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Sample records for bull semen frozen

  1. The effects of antibiotic additions to extenders on fresh and frozen-thawed bull semen.

    PubMed

    Gloria, Alessia; Contri, Alberto; Wegher, Laura; Vignola, Giorgio; Dellamaria, Debora; Carluccio, Augusto

    2014-11-10

    Reproduction in dairy cows is based around the use of cryopreserved semen. Antibiotics are utilized to control bacterial contamination and growth in cryopreserved bull semen. The antibiotic resistance of some bacteria required the evaluation of new antibiotic combinations with a high level of antibacterial effectiveness and a negligible effect on spermatozoa. In this research, we studied the effect of the fluorinate carboxyquinolone ofloxacin and the combination of ceftiofur/tylosin on bull spermatozoa and in-field bacterial growth. In Experiment 1, the toxicity of different levels of ofloxacin and ceftiofur/tylosin was tested by the incubation of bull spermatozoa and the evaluation of sperm kinetic parameters, membranes and acrosome integrity after dilution, and at 60 and 120 min after incubation. The data reported in this study reveals that both antibiotic combinations, at all concentrations, seem to have a negligible effect on spermatozoa with respect to all of the parameters examined (p>0.05). Furthermore, progressive motility was significantly higher for sperm diluted with both antibiotic combinations compared with samples without antibiotics (p<0.01). In Experiment 2, the ability of ofloxacin or ceftiofur/tylosin to control bacterial growth during bovine semen cryopreservation was compared with the combination of gentamicin/tylosin/spectinomycin/lincomycin. A significant reduction in progressive motility was found in cooled semen with respect to all of the antibiotic treatments (p<0.05). However, the membrane integrity was found to significantly rise in frozen samples with, compared to samples without, antibiotics (p<0.05). In a bull, gentamicin, tylosin, spectinomycin, and lincomycin failed to control bacterial growth in the cryopreserved sample, while no such growth was found in samples extended with ceftiofur/tylosin or ofloxacin. In conclusion, both ceftiofur/tylosin and ofloxacin can be safely added to bull seminal extenders, and both can protect

  2. Effect of thawing methods on frozen semen quality of yak (Poephagus grunniens L.) bulls

    PubMed Central

    Borah, Binod Kumar Dutta; Deka, Bharat Chandra; Biswas, Ranjan Kumar; Chakravarty, Prithiviraj; Deori, Sourabh; Sinha, Sudip; Ahmed, Kutubuddin

    2015-01-01

    Aim: To evaluate different thawing temperatures and duration on the post-thaw semen quality of Indian yaks bulls. Materials and Methods: Semen ejaculates from four different yak bulls were collected using artificial vagina method and extended with tris extender containing 6.4% glycerol at 35°C, cooled gradually from 35°C to 5°C at 1°C/3 min and equilibrated at 4-5°C for 4 h and frozen in French mini straws using a programmable bio-freezer and finally stored in liquid nitrogen. Thawing of frozen semen straws was carried out using three methods i.e., 35°C for 60 s (thawing method I), 37°C for 30 s (thawing method II) and 75°C for 9 s (thawing method III). The post-thaw semen quality parameters assessed were sperm motility, percent live sperm, hypo-osmotic swelling test (HOST)-reacted sperm, acrosomal changes, and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in the extracellular media. Results: The percent sperm motility, total incidence of acrosomal changes, and extracellular release of AST varied significantly (p<0.01) between thawing methods but live sperm and HOST-reacted sperm did not vary significantly between thawing methods. The percent sperm motility of frozen yak semen for thawing method III was significantly (p<0.05) higher than that for thawing methods I and II, the difference between thawing methods I and II being non-significant. The critical difference test revealed that the total incidence of acrosomal changes and extracellular release of AST were significantly (p<0.05) lower when thawing was done using methods I and II than in method III. Conclusion: On the basis of the present experiment, we can conclude that barring the post-thaw sperm motility, thawing of frozen yak semen in water either at 35°C for 60 s or 37°C for 30 s gives better post-thaw semen quality than at 75°C for 09 s. PMID:27047161

  3. Alpha-linolenic acid supplementation in tris extender can improve frozen-thawed bull semen quality.

    PubMed

    Kaka, A; Wahid, H; Rosnina, Y; Yimer, N; Khumran, A M; Behan, A A; Ebrahimi, M

    2015-02-01

    The study was conducted to evaluate the effects of α-linolenic acid (ALA) on frozen-thawed quality and fatty acid composition of bull sperm. For that, twenty-four ejaculates obtained from three bulls were diluted in a Tris extender containing 0 (control), 3, 5, 10 and 15 ng/ml of ALA. Extended semen was incubated at 37°C for 15 min, to allow absorption of ALA by sperm cell membrane. The sample was chilled for 2 h, packed into 0.25-ml straws and frozen in liquid nitrogen for 24 h. Subsequently, straws were thawed and evaluated for total sperm motility (computer-assisted semen analysis), membrane functional integrity (hypo-osmotic swelling test), viability (eosin-nigrosin), fatty acid composition (gas chromatography) and lipid peroxidation (thiobarbituric acid-reactive substances (TBARS)). A higher (p < 0.05) percentage of total sperm motility was observed in ALA groups 5 ng/ml (47.74 ± 07) and 10 ng/ml (44.90 ± 0.7) in comparison with control (34.53 ± 3.0), 3 ng/ml (34.40 ± 2.6) and 15 ng/ml (34.60 ± 2.9). Still, the 5 ng/ml ALA group presented a higher (p < 0.05) percentage of viable sperms (74.13 ± 0.8) and sperms with intact membrane (74.46 ± 09) than all other experimental groups. ALA concentration and lipid peroxidation in post-thawed sperm was higher in all treated groups when compared to the control group. As such, the addition of 5 ng/ml of ALA to Tris extender improved quality of frozen-thawed bull spermatozoa. PMID:25366298

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

    PubMed

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

    2016-07-01

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

  5. 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

  6. Improvement of liquid and frozen-thawed semen quality of Nili-Ravi buffalo bulls (Bubalus bubalis) through supplementation of fat.

    PubMed

    Adeel, M; Ijaz, A; Aleem, M; Rehman, H; Yousaf, M S; Jabbar, M A

    2009-05-01

    The aim of the study was to investigate the effects of dietary fat on quality of liquid and frozen-thawed semen of Nili-Ravi buffalo bulls. Adult bulls (n=21) were fed a balanced ration (Con; n=7) or the same ration either containing sunflower oil (SF-O; n=7) or whole sunflower seeds (SF-S; n=7) for 63 days. Body weight and body condition score of each bull was recorded on days 0, 30 and 60 of the experiment. Semen was collected on days 39, 46, 53 and 60, frozen by a fast method and stored at -196 degrees C for 24h. Sperm motility was assessed using a bright field microscope. Plasma membrane integrity of fresh and frozen-thawed spermatozoa was assessed using a hypo-osmotic swelling (HOS) assay. The concentration of spermatozoa and volume of semen was not different among groups on various days of collection. Sunflower-enriched diets did not affect the motility and number of HOS-positive spermatozoa in the fresh semen. Motility and HOS of post-thawed spermatozoa were higher (p<0.05) in bulls fed the sunflower-enriched diets. Similarly, diets did not affect the body condition score and body weight of bulls. In conclusion, feeding of sunflower oil or sunflower seed as fat sources can improve the quality of buffalo bull spermatozoa. PMID:19246083

  7. Sperm DNA integrity in frozen-thawed semen from Italian Mediterranean Buffalo bulls and its relationship to in vivo fertility.

    PubMed

    Serafini, Rosanna; Love, Charles C; Coletta, Angelo; Mari, Gaetano; Mislei, Beatrice; Caso, Chiara; Di Palo, Rossella

    2016-09-01

    The relationship among sperm attributes of DNA integrity, sperm motility, morphology, viability, acrosome integrity and in vivo fertility of frozen-thawed Italian Mediterranean Buffalo (IMB) sperm has not been reported. Straws of frozen-thawed semen samples from three bulls were examined. Sperm DNA assays (i.e., neutral Comet assay, Sperm Bos Halomax-SBH and Sperm Chromatin Structure Assay-SCSA) were not correlated to each other (P>0.05). Many neutral Comet assay measures were correlated to total sperm motility-TMOT (% head-H-DNA, r=0.74; Olive moment, r=-0.76; P<0.05) and coiled tails (r-values ranged from% H-DNA, r=-0.80 to tail length, r=-0.71; P<0.05). The COMP-αt was negatively correlated to viable acrosome intact (VAI) sperm, and distal droplets (r=-0.60 and -0.61; P<0.05), whereas Mean-αt and Mode-αt were positively correlated to bent midpieces (r=0.63 and 0.61; P<0.05). The SBH assay was positively correlated to non-viable acrosome damaged (NVAD) sperm (r=0.60; P<0.05) and negatively correlated to viable acrosome damaged (VAD) sperm (r=-0.63; P<0.05). The overall pregnancy rate (PR-at 30 and 45d post artificial insemination-AI) and the calving rate were 57%, 55% and 45%, respectively. Among sperm features analyzed the area under the Receiver Operating Characteristic (ROC) Curve was significant (P<0.05) for TMOT, NVAD, Standard Deviation-αt (SD-αt) and neutral comet measures (Olive tail moment and tail moment, % H- DNA and tail area) in estimating pregnancy. PMID:27421229

  8. 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

  9. 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. PMID:26515370

  10. Low density lipoproteins extracted from hen egg yolk by an easy method: cryoprotective effect on frozen-thawed bull semen.

    PubMed

    Moussa, M; Marinet, V; Trimeche, A; Tainturier, D; Anton, M

    2002-04-01

    Hen egg yolk is widely used as a cryoprotective agent in semen freezing extenders in order to protect the spermatozoa against cold shock. The protective action of yolk is largely presumed to be due to low density lipoproteins (LDL). In recent years, arguments concerning the presence of cryoprotective antagonists in egg yolk, have reinforced interest in the use of only the LDL extracted from egg yolk in the extenders. However, current methods of LDL purification do not support the use of LDL in commercial extenders because they offer a poor recovery rate. Consequently, we have developed an easy method to extract LDL from egg yolk. Several concentrations of purified LDL (between 2.5 and 20%, w/v) were tested in freezing extenders for bull semen, and compared with commercial extenders. Our extraction method reached 97% purity and about 67% yield, and is easily reproducible on an industrial scale. Analysis of sperm motility showed that the motility and characteristics of spermatozoa movement were improved with LDL in the extender, as compared to a commercial extender containing egg yolk. The optimum LDL concentration in the extender was 8%. In conclusion, we propose that an extender containing LDL extracted from egg yolk could be used as cryoprotective media with a better efficiency than present commercial extenders. PMID:12035979

  11. Artificial insemination of cranes with frozen semen

    USGS Publications Warehouse

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

    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.

  12. 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...

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

    PubMed Central

    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. PMID:25593679

  14. 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

  15. Seminal Plasma Heparin Binding Proteins Improve Semen Quality by Reducing Oxidative Stress during Cryopreservation of Cattle Bull Semen

    PubMed Central

    Patel, Maulikkumar; Gandotra, Vinod K.; Cheema, Ranjna S.; Bansal, Amrit K.; Kumar, Ajeet

    2016-01-01

    Heparin binding proteins (HBPs) are produced by accessory glands. These are secreted into the seminal fluid, bind to the spermatozoa at the time of ejaculation, favour capacitation, acrosome reaction, and alter the immune system response toward the sperm. The present study was conducted with an objective to assess the effect of purified seminal plasma-HBPs (SP-HBPs) on cross bred cattle bull sperm attributes during two phases of cryopreservation: Pre freezing and freezing-thawing. SP-HBPs were purified from pooled seminal plasma by heparin affinity chromatography. Three doses of SP-HBPs i.e. 10, 20, 40 μg/mL semen were standardized to find out the optimum dose and 20 μg/mL was found to be an optimum dose. Semen as such and treated with SP-HBPs was diluted with sodium citrate-egg yolk diluter and cryopreserved as per the standard protocol. Sperm parameters i.e. motility, viability, Hypo-osmotic swelling test (HOST), acrosome damage, in vitro capacitation and lipid peroxidation were evaluated in SP-HBP treated and untreated (control) semen at both phases of cryopreservation. A considerable variation in percent sperm motility, viability, membrane integrity (HOST), acrosome damage, acrosome reaction and lipid peroxidation was observed at both phases among the bulls irrespective of the treatment. Incubation of neat semen with 20 μg/mL SP-HBP before processing for cryopreservation enhanced the average motility, viability, membrane integrity by 7.2%, 1.5%, 7.9%, and 5.6%, 6.6%, 7.4% in pre-frozen and frozen-thawed semen in comparison to control. There was also an average increase of 4.1%/3.9% in in vitro capacitation and acrosome reaction in SP-HBPs-treated frozen-thawed semen as compared to control. However, binding of SP-HBPs to the sperm declined acrosome damage and lipid peroxidation by 1.3%/4.1% and 22.1/32.7 μM/109 spermatozoa in SP-HBP treated pre-frozen/frozen-thawed semen as compared to control, respectively. Significant (p<0.05) effects were observed only in

  16. 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. PMID:25497424

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

    PubMed Central

    Kumar, Umesh; Gawande, Ajay P.; Sahatpure, Sunil K.; Patil, Manoj S.; Lakde, Chetan K.; Bonde, Sachin W.; Borkar, Pradnyankur L.; Poharkar, Ajay J.; Ramteke, Baldeo R.

    2015-01-01

    Aim: To compare the seminal attributes of neat, pre-freeze (at equilibration), and post-freeze (24 h after freezing) semen in pure and crossbred Jersey bulls. Materials and Methods: Total 36 ejaculates (3 ejaculates from each bull) were collected from 6 pure Jersey and 6 crossbred Jersey bulls and evaluated for various seminal attributes during neat, pre-freeze, and post-freeze semen. Results: The mean (±standard error [SE]) values of neat semen characteristics in pure and crossbred Jersey bulls were recorded such as volume (ml), color, consistency, mass activity (scale: 0-5), and sperm concentration (millions/ml). The extended semen was further investigated at pre-freeze and post-freeze stages and the mean (±SE) values recorded at neat, pre-freeze, and post-freeze semen were compared between pure and crossbred Jersey bulls; sperm motility (80.55±1.70%, 62.77±1.35%, 46.11±1.43% vs. 80.00±1.80%, 65.00±1.66%, 47.22±1.08%), live sperm count (83.63±1.08%, 71.72±1.09%, 58.67±1.02% vs. 80.00±1.08%, 67.91±1.20%, 51.63±0.97%), total abnormal sperm count (8.38±0.32%, 12.30±0.39%, 16.75±0.42% vs. 9.00±0.45%, 12.19±0.48%, 18.11±0.64%), hypo-osmotic swelling (HOS) reacted spermatozoa (71.88±0.77%, 62.05±0.80%, 47.27±1.05% vs. 72.77±1.02%, 62.11±0.89%, 45.94±1.33%), acrosome integrity (89.05±0.83%, 81.33±0.71%, 71.94±0.86% vs. 86.55±0.57%, 78.66±0.42%, 69.38±0.53%), and DNA integrity (99.88±0.07%, 100, 99.66±0.11% vs. 99.94±0.05%, 100, 99.44±0.18%,). The volume, color, consistency, sperm concentration, and initial motility in pure and crossbred Jersey bulls did not differ significantly (p>0.05). The mass activity was significantly (p<0.05) higher in pure Jersey as compare to crossbred Jersey bulls. Live sperm percentage and acrosome integrity was significantly (p<0.01) higher in pure Jersey bulls as compared to crossbred Jersey bulls. However, no statistical difference (p>0.05) was observed in abnormal sperm; HOS reacted spermatozoa and DNA

  18. 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. PMID:27091815

  19. Effects of age and environmental factors on semen production and semen quality of Austrian Simmental bulls.

    PubMed

    Fuerst-Waltl, Birgit; Schwarzenbacher, Hermann; Perner, Christa; Sölkner, Johann

    2006-09-01

    More than 90% of the breeding stock of Austrian dual purpose Simmental cows is artificially inseminated. Knowledge of factors affecting sperm production and semen quality is of importance with regard to reproductive efficiency and thus genetic improvement as well as for the productivity and profitability of AI centers. Hence, semen data from two Austrian AI centres collected in the years 2000 and 2001 were evaluated. In total, 3625 and 3654 ejaculates from 147 and 127 AI bulls, respectively, were analysed regarding ejaculate volume, sperm concentration, percentage of viable spermatozoa in the ejaculate, total spermatozoa per ejaculate and motility. Effects accounted for were the bull (random), age of bull, collection interval, number of collection on collection day, bull handler, semen collector, temperature on day of semen collection, in the course of epididymal maturation (average temperature of days 1-11 before collection) and during spermatogenesis (average temperature of days 12-65 before collection). Age of bull significantly affected all traits (P<0.01 to P<0.001) except motility score in center 2. Ejaculate volume and total number of spermatozoa increased with age of bull while sperm concentration was lower in higher age classes (center 1). The collection team was also found to significantly influence semen quality traits. With increasing collection interval ejaculate volume and total number of spermatozoa increased significantly (P<0.05 to P<0.001) while collection intervals between 4-9 days and 1-6 days were superior with regard to sperm concentration and percentage of viable spermatozoa, respectively (P<0.10 to P<0.001). First ejaculates were superior with respect to ejaculate volumes, sperm concentrations and total number of spermatozoa per ejaculate (P<0.001). Temperature, either on day of semen collection or during epididymal maturation or spermatogenesis, had important but inconsistent effects on semen production and sperm quality. Overall, however

  20. Opportunities to improve liquid and frozen storage of boar semen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Artificial insemination has facilitated the utilization of superior genetics, and has reduced boar biosecurity problems and housing costs. The use of frozen semen permits the flexibility to inseminate animals at unscheduled times and to use semen of deceased boars. While good long-term storage exten...

  1. Comparative transcript profiling of gene expression of fresh and frozen-thawed bull sperm.

    PubMed

    Chen, Xiaoli; Wang, Yonggui; Zhu, Huabin; Hao, Haisheng; Zhao, Xueming; Qin, Tong; Wang, Dong

    2015-03-01

    Although frozen semen is widely used commercially in the cattle breeding industry, the resultant pregnancy rate is lower than that produced using fresh semen. Cryodamage is a major problem in semen cryopreservation; it causes changes to sperm transcripts that may influence sperm function and motility. We used suppression subtractive hybridization technology to establish a complementary DNA subtractive library, and combined microarray technology and sequence homology analysis to screen and analyze differentially expressed genes in the library, comparing fresh sperm with the frozen-thawed sperm of nine bulls. Overall, 19 positive differentially expressed unigenes were identified using microarray data and Significance Analysis of Microarrays software (|score (d)| ≥ 2, fold change > 1, and false discovery rate < 0.05). Of 15 differentially expressed unigenes exhibited high sequence homology (E-value ≤ 1 × 10(-3)), 12 were upregulated in frozen-thawed sperm, the remaining 3 were upregulated in fresh sperm, and 4 other clones were identified as unknown because of incomplete sequences or because there was no significant sequence homology (E-value > 1E(-03)) and were considered novel genes. The expression of five of these genes-RPL31, PRKCE, PAPSS2, PLP1, and R1G7-was verified by quantitative real-time reverse transcription-polymerase chain reaction. There was a significant differential expression of the RPL31 gene (P < 0.05). Our preliminary results provide an overview of differentially expressed transcripts between fresh and frozen-thawed sperm of Holstein bulls. PMID:25459024

  2. Enriching the captive elephant population genetic pool through artificial insemination with frozen-thawed semen collected in the wild.

    PubMed

    Hildebrandt, T B; Hermes, R; Saragusty, J; Potier, R; Schwammer, H M; Balfanz, F; Vielgrader, H; Baker, B; Bartels, P; Göritz, F

    2012-10-01

    The first successful AI in an elephant was reported in 1998, using fresh semen. Since then almost 40 calves have been produced through AI in both Asian and African elephants worldwide. Following these successes, with the objective of enriching the captive population with genetic material from the wild, we evaluated the possibility of using frozen-thawed semen collected from wild bulls for AI in captivity. Semen, collected from a 36-yr-old wild African savanna elephant (Loxodonta africana) in South Africa was frozen using the directional freezing technique. This frozen-thawed semen was used for four inseminations over two consecutive days, two before and two after ovulation, in a 26-yr-old female African savanna elephant in Austria. Insemination dose of 1200 × 10(6) cells per AI with 61% motility resulted in pregnancy, which was confirmed through ultrasound examination 75, 110 and 141 days after the AI procedure. This represents the first successful AI using wild bull frozen-thawed semen in elephants. The incorporation of AI with frozen-thawed semen into the assisted reproduction toolbox opens the way to preserve and transport semen between distant individuals in captivity or, as was done in this study, between wild and captive populations, without the need to transport stressed or potentially disease-carrying animals or to remove animals from the wild. In addition, cryopreserved spermatozoa, in combination with AI, are useful methods to extend the reproductive lifespan of individuals beyond their biological lifespan and an important tool for genetic diversity management and phenotype selection in these endangered mammals. PMID:22898009

  3. Unilateral intrauterine horn insemination of frozen semen in cats.

    PubMed

    Tsutsui, T; Tanaka, A; Takagi, Y; Nakagawa, K; Fujimoto, Y; Murai, M; Anzai, M; Hori, T

    2000-12-01

    Frozen feline semen was prepared using two types of extenders, egg yolk Tris-fructose citric acid (EYT-FC) and egg yolk sodium citrate solution (EYC), and the semen qualities after thawing and the conception rates obtained by unilateral intrauterine horn insemination (UIUI) were investigated. Cats used in the experiment were six males and 11 females aged 2-12 years (the number of experimental cases was 17). For preparation of frozen semen, semen collected by the artificial vagina method was adjusted to I x 10(8) sperm/m/ and 7% glycerol, put in 250 microl straws, and then frozen using a cell freezer. The mean sperm motility after thawing was 30.0+/-9.7 (SE) % in the semen prepared with EYT-FC and 30.0+/-3.3% in the semen prepared with EYC. Four of seven animals were fertilized by UIUI using two straws in both extenders, and the conception rate was 57.1%. The mean ratios of number of kits to the number of ovulations in the inseminated side were 61.1+/-24.5% and 30.5+/-3.4% for EYT-FC and EYC, respectively, showing that the ratio tended to be higher in the semen prepared with EYT-FC. The above findings, comparing the two extenders for preparation of frozen feline semen, showed that EYT-FC is slightly superior to EYC. To increase conception and fertility rates, it may be important to increase the sperm count for insemination and to inseminate both uterine horns. PMID:11193339

  4. Differential abundances of four forms of Binder of SPerm 1 in the seminal plasma of Bos taurus indicus bulls with different patterns of semen freezability.

    PubMed

    Magalhães, Marcos Jorge; Martins, Leonardo Franco; Senra, Renato Lima; Santos, Thaís Ferreira Dos; Okano, Denise Silva; Pereira, Paulo Roberto Gomes; Faria-Campos, Alessandra; Campos, Sérgio Vale Aguiar; Guimarães, José Domingos; Baracat-Pereira, Maria Cristina

    2016-08-01

    The Binder of SPerm 1 (BSP1) protein is involved in the fertilization and semen cryopreservation processes and is described to be both beneficial and detrimental to sperm. Previously, the relationship of BSP1 with freezability events has not been completely understood. The objective of this work was to determine the differential abundance of the forms of the BSP1 protein in cryopreserved seminal plasma of Bos taurus indicus bulls with different patterns of semen freezability using proteomics. A wide cohort of adult bulls with high genetic value from an artificial insemination center was used as donors of high quality, fresh semen. Nine bulls presenting different patterns of semen freezability were selected. Two-dimensional gel electrophoresis showed differential abundance in a group of seven protein spots in the frozen/thawed seminal plasma from the bulls, ranging from 15 to 17 kDa, with pI values from 4.6 to 5.8. Four of these spots were confirmed to be BSP1 using mass spectrometry, proteomics, biochemical, and computational analysis (Tukey's test at P < 0.05). The protein spot weighing 15.52 ± 0.53 kDa with a pI value of 5.78 ± 0.12 is highlighted by its high abundance in bulls with low semen freezability and its absence in bulls presenting high semen freezability. This is the first report showing that more than two forms of BSP1 are found in the seminal plasma of Nelore adult bulls and not all animals have a similar abundance of each BSP1 form. Different BSP1 forms may be involved in different events of fertilization and the cryopreservation process. PMID:27118515

  5. Detection of Neospora caninum in the semen and blood of naturally infected bulls.

    PubMed

    Ferre, Ignacio; Aduriz, Gorka; Del-Pozo, Itziar; Regidor-Cerrillo, Javier; Atxaerandio, Raquel; Collantes-Fernández, Esther; Hurtado, Ana; Ugarte-Garagalza, Carlos; Ortega-Mora, Luis Miguel

    2005-03-15

    A prospective study was designed to investigate the presence of Neospora caninum in semen and blood of eight bulls seropositive to N. caninum using nested-PCR procedures. Positive semen and blood samples were bioassayed in a BALB/c nu/nu mouse model. Specific anti-N. caninum serological and interferon-gamma (IFN-gamma) responses were also studied. In parallel, five seronegative bulls acted as non-infected controls. All bulls were located in a collaborating AI centre and monitored for 22 weeks. Six of eight seropositive bulls showed N. caninum DNA in their semen and/or blood samples at some time during the course of the study. In all positive semen samples, we consistently found Neospora-DNA in the cell fraction and not in seminal plasma. Parasite load, as determined by a real-time PCR in nested-PCR positive semen samples, ranged from 1 to 10 parasites/ml. We found no association between the presence of N. caninum DNA in semen and blood. N. caninum could not be detected in the BALB/c nu/nu mice inoculated with PCR-positive semen or blood samples. Specific IgG antibody levels in seropositive bulls fluctuated over time, at times falling below cut-off level. The response was predominantly IgG2, with significant differences compared to control bulls (P < 0.05). The overall mean specific IFN-gamma response in seropositive bulls was also higher than those observed in the control group (P < 0.05), although extensive variation in individual responses was observed among bulls and over time. No significant association was found between bulls showing Neospora DNA in semen, blood, or both, and specific IgG, IgG1, IgG2, IgM and IgA levels or IFN-gamma response. This study is the first to report the presence of Neospora DNA in semen and blood of naturally-infected bulls. Our observations indicate intermittent presence of N. caninum in blood and semen and shedding in semen in low numbers. PMID:15725454

  6. Kinetics of fertilization and development, and sex ratio of bovine embryos produced using the semen of different bulls.

