of curd firmness, and faster curd-firming instant

  9. Zooplankton Feeding on the Nuisance Flagellate Gonyostomum semen

    PubMed Central

    Johansson, Karin S. L.; Vrede, Tobias; Lebret, Karen; Johnson, Richard K.

    2013-01-01

    The large bloom-forming flagellate Gonyostomum semen has been hypothesized to be inedible to naturally occurring zooplankton due to its large cell size and ejection of long slimy threads (trichocysts) induced by physical stimulation. In a grazing experiment using radiolabelled algae and zooplankton collected from lakes with recurring blooms of G. semen and lakes that rarely experience blooms, we found that Eudiaptomus gracilis and Holopedium gibberum fed on G. semen at high rates, whereas Daphnia cristata and Ceriodaphnia spp. did not. Grazing rates of E. gracilis were similar between bloom-lakes and lakes with low biomass of G. semen, indicating that the ability to feed on G. semen was not a result of local adaptation. The high grazing rates of two of the taxa in our experiment imply that some of the nutrients and energy taken up by G. semen can be transferred directly to higher trophic levels, although the predominance of small cladocerans during blooms may limit the importance of G. semen as a food resource. Based on grazing rates and previous observations on abundances of E. gracilis and H. gibberum, we conclude that there is a potential for grazer control of G. semen and discuss why blooms of G. semen still occur. PMID:23667489

  10. Thawing of Frozen Tuna Meat

    NASA Astrophysics Data System (ADS)

    Tanaka, Takeo; Nishiwaki, Kôji; Kakuda, Kitonari; Tomimatsu, Takao

    Frozen southern bluefin tuna meat discolors easily and sometimes contracts when thawed caused by thaw rigor. These phenomenon often become problematic in the transaction or handling of this kind of frozen tuna. Frozen meat blocks of southern Bluefin tuna were thawed separately by air thawing, running water thawing and microwave thawing. Changes occurring during thawing were checked for meat color by met-myoglobin ratio determination and for contract by microscopic observation. Results are as follows : (1) Discoloration scarcely occurred in the process of running water thawing (at 10°C for 50 min, or at 0°C for 6 hr). (2) No contraction was observed during thawing with running water described above and air thawing (at 18-20°C for 6 hr). (3) Discoloration and contraction seemed to be minimized, as to latently contractile blocks, when meat temperature passed through rapidly between -10°C and -5°C, and slowly (for 5-6 hr) between -5°C and -1°C. When the block was originally not contractile, discloration was minimized by rising meat temperature rapidly from -10°C to -l°C.

  11. Chronic boron exposure and human semen parameters.

    PubMed

    Robbins, Wendie A; Xun, Lin; Jia, Juan; Kennedy, Nola; Elashoff, David A; Ping, Liu

    2010-04-01

    Boron found as borates in soil, food, and water has important industrial and medical applications. A panel reviewing NTP reproductive toxicants identified boric acid as high priority for occupational studies to determine safe versus adverse reproductive effects. To address this, we collected boron exposure/dose measures in workplace inhalable dust, dietary food/fluids, blood, semen, and urine from boron workers and two comparison worker groups (n=192) over three months and determined correlations between boron and semen parameters (total sperm count, sperm concentration, motility, morphology, DNA breakage, apoptosis and aneuploidy). Blood boron averaged 499.2 ppb for boron workers, 96.1 and 47.9 ppb for workers from high and low environmental boron areas (p<0.0001). Boron concentrated in seminal fluid. No significant correlations were found between blood or urine boron and adverse semen parameters. Exposures did not reach those causing adverse effects published in animal toxicology work but exceeded those previously published for boron occupational groups.

  12. Heritability estimate and genetic correlations of reproductive features in Nellore bulls, offspring of super precocious, precocious and normal cows under extensive farming conditions.

    PubMed

    Siqueira, J B; Oba, E; Pinho, R O; Quintino, H P; Eler, J P; Miranda Neto, T; Guimarães, S E F; Guimarães, J D

    2012-04-01

    The present work aimed to estimate heritability and genetic correlations of reproductive features of Nellore bulls, offspring of mothers classified as superprecocious (M1), precocious (M2) and normal (M3). Twenty one thousand hundred and eighty-six animals with average age of 21.29 months were used, evaluated through the breeding soundness evaluation from 1999 to 2008. The breeding soundness features included physical semen evaluation (progressive sperm motility and sperm vigour), semen morphology (major, minor and total sperm defects), scrotal circumference (SC), testicular volume (TV) and SC at 18 months of age (SC18). The components of variance, heritability and genetic correlations for and between the features were estimated simultaneously by restricted maximum likelihood, with the use of the vce software system vs 6. The heritability estimates were high for SC18, SC and TV (0.43, 0.63 and 0.54; 0.45, 0.45 and 0.44; 0.42, 0.45 and 0.41, respectively for the categories of mothers M1, M2 and M3) and low for physical and morphological semen aspects. The genetic correlations between SC18 and SC were high, as well as between these variables with TV. High and positive genetic correlations were recorded among SC18, SC and TV with the physical aspects of the semen, although no favourable association was verified with the morphological aspects, for the three categories of mothers. It can be concluded that the mother's sexual precocity did not affect the heritability of their offspring reproduction features.

  13. [Analysis and identification of Semen Glycines Nigrae and Semen Pharbitidis by infrared spectroscopy].

    PubMed

    Du, Juan; Peng, Xi-Yuan; Ma, Fang; Chen, Jian-Bo; Zhou, Qun; Jin, Zhe-Xiong; Sun, Su-Qin

    2014-09-01

    Semen Glycines Nigrae and Semen Pharbitidis containing a large amount of fats and proteins are commonly used in Chinese herbal medicine. Tri-step infrared spectroscopy was applied to fast analyze and identify the two samples. In the conventional infrared spectroscopy, the samples both have obvious characteristic absorption peaks at 1,745 cm(-1) assigned to the stretching mode of C==O in esters. Furthermore, the two kinds of herbs have the peaks at 1,656 and 1,547 cm(-1) assigned to the amide I and II bands of protein. Obviously, the infrared spectra of herbs demonstrate that protein and fat is the major component in two kinds of herbs, and the relative intensity of the peaks assigned to fat and protein indicate their relative content is different. And the result is consistent with the reported. In the second derivative spectra, Semen Pharbitidis has a peak at 1,712 cm(-1) assigned to the organic acid, however, Semen Glycines Nigrae has not this absorption peak. In addition, in the second derivative spectra, appeared more differences between the two samples in shape and intensity of the peaks. In two-dimensional correlation infrared spectra, the two samples were visually distinguished due to their significant differences in auto-peak position and intensity. In the region of 1,500-1,700 cm(-1), Semen Glycines Nigrae has two autopeaks and Semen Pharbitidis has three autopeaks. In the region of 2,800-3,000 cm(-1), the samples both have two autopeaks, but the position of the strongest autopeak is different. It was demonstrated that the Tri-step infrared spectroscopy were successfully applied to fast analyze and identify the two kinds of samples containing the same major component, and made sure the foundation for future researches.

  14. Donkey jack (Equus asinus) semen cryopreservation: studies of seminal parameters, post breeding inflammatory response, and fertility in donkey jennies.