    PubMed

    Alomar, M; Tasiaux, H; Remacle, S; George, F; Paul, D; Donnay, I

    2008-08-01

    The between bulls variation in in vitro fertility and the shift of sex ratio towards male embryos are two problems affecting the in vitro production (IVP) of bovine embryos. Our objective was to evaluate the kinetics of fertilization, embryo development and the sex ratio of the resulting embryos using the frozen/thawed semen of four different bulls. In a first experiment, the kinetics of pronucleus (PN) formation was evaluated at 8, 12 and 18 h post-insemination (hpi). Based upon the pronuclei sizes and the distance between the two pronuclei, inseminated oocytes were classified in three PN stages. Differences between bulls were observed at each time point, but were more important at 12 hpi. At 8 and 12 hpi bull III showed a significantly faster PN evolution by comparison with the three other bulls (P<0.05), while at 18 hpi, the proportion of the three PN stages was similar to those of bulls I and IV, bull II being delayed. In a second experiment, the kinetics of in vitro embryo development was compared using time-lapse cinematography. The analysis of embryos reaching the blastocyst stage revealed significant differences in the mean time of first cleavage (range of 22.7-25.6h, P<0.05), while the lengths of the subsequent three cell cycles did not differ between bulls. The early mean time of first cleavage with bull III was associated with an early blastulation and a high blastocyst rate at Day 7, in opposition to what was observed with bull II showing a later timing of first cleavage (first cleavage 22.1 hpi versus 25.5 hpi; blastulation 140.4 hpi versus 152.5 hpi; D7 blastocyst rates: 31.3% versus 21.9%; P<0.05). In a third experiment, 65-76 Day 8 blastocysts per bull were sexed by PCR. Only blastocysts obtained with bull III showed a shift in sex ratio towards male embryos (76% male embryos; P<0.05). Such shift was already observed at the 2-cell and morula stages. In conclusion, the bull influences the kinetics of PN formation, of embryo development and the sex

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

    PubMed Central

    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. PMID:25739957

  8. 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. PMID:16464545

  9. Preliminary study on effects of bovine frozen semen storage using a liquid nitrogen-independent method on the quality of post-thaw spermatozoa.

    PubMed

    Buranaamnuay, K; Seesuwan, K; Saikhun, K

    2016-09-01

    Frozen semen of eight bulls was used to assess effects of storage temperature and length of storage time on frozen-thawed bovine sperm quality. In experiment 1, 25 straws of frozen semen of each bull were allocated to 3 groups. The control was still maintained in liquid nitrogen (LN2). The rest were abruptly moved from LN2 to -80°C and -30°C mechanical freezers, respectively. After thawing, it was found that the sperm motility, vitality and membrane integrity were comparable (P>0.05) between the control and the -80°C samples and were significantly inferior (P<0.001) in the -30°C samples, irrespective of storage time (1-day, 1-week and 1-month). In experiment 2, two out of the three parts (16-18 straws) of frozen semen of each bull were rapidly removed from LN2 and further kept in the freezer (-80°C). One day before being thawed, half of the samples in the freezer were promptly put back to LN2. The results showed that the frozen-thawed sperm quality was not significantly affected (P>0.05) both by storage temperature (-196°C, -80°C and -80 & -196°C) and storage time [day-2, day-8 (1-week) and day-31 (1-month)]. At the same storage times, the quality measures at different temperatures were not significantly different from one another (P>0.05). In conclusion, a -80°C mechanical freezer was as effective as LN2 in preserving in vitro quality of frozen-thawed bovine spermatozoa throughout 1-month of storage. When required for use, frozen semen stored in the freezer could be thawed immediately or transferred to the LN2 tank for thawing elsewhere. PMID:27421230

  10. 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

  11. [Survivability and penetration capacity of bull spermatozoa frozen by 3 technics].

    PubMed

    Korvalan, P; Marinov, M F; Vodas, K

    1986-01-01

    Studied was the survival and penetration capacity of bull spermatozoa frozen after the following technologies--pellet, straw, and minitube--and the results obtained were compared via biologic experiments. The minitube technology of freezing the semen led to higher thermal resistance of the spermatozoa as against freezing in the form of straws and pallets (363.5 +/- 8.02 min, 356.25 +/- 7.79 min, and 339.00 +/- 8.44 min, respectively). The differences established were statistically highly significant (P less than 0.001). The penetration capacity of spermatozoa in an estral secretion of cows was highest at freezing in the form of minitubes (1.70 +/- 0.54 mm/min). The same was lower with straws (1.53 +/- 0.02 mm/min), and lowest--with pellets (1.52 +/- 0.04 mm/min). The differences were statistically significant (P less than 0.01). The fertilization capacity of spermatozoa frozen with the employment of the three technologies was best with the use of minitubes and straws--50.20, resp., 49.67 per cent, and lowest with the use of pellets--45.22 per cent. PMID:3811225

  12. A 31-kDa seminal plasma heparin-binding protein reduces cold shock stress during cryopreservation of cross-bred cattle bull semen.

    PubMed

    Patel, M K; Cheema, R S; Bansal, A K; Gandotra, V K

    2016-10-01

    In the present study, a 31-kDa protein, purified from cattle bull seminal plasma heparin-binding proteins (SP-HBP), was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry. Raw semen of six cross-bred bulls was treated with 31-kDa HBP before cryopreservation to observe its effect on motility, viability, hypo-osmotic swelling test, acrosome integrity, in vitro capacitation/acrosome reaction, and oxidative stress at pre-freeze and frozen-thawed phases of cryopreservation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 31-kDa protein eluted and purified from SP-HBP (separated on acrylamide gels) resulted in a single band of 40 kDa. In matrix-assisted laser desorption/ionization-time of flight analysis, 12 peptides were identified with matching significantly (P < 0.05) to interlukin-6 of bovine with a top score of 55. Addition of 25 μg/mL of fluorescein isothiocyanate-conjugated 31-kDa protein to raw semen and incubation at 37 °C for 20 minutes before cryopreservation resulted in its binding mainly to head region. Treatment of semen with 31-kDa HBP resulted in a significant (P < 0.05) average increase of 9.2%, 6.8%, and 11.7% and 5.5%, 6.5%, and 11.0% in motile, viable, hypo-osmotic swelling-responsive spermatozoa in six bulls at pre-freeze and frozen-thawed phases of cryopreservation, respectively. Percentage of spermatozoa with intact acrosomes nonsignificantly enhanced in the semen treated with 31-kDa HBP at both phases of cryopreservation. An average nonsignificant increase of 3.1% in in vitro capacitated and acrosome-reacted spermatozoa was obtained in semen supplemented with 31-kDa HBP. Addition of 31-kDa HBP also nonsignificantly reduced Malonadialdehyde (MDA) level by 10.7 and 19.3 μM/10(9) spermatozoa in prefrozen and frozen-thawed semen, respectively. The results obtained here indicate to conclude that treatment of cross-bred cattle bull semen with 31-kDa HBP protects the spermatozoa

  13. The effect of cysteine and glutathione on sperm and oxidative stress parameters of post-thawed bull semen.

    PubMed

    Tuncer, Pürhan Barbaros; Bucak, Mustafa Numan; Büyükleblebici, Serhat; Sarıözkan, Serpil; Yeni, Deniz; Eken, Ayşe; Akalın, Pınar Peker; Kinet, Hüseyin; Avdatek, Fatih; Fidan, A Fatih; Gündoğan, Mustafa

    2010-12-01

    The aim of this study was to determine the effects of antioxidants such as reduced glutathione (GSH) and cysteine in Laiciphose® extender on semen parameters, fertilizing ability, lipid peroxidation (LPO) level and glutathione peroxidise (GPx) activity of post-thawed bull semen. Totally 54 ejaculates of three bulls were used in the study. Five groups, namely; GSH (0.5 and 2 mM), cysteine (5 and 10 mM) and control group, were conducted to test the antioxidants in Laiciphose®. Insemination doses were processed that each 0.25-mL straw contained 15 x 10⁶ sperm. The addition of antioxidants did not present any significant effect on the percentages of post-thaw sperm morphology (acrosome and total abnormalities), subjective, CASA and progressive motilities, as well as sperm motility characteristics (VAP, VSL, VCL, LIN and ALH), compared to the control groups (P > 0.05). GSH 0.5mM (55.5±7.38%) and cysteine 10 mM (48±5.65%) led to lower rates of DNA damage, compared to control (P < 0.05). As regards to MDA level, cysteine at 10 mM dose gave the highest level (4.99±0.44 nmol/L) (P < 0.001). GPx activity was demonstrated to be higher level upon the addition of 5 mM cysteine when compared to the other groups (P < 0.05). With respect to fertility results based on 60-day non-returns, the supplementation of antioxidants did not present significant differences (P > 0.05). The results of this study may provide an useful information for the future studies in this area. So, further studies could be suggested to achieve better information in terms of the DNA damage and fertilizing capacity of bull sperm frozen with effective antioxidants. PMID:20951122

  14. 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

  15. 19 Beef cattle pregnancy rates following insemination with aged frozen angus semen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Artificial insemination has proven to be a valuable asset to the cattle industry. It is assumed that once good quality semen is frozen in liquid nitrogen it should remain viable indefinitely; however, semen viability has not been systematically evaluated after being stored for several decades. In th...

  16. Minimum number of spermatozoa per dose in Mediterranean Italian buffalo (Bubalus bubalis) using sexed frozen semen and conventional artificial insemination.

    PubMed

    Gaviraghi, A; Puglisi, R; Balduzzi, D; Severgnini, A; Bornaghi, V; Bongioni, G; Frana, A; Gandini, L M; Lukaj, A; Bonacina, C; Galli, A

    2013-05-01

    In buffaloes, AI with sexed semen is not fully optimized, and the procedure has only been performed using the approach currently in use for cattle. The objective of the present work was to compare the pregnancy rates in Mediterranean Italian buffalo cows inseminated with sexed frozen-thawed semen at 2, 4, 6, and 8 million sperm per dose, using the Ovsynch protocol and conventional AI at a fixed time. Fresh ejaculates from three buffalo bulls were processed according to Beltsville sperm sorting technology, and packaged in 0.25-mL straws with two total concentrations of 2 and 4 million live sorted sperm per straw. After thawing, semen was evaluated for total motility, forward motility, average path velocity, membrane and DNA integrity, and membrane fluidity. Sorting efficiency was estimated using a real time polymerase chain reaction method developed and validated in our laboratory. The artificial inseminations were conducted during the breeding season on 849 Italian Mediterranean buffalo heifers and cows distributed in 13 farms in northern and central Italy. No significant difference in quality parameters was reported between nonsexed and sexed straws produced with 2 and 4 million sperm. Lower pregnancy rate (P < 0.001) was reported when inseminating doses of sexed semen at 2 million were used (53/170; 31.2%), with respect to conventional nonsexed (78/142; 54.9%), and sexed doses at 4, 6, and 8 million spermatozoa (102/205, 49.8%; 84/175, 48.0%; and 74/157, 47.1%, respectively). No differences were evident using conventional doses and sexed semen with sperm numbers equal or higher than 4 million per dose. Pregnancies were not affected by the sire; 39/82 (47.6%), 120/270 (44.4%), and 151/355 (42.5%), respectively, for the three bulls. Variability in pregnancy rates observed in different herds was not significant. Furthermore, no significant difference was reported between pregnancies obtained with sexed semen in heifers and multiparous, respectively, 179/407 (44

  17. No detection of Besnoitia besnoiti DNA in the semen of chronically infected bulls.

    PubMed

    Esteban-Gil, A; Grisez, C; Prevot, F; Florentin, S; Decaudin, A; Picard-Hagen, N; Berthelot, X; Ronsin, P; Alzieu, J P; Marois, M; Corboz, N; Peglion, M; Vilardell, C; Liénard, E; Bouhsira, E; Castillo, J A; Franc, M; Jacquiet, P

    2014-06-01

    Bovine besnoitiosis is a chronic and debilitating disease observed in many European countries that may cause important economic losses in cattle. The recent widespread of the parasite in Europe had led the European Food Safety Authority to declare bovine besnoitiosis as a re-emerging disease in Europe. Many aspects of the epidemiology of bovine besnoitiosis such as the main routes of transmission are still unclear and need to be further studied. Among the different hypotheses, a sexual transmission has not yet been investigated. Therefore, the aim of this study was to evaluate the presence of Besnoitia besnoiti DNA in the semen of naturally infected bulls by using a highly sensitive method (real-time qPCR). Both pre-sperm and sperm fractions of 40 bulls, including seronegative (n = 11), seropositive subclinically (n = 17), and seropositive clinically (n = 12) infected animals, were collected by electroejaculation and analyzed by real-time qPCR. No B. besnoiti DNA was detected in 27 pre-sperm and 28 sperm fractions of the 40 examined bulls, suggesting that the transmission of B. besnoiti infection by the semen of chronically infected bulls is very unlikely. PMID:24802865

  18. Detection of Paratuberculosis in Breeding Bulls at Pakistani Semen Production Units: A Continuous Source of Threat

    PubMed Central

    Abbas, Muhammad; Munir, Muhammad; Khaliq, Syed Abdul; Haq, Muhammad Ikram Ul; Tanveer Khan, Muhammad; Qureshi, Zafar ul Ahsan

    2011-01-01

    Paratuberculosis is a chronic bowel disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Its secretion through semen highlights the importance of paratuberculosis-free breeding bulls. The breeding and teaser bulls at three semen production units (SPUs) located in Punjab, Pakistan, were screened for the presence of antibodies against MAP. A total of 253 samples were collected from SPUs and a commercially available indirect screen ELISA (Is-ELISA) was applied. Is-ELISA detected antibodies in 20 (24.6%), 16 (22.8%), and 17 (16.6%) samples from SPU-I, SPU-II, and SPU-III, respectively. Collectively, seroprevalence of 20.0% (47/235) in breeding bulls and 33.3% (6/18) in teaser bulls was observed, and thus it poses a potential threat of disease spread to a high number of heifers and cows through artificial insemination. Therefore, this paper highlights the presence of the disease for the first time at SPUs and triggers attempts to ascertain the prevalence of paratuberculosis throughout the country. PMID:23738098

  19. Sexual behavior and its relationship with semen quality parameters in Sahiwal breeding bulls

    PubMed Central

    Singh, Shushant; Bhakat, M.; Mohanty, T. K.; Kumar, A.; Gupta, A. K.; Chakravarty, A. K.; Singh, P.

    2015-01-01

    Aim: The study was conducted at Artificial Breeding Research Centre, NDRI, Karnal, to determine the sexual behavior and its relationship with semen quality parameters in Sahiwal breeding bulls. Materials and Methods: A total of 63 ejaculates were collected from six adult Sahiwal bulls (age ~47 mo and bwt ~466 kg), to study the relationship of sexual behavior and semen quality. The degree of association between different variables was estimated by Pearson’s correlation coefficient method. Results: The results depicted that, sexual aggressiveness showed significantly high positive correlation with libido score (LS) and sexual behavior score (SBS). Reaction time (RT) and total time taken in mounts (TTTM) had a significant negative correlation with LS and SBS. Penile erection score and penile protrusion score (PPS) both had a significant positive correlation with ejaculatory thrust score, mating ability score, and SBS. Results of correlation among seminal attributes and with sexual behavior depicted that ejaculate volume had positive significant correlation with initial progressive motility (IPM), sperm concentration (SCON), head abnormality, total abnormality, hypo-osmotic swelling test (HOST), acrosomal integrity (AI) whereas, mass activity had positive significant correlation with IPM, SCON, non-eosinophilic spermatozoa count (NESC), HOST, AI, RT and TTTM and IPM had positive significant correlation with SCON, NESC, HOST, AI, and TTTM, whereas and HOST had positive significant correlation with AI. Among seminal attributes, SCON had a positive significant correlation with PPS where as head abnormalities had a positive significant correlation with RT and TTTM. Conclusion: It can be concluded that the relationship of sexual behavior and semen quality parameters are reflecting that the sexual behavior of individual bulls is important to harvest good quality and quantity of semen as desired type of sexual preparation can be provided. PMID:27065641

  20. Reducing sperm concentration is critical to limiting the oxidative stress challenge in liquid bull semen.

    PubMed

    Murphy, C; Fahey, A G; Shafat, A; Fair, S

    2013-07-01

    Because of the short breeding season, the use of liquid bull semen is a viable option in seasonal grass-based dairy systems such as Ireland. Currently in Ireland, liquid bull semen contains approximately 5 million sperm per insemination dose and is used within 2.5d of collection. The hypothesis of this study was that reducing the sperm number per insemination dose would enable bull sperm to be stored for longer. Semen was collected at a commercial AI center and diluted to 1 (T1), 2 (T2), 3 (T3), 4 (T4), and 5 (T5) million sperm per 0.25-mL dose in caprogen diluent. On d 0.25 (6 h postcollection), 1, 2, 3, 4, and 5 postcollection, viability, oxidative stress, and mitochondrial activity were assessed using flow cytometry and the fluorescent probes propidium iodide, CM-H2DCFDA, and rhodamine 123, respectively. On the same days, glucose consumption, total antioxidant capacity, and progressive linear motility were assessed. We observed an effect of day and treatment on sperm cell viability, with the highest percentage live found in T 0005 and the lowest in T 0025 on all days. Oxidative stress in live sperm increased with duration of storage and was affected by treatment, being highest in T 0025 and lowest in T 0005 on all days (d 5: 56.4±2.76% and 28.8±1.22%, respectively; mean ± SEM). Both the total antioxidant capacity and percentage of live sperm positive for rhodamine 123 were unaffected by treatment. The concentration of glucose in caprogen declined with time and was lowest in T 0025 and highest in T 0005 on d 5. In conclusion, higher concentrations of sperm have detrimental effects on sperm cell viability and increase oxidative stress but have no effect on the mitochondrial activity of sperm. PMID:23660140

  1. 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

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

    PubMed

    Franks, Marcy K; Grandin, Temple

    2015-01-01

    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

  3. 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

  4. TEMPORARY STORAGE OF BOVINE SEMEN CRYOPRESERVED IN LIQUID NITROGEN ON DRY ICE AND REFREEZING OF FROZEN-THAWED SEMEN.

    PubMed

    Abdussamad, A M; Gauly, M; Holtz, W

    2015-01-01

    Two experiments were conducted. The purpose of Experiment 1 was to investigate whether viability of bovine semen stored in liquid nitrogen (-196°C) will be adversely affected by temporary exposure to dry ice (-79°C). It was convincingly shown that post thaw-motility was not affected, regardless whether semen was thawed immediately or after being returned to liquid nitrogen. Shipping or temporary storage on dry ice, thus, is a viable option. In Experiment 2, refreezing of frozen-thawed semen was attempted. The proportion of motile spermatozoa was reduced by a factor of ten to between 6.0 % and 7.4 %, regardless whether thawing occurred directly after removal from liquid nitrogen or after an interim period on dry ice. When semen was refrozen on dry ice before being returned to liquid nitrogen, motility rates were significantly improved (13.0 % to 17.0 %, P<0.05). In both experiments sperm cells that remained motile displayed vigorous forward movement and normal morphological appearance. PMID:26576003

  5. Semen quality parameters as fertility predictors of water buffalo bull spermatozoa during low-breeding season.

    PubMed

    Ahmed, Hussain; Andrabi, S Murtaza Hassan; Jahan, Sarwat

    2016-10-01

    The present study was carried out to assess various postthaw semen quality parameters for the prediction of fertility in buffalo bull during low-breeding season. Semen (30 ejaculates) was collected from five adult buffalo bulls with artificial vagina (42 °C). Sperm motility parameters, velocity distribution, motion kinematics, and subpopulations were analyzed by computer-aided sperm motion analyzer (CASA). Moreover, sperm visual motility, supravital plasma membrane integrity, viability/acrosome integrity, viability/mitochondrial transmembrane potential, DNA fragmentation/integrity, and morphology were analyzed by phase-contrast microscope, supravital hypoosmotic swelling test, Trypan blue/Giemsa staining, propidium iodide/"5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide" (JC-1) fluorochromes, neutral comet assay/acridine orange assay and wet mount technique, respectively. Outcome of 528 inseminations was analyzed for in vivo fertility. Pearson's correlation coefficients revealed that sperm progressive motility (%), rapid velocity (%), average path velocity (μm/s), straight line velocity (μm/s), subpopulation one (most rapid, and progressive) of motile spermatozoa (%), supravital plasma membrane integrity (%), and viable spermatozoa with intact acrosome (%) were significantly correlated with in vivo fertility (r = 0.64, P < 0.01; r = 0.57, P < 0.01; r = 0.52, P < 0.01; r = 0.56, P < 0.01; r = 0.73, P < 0.001; r = 0.74, P < 0.001; r = 0.88, P < 0.001); whereas nonviable spermatozoa with damaged acrosome or low-mitochondrial transmembrane potential and comet length (μm) of neutral comet assay were negatively associated with in vivo fertility (r = -0.79, r = -0.75, P < 0.001, and r = -0.60, P < 0.05, respectively). Multiple regression analysis reported that combination of semen quality parameters as predictor of fertility were better (R(2) adjusted = 81.30%, P < 0.001) as compared with single parameter (R(2

  6. 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. PMID:27509873

  7. 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

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

    PubMed Central

    Sharafi, Mohsen; Zhandi, Mahdi; Shahverdi, Abdolhossein; Shakeri, Malak

    2015-01-01

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

  9. Insemination of susceptible heifers with semen from a non-viraemic bull with persistent bovine virus diarrhoea virus infection localized in the testes.

    PubMed

    Niskanen, R; Alenius, S; Belák, K; Baule, C; Belák, S; Voges, H; Gustafsson, H

    2002-06-01

    Bulls shedding bovine viral diarrhoea virus (BVDV) in semen and simultaneously having a high concentration of circulating antibodies may cause reproductive problems and spread the viral infection within cattle populations. To investigate this in detail, three heifers were inseminated with BVDV-infected semen from a non-viraemic, seropositive Holstein-Friesian bull, named 'Cumulus'. One control heifer was inseminated with semen from a healthy bull that was free of BVDV. All four heifers remained clinically healthy throughout the experiment. The conception succeeded in the control animal and in two of the three heifers inseminated with semen containing BVDV. The heifer with the failed conception was the only one that became systemically infected with BVDV. This animal was deemed non-pregnant by ultrasonic examination on day 34 after insemination and showed no signs of subsequent oestrus during the entire experimental period. At slaughter, 42 days after insemination, there were no histopathological changes in the ovaries and virus was not detected in ovarian tissue. The fact that seronegative dams served with semen from persistently infected bulls have occasionally produced persistently infected calves together with the present findings and the fact that non-viraemic, seropositive bulls can constantly shed BVDV, suggest that the use of semen from such bulls in BVDV-free herds could have far-reaching consequences, especially if it led to the birth of persistently infected (P1) calves. PMID:12071892

  10. Integrated evaluation of scrotal temperature and testosteronemia after GnRH administration in young bulls with low semen production.

    PubMed

    Vencato, J; Cestaro, L; Vazzana, I; Carrer, G; Carlo, E; Dara, S; Stelletta, C

    2014-06-01

    The aim of this study was to determine the suitability of thermographic monitoring of scrotal surface temperature (SST) as a method to monitor testicular function. Yearling bulls (n = 23) with low semen production were selected. Scrotal surface temperature and serum testosterone (T) concentrations were evaluated before and after administration of 10.5 μg buserelin acetate IV. Thermographic images of scrotum were recorded at 0, 15, 30, 45 and 60 min post-GnRH, while blood sampling was only performed at 60 min post-GnRH. Bulls were divided in two groups: LowTemp bulls (n = 10) had a decreased SST at 60 min; HighTemp bulls (n = 13) had an increased SST. After 60 min, LowTemp bulls had higher T concentrations compared to HighTemp bulls: 14.32 ng/ml ± 0.53 vs 10.30 ± 1.37 ng/ml (mean ± SEM; p < 0.05), respectively. Reproductive performances in both groups improved after GnRH administration, resulting in an increased number of inseminating doses from each collection, which was higher in LowTemp bulls. Pearson correlation test showed a negative relationship between T and SST (r = -0.554). In conclusion, a decreased scrotal surface temperature 60 min after GnRH treatment was associated with improved semen production. PMID:24750418

  11. 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

  12. 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. PMID:27173954

  13. 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. PMID:23279520

  14. 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

  15. Non-surgical intrauterine artificial insemination in bitches using frozen semen.

    PubMed

    Wilson, M S

    1993-01-01

    A total of 46 bitches were inseminated directly into the uterus using non-surgical insemination procedures; the technique used in six bitches involved specially designed metal catheters and abdominal fixation of the cervix, whereas the remainder were inseminated by passing a flexible plastic catheter through the cervix using direct endoscopic visualization to facilitate the process. Twenty-seven bitches were inseminated with semen frozen at the clinic; the remainder were inseminated with imported semen. Insemination timing was based on endoscopic assessment of the vaginal mucosa, vaginal cytology and blood progesterone concentration determined using a rapid, qualitative enzyme-linked immunosorbent assay (ELISA) kit. Each bitch received between 50 x 10(6) and 200 x 10(6) total spermatozoa per insemination; post-thaw motility varied from 10 to 80%. Two inseminations were performed 48 h apart in the majority of bitches. An overall pregnancy rate of 80% (37/46) was obtained with a mean litter size of 5 +/- 3.14. Subsequent pregnancy rates were comparable for both techniques and both were considered to be effective methods of inseminating frozen semen. Considerably fewer spermatozoa were inseminated in many of these bitches than have previously been reported. In a series of seven bitches using the semen from one dog, each bitch received two inseminations of 30-35 x 10(6) live normal spermatozoa per insemination. A pregnancy rate of 85% (6/7) and a mean litter size of 7.8 was achieved. Rapid ELISA progesterone kits were used to identify the optimum time for insemination. PMID:8229942

  16. 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

  17. Fertilizing potential in vitro of semen from young beef bulls containing a high or low percentage of sperm with a proximal droplet.

    PubMed

    Amann, R P; Seidel, G E; Mortimer, R G

    2000-12-01

    Fertilizing potential of semen containing a high percentage of sperm with a proximal droplet was evaluated using IVF. Design criteria: (a) specified semen with >100 x 10(6) sperm/mL with >40% progressively motile spermatozoa, after collection via electro-stimulation; (b) designated a droplet group, bulls whose semen contained >30% spermatozoa with a proximal droplet and <25% with other morphological abnormalities, and a control group, with <25% abnormalities of any type; and (c) stipulated evaluations at 11 to 13 mo of age and again -4 wk later. At the initial evaluation, when a bull was assigned to the droplet group, the next bull meeting control criteria was designated his pair; 15 pairs in four herds were studied. Semen was extended in egg-yolk citrate, cooled to 5 degrees C over approximately 2.5 h, and held at 5 degrees C. After 20 to 44 h, spermatozoa were processed by swimup, incubated with heparin, and co-cultured with oocytes (35 to 56 oocytes/sample; 18 h). Ova were observed for cleavage approximately 42 h after co-culture, and further development was evaluated on day 8. At first evaluation, cleavage rates were 18 and 46% for droplet and control groups (P < 0.01); semen had 34 to 70% and 0 to 12% droplet spermatozoa. For 10 of 15 droplet bulls, <10% of ova were cleaved whereas cleavage rate was >15% for all control bulls. At second evaluation, only three droplet bulls still had >30% of spermatozoa with a proximal droplet. Cleavage rates increased accordingly; only four droplet bulls had <10% cleaved ova and 10 had >34% cleaved ova. Three control bulls had <10% cleaved ova and nine had > or = 34% cleaved ova. Considering all 60 ejaculates, correlation between percentage of spermatozoa with a proximal droplet and percentage of cleaved ova was -0.49 (P < 0.0 1). Correlations between percentages of motile or normal spermatozoa in field evaluations and outcome in IVF were 0.28 and 0.52. Laboratory evaluations of spermatozoa concomitant with preparation for IVF

  18. 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. PMID:27272488

  19. 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.

  20. French field results (1985-2005) on factors affecting fertility of frozen stallion semen.

    PubMed

    Vidament, M

    2005-10-01

    Results on procedures for freezing stallion semen and the subsequent fertility during 20 years are presented. The present system applied in French National Stud includes: (1) a freezing protocol (dilution in milk, centrifugation and addition of freezing extender (INRA82+egg yolk (2%, v/v)+glycerol (2.5%, v/v) at 22 degrees C, a moderate cooling rate to 4 degrees C and freezing at -60 degrees C/min in 0.5-ml straws); (2) selection of ejaculates showing post-thaw rapid motility >35%; and (3) an insemination protocol (mares examined once daily, two AI of 400 x 10(6) spermatozoa 24 h apart before ovulation, sufficient number of straws to have the possibility to perform six AI of 400 x 10(6) total spermatozoa, i.e. 2.4 x 10(9) total spermatozoa available per mare per season). This system was applied to >110 stallions per year, the average post-thaw motility of ejaculates was 50% (>1800 ejaculates) before selection. The semen freezability was defined as the number of selected ejaculates divided by the total number of ejaculates frozen. Of the stallions, 5, 4, 5, 21 and 64% had semen freezability of 0-10, 10-33, 33-60, 60-90 and over 90%, respectively. Per-cycle pregnancy rate was 45-48% (>1500 mares per year, 1.8 cycles per mare) and foaling rate 64%. In comparison, per-cycle pregnancy rate and foaling rate of mares hand-mated to stallions were 57-59% and 64%, respectively. The average number of straws used was 32-35 (1.75 x 10(9) total spermatozoa) per mare per season. According to our results and the literature, the most important factors for improving fertility of frozen equine semen include: (1) a low concentration of glycerol (2-3.5% final concentration); (2) a suitable base extender for freezing like Lactose-Glucose EDTA or INRA82; (3) a post-thaw motility >30-35%; and (4) a sufficient number of spermatozoa per mare per season (1.5-2 x 10(9) total spermatozoa for two to three cycles) divided into small units. Numbers of spermatozoa, lower than 750.10(6) total

  1. 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

  2. Effect of Various Concentrations of Caffeine, Pentoxifylline, and Kallikrein on Hyperactivation of Frozen Bovine Semen

    PubMed Central

    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. PMID:25950005

  3. Effects of the platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen (AI) with reduced sperm motility.

    PubMed

    Lecewicz, M; Kordan, W; Majewska, A; Kamiński, S; Dziekońska, A; Mietelska, K

    2016-01-01

    The aim of the study was to determine the effects of platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen with reduced sperm motility used for artificial insemination. The aim of experiment 1 was to identify the optimal concentration of the phospholipid able to preserve sperm viability. Cryopreserved semen was treated with different PAF concentrations: 1×10(-5) M, 1×10(-6) M, 1×10(-7) M, 1×10(-8) M and 1×10(-9) M. The experiment demonstrated that PAF at concentration 1×10(-9) M increased most the sperm viability parameters (motility parameters, plasma membrane integrity and mitochondrial function) after 120 min of incubation of thawed semen at 37°C. Cryopreserved bull semen with reduced sperm motility (below 70%) was supplemented with PAF in a concentration of 1×10(-9) M. A statistically significant increase in sperm motility, percentage of linear motile spermatozoa and VSL value was observed after 120 min incubation of sperm with 1×10(-9) M PAF. Sperm supplementation with PAF also had positive effects on plasma membrane integrity and percentage of spermatozoa with preserved mitochondrial transmembrane potential, but the differences were not statistically significant. The results indicated positive effects of PAF supplementation at a concentration of 1×10(-9) M on the selected sperm quality parameters in cryopreserved bull semen with reduced motility. PMID:27096799

  4. Differences in CASA output according to the chamber type when analyzing frozen-thawed bull sperm.

    PubMed

    Ibănescu, Iulian; Leiding, Claus; Ciornei, Ştefan Gregore; Roșca, Petru; Sfartz, Ioana; Drugociu, Dan

    2016-03-01

    As demonstrated by some authors, the type of analyzing chamber can greatly influence the results of computer-assisted sperm analysis (CASA). This study aimed to compare three of the disposable chamber types currently available on the market and to determine whether the CASA output may be significantly different among them. The semen from five Fleckvieh bulls was analyzed by CASA using three different disposable chambers: Leja (20μm), MofA (20μm) and Minitube (20μm), at three different time points: immediately after filling the chamber, at 6min, and also at 12min after filling. Sperm concentration was also determined using the Nucleocounter® NC-100™ device and the hemocytometer as standard methods. The results showed higher values in terms of total and progressive sperm motility for MofA compared to the other two chambers immediately after filling (p<0.05), but higher values for Leja and Minitube after 6 and 12min (p<0.05). All three disposable chambers offered lower values for sperm concentration compared to standard methods (Leja: 68.4±4.9×106/mL; MofA: 80.8±9.6×106/mL; Minitube: 67.3±5.4×106/mL; Nucleocounter: 86.5×106/mL; Hemocytometer: 84.0×106/mL). We conclude that for rapid analyses the MofA chambers provide superior results when compared to the other types that we tested. However, when the analysis requires a longer duration, the Minitube type, and especially the Leja type provide a greater degree of confidence. Further, for determining sperm concentration we think that examiners would be more accurate using the Nucleocounter or the hemocytometer and should make use of CASA only when the other methods are not available. PMID:26791331

  5. Chemical sterilisation of Bos indicus bull calves following intratesticular injection of zinc acetate: effects on semen quality and testicular changes.

    PubMed

    Cavalieri, J; Wang, M; Johnson, L

    2015-05-01

    The aim of this study was to determine the effects in Bos indicus bull calves of intratesticular administration of 1mL of either saline (n=9) or one of the two doses of zinc acetate (ZA1, 57.75mg, n=10 or ZA2, 71.75mg, n=10) on semen quality and testicular changes. Semen was collected by electroejaculation on Days 343, 524 and 783 and animals were slaughtered on Day 860. Treatment reduced median maximum number of progressively motile and morphologically normal sperm collected (P=0.001) and the percentage of animals in which sperm were recovered (saline: 100%, 9/9; ZA1: 44.9%, 4/9 and ZA2: 40.0%, 4/10; P=0.013). Compared to saline treated controls, treatment with ZA reduced the mean diameter of the testes after Day 34 of treatment (treatment×time, P=0.013) and total testicular weight at slaughter (treatment: mean±SEM; saline: 569.4±59.0g, ZA1: 249.3±72.9g, ZA2: 247.5±68.1g; P=0.004). Histological changes in testes of bulls treated with ZA were characterized by germ cell depletion, vacuolation of Sertoli cells, interstitial fibrosis, epididymal duct atrophy with variable remnants of testicular tissue and degeneration. We conclude that intratesticular administration of two doses of ZA in B. indicus calves is able to severely impair spermatogenesis and cause varying degrees of testicular degeneration and a reduction in testicular diameter and mass. Further investigation is required to determine ways of obtaining more consistent results from treatment. PMID:25752498

  6. 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...

  7. Association of heat shock protein 90 with motility of post-thawed sperm in bulls.

    PubMed

    Zhang, Xiao-Gang; Hu, Shan; Han, Cong; Zhu, Qing-Chao; Yan, Guan-Jie; Hu, Jian-Hong

    2015-04-01

    The correlation between the 90 kDa heat-shock protein (HSP90) and sperm quality following the process of freezing-thawing in bulls has not been studied clearly. Therefore, the objective of the present was to clarify the relationship between HSP90 level and semen parameters during the process of cryopreservation in bulls. Semen samples from 5 Holstein bulls were obtained by artificial vagina. Characteristics of these semen at three stages (fresh, after equilibration and frozen-thawed), including motility, plasma membrane integrity and acrosome integrity were evaluated. The mRNA expression level of HSP90 at the three stages was evaluated by using quantitative Real-Time PCR. Meanwhile, the protein level of HSP90 expression at the three stages was detected according to Western blot. The results showed that sperm parameters evaluated in fresh semen was the highest in the three groups. Sperm parameters in semen after equilibration were lower than those in fresh semen (P>0.05) and higher than those in post-thawed semen (P<0.05). Sperm parameters in frozen-thawed semen were the lowest among the three groups (P<0.05). This study indicated that HSP90 expression is proportional to sperm quality. HSP90 expression level in fresh semen was significantly higher than that in frozen-thawed semen (P<0.05). Although no significant differences in HSP90 expression were observed between fresh semen and semen after equilibration (P>0.05). Results in this study suggest that HSP90 level in bull spermatozoa was gradually declined following the process of freezing-thawing, and might be associated with sperm motility, plasma membrane integrity and acrosome integrity. PMID:25578982

  8. 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. PMID:26861956

  9. 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

  10. 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. PMID:25459425

  11. 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...

  12. Supplementing cryopreservation media with reduced glutathione increases fertility and prolificacy of sows inseminated with frozen-thawed boar semen.

    PubMed

    Estrada, E; Rodríguez-Gil, J E; Rocha, L G; Balasch, S; Bonet, S; Yeste, M

    2014-01-01

    The main aim of this work was to evaluate how supplementing freezing media with reduced glutathione (GSH) affected the 'in vivo' fertilizing ability of boar semen subjected to cryopreservation procedures. With this purpose, 12 ejaculates coming from 12 boars were cryopreserved in the presence or absence of 2 mm GSH, whereas the same number of extended ejaculates coming from the same boars was used as negative/farm controls. Eight different sperm parameters (levels of free-cysteine residues in sperm nucleoproteins, DNA fragmentation, sperm viability, acrosome-membrane integrity, intracellular peroxide and superoxide levels, and total and progressive sperm motility) were evaluated before freezing and after 30 and 240 min of thawing. In addition, a total of 180 multiparous sows were used in the field fertility trials, the females being randomly divided into three groups and inseminated with extended, frozen-thawed control or frozen-thawed semen supplemented with 2 mm GSH. The presence of GSH in the freezing media significantly (p < 0.05) increased farrowing rates and the number of total born piglets and alive born piglets, and partially counteracted the cryopreservation-induced damages inflicted on frozen-thawed spermatozoa. We can thus conclude that supplementing freezing media with 2 mm GSH greatly improves boar sperm cryopreservation technology, as it significantly improves the fertilizing ability of frozen-thawed spermatozoa. PMID:24123940

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

    PubMed

    Purdy, P H; Tharp, N; Stewart, T; Spiller, S F; Blackburn, H D

    2010-10-15

    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 influence the fertility of boar sperm. Therefore, the purpose of this study was to determine the effects of pH and storage temperature on fresh and frozen-thawed boar sperm motility end points. Semen samples (n = 199) were collected, diluted, cooled and shipped overnight to the National Animal Germplasm Program laboratory for freezing and analysis from four boar stud facilities. The temperature, pH and motility characteristics, determined using computer automated semen analysis, were measured at arrival. Samples were then cryopreserved and post-thaw motility determined. The commercial stud was a significant source of variation for mean semen temperature and pH, as well as total and progressive motility, and numerous other sperm motility characteristics. Based on multiple regression analysis, pH was not a significant source of variation for fresh or frozen-thawed boar sperm motility end points. However, significant models were derived which demonstrated that storage temperature, boar, and the commercial stud influenced sperm motility end points and the potential success for surviving cryopreservation. We inferred that maintaining cooled boar semen at approximately 16 °C during storage will result in higher fresh and frozen-thawed boar sperm quality, which should result in greater fertility. PMID:20494424

  14. Association of TNP2 Gene Polymorphisms of the bta-miR-154 Target Site with the Semen Quality Traits of Chinese Holstein Bulls

    PubMed Central

    Huang, Jinming; Zhang, Xiaojian; Qi, Chao; Li, Jianbin; Zhong, Jifeng; Li, Guorong; Wang, Changfa

    2014-01-01

    Transition protein 2 (TNP2) participates in removing nucleohistones and the initial condensation of spermatid nucleus during spermiogenesis. This study investigated the relationship between the variants of the bovine TNP2 gene and the semen quality traits of Chinese Holstein bulls. We detected three single nucleotide polymorphisms (SNPs) of the TNP2 gene in 392 Chinese Holstein bulls, namely, g.269 G>A (exon 1), g.480 C>T (intron 1), and g.1536 C>T (3′-UTR). Association analysis showed that the semen quality traits of the Chinese Holstein bulls was significantly affected by the three SNPs. The bulls with the haplotypic combinations H6H4, H6H6, and H6H8 had higher initial semen motility than those with the H7H8 and H8H4 haplotypic combinations (P<0.05). SNPs in the microRNA (miRNA) binding region of the TNP2 gene 3′-UTR may have contributed to the phenotypic differences. The phenotypic differences are caused by the altered expression of the miRNAs and their targets. Bioinformatics analysis predicted that the g.1536 C>T site in the TNP2 3′-UTR is located in the bta-miR-154 binding region. The quantitative real-time polymerase chain reaction results showed that the TNP2 mRNA relative expression in bulls with the CT and CC genotypes was significantly higher than those with the TT genotype (P<0.05) in the g.1536 C>T site. The luciferase assay also indicated that bta-miR-154 directly targets TNP2 in a murine Leydig cell tumor cell line. The SNP g.1536 C>T in the TNP2 3′-UTR, which altered the binding of TNP2 with bta-miR-154, was found to be associated with the semen quality traits of Chinese Holstein bulls. PMID:24416221

  15. Effects of extender and equilibration time on post-thaw motility and membrane integrity of cryopreserved Gyr bull semen evaluated by CASA and flow cytometry.

    PubMed

    Leite, Ticiano Guimarães; do Vale Filho, Vicente Ribeiro; de Arruda, Rubens Paes; de Andrade, André Furugen Cesar; Emerick, Lucas Luz; Zaffalon, Fabiane Gilli; Martins, Jorge André Matias; de Andrade, Venício José

    2010-07-01

    The objectives of the present study were to investigate the effects of three equilibration times (0, 2, and 4h) and two extenders (TRIS or Bioxcell) for cryopreservation of bull semen. Semen from 12 Gyr bulls was cryopreserved using an automated freezing machine. There were significant interactions between equilibration times and extenders for sperm motility and membrane integrity. The control treatment (0h equilibration) had the lowest values (P<0.05) for total (MOT) and progressive motilities (PROG), and percentage of sperm with intact plasma and acrosomal membranes (IPIA), with no significant differences between extenders. Extender TRIS had greater cryoprotective action than Bioxcell, with greater MOT, PROG, IPIA at 2 and 4h, as well as the lowest proportion of damaged plasma membrane (DPM, 72.2% vs. 85.8%) for all times. Equilibration for 4h yielded the most desirable (P<0.05) for MOT, PROG, and IPIA, and the least DPM percentage (86.5, 78.0, and 72.6% for 0, 2, and 4h, respectively). Overall, the combination of TRIS and 4h of equilibration was the most desirable semen cryopreservation method, with greatest MOT, PROG, and IPIA (TRIS-T4=26.8%; BIO-T4=18.3%) and the least DPM. In conclusion, based on objective analyses, equilibration during cryopreservation was essential for maintaining motility and integrity of sperm membranes; equilibration for 4h yielded the greatest sperm survival, independent of the extender used. PMID:20434857

  16. Liquid and Frozen Storage of Agouti (Dasyprocta leporina) Semen Extended with UHT Milk, Unpasteurized Coconut Water, and Pasteurized Coconut Water

    PubMed Central

    Mollineau, W. M.; Adogwa, A. O.; Garcia, G. W.

    2011-01-01

    This study evaluated the effects of semen extension and storage on forward progressive motility % (FPM%) in agouti semen. Three extenders were used; sterilized whole cow's milk (UHT Milk), unpasteurized (CW) and pasteurized coconut water (PCW), and diluted to 50, 100, 150, and 200 × 106 spermatozoa/ml. Experiment 1: 200 ejaculates were extended for liquid storage at 5∘C and evaluated every day for 5 days to determine FPM% and its rate of deterioration. Experiment 2: 150 ejaculates were extended for storage as frozen pellets in liquid nitrogen at −195∘C, thawed at 30∘ to 70∘C for 20 to 50 seconds after 5 days and evaluated for FPM% and its rate of deterioration. Samples treated with UHT milk and storage at concentrations of 100 × 106 spermatozoa/ml produced the highest means for FPM% and the slowest rates of deterioration during Experiment 1. During Experiment 2 samples thawed at 30∘C for 20 seconds exhibited the highest means for FPM% (12.18 ± 1.33%), 85% rate of deterioration. However, samples were incompletely thawed. This was attributed to the diameter of the frozen pellets which was 1 cm. It was concluded that the liquid storage method was better for short term storage. PMID:20871831

  17. Methodological factors affecting the results of staining frozen-thawed fertile and subfertile Japanese Black bull spermatozoa for acrosomal status.

    PubMed

    Almadaly, Essam; El-Kon, Ismail; Heleil, Bassiouni; Fattouh, El-Sayed; Mukoujima, Koushi; Ueda, Takuya; Hoshino, Youichirou; Takasu, Masaki; Murase, Tetsuma

    2012-12-01

    In the present study, some methodological factors affecting the acrosomal staining of frozen-thawed Japanese Black bull spermatozoa were investigated by examining; the effect of fixation/permeabilization procedure on intact acrosome percentage after fluorescein isothiocyanate peanut agglutinin (FITC-PNA) staining, the acrosomal staining patterns by using two types of fluorescent probes FITC-PSA (Pisum Sativum Agglutinin) and FITC-PNA and the effect of staining methods, either smear or vial, on intact acrosome percentage. Then intact acrosome percentage was compared between the samples stained by thus established method and those simply fixed with glutaraldehyde (glutaraldehyde fixation method). A possibility that FITC-PNA staining or the glutaraldehyde fixation methods could detect any difference in intact acrosome percentage or acrosomal staining patterns between fertile and subfertile bulls was also examined. The results showed that (1) 4% paraformaldehyde fixation plus 1% Triton X-100 permeabilization was better than absolute ethanol alone, (2) FITC-PNA acrosomal labeling was more specific than FITC-PSA, (3) sperm suspensions should be smeared and gently processed before acrosomal staining rather than spotted onto glass slides after staining in vial in order to avoid excessive mechanical damage of the sperm acrosome, and (4) staining spermatozoa with FITC-PNA had no major advantages over examination of simply glutaraldehyde fixed sperm samples and both failed to detect any significant difference in intact acrosome percentage between the fertile and the subfertile bulls used here. The present study demonstrates important methodological considerations which need to be taken into account in order to design a reliable and reproducible protocol for the study of the acrosome. PMID:23182469

  18. Relationship between in vitro sperm functional tests and in vivo fertility of rams following cervical artificial insemination of ewes with frozen-thawed semen.