    PubMed

    Rota, A; Panzani, D; Sabatini, C; Camillo, F

    2012-11-01

    The aims of this study were (1) to evaluate motility parameters of donkey jack (jack; Equus asinus) semen cryopreserved in INRA-96 (INRA; IMV Technologies, France, 2% egg-yolk enriched) using either glycerol (GLY) or ethylene glycol (EG) as a cryoprotector; (2) to compare in vitro the postthaw re-extension with homologous seminal plasma (SPL) or INRA; (3) to compare fertility in donkey jennies (jennies; Equus asinus) timed artificially inseminated with jack semen cryopreserved using GLY or EG, re-extended with INRA; (4) to compare fertility in jennies timed artificially inseminated with jack semen cryopreserved using GLY re-extended with SPL, INRA, or not re-extended (NN); and (5) to describe some preliminary results of the inflammatory uterine response postbreeding. Semen from two jacks was collected and frozen in an INRA-2% egg yolk extender added of either 2.2% GLY or 1.4% EG. Postthaw motility was evaluated by a computer-assisted motility analyzer. Uterine inflammatory response and fertility were evaluated after artificial insemination (AI) of 13 jennies with frozen-thawed semen, either further extended with INRA (Group GLY-INRA, 13 cycles, and EG-INRA, 8 cycles), or with SPL (Group GLY-SPL, 13 cycles), or not re-extended (GLY-NN, 5 cycles). In each cycle, jennies were bred twice with 500 × 10(6) sperm cells (250 × 10(6) from each jack), at fixed times after induction of ovulation, and uterus was flushed at 6 and 10 h after first and second breeding, respectively. Cells in the recovered fluid were counted and distinguished as polymorphonuclear neutrophils (PMN) or other cell types. Total and progressive motility did not differ between cryoprotectants, but were higher when semen samples were re-extended in INRA, compared with SPL (P < 0.05). Pregnancy was diagnosed by transrectal palpation and ultrasonography examinations at 14 and 16 days postovulation. In 7/13 (53.8%) jennies and 12/39 (30.4%) cycles postbreeding intrauterine fluid accumulation was observed

  15. 28. UPPER STATION, LOWER FLOOR, BULL WHEEL ROOM, SAFETY BRAKE ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    28. UPPER STATION, LOWER FLOOR, BULL WHEEL ROOM, SAFETY BRAKE ADJUSTING MACHINERY. - Monongahela Incline Plane, Connecting North side of Grandview Avenue at Wyoming Street with West Carson Street near Smithfield Street, Pittsburgh, Allegheny County, PA

  16. POURING IRON FROM BULL LADLE INTO MOBILE LADLES USED TO ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    POURING IRON FROM BULL LADLE INTO MOBILE LADLES USED TO FILL MOLDS ON CONVEYOR LINES AFTER FERRO-SILICON IS ADDED TO ENHANCE DUCTILITY AND FLUIDITY. - Southern Ductile Casting Company, Casting, 2217 Carolina Avenue, Bessemer, Jefferson County, AL

  17. POURING IRON FROM ELECTRIC FURNACE INTO BULL LADLE AFTER MAGNESIUM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    POURING IRON FROM ELECTRIC FURNACE INTO BULL LADLE AFTER MAGNESIUM HAD BEEN ADDED TO GENERATE DUCTILE IRON WHEN IT COOLS IN THE MOLD. - Southern Ductile Casting Company, Casting, 2217 Carolina Avenue, Bessemer, Jefferson County, AL

  18. 26. UPPER STATION, LOWER FLOOR, BULL WHEEL. Monongahela Incline ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    26. UPPER STATION, LOWER FLOOR, BULL WHEEL. - Monongahela Incline Plane, Connecting North side of Grandview Avenue at Wyoming Street with West Carson Street near Smithfield Street, Pittsburgh, Allegheny County, PA

  19. HENRY STREET SCHOOL, 1300 BLOCK BULL STREET, DETAIL OF NORTH ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    HENRY STREET SCHOOL, 1300 BLOCK BULL STREET, DETAIL OF NORTH ELEVATION - Savannah Victorian Historic District, Bounded by Gwinnett, East Broad, West Broad Street & Anderson Lane, Savannah, Chatham County, GA

  20. TRAM HOUSE INTERIOR, LOOKING SOUTHEAST. NOTE TRACTION CABLE BULL WHEEL ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    TRAM HOUSE INTERIOR, LOOKING SOUTHEAST. NOTE TRACTION CABLE BULL WHEEL AND DEPARTING BUCKET "12," STILL ON RAIL AND JUST PRIOR TO ENGAGING TRACTION CABLE. - Shenandoah-Dives Mill, 135 County Road 2, Silverton, San Juan County, CO

  1. 13. Bull Wheel Core Showing Name of Manufacturer (National Supply ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    13. Bull Wheel Core Showing Name of Manufacturer (National Supply Co.), Looking Southwest - David Renfrew Oil Rig, East side of Connoquenessing Creek, 0.4 mile North of confluence with Thorn Creek, Renfrew, Butler County, PA

  2. 17. View of disassembled reduction gear parts including bull and ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    17. View of disassembled reduction gear parts including bull and intermediate gears and pedestal bearing. - Hacienda Azucarera La Esperanza, Steam Engine & Mill, 2.65 Mi. N of PR Rt. 2 Bridge over Manati River, Manati, Manati Municipio, PR

  3. 27. UPPER STATION, LOWER FLOOR, BULL WHEEL, BRAKE AIR CYLINDER. ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    27. UPPER STATION, LOWER FLOOR, BULL WHEEL, BRAKE AIR CYLINDER. - Monongahela Incline Plane, Connecting North side of Grandview Avenue at Wyoming Street with West Carson Street near Smithfield Street, Pittsburgh, Allegheny County, PA

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2015-06-01

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

  6. Influence of fetuin and hyaluronan on the post-thaw quality and fertilizing ability of Holstein bull semen.

    PubMed

    Sarıözkan, Serpil; Bucak, Mustafa Numan; Tuncer, Pürhan Barbaros; Büyükleblebici, Serhat; Eken, Ayşe; Akay, Cemal

    2015-08-01

    It was determined that fetuin and hyaluronan supplementation did not provide any significant effect on the post-thaw subjective and CASA motility percentages and sperm motion characteristics, in comparison to the controls (P>0.05). Sperm acrosome and total abnormalities were similar in all groups (P>0.05). Groups M (hyaluronan+fetuin) and H (hyaluronan) displayed a higher rate of sperm membrane integrity, compared to that of Group C (control) (P<0.01). According to the results of the comet assay, the lowest percentage of sperm with damaged DNA was achieved in Group H, when compared to all of the experimental groups (P<0.01). Furthermore, all of the additives resulted in a lower rate of sperm with damaged DNA than that of the controls, and thus, reduced DNA damage (P<0.01). For pregnancy rates, there were no significant differences between the extender groups (P>0.05). MDA formation was found to be lower in Groups M and F (P<0.01). In Group M, SOD activity was determined to have significantly increased (23.61±5.62 U/ml) compared to the other groups (P<0.01). All experimental groups had a GSH-Px activity higher than that of the control group (P<0.01).

  7. Schofield memorial lecture. Bull semen and muscle ATP: some evidence of the dawn of medical science in ancient Egypt.

    PubMed Central

    Schwabe, C W

    1986-01-01

    The importance of animal experimentation to human and animal health is not well understood by an increasingly articulate segment of the public. This could have very unfortunate consequences for man and his domestic animals. Even veterinarians and physicians are not as conversant as they need be about the great extent to which advances in human health have depended upon animal observations and experiments. Some believe that resort to "animal models" of biomedical phenomena, including diseases--a comparative or analogical approach to medical studies--is a relatively recent event. Even medical historians often treat these subjects as occasionally recurring aberrations which began with the Greeks, thus largely overlooking the historical meaning and continuing importance of "one medicine" irrespective of species. In fact, comparative medicine has probably been basic to medical progress ever since the dawn of a medical science. Recent research indicates that this approach to biomedical mysteries began to evolve in the minds of Egypt's healer-priests long before Aristotle and the later Alexandrian Greeks made the whole process explicit. Here we examine the origins of what were possibly the first two biomedical theories profounded from inferences based upon dissections, confirmed in at least one instance by experiment, and then applied to medical practice. PMID:3530413

  8. Semen phthalate metabolites, semen quality parameters and serum reproductive hormones: A cross-sectional study in China.

    PubMed

    Wang, Yi-Xin; Zeng, Qiang; Sun, Yang; Yang, Pan; Wang, Peng; Li, Jin; Huang, Zhen; You, Ling; Huang, Yue-Hui; Wang, Cheng; Li, Yu-Feng; Lu, Wen-Qing

    2016-04-01

    Exposure to phthalates has been found to have adverse effects on male reproductive function in animals. However, the findings from human studies are inconsistent. Here we examined the associations of phthalate exposure with semen quality and reproductive hormones in a Chinese population using phthalate metabolite concentrations measured in semen as biomarkers. Semen (n = 687) and blood samples (n = 342) were collected from the male partners of sub-fertile couples who presented to the Reproductive Center of Tongji Hospital in Wuhan, China. Semen quality parameters and serum reproductive hormone levels were determined. Semen concentrations of 8 phthalate metabolites were assessed using high-performance liquid chromatography and tandem mass spectrometry. Associations of the semen phthalate metabolites with semen quality parameters and serum reproductive hormones were assessed using confounder-adjusted linear and logistic regression models. Semen phthalate metabolites were significantly associated with decreases in semen volume [mono-n-butyl phthalate (MBP), mono-(2-ethylhexyl) phthalate (MEHP), mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono(2-ethyl-5-oxohexyl) phthalate (MEOHP)], sperm curvilinear velocity [monobenzyl phthalate (MBzP), MEHP, the percentage of di-(2-ethylhexyl)-phthalate metabolites excreted as MEHP (%MEHP)], and straight-line velocity (MBzP, MEHP, %MEHP), and also associated with an increased percentage of abnormal heads and tails (MBzP) (all p for trend <0.05). These associations remained suggestive or significant after adjustment for multiple testing. There were no significant associations between semen phthalate metabolites and serum reproductive hormones. Our findings suggest that environmental exposure to phthalates may impair human semen quality.

  9. Cryopreservation of rabbit semen: comparing the effects of different cryoprotectants, cryoprotectant-free vitrification, and the use of albumin plus osmoprotectants on sperm survival and fertility after standard vapor freezing and vitrification.

    PubMed

    Rosato, Maria Pina; Iaffaldano, Nicolaia

    2013-02-01

    This study was designed to improve current freezing protocols for rabbit sperm by examining: (1) the toxicity of different permeable cryoprotectants (CPAs) used for standard vapor freezing (conventional freezing); (2) the feasibility of ultrarapid nonequilibrium freezing (vitrification) of sperm in the absence of permeating CPAs; and (3), the addition of bovine serum albumin (BSA), alone or with sucrose or trehalose as osmoprotectants. First, we evaluated the effects on sperm motility of the incubation time (5 to 60 minutes) with different final concentrations (5% to 20%) of glycerol, N-N-dimethylacetamide, dimethylsulfoxide (DMSO), ethylene glycol, propylene glycol, and methanol. N-N-dimethylacetamide (5%) and DMSO (5% and 10%) showed the least toxic effects; the use of 10% DMSO producing the best postthaw sperm motility and membrane integrity results (P < 0.05) after conventional freezing. For vitrification, semen was diluted in the absence of permeable CPAs and frozen by dropping semen directly in liquid nitrogen. However, this led to the low or null cryosurvival of sperm postvitrification (0.16 ± 0.4%, 1.8 ± 1.6%, and 94.5 ± 1.4% of motile, membrane-, and DNA-intact sperm cells, respectively). To assess the effects of albumin and osmoprotectants on sperm cryosurvival, sperm was conventionally frozen with 10% DMSO or vitrified in the absence of permeable CPAs without or with 0.5% BSA alone or combined with sucrose or trehalose (range, 0-0.25 M). In the conventional freezing procedure, the addition of BSA alone failed to improve sperm cryosurvival, however, in the presence of BSA plus either sucrose or trehalose, the postthaw motility (using 0.1 M sucrose or trehalose) and DNA integrity (using all additive concentrations) of sperm were significantly better (P < 0.05) than control. Higher numbers of motile and membrane-intact cells were observed when semen was vitrified with BSA alone or with BSA and sucrose (0.1 and 0.25 M) or BSA and trehalose (0.25 M) and a

  10. Perfluorochemicals and Human Semen Quality: The LIFE Study

    PubMed Central

    Chen, Zhen; Schisterman, Enrique F.; Kim, Sungduk; Sweeney, Anne M.; Sundaram, Rajeshwari; Lynch, Courtney D.; Gore-Langton, Robert E.; Barr, Dana Boyd

    2014-01-01

    Background: The relation between persistent environmental chemicals and semen quality is evolving, although limited data exist for men recruited from general populations. Objectives: We examined the relation between perfluorinated chemicals (PFCs) and semen quality among 501 male partners of couples planning pregnancy. Methods: Using population-based sampling strategies, we recruited 501 couples discontinuing contraception from two U.S. geographic regions from 2005 through 2009. Baseline interviews and anthropometric assessments were conducted, followed by blood collection for the quantification of seven serum PFCs (perfluorosulfonates, perfluorocarboxylates, and perfluorosulfonamides) using tandem mass spectrometry. Men collected a baseline semen sample and another approximately 1 month later. Semen samples were shipped with freezer packs, and analyses were performed on the day after collection. We used linear regression to estimate the difference in each semen parameter associated with a one unit increase in the natural log–transformed PFC concentration after adjusting for confounders and modeling repeated semen samples. Sensitivity analyses included optimal Box-Cox transformation of semen quality end points. Results: Six PFCs [2-(N-methyl-perfluorooctane sulfonamido) acetate (Me-PFOSA-AcOH), perfluorodecanoate (PFDeA), perfluorononanoate (PFNA), perfluorooctane sulfonamide (PFOSA), perfluorooctane sulfonate (PFOS), and perfluorooctanoic acid (PFOA)] were associated with 17 semen quality end points before Box-Cox transformation. PFOSA was associated with smaller sperm head area and perimeter, a lower percentage of DNA stainability, and a higher percentage of bicephalic and immature sperm. PFDeA, PFNA, PFOA, and PFOS were associated with a lower percentage of sperm with coiled tails. Conclusions: Select PFCs were associated with certain semen end points, with the most significant associations observed for PFOSA but with results in varying directions. Citation

  11. Protein changes in frozen fish.

    PubMed

    Sikorski, Z; Olley, J; Kostuch, S

    1976-09-01

    Storage of frozen fish brings about a decrease of extractability of myofibrillar proteins. There is also deterioration of the texture and functional properties of the flesh. In model systems, aggregation of myosin, actin, tropomyosin, and whole myofibrils have been described. These changes are caused by concurrent action of partial dehydration due to the freezing out of water, exposure of the proteins to inorganic salts which are concentrated in the remaining nonfrozen fluid, interactions with free fatty acids liberated from phospholipids and with lipid oxidation products, and cross-linking by formaldehyde produced in some species of fish as a result of enzymic decomposition of trimethylamine oxide. The extent of protein alterations increases with time and temperature of storage as well as with advanced disintegration of the tissues and intermixing of their components. The role played by the individual factors and the significance of different types of bonds, i.e., hydrophobic adherences, ionic bonds, and covalent cross-links in particular cases are not yet fully disclosed. Retardation of the deteriorative changes of proteins in frozen fish is possible by avoiding high storage temperatures and oxidation of lipids, removing hematin compounds and other constituents promoting cross-linking reactions, and by adding cryoprotectors like sugars, several organic acids, amino acids, or peptides.