    PubMed

    O' Meara, C M; Hanrahan, J P; Prathalingam, N S; Owen, J S; Donovan, A; Fair, S; Ward, F; Wade, M; Evans, A C O; Lonergan, P

    2008-03-01

    Several procedures have been proposed to assess structural and functional characteristics of cryopreserved ram semen but none so far have yielded consistent relationships with in vivo fertility. The objectives of this study were to evaluate several sperm function tests as potential markers of in vivo ram fertility (determined by pregnancy rate in ewes) using frozen-thawed semen. In experiment 1, frozen-thawed straws (n=3 per ram) of semen from three high and three low fertility rams were assessed using fluorescent microscopy for (1) progressive motility, (2) viability and, (3) acrosomal status. In experiment 2, frozen-thawed straws (n=3 per ram) of semen from 18 rams of known fertility were analysed using either computer-assisted sperm analysis (CASA) for eight motion characteristics or flow cytometric staining for: (1) viability and acrosomal status, (2) plasma membrane status and capacitation-like changes, and (3) live cells following an osmotic resistance test (ORT). In experiment 3, platelet-activating factor (PAF) was isolated from straws (n=2 per ram) of semen using high-pressure liquid chromatography (HPLC) and quantified using HPLC-tandem mass spectrometry for 18 rams. In experiment 1, no association was found between motility, viability (% live) or acrosomal status (% damaged, % intact and % reacted) and in vivo fertility. In experiment 2, no correlation was found between motility (CASA), viability (% live), acrosomal status (% live, % live intact and % reacted), capacitation status (% capacitated, % non-capacitated), plasma membrane stability (% dead) and % live cells following ORT and ram in vivo fertility. In experiment 3, there was no relationship between PAF content in spermatozoa and ram fertility. In conclusion, we were unable to relate the in vivo fertility of rams with in vitro functional tests of their frozen-thawed semen and suggest that the fertility of a given semen sample cannot easily be quantified using available in vitro tests. PMID

  19. Effects of Adding Sodium Nitroprusside to Semen Diluents on Motility, Viability and Lipid Peroxidation of Sperm in Holstein Bulls

    PubMed Central

    Khodaei, Hamidreza; Chamani, Mohammad; Mahdavi, Behnaz; Akhondi, Ali Asghar

    2016-01-01

    Background Nitric oxide (NO) that plays important role in all sexual activities of animals is made from the amino acid L-arginine by the enzymatic action of NO synthase (NOS). NO makes a band with sulfur-iron complexes, but due to production of steroid sexual hormones related to the enzymes involved in this complex, NO can change the activity of these enzymes. NO affects many cells including vein endothelial cells, macrophages and mast cells. These cells are also found in Leydig cells; therefore, they are important source of NO in testis tissue. Therefore, minimizing damages to sperm at the time of freezing thawing process are really important. The aim of this study was to determine the appropriate NO concentration to be added to the freezing extender to improve the quality of thawed sperm. Materials and Methods In this experimental randomized study, sperms of four Holstein bulls with an average age of 4 were collected twice a week for 3 weeks. They received sodium nitroprusside (SNP) in concentrations of 0, 10, 50 and 100 nmol/ml. Data analysis was performed using the special issue and static (SAS) 98 software. Also, mean comparison was done using Duncan’s multiple ranges test (P<0.05).This research was conducted at the laboratory of Science and Research Branch, Islamic Azad University, Tehran at spring and summer of 2013. Results All concentrations of SNP used was found to increase motility and viability of spermatozoa at 1, 2 and 3 hours after thawing, significantly (P<0.05), but there was no significant difference at zero time. Different concentrations of SNP reduced the membrane lipid peroxidation level of sperm and increased acrosome membranes integrity, implying that SNP generally improved samples membranes, especially in 50 and 100 nmol/ml concentrations. Conclusion According to the obtained results, addition of SNP to semen diluents increases motility and viability of spermatozoa. Also, it reduces membrane lipid peroxidation level that leads to improved

  20. Successful pregnancies with directional freezing of large volume buck semen.

    PubMed

    Gacitua, H; Arav, A

    2005-02-01

    Artificial insemination with frozen-thawed buck semen shows variable results which depend on many factors related to semen quality and the cryopreservation processing. We conducted experiments based on a new freezing method, directional freezing, of large volumes (8 ml). In the first experiment semen from three Saanen bucks, ages 1-2-years-old and genetically selected for milk improvement, was frozen individually. Two to three-years-old Saanen females (n = 164) were synchronized with controlled internal drug release (CIDR), pregnant mare serum gonadotrophin (PMSG) and prostaglandin. Double cervical inseminations were performed with frozen-thawed semen and fresh semen as control. In the second experiment we used pooled, washed frozen semen to examine the effect of washed seminal plasma. The motility after washing was 80-90% and after thawing was 55-65% for all bucks. The sperm concentration increased with the collections and the advance into the breeding season from 1.9 x 10(9) to 4.4 x 10(9) cell/ml average. Two inseminations were carried out at 8h intervals. The first insemination was performed at 32 h after CIDR withdrawal with fresh and frozen-thawed semen. Pregnancy rates were assessed by ultrasonography conducted 40 and 90 days post-insemination (from three bucks). Results were 58, 67, 50% with fresh semen, and for frozen semen were 33, 37 and 53%; these results were significantly different in one of the three bucks (P < 0.005). In the second experiment with pooled, washed semen the pregnancy rate was 41.6%, which compared with the average results of the frozen semen in the first experiment 38.9% no significant difference was found. We conclude that freezing buck semen in large volumes (8 ml) is possible. Cryobanking of buck semen will facilitate a genetic breeding program in goats and preservation of biodiversity. Washed semen did not improve the fertility of the semen when Andromed bull extender is used. PMID:15629809

  1. Conception rate and litter size in multiparous sows after intrauterine insemination using frozen-thawed boar semen in a commercial swine herd in Thailand.

    PubMed

    Chanapiwat, Panida; Olanratmanee, Em-On; Kaeoket, Kampon; Tummaruk, Padet

    2014-10-01

    The aim of the present study was to determine the conception rate and litter size in sows after fixed time intra-uterine insemination using frozen-thawed boar semen in a commercial swine herd in Thailand. Sixty-nine Landrace multiparous sows were randomly allocated into two groups, including control (n=36) and treatment (n=33). The control sows were inseminated with extended fresh semen (3 × 10(9) motile sperm/dose, 100 ml) at 24, 36 and 48 hr after the onset of estrus. The treatment sows were inseminated with frozen-thawed semen (2 × 10(9) motile sperm/dose, 20 ml) at 24 and 36 hr after induction of ovulation by human chorionic gonadotropin. All inseminations were carried out by using an intra-uterine insemination technique. The time of ovulation was determined by using transrectal real-time B-mode ultrasonography. The conception rate, farrowing rate, total number of piglets born/litter (TB) and number of piglets born alive/litter (BA) were evaluated. The sows inseminated with extended fresh semen yield a higher TB (10.8 versus 9.0 piglets/l, P=0.015) and tended to have a higher conception rate (88.9% versus 75.8%, P=0.150) than sows inseminated with frozen-thawed semen. In conclusion, insemination using frozen-thawed boar semen can be practiced with convinced fertility under field conditions by fixed-time intrauterine insemination with 2 × 10(9) sperm/ dose of 20 ml at 24 and 36 hr after the onset of estrus. PMID:24954517

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

    PubMed Central

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

    2014-01-01

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

  3. The g.-165 T>C Rather than Methylation Is Associated with Semen Motility in Chinese Holstein Bulls by Regulating the Transcriptional Activity of the HIBADH Gene

    PubMed Central

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

    2015-01-01

    The 3-hydroxyisobutyrate dehydrogenase (HIBADH) is regarded as a human sperm-motility marker. However, the molecular mechanisms involved in the regulation of expression of the HIBADH gene in bulls remain largely unknown. HIBADH was detected in the testis, epididymis, and sperm via reverse transcription polymerase chain reaction and Western blot analysis. It is also expressed in the seminiferous epithelium, spermatids, and the entire epididymis, as detected by immunohistochemistry. Furthermore, HIBADH was expressed in the neck-piece and mid-piece of bull spermatids, as shown in the immunofluorescence assay. Using serially truncated bovine HIBADH promoters and luciferase constructs, we discovered an 878 bp (-703 bp to +175 bp) fragment that constitutes the core promoter region. One SNP g.-165 T>C of HIBADH was identified and genotyped in 307 Chinese Holstein bulls. Correlation analysis revealed that bulls with the TT genotype had higher initial sperm motility than those with the CC genotype (P < 0.05). Furthermore, the T- or C-containing loci (designated as pGL3-T and pGL3-C) were transiently transfected into MLTC-1 to test the effect of SNP on HIBADH expression. The luciferase reporter assay showed that the pGL3-T genotype exhibited 58% higher transcriptional activity than the pGL3-C genotype (P < 0.05). The bisulfite sequencing analysis revealed that the methylation pattern of the core promoter presented hypomethylation in the ejaculated semen in high-motility and low-motility bulls. The results demonstrated for the first time that the g.-165 T>C rather than methylation in the 5'-flanking region could affect the bovine sperm motility through the regulation of HIBADH gene transcriptional activity. PMID:26133183

  4. The g.-165 T>C Rather than Methylation Is Associated with Semen Motility in Chinese Holstein Bulls by Regulating the Transcriptional Activity of the HIBADH Gene.

    PubMed

    Zhang, Shuai; Zhang, Yan; Yang, Chunhong; Ju, Zhihua; Wang, Xiuge; Jiang, Qiang; Sun, Yan; Huang, Jinming; Zhong, Jifeng; Wang, Changfa

    2015-01-01

    The 3-hydroxyisobutyrate dehydrogenase (HIBADH) is regarded as a human sperm-motility marker. However, the molecular mechanisms involved in the regulation of expression of the HIBADH gene in bulls remain largely unknown. HIBADH was detected in the testis, epididymis, and sperm via reverse transcription polymerase chain reaction and Western blot analysis. It is also expressed in the seminiferous epithelium, spermatids, and the entire epididymis, as detected by immunohistochemistry. Furthermore, HIBADH was expressed in the neck-piece and mid-piece of bull spermatids, as shown in the immunofluorescence assay. Using serially truncated bovine HIBADH promoters and luciferase constructs, we discovered an 878 bp (-703 bp to +175 bp) fragment that constitutes the core promoter region. One SNP g.-165 T>C of HIBADH was identified and genotyped in 307 Chinese Holstein bulls. Correlation analysis revealed that bulls with the TT genotype had higher initial sperm motility than those with the CC genotype (P < 0.05). Furthermore, the T- or C-containing loci (designated as pGL3-T and pGL3-C) were transiently transfected into MLTC-1 to test the effect of SNP on HIBADH expression. The luciferase reporter assay showed that the pGL3-T genotype exhibited 58% higher transcriptional activity than the pGL3-C genotype (P < 0.05). The bisulfite sequencing analysis revealed that the methylation pattern of the core promoter presented hypomethylation in the ejaculated semen in high-motility and low-motility bulls. The results demonstrated for the first time that the g.-165 T>C rather than methylation in the 5'-flanking region could affect the bovine sperm motility through the regulation of HIBADH gene transcriptional activity. PMID:26133183

  5. 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. PMID:27107033

  6. Banking North American buffalo semen.

    PubMed

    Lessard, C; Danielson, J; Rajapaksha, K; Adams, G P; McCorkell, R

    2009-04-15

    The purpose of this study was to develop a procedure to collect and preserve semen from wood bison (Bison bison athabascae) and plains bison (Bison bison bison). Semen samples from three wood and three plains bison bulls were collected by electroejaculation from June through October. In addition, sperm was collected from the cauda epididymis of seven plains bison. Semen was cryopreserved using two commercially available cryopreservation media, an egg yolk-based medium (Triladyl), and a medium free of products of animal origin (Andromed). Sperm morphology and motility were recorded on fresh and post-thawed semen samples. Total sperm motility was not different between plains and wood bison for the months of June (50%), July (69%) and October (54%). However, total sperm motility for wood bison was higher (P<0.05) than plains bison for the months of August and September (August: 80% vs 55%; September: 73% vs 40%). Plains and wood bison did not differ in mean total and mean progressive motility (35 and 15%, respectively) of frozen-thawed sperm samples. The post-thaw motility of Triladyl-treated sperm was higher (P<0.05) than Andromed-treated sperm (35% vs 13%, respectively). Interestingly, post-thawed epididymal spermatozoa had higher total motility (P<0.05) than post-thawed electroejaculated sperm when cryopreserved with a medium free of products of animal origin (Andromed; 35% vs 9%, respectively). In conclusion, we used electroejaculation to collect high quality bison semen, and cryopreserved it for future needs. PMID:19181375

  7. Influence of the uterine inflammatory response after insemination with frozen-thawed semen on serum concentrations of acute phase proteins in mares.

    PubMed

    Tuppits, U; Orro, T; Einarsson, S; Kask, K; Kavak, A

    2014-05-01

    The aim of this study was to investigate the clinical relevance of measuring blood concentrations of serum amyloid A (SAA), haptoglobin (Hp) and fibrinogen (Fib) in horse reproductive management, and changes in response to artificial insemination (AI) with frozen-thawed semen. Standardbred mares (n=18) with different reproductive status (eight healthy mares in first postpartum oestrus, five healthy barren mares and five mares with endometritis) were inseminated with frozen-thawed semen. Endometritis was evaluated during oestrus by bacteriological culture, cytology and presence of ultrasonically visible intrauterine fluid during oestrus. Concentrations of SAA, Hp and Fib were analysed in the blood in every 48h during oestrus and until 5, 6 or 7 days after AI. The day of sampling and number of blood samples varied between mares because of length of the oestrus and time of AI. Changes in concentrations of SAA, Hp and Fib were evaluated based on the day of sampling regard to AI and classification of the mares. There were no differences in SAA, Hp and Fib concentrations over time before or after AI or between the groups of mares. The insemination of mares with frozen-thawed semen did not increase the plasma concentrations of SAA, Hp and Fib above clinical threshold concentration and there were no differences between susceptible or healthy mares. PMID:24636940

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

    PubMed

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

    1984-07-01

    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

  9. 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

  10. 59 Effect of storage duration on post-thaw parameters of bull semen. Reproduction, Fertility and Development 25: 177.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    It has been proposed that once good-quality sperm are collected, extended, and cryopreserved that the post-thaw quality will remain high regardless of the duration of storage. A previous study has suggested that there is no effect on post-thaw sperm motility of bovine semen stored in liquid nitrogen...

  11. Estimation of genetic trends from 1977 to 1998 for farrowing characteristics in the French Large White breed using frozen semen.

    PubMed

    Canario, L; Rydhmer, L; Gogué, J; Tribout, T; Bidanel, J P

    2007-08-01

    The objective of the study was to estimate genetic trends from 1977 to 1998 in the French Large White (LW) breed for stillbirth and associated traits measured at farrowing using frozen semen. Two groups of pigs (G77 and G98) were obtained by inseminating LW sows with semen from LW boars born either in 1977 or in 1998. A second generation was produced by inter se mating in each group. Farrowing was thoroughly supervised through both direct observations and video recording all long farrowing on a total of 137 first- and second-parity litters produced by sows from this second generation (68 G77 and 69 G98 litters, respectively). Measurements included birth time, weight and birth characteristics (including orientation, presence of cyanosis or oedema, membrane obstruction, umbilical cord length/content) of each piglet, as well as sow traits (weight and backfat thickness, farrowing duration, litter size and within-litter variation of weights at birth). The data were analysed using linear or generalised linear mixed models, according to the definition of the trait (continuous or binary data). The importance of several effects to piglet probability of stillbirth was then quantified by computing the reduction of variance associated with the addition of each effect in the model. Litter size did not significantly differ in first parity, but was higher in G98 second-parity sows: the differences for global (including pre partum dead piglets) and total numbers of piglets born per litter were +2.3 ± 1.1 and +1.3 ± 0.6, respectively. G98 sows also had a higher number of stillbirths in both parities (+0.7 ± 0.3 stillborn per litter). Piglets from G98 litters were heavier at birth (+130 ± 40 g for birth weight adjusted for litter size), without any increase in within-litter heterogeneity of birth weight. No significant difference was detected between G77 and G88 groups for farrowing length and the distribution of time interval between piglet births. G98 stillborn piglets had

  12. 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. PMID:26588890

  13. 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

  14. Influence of seminal plasma on leucocyte migration and amount of COX-2 protein in the jenny endometrium after insemination with frozen-thawed semen.

    PubMed

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

    2013-12-01

    After mating, seminal plasma has an immuno-modulatory effect on the endometrium in some mammals. In jennies, achieving conception via artificial insemination (AI) with frozen-thawed semen is generally much more difficult than in mares. The endometrial inflammatory response is hypothesized to be a contributing factor to the lesser fertility. Following a cross-over experimental design, the uterine inflammatory response of six jennies was evaluated at 6h after AI with frozen-thawed semen (deposited in the uterine body) in the presence or absence of autologous seminal plasma (+SP or -SP). The endometrial cytology and histology of the animals were examined by uterine lavage, uterine swabbing and biopsy. The amount of cyclooxygenase-2 (COX-2) protein in endometrial cells was also evaluated. As a control (C), the same examinations were made before any AI procedure (i.e., when the jennies were in oestrus). Large numbers of polymorphonuclear neutrophils (PMN) were observed in the -SP and +SP cytology and biopsy samples; more than in the C samples. The -SP samples also had intense COX-2 labelling; less labelling was detected in the +SP and C samples (no significant difference between these latter two types). Thus, while the presence of SP does not change the post-AI number of PMNs with regard to that detected in its absence, it does reduce COX-2 protein. Further research into the complex mix of molecules in SP and its effects during AI might help increase the pregnancy rates achieved in jennies. PMID:24280633

  15. Protective effects of ascorbic acid and vitamin E on antioxidant enzyme activity of freeze-thawed semen of Qinchuan bulls.

    PubMed

    Zhao, X L; Li, Y K; Cao, S J; Hu, J H; Wang, W H; Hao, R J; Gui, L S; Zan, L S

    2015-01-01

    The aim of this study was to determine the protective effects of the combination of ascorbic acid (Vc) and vitamin E (VE) on antioxidant enzyme activity, sperm motility, viability, and acrosome integrity of Qinchuan bulls after freeze-thaw. In this study, we determined the effects of Vc and VE on the activity of the antioxidant enzyme defense system comprising glutathione peroxidase (GSH-Px), glutathione reductase (GR), catalase (CAT), and superoxide dismutase (SOD). The combination of Vc and VE had protective effects on sperm motility and viability. With respect to acrosome integrity and the activity of GR and SOD, differences were observed between the experimental groups with added Vc (7 mg/mL) and VE (0.12 IU/mL) and the control group. The activity of GSH-Px in the experimental group (1400 IU/mL Vc and 0.12 IU/mL VE) was not different (P > 0.05) compared with that in the control group, while the activity of CAT showed a significant difference between the 2 groups (P < 0.05). Therefore, we inferred that the combination of Vc (1400 IU/mL) and VE (0.12 IU/mL) protected the sperm quality in the freeze-thaw process. PMID:25867404

  16. GnRH analogue treatment on LH surge day 0 followed by single transvaginal artificial insemination with frozen semen on day 5 in bitches

    PubMed Central

    OHTAKI, Tadatoshi; KOGA, Yasuna; ONO, Mamiko; WATANABE, Gen; TAYA, Kazuyoshi; TSUMAGARI, Shigehisa

    2014-01-01

    ABSTRACT Reproductive parameters were evaluated in 19 and 14 estrous beagles that received 100 µg of gonadotropin-releasing hormone (GnRH) and saline treatment, respectively, on the day of luteinizing hormone (LH) surge (Day 0; estimated by serial progesterone assay) and balloon catheter-aided single transvaginal artificial insemination of frozen semen on Day 5. Although the conception rate and litter size were similar between the GnRH and saline groups, the concentration of LH peak was significantly higher in GnRH-treated bitches (P<0.01). In addition, the actual LH surge did not occur on the estimated Day 0 in one saline-treated bitch. In clinical practice that daily progesterone assay is difficult, administration of GnRH on estimated Day 0 would be recommended to induce or enhance the LH surge for timely and successful insemination. PMID:25311914

  17. GnRH analogue treatment on LH surge day 0 followed by single transvaginal artificial insemination with frozen semen on day 5 in bitches.

    PubMed

    Ohtaki, Tadatoshi; Koga, Yasuna; Ono, Mamiko; Watanabe, Gen; Taya, Kazuyoshi; Tsumagari, Shigehisa

    2015-01-01

    Reproductive parameters were evaluated in 19 and 14 estrous beagles that received 100 µg of gonadotropin-releasing hormone (GnRH) and saline treatment, respectively, on the day of luteinizing hormone (LH) surge (Day 0; estimated by serial progesterone assay) and balloon catheter-aided single transvaginal artificial insemination of frozen semen on Day 5. Although the conception rate and litter size were similar between the GnRH and saline groups, the concentration of LH peak was significantly higher in GnRH-treated bitches (P<0.01). In addition, the actual LH surge did not occur on the estimated Day 0 in one saline-treated bitch. In clinical practice that daily progesterone assay is difficult, administration of GnRH on estimated Day 0 would be recommended to induce or enhance the LH surge for timely and successful insemination. PMID:25311914

  18. 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

  19. Ewe breed differences in fertility after cervical AI with frozen-thawed semen and associated differences in sperm penetration and physicochemical properties of cervical mucus.

    PubMed

    Richardson, Lorraine; Hanrahan, J P; O'Hara, Lydia; Donovan, Anne; Fair, Sean; O'Sullivan, Michael; Carrington, Stephen D; Lonergan, Pat; Evans, A C O

    2011-11-01

    The objective of this study was to examine sperm penetration through cervical mucus and associated physicochemical properties of cervical mucus from Belclare and Suffolk ewes - two breeds with divergent pregnancy rate following cervical AI using frozen-thawed semen. In Experiment 1, sperm penetration through cervical mucus was assessed in 15 Belclare and 15 Suffolk ewes at 30, 48 and 57h post sponge removal. In Experiment 2, rheological properties of mucus from 17 Belclare and 19 Suffolk ewes at 48 and 57h post sponge removal were determined. In Experiment 3, 20 Belclare and 20 Suffolk ewes were used to assess mucus ferning and pH collected at 42, 48, 57 and 65h post sponge removal. In Experiment 1, a higher number of sperm penetrated cervical mucus from Belclare ewes at 48h, reflected by a breed by time interaction (P=0.05). In Experiment 2, mucus from Suffolk ewes tended to have higher elastic and complex moduli than that from Belclare ewes (P=0.06) regardless of time of collection. There was no effect of ewe breed on the viscous modulus. In Experiment 3, there was a significant effect of time post sponge removal on ferning (P<0.01), but there was no effect of breed. There was no effect of time or breed on mucus pH. It is concluded that breed differences in the rheological properties of cervical mucus affect the ability of sperm to swim through cervical mucus and this may explain breed differences in fertility observed after cervical AI using frozen-thawed semen. PMID:22115522

  20. Cryopreservation of crane semen

    USGS Publications Warehouse

    Gee, G.F.

    1991-01-01

    The method for the cryopreservation of crane semen at Patuxent Wildlife Research Center is described in detail. Cryopreservation is useful for the long-term storage of crane semen and for specialized propagation needs. A 50% fertility rate from most sandhill cranes, Grus canadensis, inseminated with frozen-thawed semen can be expected. Additional research should improve the fertility rate and determine how applicable the technique is to other crane species.

  1. 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. PMID:25744822

  2. Antioxidative effects of melatonin on kinetics, microscopic and oxidative parameters of cryopreserved bull spermatozoa.

    PubMed

    Ashrafi, Iraj; Kohram, Hamid; Ardabili, Farhad Farrokhi

    2013-06-01

    Reactive oxygen species generated during the freeze-thawing process may reduce sperm quality. This study evaluates the effects of melatonin supplementation as an antioxidant in the semen extender on post-thaw parameters of bull spermatozoa. Melatonin was added to the citrate-egg yolk extender to yield six different final concentrations: 0, 0.1, 1, 2, 3 and 4mM. Ejaculates were collected from six proven Holstein bulls. Semen was diluted in the extender packaged in straws, which was frozen with liquid nitrogen. The semen extender supplemented with various doses of melatonin increased (p<0.05) total motility, progressive motility, linearity, sperm track straightness, lateral head displacement, viability, integrity of the sperm membrane and total normal morphology of sperm after the freeze-thawing process. The most effective concentration of melatonin in microscopic evaluations of the bull sperm freezing extender was 2mM. The highest (p<0.05) value of total antioxidant capacity (48.9±2.7) and the lowest value of lipid peroxidation (2.7±0.8) were achieved by inclusion of 3mM concentration of melatonin in the semen extender and the highest activity of catalase (0.7±0.1) was obtained by 2mM melatonin. Four millimolar concentration of melatonin were reduced (p<0.05) the progressive motility and straight linear velocity. In conclusion, supplementation of 2 or 3mM concentration of melatonin in the semen extender improved the quality of post-thawed semen, which may associate with a reduction in lipid peroxidation as well as an increase in the total antioxidant capacity and antioxidant enzyme activity. PMID:23664651

  3. Prediction of the optimal time for insemination using frozen-thawed semen in a multi-sire insemination trial in bitches.

    PubMed

    Steckler, D; Nöthling, J O; Harper, C

    2013-11-30

    The aims of the study were to determine which of Days 5, 6 or 7 after the blood plasma progesterone concentration (PPC) of bitches first reached 6-9 nmol/L (Day 0) yield the highest fertility and whether day of insemination affects the gender ratio of conceptuses. Six bitches were inseminated on Days 5 and 6 and 6 on Days 6 and 7. Ten million progressively motile frozen-thawed sperm from each of 5 dogs were pooled for the first insemination. The same number of sperm from 5 other dogs were pooled for the second insemination. Only one batch of semen from each dog was used on all bitches, which largely prevented any effect of male and semen. Twenty-three autosomal microsatellites and the amelogenin gene were used to determine the paternity and gender of the conceptuses. Pregnancy rate was 100%. Out of 103 ovulations 66 conceptuses were conceived (conception rate: 64%). The proportion of available oocytes fertilised was 0.11, 0.56, and 0.27 for Days 5, 6, and 7, respectively. The odds of fertilisation was 16.7 and 4.2 times higher from insemination on Day 6 compared to Day 5 (P<0.001) and Day 7 (P = 0.013), respectively. The numbers of male- and female conceptuses were equal (33 each) and gender was independent of insemination day (P = 0.18). This study suggests that intrauterine insemination of bitches should best be done 6 days after PPC first reaches a value between 6 and 9 nmol/L with a second insemination one day later. PMID:24128644

  4. 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. PMID:26725319

  5. Comparative evaluation of the fluorescent antibody test and microtiter immunoperoxidase assay for detection of bovine viral diarrhea virus from bull semen.