  12. Tritrichomonas foetus: prevalence study in naturally mating bulls in Switzerland.

    PubMed

    Bernasconi, Ch; Bodmer, M; Doherr, M G; Janett, F; Thomann, A; Spycher, C; Iten, C; Hentrich, B; Gottstein, B; Müller, N; Frey, C F

    2014-03-01

    Switzerland is officially free from bovine Tritrichomonas foetus. While bulls used for artificial insemination (AI) are routinely examined for this pathogen, bulls engaged in natural mating, as well as aborted fetuses, are only very sporadically investigated, indicating that the disease awareness for bovine tritrichomoniasis is low. Natural mating in cattle is becoming increasingly popular in Switzerland. Accordingly, a re-introduction/re-occurrence of T. foetus in cattle seems possible either via resurgence from a yet unknown bovine reservoir, or via importation of infected cattle. The low disease awareness for bovine tritrichomoniasis might favor an unnoticed re-establishment of T. foetus in the Swiss cattle population. The aim of our study was thus to search for the parasite, and if found, to assess the prevalence of bovine T. foetus in Switzerland. We included (1) bulls over two years of age used in natural mating and sent to slaughter, (2) bulls used for natural service in herds with or without fertility problems and (3) aborted fetuses. Furthermore, the routinely examined bulls used for AI (4) were included in this study. In total, 1362 preputial samples from bulls and 60 abomasal fluid samples of aborted fetuses were analyzed for the presence of T. foetus by both in vitro cultivation and molecular analyses. The parasite could not be detected in any of the samples, indicating that the maximal prevalence possibly missed was about 0.3% (95% confidence). Interestingly, in preputial samples of three bulls of category 1, apathogenic Tetratrichomonas sp. was identified, documenting a proof-of-principle for the methodology used in this study.

  13. Effect of semen extender and storage temperature on ram sperm motility over time

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Storage of ram semen for long period of time depends on a number of factors, including type of extender and storage temperature. A study compared the effect of semen extender and storage temperature on motility of ram semen stored for 72 h. Semen collected via electroejaculator from 5 mature Katahd...

  14. The uses of infrared thermography to evaluate the effects of climatic variables in bull's reproduction.

    PubMed

    Menegassi, Silvio Renato Oliveira; Pereira, Gabriel Ribas; Dias, Eduardo Antunes; Koetz, Celso; Lopes, Flávio Guiselli; Bremm, Carolina; Pimentel, Concepta; Lopes, Rubia Branco; da Rocha, Marcela Kuczynski; Carvalho, Helena Robattini; Barcellos, Júlio Otavio Jardim

    2016-01-01

    The objective of this study was to evaluate the seasonal effects of the environment on sperm quality in subtropical region determined by temperature and humidity index (THI). We used 20 Brangus bulls (5/8 Angus × 3/8 Nellore) aged approximately 24 months at the beginning of the study. Semen evaluations were performed twice per season during 1 year. Climate THI data were collected from an automatic weather station from the National Institute of Meteorology. Infrared thermography images were used to determine the temperature of the proximal and distal poles of the testis to assess the testicular temperature gradient (TG). The seasonal effects on seminal and climatic variables were analyzed with ANOVA using MIXED procedure of SAS. Sperm motility in spring (60.1%), summer (57.6%), and autumn (64.5%) showed difference compared to winter (73.0%; P < 0.01). TG was negatively correlated with THI at 18 days (spermiogenesis) (-0.76; P < 0.05) and at 12 days (epididymal transit) (-0.85; P < 0.01). Ocular temperature (OcT) had a positive correlation with THI at 18 days (0.78; P < 0.05) and at 12 days (0.84; P < 0.01). Motility showed a negative correlation with THI only at 18 days (-0.79; P < 0.05). During spermiogenesis, the TG had higher negative correlation compared to OcT (-0.97; P < 0.01) and rectal temperature (-0.72; P < 0.05). Spermatozoa with distal midpiece reflex were correlated with THI during transit epididymis (0.72; P < 0.05). Seminal parameters are not affected when THI reaches 93.0 (spermiogenesis) and 88.0 (epididymal transit). We concluded that infrared thermography can be adopted as an indirect method in order to assess the effect of environmental changes in TG and OcT of Brangus bulls.

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

    PubMed

    Hutchison, J L; Cole, J B; Bickhart, D M

    2014-05-01

    The availability of genomic evaluations since 2008 has resulted in many changes to dairy cattle breeding programs. One such change has been the increased contribution of young bulls (0.8 to 3.9 yr old) to those programs. The increased use of young bulls was investigated using pedigree data and breeding records obtained from the US national dairy database (Beltsville, MD). The adoption of genotyping was so rapid that by 2009, >90% of all Holstein artificial insemination (AI) service sires and 86% of Jersey AI service sires were genotyped, regardless of age. The percentage of sons sired by young bulls increased by 49 percentage points (10% in 2008 compared with 59% in 2012) due to the onset of genomic evaluations for Holsteins and by 46 percentage points for Jerseys (11 and 57%, respectively). When limiting these data to sons retained for breeding purposes through AI, the increase was even more dramatic, increasing approximately 80 percentage points from 2008 to 2012 for both Holsteins and Jerseys (1, 5, 28, 52, and 81% for Holsteins and 3, 4, 43, 46, and 82% for Jerseys from 2008 through 2012). From US breeding records from 2007 through 2012, 24,580,793 Holstein and 1,494,095 Jersey breedings were examined. Young bulls accounted for 28% and 25% of Holstein and Jersey breedings in 2007, respectively. These percentages increased to 51% of Holstein and 52% of Jersey breedings in 2012, representing 23- and 27-percentage-unit increases, respectively. Matings to genotyped young bulls have rapidly increased while the use of nongenotyped bulls has diminished since the onset of genomics. Mean sire age for Holstein male progeny born in 2012 was 2.7 yr younger than males born in 2006, and 1.3 yr younger for females; corresponding values for Jerseys were 2.3 and 0.9 yr. Holstein male offspring had an increase of 281 kg between 2006 and 2012, compared with 197 kg between 2000 and 2006 for parent averages (PA) for milk, an increase of 84 kg between the 2 periods. Jersey male

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

    USGS Publications Warehouse

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

    2015-01-01

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

  17. The relationship of bull fertility to sperm nuclear shape

    USGS Publications Warehouse

    Ostermeier, G.C.; Sargeant, G.A.; Yandell, B.S.; Parrish, J.J.

    2001-01-01

    group had a linear relationship (r .89, P .05) with fertility. To construct a plot of mean sperm shapes, a novel technique to automatically orient and identify the anterior tip of the sperm head was developed. The mean nuclear shape of high-fertility sperm was more elongated and tapered than those of lower fertility. A discriminant function (P .05) was also constructed that separated the 6 bulls into 2 groups based only on the harmonic amplitudes or sperm nuclear shape. The bulls were correctly classified into the 2 fertility groups. A comparison of sperm chromatin structure analysis (SCSA) and harmonic amplitudes found that overall size variance, anterior roundness, and posterior taperedness of sperm nuclei were related to chromatin stability (P .05). Some of the differences observed in sperm nuclear shape between the high- and lower-fertility bulls may be explained by varying levels of chromatin stability. However, sperm nuclear shape appears to contain additional information from chromatin stability alone. In this particular study, with 6 bulls, all with good chromatin quality, sperm nuclear shape was a better predictor of bull fertility.

  18. Behavioral aspects of electronic bull separation and mate allocation in multiple-sire mating paddocks.

    PubMed

    Lee, C; Prayaga, K C; Fisher, A D; Henshall, J M

    2008-07-01

    Controlling spatial positioning of cattle through use of electronic collars could provide new ways to farm under extensive conditions. This study examined the potential for bulls to be controlled during mating using mild electric shocks delivered through radio-controlled collars. Eighteen Belmont Red bulls were fitted with collars containing the Global Positioning System and that were able to emit a mild electric shock (500 mW) at the top of the neck behind the poll. Eighteen Belmont Red cows were fitted with Global Positioning System collars only. The experiment was replicated 3 times in 3 paddocks. Each paddock contained 2 bulls and 1 cow in induced estrus. On d 1, the bulls were either assigned to the cow or not assigned to the cow, and on d 2, the assignments were reversed, and bulls received the other treatment using a new cow. Treatments were applied for 2 h on each day. The nonassigned bull received a mild electric shock on approach to either the cow or to a bull, whereas the assigned bull received a mild electric shock on approach to the other bull only. The electric shock was applied when the bulls were within approximately 10 m and moving toward the nonallowed animal. The electric shock was terminated when the animal responded by stopping movement toward the nonallowed animal. In the first 10 min, nonassigned bulls spent less time within 5 m of the cow (P = 0.03) than assigned bulls. Assigned bulls spent more time close to the cow during the entire 120 min on d 1 than on d 2 (P = 0.014). On d 1, the assigned bulls moved more toward the cow and the nonassigned bull than they did on d 2 (P = 0.02). Assigned bulls displayed more sexual behaviors than nonassigned bulls (P = 0.004). Nonassigned bulls were sometimes observed not to approach the cow despite a change in its location. This suggests that the bull associated the electric shock with the cow and not with the location in which it received the electric shock. Instances were observed in which the cow

  19. Environmental Exposure to Triclosan and Semen Quality.

    PubMed

    Zhu, Wenting; Zhang, Hao; Tong, Chuanliang; Xie, Chong; Fan, Guohua; Zhao, Shasha; Yu, Xiaogang; Tian, Ying; Zhang, Jun

    2016-02-17

    Triclosan (2,4,4'-trichloro-2'-hydroxy-diphenyl ether, TCS) is widely used in personal care, household, veterinary and industrial products. It was considered as a potential male reproductive toxicant in previous in vitro and in vivo studies. However, evidence from human studies is scarce. Our study aims to investigate the relationship between TCS exposure and semen quality. We measured urinary TCS concentrations in 471 men recruited from a male reproductive health clinic. TCS was detected in 96.7% of urine samples, with a median concentration of 0.97 ng (mg·creatinine)(-1) (interquartile range, 0.41-2.95 ng (mg·creatinine)(-1)). A multiple linear regression analysis showed a negative association between natural logarithm (Ln) transformed TCS concentration (Ln-TCS) and Ln transformed number of forward moving sperms (adjusted coefficient β = -0.17; 95% confidence interval (CI) (-0.32, -0.02). Furthermore, among those with the lowest tertile of TCS level, Ln-TCS was negatively associated with the number of forward moving sperms (β = -0.35; 95% CI (-0.68, -0.03)), percentage of sperms with normal morphology (β = -1.64; 95% CI (-3.05, -0.23)), as well as number of normal morphological sperms, sperm concentration and count. Our findings suggest that the adverse effect of TCS on semen quality is modest at the environment-relevant dose in humans. Further studies are needed to confirm our findings.

  20. 7 CFR 58.349 - Frozen cream.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Frozen cream. 58.349 Section 58.349 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards... Products Bearing Usda Official Identification § 58.349 Frozen cream. The flavor shall be sweet,...

  1. Persistent Organic Pollutants and Semen Quality: The LIFE Study

    PubMed Central

    Mumford, Sunni L.; Kim, Sungduk; Chen, Zhen; Gore-Langton, Robert E.; Barr, Dana Boyd; Buck Louis, Germaine M.

    2014-01-01

    Growing evidence suggests that persistent environmental chemicals such as polychlorinated biphenyls may adversely affect human fecundity. The purpose of this study was to evaluate associations between persistent environmental chemicals and semen quality among 501 male partners of couples discontinuing contraception for purposes of becoming pregnant. Men provided a blood specimen and two fresh semen samples collected approximately a month apart that underwent next day analysis for 35 semen quality endpoints. Serum samples were analyzed for 36 polychlorinated biphenyls (congeners #18, 28, 44, 49, 52, 66, 74, 87, 99, 101, 114, 118, 128, 138, 146, 149, 151, 153, 156, 157, 167, 170, 172, 177, 178, 180, 183, 187, 189, 194, 195, 196, 201, 206, 209); 1 polybrominated biphenyl (#153); 9 organochlorine pesticides; and 10 polybrominated diphenyl ethers (congeners #17, 28, 47, 66, 85, 99, 100, 153, 154, 183) using high resolution mass spectrometry. To estimate the effect of chemicals on semen quality, we regressed each semen marker on each chemical while adjusting for research site, age, body mass index, serum lipids, and cotinine levels. Males with chemical concentrations in the fourth quartile, as compared to the first quartile, showed significant associations for several individual chemicals in each chemical class and type of semen quality parameter indicating negative and positive associations with semen quality. Polybrominated diphenyl ethers in particular were associated with several measures of increased abnormal morphology. These exploratory results highlight the role of environmental influences on male fecundity, and are of particular interest given the ubiquitous exposures to these compounds. PMID:25441930

  2. Testing the frozen flow approximation

    NASA Technical Reports Server (NTRS)

    Lucchin, Francesco; Matarrese, Sabino; Melott, Adrian L.; Moscardini, Lauro

    1993-01-01

    We investigate the accuracy of the frozen-flow approximation (FFA), recently proposed by Matarrese, et al. (1992), for following the nonlinear evolution of cosmological density fluctuations under gravitational instability. We compare a number of statistics between results of the FFA and n-body simulations, including those used by Melott, Pellman & Shandarin (1993) to test the Zel'dovich approximation. The FFA performs reasonably well in a statistical sense, e.g. in reproducing the counts-in-cell distribution, at small scales, but it does poorly in the crosscorrelation with n-body which means it is generally not moving mass to the right place, especially in models with high small-scale power.

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

    PubMed

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

    1992-02-15

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

  4. Perineal Bull Gore with Urinary Bladder Perforation and Pneumoperitoneum

    PubMed Central

    R, Santhosh; Barad, Arun Kumar; Ghalige, Hemanth Sureshwara; K, Sridartha; Sharma M, Birkumar

    2013-01-01

    Animal related injuries are frequently reported in India and other countries, where bulls are used for sporting events as well as in places where farming and livestock rearing is practised. The presentation is, many times, atypical and misleading as well. They have unique mechanics of injury. The patterns of the injury are reviewed. An intra-peritoneal urinary bladder injury which is caused by a perineal bull gore with a pneumoperitoneum is unusual and it has not been reported in the literature which was reviewed. We are reporting a successfully treated 25 years old male patient from the slopes of the southern district of Manipur, India, who had presented 40 hours after he was injured. The identification and prompt exploration, keeping in mind the mechanics of bull goring, helps the surgeons to adequately deal such atypical injuries, for optimal outcomes. PMID:23814738

  5. Zinc and magnesium in bull and boar spermatozoa.

    PubMed

    Arver, S; Eliasson, R

    1980-11-01

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

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

  8. Pit-1 gene polymorphism, milk yield, and conformation traits for Italian Holstein-Friesian bulls.

    PubMed

    Renaville, R; Gengler, N; Vrech, E; Prandi, A; Massart, S; Corradini, C; Bertozzi, C; Mortiaux, F; Burny, A; Portetelle, D

    1997-12-01

    The growth hormone factor-1/pituitary-specific transcription factor Pit-1 is responsible for the expression of growth hormone in mammals. Mutations in Pit-1 have been found in growth hormone disorders of mice and humans. We studied the eventual association between Pit-1 polymorphism using the HinfI enzyme and the milk yield and conformation traits of 89 Italian Holstein-Friesian bulls. A strategy employing polymerase chain reaction was used to amplify a 451-bp fragment from semen DNA. Digestion of polymerase chain reaction products with HinfI revealed two alleles: allele A was not digested (451-bp fragment), and allele B was cut at one restriction site, generating two fragments of 244 and 207 bp. Three patterns were observed; frequencies were 2.2, 31.5, and 66.3% for AA, AB, and BB, respectively. Fixed and mixed linear models were fitted on daughter yield deviations for milk yields and on deregressed proofs for conformation traits. Predictions were weighted using the inverse of the estimated variance of records. The models used contained mean and gene substitution effects for Pit-1 A allele as fixed effects and random sire effect for the mixed model. The A allele was found to be superior for milk and protein yields, inferior for fat percentage, and superior for body depth, angularity, and rear leg set, which is difficult to explain. A canonical transformation revealed that Pit-1 had three actions, one linked to milk yield traits and angularity, a second linked to body depth and rear leg set, and a third linked to lower fat yields and to higher angularity.