    PubMed Central

    Afshar, A; Dulac, G C; Dubuc, C; Howard, T H

    1991-01-01

    An indirect immunoperoxidase staining technique (IP) is described for the detection of bovine viral diarrhea virus (BVDV) in bovine semen. The performance of the IP was compared to the reference immunofluorescent staining test in its ability to detect BVDV in 23 coded field semen samples. The IP assay which can be applied with ease to a large number of samples and does not require expensive fluorescence microscope equipment, appears to be an alternative method for BVDV detection. The IP assay can be strongly recommended for certification of BVDV-free bovine semen for artificial insemination and trading purposes and for laboratories which are not equipped for performing the immunofluorescent test. PMID:1653102

  6. 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

  7. Effect of different thawing temperatures on the viability, in vitro fertilizing capacity and chromatin condensation of frozen boar semen packaged in 5 ml straws.

    PubMed

    Córdova-Izquierdo, A; Oliva, J H; Lleó, B; García-Artiga, C; Corcuera, B D; Pérez-Gutiérrez, J F

    2006-03-01

    The effect of two different thawing temperatures on frozen boar semen viability, in vitro fertilizing capacity and chromatin condensation and stability was studied. Freeze-thaw motility, normal apical ridge (NAR), in vitro fertilizing (IVF) capacity and chromatin condensation and stability were evaluated after thawing at 42 degrees C, 40s and 50 degrees C, 40s. Chromatin condensation degree was determined by flow cytometry, using propidium iodide as fluorochrome intercalating agent, and chromatin stability was evaluated by the same procedure after inducing sperm chromatin decondensation with ethylene diamine tetraacetic acid (EDTA) and sodium dodecyl sulfate (SDS). The results showed that thawing straws at 42 degrees C, 40s significantly reduced motility compared to straws thawed at 50 degrees C, 40s. NAR, penetration, monospermy and polyspermy were not different between the two groups of samples thawed at different temperatures. Chromatin was significantly more compact when thawing was performed at 50 degrees C, but its stability did not show any difference relative to thawing at 42 degrees C. It is suggested that the interactions involved in chromatin overcondensation had a non-covalent nature. PMID:15975744

  8. Pregnancy rate and birth rate of calves from a large-scale IVF program using reverse-sorted semen in Bos indicus, Bos indicus-taurus, and Bos taurus cattle.

    PubMed

    Morotti, F; Sanches, B V; Pontes, J H F; Basso, A C; Siqueira, E R; Lisboa, L A; Seneda, M M

    2014-03-15

    Obtaining sexed sperm from previously frozen doses (reverse-sorted semen [RSS]) provides an important advantage because of the possibility of using the semen of bulls with desired genetic attributes that have died or have become infertile but from whom frozen semen is available. We report the efficiency of RSS on the pregnancy rate and birth rate of calves in a large-scale program using ovum pick-up and in vitro embryo production (IVEP) from Bos indicus, Bos indicus-taurus, and Bos taurus cattle. From 645 ovum pick-up procedures (Holstein, Gir, and Nelore), 9438 viable oocytes were recovered. A dose of frozen semen (Holstein, Nelore, Brahman, Gir, and Braford) was thawed, and the sperm were sex-sorted and cooled for use in IVF. Additionally, IVF with sperm from three Holstein bulls with freeze-thawed, sex-sorted (RSS) or sex-sorted, freeze-thawed (control) was tested. A total of 2729 embryos were produced, exhibiting a mean blastocyst rate of 29%. Heifers and cows selected for adequate body condition, estrus, and health received 2404 embryos, and 60 days later, a 41% average pregnancy rate was observed. A total of 966 calves were born, and 910 were of a predetermined sex, with an average of 94% accuracy in determining the sex. Despite the lower blastocyst rate with freeze-thawed, sex-sorted semen compared with sex-sorted semen, (P < 0.05), the pregnancy rate (bull I, 45% vs. 40%; II, 35% vs. 50%; and III, 47% vs. 48% for RSS and control, respectively; P > 0.05) and sex-sorted efficiency (bull I, 93% vs. 98%; II, 96% vs. 94%; and III, 96% vs. 97% for RSS and control, respectively; P > 0.05) were similar for each of the three bulls regardless of the sperm type used in the IVF. The sexing of previously frozen semen, associated with IVEP, produces viable embryos with a pregnancy rate of up to 40%, and calves of the desired sex are born even if the paternal bull has acquired some infertility, died, or is located a long distance from the sexing laboratory. Furthermore

  9. 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...

  10. Evaluation of frozen thawed cauda epididymal sperms and in vitro fertilizing potential of bovine sperm collected from the cauda epididymal

    PubMed Central

    Chaveiro, A; Cerqueira, C; Silva, J; Franco, J; Moreira da Silva, F

    2015-01-01

    In the present study, the fertilizing potential of semen recovered from slaughtered bulls epididymis was evaluated after cryopreservation, by conventional techniques and flow cytometry methods. The cauda epididymal was dissected and sperm were recovered and evaluated for volume, sperm concentration, and membrane and acrosome integrity using a flow cytometer. Sperm fertility potential was tested by in vitro fertilization (IVF). For each bull, three trials of IVF were performed. Before freezing, on average, the sperm concentration was 216 ± 27.5 × 106 sperm/ml. Sperm viability averaged 86.5 ± 4%. The mean percentage of sperm with intact plasma membrane and acrosome before and after cryopreservation was 90.7 ± 2.9% and 90.8 ± 1.9% (P≥0.05), respectively. The fertilization rate using frozen/thawed epididymal semen averaged 64.1 ± 3.9% fertilization with no significant differences between bulls (P>0.05). For the bull considered as control, the fertilization rate was 72.2 ± 4.5%, differing significantly (P>0.05) from the frozen/thawed epididymal semen’s fertilization rate. In conclusion, it is possible to use in vitro techniques with cryopreserved spermatozoa obtained from bull’s epididymis using a controlled rate freezing method with a predetermined freezing curve, and with assessment of sperm’s viability by conventional techniques and flow cytometry methods, together with the fertilizing ability of cryopreserved epididymal spermatozoa. PMID:27175174

  11. Characterization and usage of sexed semen from US field data

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objectives were to characterize sexed semen available and its usage from US field data. This included investigating active Holstein proven bulls with sexed semen available, as well as percentages and frequencies of sexed semen matings for heifers and cows. Herds were also characterized for the...

  12. Avian artificial insemination and semen preservation

    USGS Publications Warehouse

    Gee, G.F.

    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

  13. 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.

  14. Subfertility Problems Leading to Disposal of Breeding Bulls

    PubMed Central

    Khatun, Marzina; Kaur, Simarjeet; Kanchan; Mukhopadhyay, C. S.

    2013-01-01

    Subfertility problems are encountered frequently in the cattle and buffalo bulls commercially maintained for semen production in dairy farms and under field conditions for natural insemination. Reports are scarce on the incidence of subfertility in breeding bulls, especially in India. The objective of the present study was to assess the incidence of the male reproductive anomalies leading to disposal of bovine bulls at GADVASU dairy farm, Ludhiana, Punjab (India). Data on frequency of various subfertility and disposal pattern of bulls maintained at the dairy farm, GADVASU, were collected for 12 yrs (1999 to 2010) and compiled from different record registers. Percentage of bulls that produced freezable semen (out of reserved ones) was less in cattle (25.641%) as compared to that of buffalo (30.4%). Various subfertility traits like poor libido and unacceptable seminal profile were found to be the significant reasons (p<0.01) for culling of the breeding bulls. Inadequate sex drive and poor semen quality were the main contributing factors for bull disposal in cattle whereas poor semen freezability was most frequently observed in buffalo bulls. All the male reproductive traits were significantly different (p<0.05) for the periods of birth, except for semen volume, initial motility (IM), age at last semen collection (ALSC) and age at disposal. The ages at first and last semen collection as well as freezing (i.e. AFSC, ALSC and AFSF, ALSF, respectively) and age at disposal (AD) were higher in buffalo. The spermatological parameters and semen production period (SPP) were higher in cattle. The age at first semen donation and breeding period could be reduced by introducing the bulls to training at an early age. The results revealed an increasing trend in individual motility (IM) while semen volume, AFSC, AFSF, AD, FSPP, SPP, ALSC and ALSF showed a decreasing, however, not a definite trend, over the periods. The semen donation traits like, AFSF, of the cattle and buffalo

  15. 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. PMID:26503476

  16. HSP90 expression correlation with the freezing resistance of bull sperm.

    PubMed

    Wang, Peng; Wang, Yan-Feng; Wang, Hong; Wang, Chun-Wei; Zan, Lin-Sen; Hu, Jian-Hong; Li, Qing-Wang; Jia, Yong-Hong; Ma, Guo-Ji

    2014-05-01

    To date, there has been little improvement in cryopreservation of bull sperm due to lack of understanding of the freezing mechanisms. Therefore, this study set out to investigate expression levels of fertility-associated proteins in bull sperm, and in particular the relationship between the 90 kDa heat-shock protein (HSP90) and the sperm characteristics after freezing-thawing. Semen was collected from eight Holstein bulls by artificial vagina. Characteristics of these fresh semen, including sperm motility, morphology, viability and concentration, were evaluated. Sperm quality was also assessed after freezing-thawing. Eight ejaculates were divided into two groups based on freezing resistance and sperm motility. Sperm proteins were extracted and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and western blotting were performed. SDS-PAGE results showed that there was substantial diversity in 90 kDa proteins in the frozen-thawed sperm and HSP90 was confirmed as one of the 90 kDa proteins by western blot. This study indicated that HSP90 expression correlated positively with sperm quality. The amount of expressed 90 kDa proteins in the high freezing resistance (HFR) group was significantly higher than that in the low freezing resistance (LFR) group (P < 0.05). Thus, higher expression of HSP90 could probably lead to the higher motility and freezing resistance of sperm found after freezing-thawing. Therefore, we concluded that level of HSP90 expression could be used to predict reliably and simply the freezing resistance of bull sperm. PMID:23506739

  17. The Knobbed Acrosome Defect in Beef Bulls

    PubMed Central

    Barth, Albert D.

    1986-01-01

    The knobbed acrosome defect was found at levels of 25 to 100 percent of spermatozoa from 16 of 2054 beef bulls. The incidence of this defect appeared to be particularly high in the Charolais breed. Pedigree analysis of some of the affected Charolais bulls indicated there may be a genetic predisposition for this sperm defect. In eosin-nigrosin stained semen smears the most common form of the abnormality was a flattened or indented apex of the sperm head. A refractile bead at the apex of the sperm head was seen less commonly. Electron microscopy of the spermatozoa from one bull showed that the abnormality was similar to the knobbed sperm defect previously described in Friesian bulls. A breeding trial confirmed that bulls producing spermatozoa with a high incidence of knobbed acrosomes are infertile. ImagesFigure 2 and 3.Figure 4.Figure 5.Figure 6 and 7.Figure 8.Figure 9.Figure 10. PMID:17422706

  18. 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. PMID:26925808

  19. 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

  20. Does bovine besnoitiosis affect the sexual function of chronically infected bulls?

    PubMed

    Esteban-Gil, A; Jacquiet, P; Florentin, S; Decaudin, A; Berthelot, X; Ronsin, P; Grisez, C; Prevot, F; Alzieu, J P; Marois, M; Corboz, N; Peglion, M; Vilardell, C; Liénard, E; Bouhsira, E; Castillo, J A; Franc, M; Picard-Hagen, N

    2016-09-15

    Bovine besnoitiosis is a reemerging disease in Europe. The clinically Besnoitia besnoiti infection in bulls is characterized by fever, nasal discharge, and orchitis in the acute phase and by scleroderma in the chronic phase. However, in many bulls, B besnoiti infection remains at a subclinical stage. Bull infertility is an economically relevant consequence of besnoitiosis infection. It is not clear, however, if semen quality returns to normal levels when infected animals have clinically recovered. The aim of this study was to examine the relationship between chronic besnoitiosis and bull sexual function in a region of eastern France, where the disease is reemerging, by comparing semen quality and genital lesions in 11 uninfected, 17 subclinically infected, and 12 clinically infected bulls. The presence of anti-B besnoiti antibodies was detected by Western blot test. Semen was collected by electroejaculation. Bulls clinically infected with B besnoiti showed significantly more genital tract alterations than uninfected or subclinically infected bulls. No relationship was evidenced between besnoitiosis infectious status and semen quality, whereas a significant relationship was noted between genital lesions and semen score. This means that in the absence of moderate to severe genital lesions, chronic bovine besnoitiosis is unlikely to alter semen quality. However, as the presence of infected animals could lead to spread of the disease, culling or separation of clinically infected bulls from the remaining healthy animals is strongly recommended. PMID:27264738

  1. Association between ETFA genotype and activity of superoxide dismutase, catalase and glutathione peroxidase in cryopreserved sperm of Holstein-Friesian bulls.

    PubMed

    Hering, D M; Lecewicz, M; Kordan, W; Kamiński, S

    2015-02-01

    The aim of this study was to determine whether C/T missense mutation within the ETFA gene is associated with sperm antioxidant enzymatic activity. One hundred and twenty Holstein-Friesian bulls were genotyped by the PCR-RFLP technique (MwoI). Commercial straws of frozen-thawed semen were used to evaluate the activity of three antioxidant enzymes: superoxide dismutase, catalase and glutathione peroxidase. Among all bulls investigated, genotype CT was the most frequent (44.2%), in comparison with CC (42.5%) and TT (13.3%). Significant differences in glutathione peroxidase activity were observed between homozygous individuals (CC vs TT) with heterozygous CT having intermediate values. Dismutase activity was significantly associated with ETFA genotype, although only bulls with the CT genotype were significantly different from bulls carrying the CC genotype. The activity of catalase showed a similar trend (but was not statistically significant). In conclusion, we found that bulls with the ETFA TT genotype produce sperm with the highest glutathione peroxidase activity and can therefore be more efficiently protected from reactive oxygen. The mechanism of this interaction needs to be elucidated in future research. PMID:25472694

  2. 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. PMID:10735086

  3. 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.

  4. Semen analysis

    MedlinePlus

    ... The following things will be evaluated: How the semen thickens into a solid and turns to liquid Fluid thickness, acidity, and sugar content Resistance to flow (viscosity) Movement of the sperm (motility) Number and structure ...

  5. Semen Analysis

    MedlinePlus

    ... Email Other share options Semen analysis is a test on the fluid that is released when a man has an orgasm ©1996 - 2016 SART, Society for Assisted Reproductive Technology . All Rights Reserved. ASRM/SART Nondiscrimination Policy ASRM/ ...

  6. 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 (...

  7. 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

  8. Evaluation of the semen swim-up method for bovine sperm RNA extraction.

    PubMed

    Han, C M; Chen, R; Li, T; Chen, X L; Zheng, Y F; Ma, M T; Gao, Q H

    2016-01-01

    Isolation of high-quality RNA is important for assessing sperm gene expression, and semen purification methods may affect the integrity of the isolated RNA. This study evaluated the effectiveness of the sperm swim-up method for seminal RNA isolation. Frozen semen samples in straws from three bulls of proven fertility were purified by the swim-up method. RNA extraction was carried out using the E.Z.N.A.(TM) Total RNA kit II, with non-swim-up sperm as a control. Total sperm RNA was analyzed by UV spectrophotometry, reverse transcription polymerase chain reaction (RT-PCR), and agarose gel electrophoresis, and expression of the sex-determining region on the Y chromosome (SRY), leptin (LEP), and ribosomal protein subunit 23 (RPS23) genes, were determined. 18S RNA was used as a positive control. Fewer somatic cells were found in sperm swim-up samples than in the non-swim-up counterparts (0 x 10(3) vs 17.33 ± 2.52 x 10(3) sperm, P < 0.05). In addition, high-quality RNA was obtained in about 2 h, with no significant difference between groups. Interestingly, the yields of RNA fragments containing ≥200 nucleotides were significantly reduced in sperm swim-up samples (0.92 ± 0.41 x 10(7) sperm) compared with the non-swim-up samples (1.36 ± 0.33 x 10(7) sperm, P < 0.05). After RT-PCR, clear bands representing SRY, LEP, and RPS23 in sperm cDNA were observed on agarose gel electrophoresis. Finally, no bands corresponding to 18S RNA were found in RNA samples from the sperm swim-up group. Our findings suggest that small amounts of sperm RNA can be efficiently extracted from frozen straw semen samples using the swim-up technique. PMID:27173315

  9. 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...

  10. Relationship of nonreturn rates of dairy bulls to binding affinity of heparin to sperm

    SciTech Connect

    Marks, J.L.; Ax, R.L.

    1985-08-01

    The binding of the glycosaminoglycan (3H) heparin to bull spermatozoa was compared with nonreturn rates of dairy bulls. Semen samples from five bulls above and five below an average 71% nonreturn rate were used. Samples consisted of first and second ejaculates on a single day collected 1 d/wk for up to 5 consecutive wk. Saturation binding assays using (TH) heparin were performed to quantitate the binding characteristics of each sample. Scatchard plot analyses indicated a significant difference in the binding affinity for (TH) heparin between bulls of high and low fertility. Dissociation constants were 69.0 and 119.3 pmol for bulls of high and low fertility, respectively. In contrast, the number of binding sites for (TH) heparin did not differ significantly among bulls. Differences in binding affinity of (TH) heparin to bull sperm might be used to predict relative fertility of dairy bulls.

  11. Homeopathic treatment for infertility in a prize Nelore bull.

    PubMed

    Lobreiro, J

    2007-01-01

    Treatments for infertility in bulls are not described in homeopathic literature. A few treatments, such as changing the protein content of the diet, giving extra minerals, etc have been proposed. This case report describes homeopathic treatment for infertility in a prize bull. A Nelore bull, considered infertile for 3 years, was treated with homeopathic Pulsatilla nigricans 200 CH. Decreased total sperm defects, increased sperm motility and a very impressive increased number of doses of semen produced were observed. The bull relapsed after treatment was withdrawn, but again responded when it was resumed. Since only one animal was observed one cannot assume that the observed changes were due only to this treatment. Further studies may establish the real benefits of a homeopathic medicine in bull infertility. PMID:17227749

  12. 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

  13. 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...

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

    PubMed

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

    1989-07-01

    Three groups of 1/2 Simmental X 1/4 Brahman X 1/4 Hereford bull calves were used during two different years to study effects of zeranol on sexual development. At 154 d of age, half the calves were implanted with 36 mg zeranol and half, not implanted, served as controls. Implanted calves were reimplanted at 90-d intervals throughout the trial (9 mo) each year. Trial 1 was conducted with 24 calves and Trial 2 was conducted the following year with 10 bulls. Twenty-four days after weaning (200 d of age) and at 28-d intervals thereafter, bulls in drylot in Trial 1 were weighted, scrotal circumference (SC) was measured and an ejaculate of semen was collected by electroejaculation to determine puberty. At these times, bulls were given 200 micrograms of GnRH i.m. and blood was collected at 0, 1, 2, 3, 4 and 5 h after GnRH. Serum concentrations of LH and testosterone (TEST) were determined. At slaughter, testis weight, length and circumference and pubertal status were recorded. Bulls implanted with zeranol had smaller SC than control bulls during the entire 9-mo period (P less than .0001). More control bulls reached puberty than did implanted bulls (82.4 vs 23.5%, respectively; P less than .001). Control bulls had larger testis measurements at slaughter (P less than .0001). Implants did not alter total weight gain or ADG (P greater than .10).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2768123

  15. Effect of egg yolk powder on freezability of Murrah buffalo (Bubalus bubalis) semen

    PubMed Central

    Kumar, N.; Lone, S. A.; Prasad, J. K.; Jan, M. H.; Ghosh, S. K.

    2016-01-01

    Aim: The aim of this study was to investigate the effect of commercial egg yolk powder as an alternative to fresh egg yolk on freezability of Murrah buffalo semen. Materials and Methods: Semen samples (12) from 3 Murrah buffaloes (4 from each bull) with mass motility (≥3+) and total motility (70% and above) were utilized in this study. Immediately after collection, each sample was divided into four groups. Groups I was diluted up to 60×106 sperm/ml with tris extender containing 10% fresh egg yolk and Groups II, III, and IV were diluted up to 60×106 sperm/ml with tris extender containing 2%, 4%, and 6% egg yolk powder, respectively. Semen samples were processed and cryopreserved followed by examination of frozen semen samples after 24 h. Semen samples from each group were evaluated for total motility, viability, acrosomal integrity, abnormality, and hypo-osmotic swelling test (HOST) response after dilution, pre-freeze, and post-thaw stage. Results: Pre-freeze total motility was significantly (p<0.05) higher in Groups III and IV as compared to Groups I and II, and post-thaw total motility was significantly (p<0.01) higher in Group III as compared to other three groups. Viability was significantly (p<0.05) higher in Groups II, III, and IV than Group I at the pre-freeze stage. Significantly (p<0.01) higher viability and acrosomal integrity were recorded in Group III as compared to other three groups at the post-thaw stage. Abnormality was significantly (p<0.05) higher in Group IV than other three groups. HOST response was significantly (p<0.05) higher in Groups II and III than Groups I and IV at the pre-freeze and post-thaw stages. Conclusion: Addition of egg yolk powder at 4% level yielded significantly better results in terms of post-thaw semen quality as compared to the fresh egg yolk and other concentrations of egg yolk powder (2% and 6%). PMID:27397983

  16. Microsatellite analysis of cryopreserved stallion semen stored on FTA paper.

    PubMed

    Schulman, M L; Harper, C K; Bell, E; Nel, A; Guthrie, A J

    2002-12-01

    The aim of this study was to establish and validate a method to permit microsatellite analysis of DNA profiles obtained from frozen-thawed stallion sperm cells. This would provide reliable and accurate verification of the identification of a semen donor. Ejaculates from 5 pony stallions were collected, processed and frozen in 0.5 ml plastic straws. Aliquots of 100 microl of the frozen-thawed semen thus obtained were either placed directly, or diluted (1:10; 1:100; and 1:1000) and placed on slides of FTA paper. Similarly, blood samples obtained from each of the stallions were placed onto slides of FTA paper. A punch was removed from each sample after drying Each sample was mixed with FTA purification reagent, Dithiothreitol and Proteinase K before incubation and processing. All samples were processed with a set of 13 microsatellite markers. Further analysis permitted a comparison of the DNA profiles of the frozen-thawed semen and the blood samples. A full profile of markers was obtained from the 1:10 and 1:100 dilutions of the frozen-thawed semen samples as well as from the blood samples. The DNA profiles from the frozen-thawed semen and blood samples obtained from the stallions matched in all cases. PMID:12665139

  17. Semen collection methods affect the bacterial composition of post-thawed semen of silver barb (Barbodes gonionotus).

    PubMed

    Boonthai, Traimat; Khaopong, Weerasith; Sangsong, Jumlong; Sooksawat, Treerat; Nimrat, Subuntith; Vuthiphandchai, Verapong

    2016-03-01

    Biosafety issue associated with the risk of pathogenic contamination of cryopreserved semen is a common concern because of associated declines in sperm quality, storage period and disease transmission. This study was conducted to evaluate the effects of methods of semen collection on sperm quality and bacterial composition of post-thawed semen of silver barb (Barbodes gonionotus). Semen collection methods consisted of four treatments: (1) hand-stripping of abdomen without rinsing of urogenital area with water, (2) hand-stripping of abdomen after rinsing of urogenital area with water, (3) catheterization without rinsing of urogenital area with water and (4) catheterization after rinsing of urogenital area with water. Semen diluted with calcium-free Hank's balanced salt solution containing 10% dimethylsulfoxide (DMSO) was frozen at a freezing rate of -8°Cmin(-1) before plunging in liquid nitrogen. Post-thawed semen collected by catheterization after rinsing urogenital area had the lowest bacterial number, about 2-log reduction of total heterotrophic, Gram negative and pseudomonad bacteria, compared with the other three collection treatments. However, percentages of motile and viable sperm were not significantly (P>0.05) different among treatments. This method eliminated Flavobacterium aquatile, Bacillus megaterium, Kocuria varians, Staphylococcus haemolyticus and Aeromonas media in cryopreserved semen. This is the first report demonstrating the effects of semen collection methods on bacteriological quality of frozen-thawed fish semen. PMID:26778122

  18. Replication of bovine viral diarrhoea virus in the bovine reproductive tract and excretion of virus in semen during acute and chronic infections.