  9. Semen cryopreservation and radical reduction capacity of seminal fluid in captive African lion (Panthera leo).

    PubMed

    Luther, I; Jakop, U; Lueders, I; Tordiffe, A; Franz, C; Schiller, J; Kotze, A; Müller, K

    2017-02-01

    Optimizing cryopreservation protocols for nondomestic felids contributes to the successful development of assisted reproduction techniques and genetic resource banking. In this study, we describe a simple cryopreservation procedure for African lion (Panthera leo) ejaculates, which was tested with different packaging options and different sperm numbers per dose. By applying urethral catheterization and electroejaculation, 17 ejaculates with greater than 20% motile and greater than 5% progressively motile sperm were collected. A lyophilized extender (a modified egg yolk-Tes-Tris-fructose-glycerol medium) was rehydrated and added in one step at ambient temperature (∼25 °C) to semen, which was prediluted in cell culture medium M199. After slow cooling of insulated samples to 15 °C in a refrigerator (4 °C), the samples were fast frozen over the surface of liquid nitrogen or in a dry shipper. Aliquots of 300 μL containing 20 × 10(6) sperm were frozen in cryovials and in 0.5-mL straws. Differences were observed in the total motility after thawing between vial (31.5 ± 14.1%) and straw freezing (20.1 ± 8.6%). However, the subpopulations of vital (22.7 ± 7.8% for vial and 19.8 ± 8.5% for straw) and progressively motile (10.0 ± 7.9% for vial and 10.0 ± 6.4% for straw) sperm after washing and 1 hour incubation at 38 °C were of similar magnitude, velocity, and linearity for both packaging options. After freezing of five ejaculates with 20, 60, and 100 × 10(6) sperm per dose, best results were achieved at the lowest concentration. In general, post-thaw results were highly variable (2.2% and 56.5% total motility) and not correlated to motility or morphology of the fresh semen. To further characterize semen quality, we assessed the protective potential of seminal fluid against oxidative stress, which might be challenged on freeze thawing. The capacity of seminal fluid to reduce radicals was measured in 10 semen samples by electron spin resonance

  10. 75 FR 13715 - Endangered and Threatened Wildlife and Plants; Revised Designation of Critical Habitat for Bull...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-23

    ...; Revised Designation of Critical Habitat for Bull Trout in the Coterminous United States AGENCY: Fish and... of critical habitat for the bull trout (Salvelinus confluentus) ] under the Endangered Species Act of... January 14, 2010, we published our proposed revised designation of critical habitat for bull trout in...

  11. The influence of temperament, transportation stress, and endotoxin challenge on body composition traits in Brahman bulls

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was designed to determine the influence of temperament of bulls on ultrasound body composition traits in response to transportation and an endotoxin challenge. Purebred Brahman bulls were selected from a pool of 60 bulls based on temperament scores measured at weaning (n=8 each: calm, int...

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

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-03

    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Red Bull Flugtag Miami... during the Red Bull Flugtag. The Red Bull Flugtag is scheduled to take place on Saturday, November 3... to the water below. 150 spectator vessels are expected to attend the event. The special...

  13. 78 FR 57061 - Special Local Regulation; Red Bull Flugtag Miami, Biscayne Bay; Miami, FL

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-17

    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulation; Red Bull Flugtag Miami..., during the Red Bull Flugtag. The Red Bull Flugtag is scheduled to take place on September 21, 2013. The... foot ramp to the water below. Approximately 100 spectator vessels are anticipated to be at the...

  14. 78 FR 40079 - Special Local Regulations; Red Bull Flugtag Miami, Biscayne Bay; Miami, FL

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-03

    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Red Bull Flugtag Miami... Park, in Miami, Florida, during the Red Bull Flugtag event. The Red Bull Flugtag is scheduled to take... flying objects from a 30 foot ramp to the water below. The special local regulation is necessary...

  15. Environmental Exposure to Triclosan and Semen Quality

    PubMed Central

    Zhu, Wenting; Zhang, Hao; Tong, Chuanliang; Xie, Chong; Fan, Guohua; Zhao, Shasha; Yu, Xiaogang; Tian, Ying; Zhang, Jun

    2016-01-01

    Triclosan (2,4,4′-trichloro-2′-hydroxy-diphenyl ether, TCS) is widely used in personal care, household, veterinary and industrial products. It was considered as a potential male reproductive toxicant in previous in vitro and in vivo studies. However, evidence from human studies is scarce. Our study aims to investigate the relationship between TCS exposure and semen quality. We measured urinary TCS concentrations in 471 men recruited from a male reproductive health clinic. TCS was detected in 96.7% of urine samples, with a median concentration of 0.97 ng (mg·creatinine)−1 (interquartile range, 0.41–2.95 ng (mg·creatinine)−1). A multiple linear regression analysis showed a negative association between natural logarithm (Ln) transformed TCS concentration (Ln-TCS) and Ln transformed number of forward moving sperms (adjusted coefficient β = −0.17; 95% confidence interval (CI) (−0.32, −0.02). Furthermore, among those with the lowest tertile of TCS level, Ln-TCS was negatively associated with the number of forward moving sperms (β = −0.35; 95% CI (−0.68, −0.03)), percentage of sperms with normal morphology (β = −1.64; 95% CI (−3.05, −0.23)), as well as number of normal morphological sperms, sperm concentration and count. Our findings suggest that the adverse effect of TCS on semen quality is modest at the environment-relevant dose in humans. Further studies are needed to confirm our findings. PMID:26901211

  16. Indian story on semen loss and related Dhat syndrome

    PubMed Central

    Prakash, Om; Kar, Sujit Kumar; Sathyanarayana Rao, T. S.

    2014-01-01

    India is a country of many religions and ancient cultures. Indian culture is largely directed by the Vedic culture since time immemorial. Later Indian culture is influenced by Buddhism, Islam, and Christianity. Indian belief system carries the footprints of these cultures. Every culture describes human behaviors and an interpretation of each human behavior is largely influenced by the core cultural belief system. Sexuality is an important domain which is colored by different cultural colors. Like other cultures, Indian culture believes “semen” as the precious body fluid which needs to be preserved. Most Indian beliefs consider loss of semen as a threat to the individual. Ancient Indian literature present semen loss as a negative health related event. Dhat syndrome (related to semen loss) is a culture-bound syndrome seen in the natives of Indian subcontinent. This article gathers the Indian concepts related to semen loss. It also outlines belief systems behind problems of Dhat syndrome. PMID:25568479

  17. Risk factors for bacterial contamination during boar semen collection.

    PubMed

    Goldberg, Ana Maria G; Argenti, Laura E; Faccin, Jamil E; Linck, Lídia; Santi, Mônica; Bernardi, Mari Lourdes; Cardoso, Marisa R I; Wentz, Ivo; Bortolozzo, Fernando P

    2013-10-01

    The aim of this study was to evaluate the influence of multiple factors on bacterial contamination in 213 ejaculates from four boar studs. Semen contamination by aerobic mesophiles increased in ejaculates where the preputial fluid flowed into the collection container, collection glove was dirty, preputial hair was long (>1.0 cm), the collection lasted >7 min and boars were older than 18 months. An increase in coliforms occurred when preputial fluid dripped into the collection container, collections lasted >7 min or when penis escaped during collection. Semen contamination increased when two or more factors related to hygiene (poor hygiene of the boar, dirty preputial ostium, large preputial diverticulum, long preputial hair, dirty gloves, preputial liquid trickling from the hand of the technician into the semen container and penis escaping) were present. A vigilant protocol of collection must be followed to minimize bacterial contamination, especially avoiding dripping of preputial liquid into the semen container.