    PubMed

    Kirkland, P D; Richards, S G; Rothwell, J T; Stanley, D F

    1991-06-22

    Five mature bulls were studied during an acute transient infection with bovine viral diarrhoea virus (BVDV). The bulls had been infected experimentally by the intranasal instillation of blood and serum from a cow which was a persistent carrier of the virus. Infection was confirmed by the demonstration of a low titred viraemia in four of the five animals and by the seroconversion of all five. Semen samples were collected from each bull on four occasions between seven and 14 days after infection. The virus was isolated from the semen of three of the five bulls and from nine of 12 batches of semen from them. In contrast to other studies of the infection of semen, BVDV was isolated with similar efficiency from raw, unprocessed semen and from diluted, extended semen. The titres of virus in the semen ranged from 5 to 75 TCID50/ml. The infection did not appear to affect the quality of the semen. Shedding of virus continued after the end of the period of viraemia and appeared to be a consequence of the replication of the virus in the reproductive tract and its subsequent excretion in the seminal fluid. Virological studies of the reproductive tracts of these bulls suggested that the most productive sites of virus replication were the seminal vesicles and the prostate gland. Concurrent studies in a persistently infected bull supported these findings. PMID:1654660

  19. 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.

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

    PubMed

    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

  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. The effect of nocturnal sampling on semen quality and the efficiency of collection in bovine species.

    PubMed

    Yates, Jennifer H; Chandler, John E; Canal, Anita L; Braden Paul, J

    2003-12-01

    This study evaluated night and day semen collection regimes in Holstein and Brahman bulls (four bulls of each breed) that were collected weekly, each during a morning and a night collection. Ejaculates (n=64) were obtained via artificial vagina over 4 weeks. The first collection of each week alternated between night and day. Two collection teams were employed. Bull behavior parameters included reaction time to first mount, time to ejaculation, a refractory period test, and a thrust intensity test. The numbers of interruptions were counted as a managerial parameter. Pre-freeze semen parameters included total volume, initial motility and concentration. Post-freeze semen parameters measured were: 0- and 3-h post-thaw motility; percent intact acrosomes; and percent sperm abnormalities. Data were analyzed by least squares methods. The bull within breed effect differed (P<0.05) for behavior parameters. The bull within breed effect for total motile sperm harvested was not significant. The bull within breed response was mixed for post-freeze semen viability parameters. Bull within breed was not significant for sperm abnormalities. The night versus day treatment was significant for the managerial parameter (P=0.002). Although a different collection schedule for Bos indicus cattle was not warranted, the efficiency of the collection process was affected by extraneous environmental conditions. PMID:14580649

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

    PubMed

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

    2015-08-01

    Sperm morphometry has some value as an indicator of reproductive capacity in males. In laboratory practice a variety of slide-staining methods are used during morphological evaluation of semen to predict male fertility. The aim of this study was to determine the effect of staining of semen using four different techniques on the morphometry of the bull sperm cell. The material for the study consisted of semen collected from test bulls of the Black-and-White variety of Holstein-Friesians. The results obtained in the study indicate differences in the dimensions of bull sperm heads when different slide staining techniques were used. The most similar results for sperm head dimensions were obtained in the case of SpermBlue(®) and eosin+gentian violet complex, although statistically significant differences were found between all the staining techniques. Extreme values were noted for the other staining techniques - lowest for the Papanicolaou and highest for silver nitrate, which may indicate more interference in the cell by the reagents used in the staining process. However, silver nitrate staining was best at identifying the structures of the sperm cell. Hence it is difficult to determine which of the staining methods most faithfully reveals the dimensions and shape of the bull sperm. PMID:26149220

  4. 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

  5. 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

  6. Optimizing age of bull at first use in relation to fertility of Murrah breeding bulls

    PubMed Central

    Mir, M. A.; Chakravarty, A. K.; Gupta, A. K.; Naha, B. C.; Jamuna, V.; Patil, C. S.; Singh, A. P.

    2015-01-01

    Aim: The aim of the present investigation was to optimize the age at first use (AAFU) of semen of Murrah breeding bulls, which will help in early selection of bulls under progeny testing program for improving the reproductive performance in the herd. Materials and Methods: The data on AAFU, conception rate based on first A.I. (CRFAI), overall conception rate (OCR), and birth weight (B.WT) of 57 Murrah bulls during 1993-2014 at NDRI center pertaining to 14 sets of Network Project on Buffalo Improvement at ICAR-National Dairy Research Institute, Karnal, Haryana, India were adjusted for significant environmental influences and subsequently analyzed. Simple and multiple regression models were used for prediction of CRFAI and OCR of Murrah breeding bulls. Comparative evaluation of three developed models (I-III) showed that Model III, having AAFU and B.WT, fulfill the accuracy of model as revealed by high coefficient of determination, low mean sum of squares due to error, low conceptual predictive value, and low Bayesian information criterion. Results: The results revealed that the average predicted CRFAI was highest (39.95%) at <3.5 years and lowest (34.87%) at >4.5 years of age at first A.I/use. Similarly, average predicted OCR was highest (41.05%) at <3.5 years and lowest (39.42%) at >4.5 years of age at first A.I/use of Murrah bulls. Conclusion: In organized herd under progeny testing program, Murrah bulls should be used at young age, i.e. prior to 3.5 years, which is expected to result in 5.08% better CRFAI and 1.63% better OCR in comparison to Murrah bulls used after 4.5 years of age. PMID:27047126

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

    PubMed

    Geary, T W; Kelly, W L; Spickard, D S; Larson, C K; Grings, E E; Ansotegui, R P

    2016-05-01

    Objectives were to determine if 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 peripubertal bulls were blocked by age and scrotal circumference and assigned to one of five treatments: (1) 1x sulfate form (1S); (2) 1x complexed form (1C); (3) 1S+1C (2SC); (4) 1S + 2 × 1 C (3SCC); and (5) 3 × 1S (3S). Each 1x supplementation level contained 360 mg Zn, 125 mg Cu, 200mg Mn and 12.5mg Co. Liver biopsies were collected on d -21 and 100, and scrotal circumference, semen, and blood samples were collected on d -14, 14, 42, 70, and 98. All bulls were deficient in Cu yet adequate in Zn on d -21. Following 100 d on treatment, liver Zn concentrations decreased (P<0.01) and liver Cu concentrations increased (P<0.01) in bulls regardless of treatment. Day 100 liver Zn concentrations were similar (P=0.50) across treatments, but liver Cu concentrations were greater (P=0.07) in 3SCC and 3S bulls compared to 1C and 1S bulls, whereas 2SC bulls were intermediate. Bulls fed complexed minerals tended to reach puberty after fewer (P=0.11) days on treatment (43.9 ± 5.7 d) than bulls fed only sulfate minerals (58.5 ± 6.7 d). Supplementing complexed Cu and Zn to prepubertal bulls may lower the age at puberty, however, no differences (P ≥ 0.40) in semen characteristics or scrotal measurements (P ≥ 0.11) were observed. PMID:26968246

  8. Semen preservation and artificial insemination in domesticated South American camelids.

    PubMed

    Bravo, P Walter; Alarcon, V; Baca, L; Cuba, Y; Ordoñez, C; Salinas, J; Tito, F

    2013-01-10

    Semen preservation and artificial insemination in South American camelids are reviewed giving emphasis to work done in Peru and by the authors. Reports on semen evaluation and the preservation process indicate that semen of alpacas and llamas can be manipulated by making it liquid first. Collagenase appears to be the best enzyme to eliminate viscosity. Tris buffer solution maintains a higher motility than egg-yolk citrate, phosphate buffered saline (PBS), Triladyl, and Merck-I extenders. Cooling of semen took 1h after collected, and equilibrated with 7% glycerol presented a better motility and spermatozoa survival at 1, 7, 15 and 30days after being slowly frozen in 0.25mL plastic straws. Trials of artificial insemination with freshly diluted semen and frozen-thawed semen are encouraging and needs to be tested extensively under field conditions. Recently, fertility rates varied from 3 to 67%. Semen preservation and most important, artificial insemination appear to be a reality, and could be used to improve the genetic quality of alpacas and llamas. PMID:23153624

  9. Blood in the semen

    MedlinePlus

    Semen - bloody; Blood in ejaculation ... Most of the time, blood in the semen is caused by swelling or infection of the prostate or seminal vesicles. The problem may occur after a prostate biopsy . Blood in the ...

  10. 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.

  11. Adapting Bulls to Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  12. Cytosine methylation of sperm DNA in horse semen after cryopreservation.

    PubMed

    Aurich, Christine; Schreiner, Bettina; Ille, Natascha; Alvarenga, Marco; Scarlet, Dragos

    2016-09-15

    Semen processing may contribute to epigenetic changes in spermatozoa. We have therefore addressed changes in sperm DNA cytosine methylation induced by cryopreservation of stallion semen. The relative amount of 5-methylcytosine relative to the genomic cytosine content of sperm DNA was analyzed by ELISA. In experiment 1, raw semen (n = 6 stallions, one ejaculate each) was shock-frozen. Postthaw semen motility and membrane integrity were completely absent, whereas DNA methylation was similar in raw (0.4 ± 0.2%) and shock-frozen (0.3 ± 0.1%) semen (not significant). In experiment 2, three ejaculates per stallion (n = 6) were included. Semen quality and DNA methylation was assessed before addition of the freezing extender and after freezing-thawing with either Ghent (G) or BotuCrio (BC) extender. Semen motility, morphology, and membrane integrity were significantly reduced by cryopreservation but not influenced by the extender (e.g., total motility: G 69.5 ± 2.0, BC 68.4 ± 2.2%; P < 0.001 vs. centrifugation). Cryopreservation significantly (P < 0.01) increased the level of DNA methylation (before freezing 0.6 ± 0.1%, postthaw G 6.4 ± 3.7, BC 4.4 ± 1.5%; P < 0.01), but no differences between the freezing extenders were seen. The level of DNA methylation was not correlated to semen motility, morphology, or membrane integrity. The results demonstrate that semen processing for cryopreservation increases the DNA methylation level in stallion semen. We conclude that assessment of sperm DNA methylation allows for evaluation of an additional parameter characterizing semen quality. The lower fertility rates of mares after insemination with frozen-thawed semen may at least in part be explained by cytosine methylation of sperm-DNA induced by the cryopreservation procedure. PMID:27242182

  13. Laboratory diagnosis and transmissibility of bovine viral diarrhea virus from a bull with a persistent testicular infection.

    PubMed

    Newcomer, Benjamin W; Toohey-Kurth, Kathy; Zhang, Yan; Brodersen, Bruce W; Marley, M Shonda; Joiner, Kellye S; Zhang, Yijing; Galik, Patricia K; Riddell, Kay P; Givens, M Daniel

    2014-06-01

    Recently, in the United States, a dairy bull was diagnosed as the second confirmed case of persistent testicular infection (PTI) with bovine viral diarrhea virus (BVDV). The first objective of this study was to evaluate the testing methodologies currently used by the artificial insemination industry in order to improve the detection of bulls with PTI. This study evaluated the impact of multiple factors ([1] sample tested, [2] sample handling, [3] assay used, and [4] assay methodology) on the sensitivity of detection of BVDV. The second objective of this study was to evaluate the transmissibility of BVDV from the bull through casual or sexual contact. Results from this study indicate that straws of semen should be transported to the diagnostic laboratory in liquid nitrogen dry shippers. PCR proved to be a more sensitive assay than virus isolation; however, certain PCR protocols exhibited greater diagnostic sensitivity than others. Insemination with cryopreserved semen from this infected bull caused viral transmission to a seronegative heifer resulting in viremia and seroconversion. After 42 months of age, the bull appeared to clear the infection. In conclusion, this bull validates that natural exposure to a 1a strain of BVDV can result in a unique PTI causing contamination of semen with detectable infectious virus. Appropriate handling and testing of samples is necessary in order to detect bulls exhibiting PTI. Additionally, PTI with BVDV may potentially be cleared after an extended duration. PMID:24656648

  14. Cryopreservation of buffalo (Bubalus bubalis) semen in Bioxcell extender.

    PubMed

    Akhter, S; Ansari, M S; Rakha, B A; Andrabi, S M H; Iqbal, S; Ullah, N

    2010-10-01

    This study was designed to compare commercially available extender Bioxcell with tris-citric egg yolk extender for post thaw quality and in vivo fertility of buffalo semen. For comparison of post thaw semen quality: semen was collected from five adult Nili-Ravi buffalo (Bubalus bubalis) bulls of similar age group with artificial vagina (at 42 degrees C) for three weeks (replicates). Qualifying ejaculates having motility >60% from each buffalo bull were divided in two aliquots and diluted (at 37 degrees C having 50 x 10(6) spermatozoa/ml) in tris-citric egg yolk or Bioxcell extender. Diluted semen was cooled to 4 degrees C in 2 hours, equilibrated for 4 hours and filled in 0.5 ml straws. Semen straws were kept over liquid nitrogen vapors (5 cm) for 10 minutes. Straws were then plunged and stored in liquid nitrogen (-196 degrees C). After 24 hours of storage, semen straws were thawed at 37 degrees C for 30 seconds to assess sperm motility, viability, plasma membrane integrity, normal apical ridge, and abnormalities (head, mid piece, and tail). For comparison of in vivo fertility: semen from two buffalo bulls of known fertility was cryopreserved in tris-citric egg yolk and Bioxcell as described earlier, and used for inseminations under field conditions. Post-thaw percentage of sperm motility (45.3 +/- 1.1, 45.0 +/- 1.4), viability (66.2 +/- 1.1, 64.4 +/- 1.3) plasma membrane integrity (60.4 +/- 1.2, 59.2 +/- 1.4) and normal apical ridge (82.9 +/- 0.5, 80.7 +/- 0.5) did not differ (P > 0.05) in tris-citric egg yolk and Bioxcell extender, respectively. Similarly, sperm abnormalities of head (1.20 +/- 0.1, 1.20 +/- 0.1), mid piece (0.67 +/- 0.1, 0.87 +/- 0.1) and tail (11.7 +/- 0.2, 11.6 +/- 0.3) remained similar (P > 0.05) in tris-citric egg yolk and Bioxcell extender, respectively. In vivo fertility rates of buffalo semen cryopreserved in tris-citric egg yolk and Bioxcell also remained similar (44% vs. 47%). It is concluded that commercially available Bioxcell may be

  15. Effect of age on spermiogram of Holstein Friesian × Sahiwal crossbred bulls.

    PubMed

    Mandal, D K; Kumar, M; Tyagi, S

    2010-04-01

    This study was conducted on 94 Frieswal (5/8 Holstein Friesian 3/8 Sahiwal) crossbred bulls of three different grades, categorized based on their semen freezability visualising Group 1 (consistently freezable semen producer bulls, N = 11), Group 2 (inconsistent freezable, N = 16) and Group 3 (Non freezable, N = 67). Each group was further divided into two classes that is young (up to 30 months) and adult (31 to 70 months) bulls depending upon their age. Sperm morphology was studied by using the eosin-nigrosin staining technique. Bulls age significantly (P < 0.01) affected semen quality and sperm morphology. In adult bulls, semen volume, mass activity and sperm concentration were 36%, 17.56% and 19.6%, respectively, higher than young. Initial progressive motility (%) and livability showed significant (P < 0.01) improvement with the advancement of age (43.37 ± 1.21 and 67.71 ± 1.11, respectively, in young; 53.02 ± 1.11 and 74.17 ± 1.03, respectively, in adult). In young bulls, sperm head, mid piece, tail abnormality and total abnormal sperm percent (12.38 ± 0.92, 4.87 ± 0.24, 11.01 ± 0.60 and 28.26 ± 1.34, respectively) were 1.85, 1.27, 1.20 and 1.44 folds higher than that of their mature stage (6.69 ± 0.64, 3.82 ± 0.32, 9.14 ± 0.64 and 19.66 ± 1.31, respectively). Significant reduction (P < 0.01) in micro cephalic sperm, free heads, bent mid piece, looped mid piece and proximal protoplasmic droplets were observed at mature age as compared with their younger stage. In bulls of consistent freezing category, abnormal sperm heads significantly decreased from 4.40 ± 0.31% to 3.28 ± 0.02% on maturity. Similarly, in inconsistent freezing grade bulls sperm head abnormality (9.28 ± 0.75% to 5.13 ± 1.20%) and total abnormal sperm percent (24.89 ± 1.43 to 18.73 ± 3.40) was decreased over the age. On the contrary, in non-freezing category bulls' sperm morphology did not show significant (P > 0.05) improvement with age advancement, rather some abnormalities

  16. Breed and other effects on reproductive traits and breeding soundness categorization in young beef bulls in Florida.

    PubMed

    Chenoweth, P J; Chase, C C; Thatcher, M J; Wilcox, C J; Larsen, R E

    1996-11-01

    Yearling, grass-fed, beef bulls at the USDA Subtropical Agricultural Research Station, Brooksville, Florida, were assessed for physical and semen traits in January, April, July and October of 1991 (Trial 1) and 1992 (Trial 2). Bulls were given a breeding soundness evaluation (BSE) using revised semen and scrotal circumference (SC) criteria. In Trial 1, the bulls consisted of Angus (n = 15), Brahman (n = 14), Hereford (n = 15) and Senepol (n = 14). In Trial 2, the breeds were Angus (n = 15), Brahman (n = 16), Romosinuano (n = 13) and Nellore x Brahman (n = 9). Trial bulls generally showed delayed growth compared with grain-fed bulls in temperate environments. Breed influenced semen traits (percentage sperm motility, normal spermatozoa and those with primary abnormalities) in both trials. Temperate Bos taurus breeds (Angus, Hereford) were generally superior to Bos indicus breeds (Brahman, Nellore x Brahman). Tropically-adapted Bos taurus breeds (Senepol, Romosinuano) were intermediate for those traits tested. In general, tropically-adapted Bos taurus breeds were more similar in reproductive development to temperate Bos taurus than to Bos indicus breeds. Breed by test period interactions occurred and were mainly influenced by delayed sexual maturity of Bos indicus bulls. Qualitative semen traits increased with bull age, particularly from 12 to 18 mo. Scrotal circumference development was slower in the Bos indicus breeds. Bulls of satisfactory BSE status at 18.1 to 22 mo of age were 73.9% in Trial 1 and 58.5% in Trial 2. Brahman bulls had the least satisfactory BSE scores in both years (Trial 1, 44.4%; Trial 2, 22.2%). Most bulls failed to achieve satisfactory BSE status due to a small SC relative to age (Trial 1, 66%; Trial 2, 72%). The most efficacious use of the BSE was > or = 15 mo in Bos taurus bulls and > 18 mo for Bos indicus bulls. Although the BSE has proven to be useful for the assessment of young, pasture-raised bulls in semi-tropical environments, use of SC

  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. Mechanical bull thumb

    SciTech Connect

    Ginthner, T.P.; Schabel, S.I.

    1981-11-01

    Two patients suffered intra-articular fractures of the thumb while attempting to ride a mechanical bull. The mechanism of injury in these patients and the riding technique which leads to fracture are discussed.

  19. 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 ...

  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. PMID:26081351

  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. PMID:25088062

  2. Artificial insemination and cryopreservation of semen from nondomestic birds

    USGS Publications Warehouse

    Gee, G.F.

    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.

  3. 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...

  4. Frozen shoulder

    MedlinePlus

    Frozen shoulder is a condition in which the shoulder is painful and loses motion because of inflammation. ... The capsule of the shoulder joint has ligaments that hold the shoulder bones to each other. When the capsule becomes inflamed, the shoulder bones are ...

  5. Relative efficacy of egg yolk and soya milk-based extenders for cryopreservation (−196°C) of buffalo semen

    PubMed Central

    Chaudhari, D.V.; Dhami, A. J.; Hadiya, K. K.; Patel, J. A.

    2015-01-01

    Aim: The aim was to compare commercially available soybean milk-based extenders, viz. Bioxcell® and Optixcell® (IMV, France) with standard Tris-citrate-fructose-egg yolk-glycerol (TFYG) extender for cryopreservation of buffalo semen. Materials and Methods: Semen was collected twice a week in artificial vagina from six sexually mature, 4-6 years old, healthy breeding bulls of Surti buffalo breed. In all 48 qualifying ejaculates (8 per bull) having initial motility >70% were split into three equal aliquots and were diluted (at 34°C keeping 100×106 sperm ml−1) in TFYG, Bioxcell and Optixcell extenders. The French mini straws filled from each aliquot were gradually cooled to 4-5°C, equilibrated at 4°C for 4 h and frozen in liquid nitrogen 2 vapor using programmable biofreezer. Just before freezing (post-equilibration) and 24 h after frozen storage, the samples were evaluated for various sperm quality parameters using standard protocols. Frozen semen straws were thawed in a water bath at 37°C for 30 s. The post-thaw incubation survival (37°C for 1 h) was assessed through motility rating at 0, 30 and 60 min of incubation. Results: The mean percentages of prefreeze sperms in TFYG, Bioxcell and Optixcell extenders in terms of progressive motility (69.48±0.37, 68.02±0.49, 70.94±0.38), viability (79.21±0.39, 77.38±0.48, 81.58±0.38), total abnormalities (7.90±0.14, 8.60±0.16, 7.08±0.15), intact acrosome (89.54± 0.18, 88.58±0.22, 90.52±0.21) and hypoosmotic swelling (HOS) reactivity (67.96±0.32, 65.65±0.42, 70.23±0.37) varied significantly (p<0.05) between extenders. Similar pattern of significant (p<0.05) variations between these extenders for post-thaw sperm progressive motility (47.71±0.79, 44.38±0.85, 49.90±0.90), viability (57.19±0.79, 53.85±0.84, 59.67±0.91), total abnormalities (12.33±0.17, 12.75±0.21, 11.27±0.18), intact acrosome (76.83±0.23, 75.90± 0.27, 78.50±0.25) and HOS reactivity (45.02±0.84, 42.31±0.82, 47.81±0.90) was

  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. PMID:17524303

  7. The effect of cryopreservation on goat semen characteristics related to sperm freezability.

    PubMed

    Dorado, J; Muñoz-Serrano, A; Hidalgo, M

    2010-08-01

    Seminal quality parameters were used to evaluate the effect of freeze-thawing procedure on goat sperm characteristics, and to relate possible changes in sperm parameters to cryopreservation success. Semen samples (n=110) were frozen with TRIS and milk-based extenders and thawed. Sperm quality parameters (motility, morphology and acrosome) were compared between fresh and frozen-thawed samples. Sperm freezability was judged by classifying the semen samples as "suitable" or "not suitable" according to the sperm quality parameters assessed before and after thawing. Fertility data was obtained after cervical insemination with frozen semen doses. The ejaculates were grouped into two categories according to their fertility results. In experiment 1, significant differences were found between semen extenders (P<0.001), bucks (P<0.05) and ejaculates within the same male (P<0.05) in terms of sperm quality. There was no seasonal effect (P>0.05) on the majority of the sperm parameters assessed after thawing. Moreover, significant differences (P<0.001) in semen parameters assessed in fresh semen and frozen-thawed samples were found between groups. The effect of the freeze-thawing procedure on sperm quality parameters was also different (P<0.05) between extenders within the same group. The number of sperm quality parameters that had changed after cryopreservation was lower in "suitable" semen samples before and after thawing. In experiment 2, no differences (P>0.05) in semen parameters assessed in fresh semen and frozen-thawed samples were found between groups. The effect of freezing and thawing on sperm quality parameters were different (P<0.05) between extenders within the same group. Only mean beat cross frequency (BCF) values were significantly higher (P<0.05) in TRIS diluted samples that led to successful pregnancies after artificial insemination. In conclusion, CASA-derived motility parameters, together with traditional semen assessment methods, give valuable information on

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

    PubMed

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

    2008-11-01

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

  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. 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. PMID:25218757

  11. Factors affecting economics of using sexed semen in dairy cattle.

    PubMed

    McCullock, Katelyn; Hoag, Dana L K; Parsons, Jay; Lacy, Michael; Seidel, George E; Wailes, William

    2013-10-01

    The use of sexed semen in the dairy industry has grown rapidly. However, high costs and low fertility have limited the use of this potentially valuable tool. This study used simulation to evaluate 160,000 combinations of key variables in 3 spheres of influence related to profit feasibility: (1) market (e.g., milk and calf prices), (2) dairy farm management (e.g., conception rates), and (3) technology (e.g., accuracy of sexing). These influential variables were used to determine the most favorable circumstances in which managers or technicians can effect change. Three distinct scenarios were created to model 3 initiatives that a producer might take with sexed semen: (1) using sexed semen on heifers, (2) using sexed semen on heifers and a fraction of the genetically superior cows, and (3) using sexed semen on heifers and a fraction of the genetically superior cows, and breeding all other cows with beef semen. Due to the large number of management, market, and technology combinations, a response surface and interpretive graphs were created to map the scope of influence for the key variables. Technology variables such as the added cost of sexed semen had relatively little effect on profitability, defined as net present value gain per cow, whereas management variables such as conception rate had a significant effect. Milk price had relatively little effect within each scenario, but was important across scenarios. Profitability was very sensitive to the price of dairy heifer calves, relative to beef and dairy bull calves. Scenarios 1 and 2 added about $50 to $75 per cow in net present value, which ranged from $0 to $200 and from $100 to $300, respectively. Scenario 3 usually was not profitable, primarily because fewer excess dairy replacement heifers were available for sale. Dairy heifer price proved to be the most influential variable, regardless of scenario. PMID:23932128

  12. 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. PMID:18638144

  13. Relationship between scrotal infrared temperature patterns and natural-mating fertility in beef bulls.

    PubMed

    Lunstra, D D; Coulter, G H

    1997-03-01

    The infrared temperature pattern (IRT) of the scrotal surface was recorded for 73 yearling beef bulls and a color video thermogram of the pattern of each bull was recorded. The average scortal surface temperature, temperature at the top and bottom of the scrotum, scortal temperature gradient, and thermal class (normal, questionable, or abnormal scortal surface thermal pattern) were recorded for each thermogram. Thirty-seven bulls had a normal temperature pattern (51%), 20 had a questionable pattern (27%), and 16 had an abnormal temperature pattern (22%). Bulls exhibiting abnormal scrotal temperature patterns had lower (P < .05) percentages of sperm exhibiting normal head and tail morphology and had a higher (P < .01) percentage of sperm with proximal droplets than did bulls with normal or questionable thermogram patterns. Thirty bulls with acceptable testis size and semen quality and representing the three thermal classes were each exposed single-sire to approximately 18 heifers during a 45-d pasture breeding period. Pregnancy rate was lower (P < .01) for bulls with abnormal scrotal temperature patterns (68 +/- 4%, n = 8) than for bulls with normal (83 +/- 4%, n = 13) and questionable temperature patterns (85 +/- 4%, n = 9), and pregnancy rate was related significantly to all four major characteristics (surface, top, and bottom temperatures and temperature gradient) of scortal thermograms. Data indicated that bulls with abnormal scortal temperature patterns exhibited a reduced ability to maintain an effective thermal gradient from top to bottom of the testes and that bulls with abnormal scrotal temperature patterns achieved reduced pregnancy rates when used for natural mating. PMID:9078495

  14. 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. PMID:26641162

  15. Prediction of bull fertility.

    PubMed

    Utt, Matthew D

    2016-06-01

    Prediction of male fertility is an often sought-after endeavor for many species of domestic animals. This review will primarily focus on providing some examples of dependent and independent variables to stimulate thought about the approach and methodology of identifying the most appropriate of those variables to predict bull (bovine) fertility. Although the list of variables will continue to grow with advancements in science, the principles behind making predictions will likely not change significantly. The basic principle of prediction requires identifying a dependent variable that is an estimate of fertility and an independent variable or variables that may be useful in predicting the fertility estimate. Fertility estimates vary in which parts of the process leading to conception that they infer about and the amount of variation that influences the estimate and the uncertainty thereof. The list of potential independent variables can be divided into competence of sperm based on their performance in bioassays or direct measurement of sperm attributes. A good prediction will use a sample population of bulls that is representative of the population to which an inference will be made. Both dependent and independent variables should have a dynamic range in their values. Careful selection of independent variables includes reasonable measurement repeatability and minimal correlation among variables. Proper estimation and having an appreciation of the degree of uncertainty of dependent and independent variables are crucial for using predictions to make decisions regarding bull fertility. PMID:26791329

  16. 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

  17. 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

  18. 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...

  19. Addition of oxytocin to semen extender improves both sperm transport to the oviduct and conception rates in pigs following AI.

    PubMed

    Okazaki, Tetsuji; Ikoma, Erena; Tinen, Tukasa; Akiyoshi, Teiichi; Mori, Manabu; Teshima, Hisanori

    2014-01-01

    Oxytocin (OXT) contained in boar semen is known to produce uterine contraction; therefore, we hypothesized that the co-injection of OXT with sperm would improve artificial insemination (AI) using liquid or frozen-thawed boar sperm. We initially examined whether OXT added to semen extender improved sperm transport to the oviduct. Although the addition of OXT did not affect the fresh or frozen-thawed sperm motility or acrosomal integrity, it significantly increased the number of sperm in the oviduct at 6 h after AI injection with OXT, as compared with the control (P < 0.05). Moreover, some sperm were observed in the sperm reservoir of the isthmus in the OXT treatment group, whereas few sperm were observed in the control. When OXT was added to the semen extender immediately prior to AI, the conception rates were significantly higher in both fresh semen and frozen-thawed semen than in the control group (P < 0.05: liquid, 87.5% vs. 70.5%; frozen-thawed, 89.8% vs. 75.0%). From these results, we concluded that the addition of OXT to the semen extender assisted in sperm transportation from the uterus to the oviduct, which resulted in improved reproductive performance. PMID:23829601

  20. Omeprazole and Semen Quality.

    PubMed

    Banihani, Saleem A

    2016-03-01

    A number of studies have linked omeprazole, a commonly used acid reducer under the brand name Prilosec, with semen quality. This MiniReview systematically addresses and summarizes the effect of omeprazole on semen quality, and male infertility. We searched the MEDLINE electronic database for English-language articles using the keywords 'omeprazole' versus 'sperm' and 'testosterone' and the references from selected articles were reviewed, if relevant. In summary, omeprazole does not appear to change semen quality. This may be because, at least in part, it does not alter the basal levels of pituitary-gonadal hormones; in addition, it counteracts the damaging effect of reactive oxygen species. However, further research is still required to confirm this effect. PMID:26572503

  1. Associations between sperm abnormalities, breed, age, and scrotal circumference in beef bulls

    PubMed Central

    Menon, Ajitkumar G.; Barkema, Herman W.; Wilde, Randy; Kastelic, John P.; Thundathil, Jacob C.

    2011-01-01

    The objectives of this study were to determine the associations of breed, age, and scrotal circumference (SC), and their interaction, on the prevalence of sperm abnormalities in beef bulls in Alberta, Canada, and the percentage of satisfactory potential breeders identified during breeding soundness examination solely due to normal sperm morphology. Eosin-nigrosin stained semen smears and evaluation reports of 1642 bull breeding soundness evaluations were procured from 6 veterinary clinics in Alberta. Sperm morphology was determined for at least 100 sperm per bull. The most common defects were detached head [4.86% ± 5.71%; mean ± standard deviation (s)], distal midpiece reflex (6.19% ± 9.13%), and bent tail (1.01% ± 1.54%). Although breed, age, and SC did not significantly affect the prevalence of head or midpiece defects, morphologically normal or abnormal sperm, tail defects were more prevalent in Angus and Hereford bulls compared with other breeds. Overall, solely on the basis of sperm morphology, 1363 (83.0%) bulls were classified as satisfactory potential breeders and the remainder 279 (17.0%) as unsatisfactory (> 30% abnormal sperm, > 20% defective heads, or both). Although not significantly different, the breed with the highest percentage of satisfactory potential breeders was Limousin (90.6%) and the lowest was Hereford (78.8%). That 17% of bulls subjected to breeding soundness evaluation were designated as unsatisfactory solely on the basis of sperm morphology highlights its importance. PMID:22468020

  2. 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. PMID:26660630

  3. The use of integer programming to select bulls across breeding companies with volume price discounts.

    PubMed

    McConnel, M B; Galligan, D T

    2004-10-01

    Optimization programs are currently used to aid in the selection of bulls to be used in herd breeding programs. While these programs offer a systematic approach to the problem of semen selection, they ignore the impact of volume discounts. Volume discounts are discounts that vary depending on the number of straws purchased. The dynamic nature of volume discounts means that, in order to be adequately accounted for, they must be considered in the optimization routine. Failing to do this creates a missed economic opportunity because the potential benefits of optimally selecting and combining breeding company discount opportunities are not captured. To address these issues, an integer program was created which used binary decision variables to incorporate the effects of quantity discounts into the optimization program. A consistent set of trait criteria was used to select a group of bulls from 3 sample breeding companies. Three different selection programs were used to select the bulls, 2 traditional methods and the integer method. After the discounts were applied using each method, the integer program resulted in the lowest cost portfolio of bulls. A sensitivity analysis showed that the integer program also resulted in a low cost portfolio when the genetic trait goals were changed to be more or less stringent. In the sample application, a net benefit of the new approach over the traditional approaches was a 12.3 to 20.0% savings in semen cost. PMID:15377634

  4. 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.

  5. 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

  6. The relationship between fertility potential measurements on cryobanked semen and fecundity of sperm donors.

    PubMed

    Navarrete, T; Johnson, A; Mixon, B; Wolf, D

    2000-02-01

    Sperm penetration assay (SPA) scores obtained from cryobanked semen were correlated with therapeutic insemination (TI) fecundity in a group of established sperm donors, thereby evaluating the efficacy of the SPA in screening donors for sperm banking. While the SPA has been used to separate fertile from infertile males, we altered assay conditions to use frozen semen and to distinguish performance among fertile donors. Three frozen ejaculates from 11 pregnancy-proven donors were analysed. Of 905 TI cycles, 275 recipients achieved 95 pregnancies. There were no significant relationships between fecundity and donor semen, washed sperm parameters, sperm recoveries or recipient age. A significant relationship was revealed between mean SPA scores (range 8.7-66.6 penetrations/ovum) and donor fecundity (range 0.04-0.16, P < 0.03). Sperm concentration was varied in an effort to establish the most sensitive test condition. Using 0.25x10(6) motile spermatozoa/ml, a highly significant relationship was observed (P < 0.002). The four donors with the lowest SPA scores achieved the four lowest fecundities. It is concluded that a modified SPA can be used on frozen donor semen to estimate donor fertility potential. If applied routinely in donor semen banking, poor quality applicants could be excluded, thereby increasing pregnancy rates while decreasing donor screening costs. PMID:10655306

  7. Evaluation of seasonal variations of semen freezability in Leccese ram.

    PubMed

    D'Alessandro, A G; Martemucci, G

    2003-11-20

    The experiment was carried out in Southern Italy (41 degrees N latitude) to examine the effects of seasonal variations of semen freezability in Leccese ram. Semen from five rams, collected every 2 weeks for a whole year, was frozen in straws, using a system based on Tris-fructose egg yolk as extender to constitute semen doses of 100x10(6) spermatozoa. Post-thaw survival and acrosomal status of cells were assessed by dual staining by Hoechst 33258 and FITC-PSA. Three different forms of fluorescence distribution were displayed indicating sperm without acrosome (unstained cells), sperm with damaged acrosome (cells with incomplete fluorescence over the head), sperm with widespread fluorescence (cells completely fluorescent). Motility and kinetic rating at thawing and after 1 and 3h incubation (37 degrees C) were also assessed. Semen frozen in summer and autumn, corresponding to the breeding season, showed the highest (P<0.01) post-thaw survival of spermatozoa (41.7%) and the lowest (P<0.01) incidence of spermatozoa with damaged acrosome. The positive influence of the summer-autumn period was expressed also on motility and kinetic rating of spermatozoa at thawing. The integrity of the acrosomal membrane was positively correlated (P<0.01) with sperm viability before processing (r=0.32) and after thawing (r=0.51). In conclusion, the results show that season exerts a significant influence on semen freezability in Leccese ram, with the best performance occurring the summer and autumn period, corresponding to the reproductive season in temperate zones. PMID:12853182

  8. 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.

  9. Predicting Breed Composition Using Breed Frequencies of 50,000 Markers from the U.S. Meat Animal Research Center 2,000 Bull Project

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Our objective was to evaluate whether breed composition of crossbred cattle could be predicted using reference breed frequencies of SNP markers on the BovineSNP50 array. Semen DNA samples of over 2,000 bulls from 16 common commercial beef breeds were genotyped using the array and used to estimate cu...

  10. 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...

  11. Surgical intrauterine insemination with cat semen cryopreserved with Orvus ES paste or sodium lauryl sulfate.

    PubMed

    Tsutsui, Toshihiko; Mizutani, Tatsuji; Matsubara, Yuka; Toyonaga, Mari; Oba, Hiromichi; Hori, Tatsuya

    2011-02-01

    The mean post-thaw sperm motilities of feline frozen semen prepared with 1% OEP or 3 g/ml SLS as a cryoprotective agent, in addition to 7% glycerin, were 35.0 ± 2.4 and 37.0 ± 2.5%, respectively, showing no significant difference. On unilateral intrauterine insemination (UIUI) using these semen samples at a sperm number of 40 × 10(6), the conception rate was 70.0% (7/10) in the OEP group and 30% (3/10) in the SLS group, showing that the rate was higher in the OEP group, but the difference was not significant. It was suggested that sperm in frozen semen showing the above qualities were transferred to the contralateral uterine horn on UIUI. PMID:20948170

  12. 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

  13. Growth, puberty, and carcass characteristics of Brahman-, Senepol-, and Tuli-sired F1 Angus bulls.

    PubMed

    Chase, C C; Chenoweth, P J; Larsen, R E; Hammond, A C; Olson, T A; West, R L; Johnson, D D

    2001-08-01

    Postweaning growth, sexual development, libido, and carcass data were collected from two consecutive calf crops using 31 Brahman x Angus (B x A), 41 Senepol x Angus (S x A), and 38 Tuli x Angus (T x A) F1 bulls. Following weaning (by mid-September) and preconditioning, at the start of the study (late September) bulls were fed concentrate (three times each week at a rate equivalent to 4.5 kg/d) on bahiagrass pasture for approximately 250 d. At the start of the study and at 28-d intervals, BW, hip height, and scrotal circumference (SC) were measured. Concurrently at 28-d intervals, when the SC of a bull was > or = 23 cm, semen collection was attempted using electroejaculation. Ejaculates were evaluated for presence of first spermatozoa (FS), 50 x 10(6) sperm with at least 10% motility (PU), and 500 x 10(6) sperm with at least 50% motility (PP). After all bulls reached PP they were subjected to two libido tests. Carcass data were collected on all bulls (n = 110) and Warner-Bratzler shear (WBS) force values were assessed on a subset (n = 80). For both years, B x A bulls were heavier (P < 0.05) and taller (P < 0.05) than S x A and T x A bulls at the start and end of the study. However, breed type did not influence (P > 0.10) gain in BW or hip height during the study. Scrotal circumference of T x A bulls was larger (P < 0.05) than that of B x A or S x A bulls at the start of the study, but there was no effect (P > 0.10) of breed type by the end of the study. At PU and PP, B x A bulls were older (P < 0.05), heavier (P < 0.05), and taller (P < 0.05) and had larger (P < 0.05) SC than S x A and T x A bulls. Tuli x Angus bulls were younger (P < 0.05) than S x A bulls at PU and PP but had similar SC. Libido scores tended (P < 0.10) to be lower for B x A than for S x A and T x A bulls. Breed type affected (P < 0.05) carcass traits; B x A bulls had the heaviest (P < 0.05) hot carcass weight, greatest (P < 0.05) dressing percentage, larger (P < 0.05) longissimus muscle area than

  14. 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. PMID:27570305

  15. 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. PMID:26872498

  16. Adjusting the number of spermatozoa in mini-straws by determination of the operative volume of bovine semen.

    PubMed

    Dumont, Pascal

    2002-04-01

    The importance of the number of sperm per insemination on fertility has been well demonstrated in cattle. This number is usually calculated from the concentration in the extended semen and a theoretical value for the operative volume of semen delivered during insemination. The objective of this experiment was to investigate the usefulness of the measurement of the delivered volume of semen when estimating sperm numbers from frozen-thawed mini-straws, by comparing the results obtained with an analytical balance to the theoretical volume. The density of semen extended with Biociphos Plus and Triladyl was determined to be 1.033 g/ml using the gamma sphere method. This value was used to convert semen weight into operative volume. The effect of semen temperature at the time of weighing (37 degrees C versus 20 degrees C) was investigated on six semen batches, two technicians measuring the operative volume of 50 straws for each combination of temperature and semen batch (a total of 1200 weighings). The temperature effect was found to be insignificant, which allowed warm semen to be weighed before motility was assessed during routine quality control. The operative volume was then measured in straws routinely produced at 17 bovine Al centers (12-105 semen batches per center, mostly three straws from each batch, a total of 1912 measurements). The observed volumes were normally distributed around 198.7 microl, 98% measuring between 180 and 210 microl. The operative volume was significantly different among centers (from 192 to 205 microl, P < 0.0001) and among batches within centers (P < 0.0001). The S.D. among straws within batches was 3.4 microl. Some centers showed high variability in straw volume whereas others were more consistent. Determination of the operative volume of frozen-thawed mini-straws by weighing the delivered contents is an accurate method for estimation of the number of sperm per dose. PMID:12035983

  17. Pellet-freezing of Damascus goat semen in a chemically defined extender.

    PubMed

    Khalifa, T A A; El-Saidy, B E

    2006-07-01

    During the breeding season of goats (12 bucks and 64 does) in Egypt, five experiments were conducted using a chemically defined cryoextender (CDE) to investigate: (1) the influence of rates of semen dilution (1:2, 1:4 and 1:19) and methods of thawing of frozen semen pellets (dry thawing versus wet thawing) on sperm progressive motility (SPM), sperm acrosome abnormalities (SAA) and rate of lipid peroxidation in semen as measured by malonaldehyde (MAL) production, and (2) the effect of insemination of does in natural (n = 38) and cloprostenol-synchronized (n = 26) estrus with frozen semen on their kidding rates and prolificacy. Semen (two successive ejaculates/buck) was collected twice a week via an AV and only ejaculates of >2500 x 10(6) sperm/ml and 70% SPM were diluted in one step at 30 degrees C with the CDE, cooled to 5 degrees C over a 4h-period, frozen in the form of 0.30 ml pellets and stored in liquid nitrogen for 72 h. The results revealed that post-thaw SPM of semen diluted at a rate of 1:4 was significantly (P < 0.01) higher than that of semen diluted at the other rates. Dilution of semen at a rate of 1:19 (< or =151 x 10(6) sperm/ml) not only minimized (P < 0.01) pre-freeze and post-thaw SPM, but also augmented (P < 0.01) pre-freeze and post-thaw rates of lipid peroxidation as evidenced by the high level of MAL production and the ability of antioxidants (1mg/ml EDTA, 200 U/ml bovine liver catalase, 0.61 mg/ml reduced glutathione and 0.11 mg/ml sodium pyruvate) to restore (P < 0.01) pre-freeze and post-thaw SPM. Frozen semen pellets exposed to dry thawing had a greater percentage of SPM (P < 0.01) as well as lower values of SAA and MAL (P < 0.01) than those exposed to wet thawing. Although the kidding rates did not vary significantly among does in natural (55.26%) and synchronized (53.85%) estrus, a higher (P < 0.05) prolificacy was obtained after their insemination in natural (1.81+/-0.16) rather than in synchronized (1.22+/-0.11) estrus. PMID:16169690

  18. 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

  19. Prediction of breeding values for tenderness of market animals from measurements on bulls.

    PubMed

    Barkhouse, K L; Van Vleck, L D; Cundiff, L V; Koohmaraie, M; Lunstra, D D; Crouse, J D

    1996-11-01

    Data were tenderness measures on steaks from 237 bulls (Group II) slaughtered after producing freezable semen and on 1,431 related steers and heifers (market animals, Group I) from Angus, Hereford, Pinzgauer, Brahman, and Sahiwal crosses from the Germ Plasm Evaluation project at the U.S. Meat Animal Research Center. Tenderness was assessed through Warner-Bratzler Shear Force (SF), taste panel tenderness (TPT), marbling score (MS), and myofibrillar fragmentation index (MFI). For all traits, as fraction Bos indicus inheritance increased, implied tenderness decreased. Heritability estimates were generally not significantly different from zero. Genetic correlations generally indicated favorable associations among the traits. The range in predicted breeding values of bulls for market animal tenderness was small and from -.34 to .32 kg for market animal shear force. Because of low estimates of heritability for SF or TPT, results from this experiment indicate that selection based on tenderness of steaks sampled from intact or late castrate males slaughtered following collection of freezable quality semen would not be very effective in improving average tenderness of steaks from steers of heifer progeny. If a mean of heritability estimates reported in the literature of .27 for shear value was assumed for market steer and heifer progeny instead of .02 as found in the present study, then selection based on estimates of shear force in young bulls would be relatively more effective in improving shear force of market progeny. PMID:8923175

  20. A sucrose-DMSO extender for freezing rabbit semen.

    PubMed

    Vicente, J S; Viudes-de-Castro, M P

    1996-01-01

    The aim of this study was to define a simple extender for freezing rabbit semen from selected males used to inseminate selected doses to obtain embryos for an embryo bank. Four experiments were carried out. In the first experiment, freezing extender was defined on the basis of the results of the post-thawing motility rate. Three factors and their interactions were studied: final concentration of dimethyl sulphoxide (DMSO) (1, 1.25, 1.5 and 1.75 M), egg yolk (0% of 10 v/v) and sugar (none, or 0.5 M of glucose, lactose, sucrose, or maltose). The sucrose and 1.75 DMSO improved significantly the post-thawing motility rate (sucrose 1.75 M DMSO extender: 42 +/- 3%). In the second experiment, this freezing extender was used to freeze semen from three lines. The post-thawing motility and normal acrosome rates were similar among the lines when the covariates, fresh motility and normal acrosome rates, respectively, were used in the analysis (52 +/- 1 and 66 +/- 1%, respectively). In the third experiment, frozen semen from the White New Zealand line (NZ) was tested by morphological normality and viability of embryos recovered. Recovery data from NZ does inseminated with fresh and frozen semen were compared. No significant differences were found in the number of normal embryos obtained per donor dose (8.9 +/- 0.5) and in their survival after vitrification (52% of live foetuses at 29 days of gestation). The sucrose-DMSO extender and freezing procedure used in this work can offer satisfactory results to apply in conservation and genetic programmes. PMID:8987100

  1. Exposure of prepubertal beef bulls to cycling females does not enhance sexual development.

    PubMed

    Miller, N A; Fike, K E

    2014-08-01

    The objective of this study was to determine whether continuous, long-term, fenceline exposure of prepubertal beef bulls to cycling beef females reduced age at puberty and influenced the percentage of bulls that passed an initial breeding soundness examination (BSE). Bulls (Angus, n = 37; Simmental, n = 22; Hereford, n = 10; Simmental × Angus, n = 8) at an average age of 202 ± 21.5 days were given either continuous fenceline and visual exposure to cycling females (exposed, n = 41) or no exposure (control, n = 36). Estrus was induced in cycling beef females so at least three females were in standing estrus each week during the 182 days of exposure to bulls. Scrotal circumference (SC), body weight, and blood samples were collected every 28 days. When bulls had SC of 26 cm or more, semen samples were obtained monthly via electroejaculation until puberty was achieved (≥50 × 10(6) sperm/mL with at least 10% progressive motility). Behavioral observations were conducted twice monthly: once when females were in estrus and once during diestrus. Homosexual mounting, flehmen responses, and number of times near penned females were recorded for each observation period. Breeding soundness examinations were conducted when the average age of bulls was 364 ± 21.5 days. Normal sperm morphology of at least 70% and sperm motility of at least 30% were required to pass the BSE. Age, body weight, and SC at puberty did not differ between exposed and control bulls (320 ± 28 and 311 ± 29 days; 466.2 ± 12.2 and 437.7 ± 13.5 kg; and 34.4 ± 2.5 and 34.9 ± 2.5 cm, respectively). Percentage of bulls passing their initial BSE did not differ between treatments (exposed, 87.8%; control, 75.0%). Treatment, month, and female estrous stage interacted (P = 0.05) to affect the number of mount attempts and flehmen responses. Exposed bulls entered the cow area more times (P < 0.001) during estrus than diestrus in Months 1, 2, and 3. We concluded that bulls given continuous, long

  2. 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

  3. The cryopreservation of donor semen by a simplified method: use in an IVF and GIFT programme.

    PubMed

    Morroll, D R; Matson, P L; Troup, S A; Izzard, H; Prior, J R; Burslem, R W; Lieberman, B A

    1990-10-01

    The cryopreservation of semen used in assisted reproduction procedures was carried out exclusively by a simplified method in which a mixture of semen and cryoprotectant was contained in 1-ml tuberculin syringes and plunged directly into liquid nitrogen. Donor semen samples halved and frozen in syringes and in straws in a controlled-rate freezer showed no significant difference in post-thaw motility (P = 0.217) or survival (P = 0.217) after 30 min. However, after 180 min the survival rate showed a significant reduction in syringes (P = 0.045). A significant difference (P less than 0.00008) in the rate of fertilization of oocytes was seen in IVF cycles using frozen-thawed donor sperm (58/142, 42%) when compared to fresh sperm from husbands (2315/3926, 59%). A significant reduction (P less than 0.00005) in fertilization rate was also observed in the case of supernumerary oocytes in GIFT cycles with the cryopreserved donor sperm (29/132, 22%) compared to the husbands' sperm (239/514, 46%). However, the pregnancy rate following IVF and embryo replacement was the same after fertilization with fresh sperm (75/351, 21%) as opposed to frozen sperm (3/14, 21%). Furthermore, a higher pregnancy rate was observed in GIFT with frozen donor sperm (9/19, 47%) than with fresh sperm from husbands (28/103, 27%), though this was not statistically significant (P = 0.079). These results show this simplified methods of semen cryopreservation to be effective when used in an IVF and GIFT programme, giving pregnancy rates comparable to fresh normospermic semen samples. The method is simple, quick and inexpensive. PMID:2283181

  4. 29 CFR 1918.84 - Bulling cargo.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... shall be done with the bull line led directly from the heel block. However, bulling may be done from the..., falling, or being pulled from their stationary attachment. (e) Falls led from cargo booms of vessels...