  18. 78 FR 2946 - United States Standards for Grades of Frozen Vegetables

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-15

    ... standards covered by these proposed revisions are: frozen asparagus, frozen lima beans, frozen speckled butter beans, frozen cooked squash, frozen summer squash, frozen sweet potatoes, frozen turnip greens... document. United States Standards for Grades of Frozen Lima Beans Update address for AMS. Change...

  19. Semen Quality and Time-to-Pregnancy, the LIFE Study

    PubMed Central

    Buck Louis, Germaine M.; Sundaram, Rajeshwari; Schisterman, Enrique F.; Sweeney, Anne; Lynch, Courtney D.; Kim, Sungduk; Maisog, José M.; Gore-Langton, Robert; Eisenberg, Michael L.; Chen, Zhen

    2013-01-01

    Objective To assess semen parameters and couple fecundity as measured by time-to-pregnancy (TTP). Design Observational prospective cohort with longitudinal measurement of TTP. Setting 16 Michigan/Texas counties Participants 501 couples discontinuing contraception were followed for one year while trying to conceive; 473 (94%) men provided one semen sample and 80% provided two samples. Interventions None Main Outcome Measures Using prospectively measured TTP, fecundability odds ratios (FORs) and 95% confidence intervals (CIs) were estimated for 36 individual semen quality parameters accounting for repeated semen samples, time off contraception, abstinence, enrollment site, and couples’ ages, body mass indices (BMI), and serum cotinine concentrations. Results In adjusted models, semen quality parameters were associated with significantly shorter TTP as measured by FORs >1: percent motility, strict and traditional morphology, sperm head width, elongation factor, and acrosome area. Significantly longer TTPs or FORs <1 were observed for morphologic categories amorphous and round sperm heads neck/midpiece abnormalities. No semen quality parameters achieved significance when simultaneously modeling all other significant semen parameters and covariates, except for percent coiled tail when adjusting for sperm concentration (FOR=0.99; 95% CI 0.99, 1.00). Male age was consistently associated with reduced couple fecundity (FOR=0.96; 95% CI 0.93–0.99) reflecting a longer TTP across all combined models. Female but not male BMI also conferred a longer TTP (FOR=0.98; 95% CI 0.96–0.99). Conclusions Several semen measures were significantly associated with TTP when modeled individually, but not jointly and in the context of relevant couple based covariates. PMID:24239161

  20. [Quality of semen Hydnocarpi anthelminiticae introduced to various places].

    PubMed

    Lin, L; Xu, H; Chen, J; Chen, Y

    1996-02-01

    The result shows that the Semen Hydnocarpi Anthelminticae introduced in Guangdong, Hainan and Yunnan Provinces is similar to the control medicinal materials in fatty oil content, index of refraction, effective component content and trace element content. But the saponification value of the introduced products is higher. The quality of the Semen Hydnocarpi Anthelminticae introduced to the above places is up to the level of the control medicinal materials.

  1. Training Tribal Lay Advocates at Sitting Bull College

    ERIC Educational Resources Information Center

    Shelley, W. L.

    2015-01-01

    Students in Sitting Bull College's lay advocate program develop a well-rounded understanding of the law, enabling them to represent defendants in tribal courts. The program offers legal training for its students--and illustrates how American Indian nations can broaden legal representation for Native defendants in tribal courts. It is one of only…

  2. The Case for Bull Dogs and Mother Hens.

    ERIC Educational Resources Information Center

    Neugebauer, Bonnie; Neugebauer, Roger

    1996-01-01

    Describes traits of effective child care team members: instigator--develops ideas; day-dream believer--suggests solutions; jester--relieves tension; mother hen--ensures fair treatment; nervous Nellie--critiques ideas; keeper of the faith--focuses on center's mission; bull dog--keeps on task; compromiser--preserves unity; and mover and…

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

    ERIC Educational Resources Information Center

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

    2008-01-01

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

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

    ERIC Educational Resources Information Center

    Miles, William F. S.

    2009-01-01

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

  5. TRAM HOUSE INTERIOR, LOOKING SOUTH. NOTE HORIZONTAL, IDLER, BULL WHEEL ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    TRAM HOUSE INTERIOR, LOOKING SOUTH. NOTE HORIZONTAL, IDLER, BULL WHEEL TO REVERSE DIRECTION OF TRACTION CABLE. TRAM CABLE DRIVE MOTOR (NO LONGER EXTANT) WAS AT MINE END OF TRAM. - Shenandoah-Dives Mill, 135 County Road 2, Silverton, San Juan County, CO

  6. Cultural Connections: Bowl with Frieze of Lions Attacking Bulls

    ERIC Educational Resources Information Center

    School Arts: The Art Education Magazine for Teachers, 2004

    2004-01-01

    This article gives a brief description of the piece of art titled "Bowl with Frieze of Lions Attacking Bulls" which is thought to be the product of a court or palace of the Neo-Assyrian period and dates to the late seventh to eighth century BC, between the reigns of Sargon and Ashurbanipal. The article highlights the piece's most notable…

  7. Enumeration of semen leucocytes by fluorescence in situ hybridisation technique

    PubMed Central

    Conte, R A; Luke, S; Verma, R S

    1995-01-01

    Aim—To determine whether the fluorescent in situ hybridisation technique (FISH) using a total human DNA genomic probe can be used to enumerate semen leucocytes. Methods—Semen samples from five donors were subjected to a mild KC1 solution. These samples were then biotin labelled under FISH conditions using a total human DNA genomic probe and the leucocyte counts were determined. To check the accuracy of the technique a monoclonal antibody against the common leucocyte antigen CD45 [KC56(T-200)] served as a control. An isotypic control for [KC56(T-200)], the immunoglobulin [MsIgG1], served as a secondary control. Results—Semen leucocytes stained by the FISH technique were easily detected because of their distinct bright yellow colour, while the sperm cells were red. The leucocyte count ranged from 0·5 to 4·9 × 106 per ml of semen. KC56(T-200) and its isotypic control MsIgG1, which served as control for the FISH technique, accurately identified 94% and 97% of the semen leucocytes of a control donor, respectively. Conclusions—The FISH technique using a total human DNA probe can accurately and effectively enumerate the overall leucocyte population in semen. Images PMID:16696031

  8. Relationships among temperament, behavior, and growth during performance testing of bulls.

    PubMed

    Lockwood, S A; Kattesh, H G; Krawczel, P D; Kirkpatrick, F D; Saxton, A M; Rhinehart, J D; Wilkerson, J B

    2015-12-01

    Excitable cattle are dangerous to personnel and have reduced individual performance. The aim of this study was to 1) identify objective criteria for evaluating bull temperament and 2) examine relationships among temperament, behavior, and performance of bulls during an 84-d performance test. Angus bulls ( = 60) were reared in 6 pens based on BW and age. Pen scores (PS; 1 = docile and 5 = very aggressive) were assigned on d -1, 27, 55, and 83. Exit velocity (EV), BW, time to exit the chute, and order through the chute were recorded on d 0, 28, 56, and 84. The ADG was calculated for the 84-d test period, and ultrasound data and frame score calculations were recorded on d 84. Dataloggers measured steps taken, lying time, number of lying bouts, and lying bout duration of bulls ( = 27; 3 pens) from d 3 to 28 and d 59 to 84. Bulls with a d -1 PS of 1 or 2 were categorized as calm (PScalm; = 40), whereas bulls with a PS of 3 or 4 were categorized as excitable (PSexcitable; = 20). Bulls were separated into 2 groups based on the bottom 20 EV (EVcalm) and top 20 EV (EVexcitable) on d 0. Mixed model ANOVA (SAS 9.3) was used to compare groups for the two temperament assessment methods, behavior, and growth performance. Mean EV decreased ( < 0.05) by d 84. Total lying time from d 3 to 28 was greater ( < 0.05) for PScalm bulls when compared with PSexcitable bulls. However, total lying time from d 59 to 84 was greater ( < 0.05) for EVexcitable bulls when compared with EVcalm bulls. Regardless of initial contemporary group assignment, all bulls exited the chute slower ( < 0.001) on d 84 than on d 0. The PSexcitable bulls had greater ( < 0.01) frame scores and greater ADG than PScalm bulls. The PSexcitable bulls had more ( < 0.01) backfat than PScalm bulls. However, ribeye area was smaller ( < 0.01) in EVexcitable bulls than EVcalm bulls. Based on these results, bulls appeared to have habituated over the testing period. Additionally, the potential lack of innate temperament

  9. Assessment of semen quality, sperm cryopreservation and heterologous IVF in the critically endangered Iberian lynx (Lynx pardinus).

    PubMed

    Gañán, Natalia; González, Raquel; Garde, J Julián; Martínez, Fernando; Vargas, Astrid; Gomendio, Montserrat; Roldan, Eduardo R S

    2009-01-01

    Semen traits and factors affecting sperm cryopreservation were assessed in the Iberian lynx (Lynx pardinus), a species regarded as the most endangered felid in the world. For cryopreservation, semen was washed, resuspended in a Tes-Tris-based diluent (TEST) or a Tris-based diluent (Biladyl), both with 20% egg yolk and 4% glycerol, loaded into straws, cooled to 5 degrees C using an automated programmable system and frozen on nitrogen vapour. Heterologous IVF of in vitro-matured domestic cat oocytes was used to test the fertilising ability of cryopreserved spermatozoa. Electroejaculates from five males were obtained. Characterisation of the electroejaculates revealed mean (+/- s.e.m.) values of 3.3 +/- 0.6 x 10(6) total spermatozoa, 73.6 +/- 4.6% motile spermatozoa, 23.7 +/- 4.0% morphologically normal spermatozoa and 40.7 +/- 2.3% spermatozoa with intact acrosomes. After thawing a higher percentage of motile spermatozoa was seen in TEST than in Biladyl (34.0 +/- 6.2% v. 7.5 +/- 4.8%, respectively; P < 0.05); however, there were no differences in the percentage of intact acrosomes between the two diluents. Iberian lynx spermatozoa fertilised domestic cat oocytes in vitro, with higher fertilisation rates observed for spermatozoa cryopreserved in TEST than in Biladyl, although the difference did not reach statistical significance (20.5 +/- 4.5% v. 11.5 +/- 6.8%, respectively). There were positive significant relations between the fertilisation rates and both the percentage of normal spermatozoa and the percentage of spermatozoa with an intact acrosome before cryopreservation (P = 0.04). This first report of the collection and cryopreservation of Iberian lynx semen and analysis of fertilising ability is an important step in the development of assisted reproductive techniques for this critically endangered felid species.

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

    PubMed

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

    2015-01-01

    The aim of this study was to investigate the effects of storage of semen in different commercial extenders on the pre-freezing and post-thawing quality of boar spermatozoa. Semen was diluted in BTS, Androhep (AH) and Gedil (GD), stored for 24 h at 17°C, and then frozen in accordance with the cryopreservation protocol. Analyses of the quality of spermatozoa included: motility, normal apical ridge (NAR) acrosome, plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), measurements of ATP content and activity of superoxidase dismutase (SOD) and glutathione peroxidase (GPx). Prior to the freezing process, no significant effect of the extender on the sperm quality parameters was noted. After thawing the spermatozoa it was demonstrated that the type of extender used influenced PMI, MMP, ATP content and activity of GPx. In the AH extender the percentage of spermatozoa with PMI and ATP content in spermatozoa was significantly higher (P<0.05) as compared to the BTS or GD extenders. In addition, semen stored in the AH was characterised by a statistically higher (P<0.05) percentage of spermatozoa with MMP and increased activity of GPx as compared with the BTS. The results obtained indicate that for the cryopreservation process, boar spermatozoa stored for 24 hours in liquid state can be used. However, the type of extender used prior to freezing may have a significant effect on the post-thawing quality of the spermatozoa. The AH extender better secured the quality of thawed boar spermatozoa as compared with the BTS or GD.

  11. Use of frozen section in genitourinary pathology.

    PubMed

    Shen, Steven S; Truong, Luan D; Ro, Jae Y; Ayala, Alberto G

    2012-08-01

    Frozen section diagnosis provides critical information for immediate surgical management decision making. Over the last several years, there have been some significant advances in treatment of genitourinary cancer, particularly with regard to surgical techniques. These changes in turn impact the type and frequency of intraoperative frozen section requests. In this review, we describe the main indications and diagnostic challenges of frozen section diagnosis during surgeries of each genitourinary organ system including prostate, kidney, bladder, testis, and penis. The pitfalls and approaches to different diagnostic situations are discussed. It is also stressed that pathologists must not only be familiar with the histological diagnosis, but also understand the limitations of frozen section diagnosis and communicate with urologists during the intraoperative treatment decision making process.

  12. Chemical Effects during Storage of Frozen Foods.

    ERIC Educational Resources Information Center

    Powrie, W. D.

    1984-01-01

    Discusses (1) characteristics, interrelationships, and distribution of food constituents (including water) in unfrozen food systems; (2) the freezing process; and (3) chemical changes in food during frozen storage. Protein alterations and lipid oxidation are emphasized. (JN)

  13. The use of crossbreeding with beef bulls in dairy herds: effects on calving difficulty and gestation length.

    PubMed

    Fouz, R; Gandoy, F; Sanjuán, M L; Yus, E; Diéguez, F J

    2013-02-01

    This study was designed to analyse the evolution in the use of beef bull semen for dairy cattle insemination and, mainly, to assess calving difficulty, gestation length and proportion of stillbirths after breeding pure Holsteins or crossbreeding. Data were collected during 2004 to 2011 for 552 571 Holstein calvings (457 070 Holstein × Holstein, 43 384 Holstein × Limousine, 32 174 Holstein × Belgian Blue and 19 943 Holstein × Galician Blonde). The highest calving difficulty, compared with pure Holsteins was for crosses with Belgian Blue followed by Limousine and Galician Blonde. The Holstein × Limousine and Holstein × Galician Blonde crossbred calves had significantly longer gestation lengths than Holstein × Holstein and Holstein × Belgian Blue calves. Between the latter two, pure Holstein had the shortest gestation length. Calving difficulty and gestation length decreased as the age of the dam advanced. The most difficult calvings were observed in twin calvings, followed by the calvings of male calves and female calves. The gestations leading to the birth of male calves were longer than those leading to female calves and twin calves. Stillbirths were not related to the breed used for mating. Through examining these parameters, sire breed should be considered when selecting a beef breed for the insemination of milk-producing dams.

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

    USGS Publications Warehouse

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

    2015-01-01

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

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

    USGS Publications Warehouse

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

    2016-01-01

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

  16. 21 CFR 160.190 - Frozen egg yolks.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Frozen egg yolks. 160.190 Section 160.190 Food and... CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.190 Frozen egg yolks. (a) Frozen egg yolks, frozen yolks is the food prepared by freezing egg yolks...

  17. 21 CFR 160.190 -