  5. Laparoscopic cryptorchidectomy in standing bulls

    PubMed Central

    KANEKO, Yasuyuki; TORISU, Shidow; KITAHARA, Go; HIDAKA, Yuichi; SATOH, Hiroyuki; ASANUMA, Taketoshi; MIZUTANI, Shinya; OSAWA, Takeshi; NAGANOBU, Kiyokazu

    2015-01-01

    Laparoscopic cryptorchidectomy without insufflation was applied in 10 standing bulls aged 3 to 15 months. Nine bulls were preoperatively pointed out intra-abdominal testes by computed tomography. Preoperative fasting for a minimum of 24 hr provided laparoscopic visualization of intra-abdominal area from the kidney to the inguinal region. Surgical procedure was interrupted by intra-abdominal fat and testis size. It took 0.6 to 1.5 hr in 4 animals weighing 98 to 139 kg, 0.8 to 2.8 hr in 4 animals weighing 170 to 187 kg, and 3 and 4 hr in 2 animals weighing 244 and 300 kg to complete the cryptorchidectomy. In conclusion, standing gasless laparoscopic cryptorchidectomy seems to be most suitable for bulls weighing from 100 to 180 kg. PMID:25715955

  6. Simple and effective methods of freezing capercaillie (Tetrao urogallus L.) semen.

    PubMed

    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 x 10⁶ 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

  7. 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

  8. Relationship of conventional and fluorescent microscopic technique to assess in vitro semen quality status of Murrah buffalo males

    PubMed Central

    Shivahre, P. R; Gupta, A. K; Panmei, A; Yadav, B. R; Bhakat, M; Mohanty, T. K; Kumaresan, A; Kumar, V; Dash, S. K; Singh, S

    2015-01-01

    In vitro fertility assessment using fluorescent technique is a better predictor of fertility status of bulls as compared to traditional semen quality assessment techniques, therefore, the study was planned to assess in vitro fertility status of bulls based on conventional and fluorescent techniques. Seventy-three ejaculates were collected from 12 Murrah buffalo bulls maintained at Artificial Breeding Research Centre, NDRI, Karnal, India for the experiment and subjected to statistical analysis using SYSTAT. The mean values of ejaculate volume (ml), mass activity, individual motility (%), sperm concentration (millions/ml), live sperm (%), total abnormalities (%), HOST (%) and acrosomal integrity (%) were 2.70 ± 0.28, 2.8 ± 0.14, 63.8 ± 2.16, 1749.7 ± 122.24, 77.3 ± 2.48, 6.2 ± 0.51, 75.1 ± 1.81 and 84.5 ± 2.26, respectively. The repeatability estimates were significant (P<0.05) for ejaculate volume (0.34 ± 0.137), acrosomal integrity (0.29 ± 0.134) and live percentage (0.28 ± 0.133), indicating sufficient bull to bull variation for the parameters. The mean values of seminal attributes of fluorescent based criteria of CMA3 (Chromomycin A3), SYBR-PI and FITC-PNA (fluorescent isothiocynate-conjugated peanut agglutinin) were 5.25 ± 0.41, 67.91 ± 1.24 and 82.00 ± 1.25 percent, respectively. Bulls were ranked on the basis of expected producing ability (EPA) for semen characteristics assessed by conventional and fluorescent criteria. Rank correlations were found to be significant for FITC with most of the parameters evaluated by conventional methods. In conclusion, among the conventional criteria, individual motility (%) revealed ranking of bulls almost similar to that of fluorescent criteria. PMID:27175204

  9. Changes in the use of young bulls

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Availability of genomic information since 2008 has increased accuracy of genetic evaluations for young bulls in Holstein (HO), Jersey (JE), and Brown Swiss (BS). As a result, AI organizations have been aggressively promoting young bulls and producers have been using young bulls more extensively. Num...

  10. Effects of different cryoprotectants and freezing methods on post-thaw boar semen quality.

    PubMed

    Yang, Chung-Hsun; Wu, Ting-Wen; Cheng, Feng-Pang; Wang, Jiann-Hsiung; Wu, Jui-Te

    2016-03-01

    The current study aimed to investigate the effects of different concentrations of glycerol (0%, 1%, 2%, 3%, and 5%) and dimethylacetamide (DMA: 0%, 1%, 3%, and 5%) on post-sperm quality characteristics following semen freezing in dry ice (D) or liquid nitrogen (N). Semen was collected from Duroc boars and was allocated to 32 treatment groups for cryopreservation. Analysis of post-thaw semen quality and fertility after artificial insemination (AI) was used to examine the combinatorial effects of different treatments. The best scores for post-thaw sperm motility, sperm viability, and sperm acrosomal integrity were observed in semen frozen in: (a) dry ice in the presence of 5% glycerol and no DMA (16D-treatment); (b) dry ice in the presence of 3% glycerol and no DMA (9D-treatment); and (c) liquid nitrogen in the presence of 3% glycerol and 1% DMA (10N-treatment), with no significant difference observed among these three treatments. The farrowing rates after AI with post-thawed semen after 9D- and 10N-treatments were 33% and 50%, respectively. To summarize, the results of the present study indicated that the freezing extender containing 3% glycerol in combination with the straw-freezing method using dry ice produced the best post-thaw quality parameters of boar semen. Combinations of glycerol and DMA did not enhance the cryosurvival of boar spermatozoa. PMID:26952752

  11. Effect of ruminally undegradable protein from fish meal on growth and reproduction of peripuberal Brahman bulls.

    PubMed

    Rocha, A; Carpena, M; Triplett, B; Forrest, D W; Randel, R D

    1995-04-01

    Thirty-nine Brahman bulls (301.7 +/- 4.1 d; 202.7 +/- 4.7 kg) were allotted to one of two treatments and fed soybean meal (SBM)- or fish meal (FIS)-based supplements and hay to examine the effects of source of protein on growth and reproductive development. The fish meal supplement had 72% ruminally undegradable protein (RUP) and the soybean meal supplement had 47% RUP. Bulls assigned to the FIS treatment had higher (P < .01) total weight gain (81.2 +/- 1.4 vs 71.2 +/- 2.2 kg), higher (P < .01) ADG (.97 +/- .02 vs .85 +/- .03 kg), and better (P < .05) feed:gain ratio (7.6 +/- .1 vs 8.6 +/- .1 feed/BW gain for FIS vs SBM, respectively). Age at first motile spermatozoa was not affected (P > .05) by source of protein (429.9 +/- 9.6 vs 427.2 +/- 9.5 d, for bulls receiving FIS or SBM supplements, respectively). Likewise, age at puberty (473.3 +/- 21.7 d vs 465.9 +/- 12.9 d for bulls receiving FIS and SBM supplements, respectively) was similar for both treatment groups. There were no differences between treatments in scrotal circumference at those stages. At puberty semen quality was similar for bulls receiving FIS or SBM treatments, and no differences existed in LH and testosterone concentrations between treatments. We conclude that fish meal supplement increased growth but did not alter reproductive parameters in Brahman bulls. PMID:7628971

  12. Gobbledygook and the golden bull.

    PubMed

    1987-12-19

    Once again the Department of Health and Social Security is among the main contenders for this year's Golden Bull Awards. This is not an acknowledgement of its [Illegible word] entrepreneurialism, or even a rosette for being on target. It's a booby prize for producing gobbledygook. PMID:27319524

  13. 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...

  14. Long-term Effects of Pyrethrin and Cyfluthrin, a Type II Synthetic Pyrethroid, Insecticide Applications on Bull Reproductive Parameters.

    PubMed

    Stewart, J L; Shipley, C F; Ireland, F A; Jarrell, V L; Timlin, C L; Shike, D W; Felix, T L

    2016-10-01

    The objectives of this study were to determine effects of cyfluthrin and pyrethrin spray products, used in combination with cyfluthrin topical and ear tag applications, on bull reproductive parameters over 18 weeks. Angus or Angus x Simmental bulls were randomly assigned to one of three treatment groups: (i) no exposure to pyrethrins/cyfluthrin (CONT; n = 10), (ii) cyfluthrin ear tag and topical applications (ET; n = 10), or (iii) cyfluthrin ear tag, topical, premise spray and pyrethrin fog spray applications (ET+S; n = 8). Bull body weight was measured every 3 week, and body condition score and scrotal circumference were recorded on weeks 0, 9 and 18. Semen and serum were collected every 3 weeks for sperm evaluation and testosterone measurement, respectively. There was a treatment × week interaction (p < 0.01) for sperm with primary defects; bulls in CONT group had a greater (p = 0.01) percentage of sperm with primary defects than bulls treated with insecticides at week 18. Overall and progressive sperm motility, normal sperm morphology, secondary sperm defects and serum testosterone concentrations changed (p < 0.01) over time in all bulls; however, treatment did not affect (p ≥ 0.13) any of these parameters. There were also no treatment effects (p ≥ 0.08) on bull body weight, body condition score or scrotal circumference. The use of pyrethrin- and cyfluthrin-based insecticides, regardless of application, did not negatively affect reproductive parameters in beef bulls when administered over 18 weeks. PMID:27411861

  15. 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...

  16. Noncytopathic bovine viral diarrhea virus can persist in testicular tissue after vaccination of peri-pubertal bulls but prevents subsequent infection.

    PubMed

    Givens, M Daniel; Riddell, Kay P; Walz, Paul H; Rhoades, Jim; Harland, Richard; Zhang, YiJing; Galik, Patricia K; Brodersen, Bruce W; Cochran, Anna M; Brock, Kenny V; Carson, Robert L; Stringfellow, David A

    2007-01-15

    The objectives of this research were to evaluate the risk of prolonged testicular infection as a consequence of vaccination of peri-pubertal bulls with a modified-live, noncytopathic strain of BVDV and to assess vaccine efficacy in preventing prolonged testicular infections after a subsequent acute infection. Seronegative, peri-pubertal bulls were vaccinated subcutaneously with an approximate minimum immunizing dose or a 10x standard dose of modified-live, noncytopathic BVDV or were maintained as unvaccinated controls. Forty-nine days after vaccination, all bulls were intranasally inoculated with a noncytopathic field strain of BVDV. Semen and testicular biopsies collected after vaccination and challenge were assayed for BVDV using virus isolation, reverse transcription-nested PCR, or immunohistochemistry and the identity of viral strains was determined by nucleotide sequencing of PCR products. The vaccine strain of BVDV was detected in testicular tissue of vaccinated bulls as long as 134 days after immunization. Prolonged testicular infections with the challenge strain were detected only in unvaccinated bulls as long as 85 days after challenge. Whereas vaccination caused prolonged testicular infection in some bulls, it did prevent subsequent infection of testicular tissue with the challenge strain. This research demonstrates that subcutaneous vaccination of naïve, peri-pubertal bulls with a noncytopathic, modified-live strain of BVDV can result in prolonged viral replication within testicular tissue. The risk for these prolonged testicular infections to cause venereal transmission of BVDV or subfertility is likely to be low but requires further investigation. PMID:17005300

  17. 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. PMID:25078112

  18. Semen residual viral load and reproductive outcomes in HIV-infected men undergoing ICSI after extended semen preparation.

    PubMed

    Zamora, Maria Jose; Obradors, Albert; Woodward, Bryan; Vernaeve, Valerie; Vassena, Rita

    2016-06-01

    The aim of this study was to evaluate the residual presence of the human immunodeficiency virus (HIV) following a triple gradient extended semen wash from ejaculates of serodiscordant couples, and analyse their reproductive outcomes after intracytoplasmic sperm injection (ICSI). For this purpose, a retrospective analysis of our database was performed in serodiscordant couples, with HIV-infected men and non-infected women, using fresh or frozen sperm with ICSI in oocytes from either the patients or donors from January 2006 to September 2013. Overall, the rate of positive HIV test after semen washing was 1.86%. The positive beta human chorionic gonadotrophin, clinical and ongoing pregnancy rates in patients with their own oocytes were 47.1%, 37.5% and 30.8%, respectively, and 58.6%, 50.8% and 39.1%, respectively, in oocyte donation cycles. To summarize, the described method of sperm washing based on triple gradient sperm selection coupled with extensive centrifugations is a highly reliable technique for HIV removal, as it provides lower than reported post-wash positive tests while maintaining high pregnancy rates in assisted reproduction cycles. Despite extensive personnel training and effectiveness of the washing protocol, post-wash HIV test on semen is recommended to identify residual positive samples. PMID:26995657

  19. Neuroretinitis following bull ant sting.

    PubMed

    Ullrich, Katja; Saha, Niladri; Lake, Stewart

    2012-01-01

    Cat scratch disease causes the majority of cases of neuroretinitis. Neuroretinitis is characterised by clinical features of papillitis, macular oedema and macular star. We report a case study of infection with Bartonella henselae most likely transmitted by a bull ant sting. The patient presented with blurred vision and reduced visual acuity after being stung by an ant in her garden some 7 days earlier. Further testing revealed positive serology to B henselae and the patient improved with appropriate treatment. PMID:22865803

  20. 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.

  1. Impact of pig insemination technique and semen preparation on profitability.

    PubMed

    Gonzalez-Peña, D; Knox, R V; Pettigrew, J; Rodriguez-Zas, S L

    2014-01-01

    Artificial insemination technique and semen preparation impact boar utilization efficiency, genetic dissemination, and biosecurity. Intrauterine (IUI) and deep intrauterine (DUI) AI techniques require lower number of spermatozoa per dose compared to conventional (CON) AI. Frozen semen (FRO) has been associated with lower reproductive performance compared to fresh semen (FRE) preparation. The combined effects of 3 AI techniques (CON, IUI, and DUI) and 2 semen preparations (FRE and FRO) on the financial indicators of a pig crossbreeding system were studied. A 3-tier system was simulated in ZPLAN and the genetic improvement in a representative scenario was characterized. The cross of nucleus lines B and A generated 200,000 BA sows at the multiplier level. The BA sows were inseminated (CON, IUI, or DUI) with FRE or FRO from line C boars at the commercial level. Semen preparation and AI technique were represented by distinct sow:boar ratios in the C × BA cross. A range of farrowing rates (60 to 90%) and litter sizes (8 to 14 liveborn pigs) were tested. Genetic improvement per year for number born alive, adjusted 21-d litter weight, days to 113.5 kg, backfat, and ADG were 0.01 pigs per litter, 0.06 kg, -0.09 d, -0.29 mm, and 0.88 g, respectively. On average, the net profit for FRE (FRO) increased (P-value < 0.0001) from CON to IUI and DUI by 2.2 (3.2%) and 2.6% (4%), respectively. The differences in profit between techniques were driven by differences in costs. Differences in fixed costs between IUI and DUI relative to CON were -2.4 (-5.2%) and -3.4% (-7.4%), respectively. The differences in total costs between FRE and FRO were lower than -5%. The difference in variable costs between FRE and FRO ranged from -5.3 (CON) to -24.7% (DUI). Overall, insemination technique and semen preparation had a nonlinear effect on profit. The average relative difference in profit between FRE and FRO was less than 3% for the scenarios studied. PMID:24352964

  2. Expanding the dairy herd in pasture-based systems: the role of sexed semen use in virgin heifers and lactating cows.

    PubMed

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

    2013-10-01

    A model was developed to examine the effects of sexed semen use in virgin heifers and lactating cows on replacement heifer numbers and rate of herd expansion in a seasonal dairy production system. Five separate herds were established according to the type of semen used: conventional frozen-thawed (Conv), sexed fresh semen used in lactating cows for the first 3 wk of the breeding season (SFre1), sexed frozen-thawed semen used in lactating cows for the first 3 wk of the breeding season (SFro1), sexed fresh semen used in lactating cows for the first 6 wk of the breeding season (SFre2), or sexed frozen-thawed semen used in lactating cows for the first 6 wk of the breeding season (SFro2). In the SFro1, SFre1, SFro2, and SFre2 herds, sexed semen was used for the first and second artificial insemination in virgin heifers. Pregnancy rates achieved with sexed fresh and sexed frozen-thawed semen were assumed to be 94 and 75% of those achieved with conventional frozen-thawed semen, respectively. Initial herd size was 100 cows, which was maintained for the first 2 yr of the 15-yr simulation, after which all available replacement heifers were retained to facilitate herd expansion. Two different scenarios of land availability were examined for each of the 5 herds: land available allowed expansion to a maximum herd size of 150 cows (S1), or land available allowed expansion to a maximum herd size of 300 cows (S2). Once maximum herd size was reached, sexed semen use was discontinued and all excess heifer calves were sold at 1 mo old. All capital expenditure associated with expansion was financed with a 15-yr loan. Each of the 10 different options was evaluated in terms of annual farm profit, annual cash flow, and total discounted net profit. The use of fresh sexed semen generated more replacement heifers, leading to faster herd expansion compared with frozen-thawed sexed semen and conventional frozen-thawed semen. Maximum herd size under S1 was reached in yr 5, 5, 4, 5, and 7 for

  3. Frozen shoulder - aftercare

    MedlinePlus

    Adhesive capsulitis - aftercare; Frozen shoulder syndrome - aftercare ... Krabak BJ, Banks NL. Adhesive capsulitis. In: Frontera WR, Silver JK, eds. Essentials of Physical Medicine and Rehabilitation . 2nd ed. Philadelphia, PA: Elsevier Saunders;2008: ...

  4. Effect of different antioxidant additives in semen diluent on cryopreservability (−196°C) of buffalo semen

    PubMed Central

    Patel, Hardik A.; Siddiquee, G. M.; Chaudhari, Dinesh V.; Suthar, Vishal S.

    2016-01-01

    Aim: The aim of this study was to evaluate the effect of different antioxidant additives in standard tris-fructose-egg yolk-glycerol (TFYG) extender on the cryopreservability of buffalo semen. Materials and Methods: Semen collection using artificial vagina, twice weekly for 5 weeks from three pedigreed health breeding bulls of Mehsani breed, aged between 6 and 8 years. Immediately after initial evaluation all 30 qualifying ejaculates (10/bull) were split into three aliquots and diluted at 34°C keeping the concentration of 100 million spermatozoa/ml with standard TFYG extender as control and TFYG having two antioxidant additives - Cysteine HCl at 1 mg/ml and ascorbic acid at 0.2 mg/ml to study their comparative performance. Semen filled in French Mini straws using IS-4 system and gradually cooled to 4°C and equilibrated for 4 h in cold handing cabinet. After completion of equilibration, straws were cryopreserved in LN2 by Programmable Bio-freezer. Semen was examined at post-dilution, post-equilibration, and post-thaw stages for sperm quality parameters, and at each stage plasma was separated for enzymatic analysis of aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and alkaline phosphatase (AKP). Results: The mean percentage of sperms in TFYG, TFYG + cysteine HCl and TFYG + ascorbic acid diluents at post-thaw stage in terms of progressive motility (52.83±0.52, 57.83±0.52, 57.83±0.52), livability (78.70±0.21, 82.33±0.23, 81.73±0.22), and abnormality (5.43±0.21, 5.03±0.17, 5.23±0.18) varied significantly (p<0.05) between control TFYG and TFYG having antioxidant additives. The mean U/L activities of AST (78.70±0.47, 72.80±0.48, 73.30±0.54), LDH (172.70±0.41, 155.78±0.42, 156.33±0.41), and AKP (103.61±0.34, 90.20±0.34, 91.03±0.34) in semen diluted with TFYG, TFYG + cysteine HCl and TFYG + ascorbic acid diluents at post-thaw stage, respectively, which showed significantly (p<0.05) higher leakage of enzymes in control TFYG than TFYG

  5. Cloned embryos from semen. Part 1: in vitro proliferation of epithelial cells on embryonic fibroblasts after isolation from semen by gradient centrifugation.

    PubMed

    Nel-Themaat, Liesl; Gómez, Martha C; Pope, C Earle; Lopez, Monica; Wirtu, Gemechu; Cole, Alex; Dresser, Betsy L; Lyons, Leslie A; Bondioli, Kenneth R; Godke, Robert A

    2008-03-01

    Although epithelial-like somatic cells have been previously isolated from semen, cell proliferation rates were low. Culture of whole semen samples resulted in loss of potentially valuable spermatozoa. The aims of the present study were to: (1) isolate somatic cells from semen, while preserving sperm viability, and (2) optimize in vitro culture conditions for semen-derived epithelial cells. Density gradient centrifugation of washed ejaculates of two rams (Ovis aries) (n = 24) and one eland bull (Taurotragus oryx) (n = 4) was performed using a three-layer discontinuous Percoll column consisting of 90% (P-90), 50% (P-50), and 20% (P-20) Percoll. In vitro culture and Trypan Blue staining indicated that live somatic cells settled in the P-20 layer. Nonmotile spermatozoa were recovered at the P-50 and P-90 interfaces, whereas motile spermatozoa were collected in the pellet from the P-90 layer. Subsequently, somatic cells isolated from the P-20 layer were plated either on inactivated 3T3 mouse embryonic fibroblast feeder layers, collagen-coated plates with 3T3 feeder cell inserts, or on collagen-coated plates. Initial somatic cell plating was similar among treatments, but proliferation significantly increased when cocultured with 3T3 cells (feeder or insert). Furthermore, two different types of epithelial cells were obtained. The exact origin of the cells in the male reproduction system is uncertain and probably variable. The present method of cell isolation and in vitro culture may be of value for preserving endangered species. Specifically, cells isolated and cultured from cryopreserved semen of nonliving males could be used for producing embryos by somatic cell nuclear transfer. PMID:18241128

  6. 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%. PMID:17085891

  7. 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

  8. Stainless steel welding and semen quality

    SciTech Connect

    Jelnes, J.E.; Knudsen, L.E. )

    1988-01-01

    Questionnaire studies of patients from fertility clinics suggest that welders may have an increased risk of reduced semen quality. In this study, welders and nonwelders from the same plants were asked to provide blood, urine, and semen samples. Urine was analyzed for chromium and nickel, and for mutagenic activity and metal concentration; blood for metal concentrations, immunoglobulin G, total protein, and measures of genotoxicity in lymphocytes; and semen was evaluated by standard semen analysis. Results of the semen evaluation, presented here, showed no difference in semen quality between welders and nonwelders. Because the metal dust exposure of nonwelders in the plant may be higher than that in the general population, welders were also compared to referents not working in the metal industry. Again, no decrease in semen quality associated with welding was demonstrated.

  9. 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. PMID:26282524

  10. Does toxic fescue decrease bull fertility?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Knowledge of the detrimental effects of toxic tall fescue on bull reproductive performance is minimal. In natural breeding, reduced bull performance could decrease the pregnancy rate of the cowherd. Scientists from ARS in Booneville, AR, and the University of Arkansas investigated the influence of f...

  11. 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...

  12. Impact of bull development on reproductive success

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The herd bull influences overall herd fertility more than any other single animal. However, a bull must be developed properly and have reached puberty to be fertile. Puberty is defined by an ejaculate containing a minimum of 50 x 106 total sperm with at least 10% progressive motility; however, this ...

  13. 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. PMID:27324452

  14. Expanding the dairy herd in pasture-based systems: The role of sexed semen within alternative breeding strategies.

    PubMed

    Murphy, C; Shalloo, L; Hutchinson, I A; Butler, S T

    2016-08-01

    A simulation model was developed to determine the effects of sexed semen use in heifers and lactating cows on replacement heifer numbers and rate of herd expansion in a seasonal dairy production system. Five separate artificial insemination (AI) protocols were established according to the type of semen used: (1) conventional frozen-thawed semen (CONV); (2) sexed semen in heifers and conventional semen used in cows (SS-HEIFER); (3) sexed semen in heifers and a targeted group of cows (body condition score ≥3 and calved ≥63 d), with conventional semen used in the remainder of cows (SS-CONV); (4) sexed semen in heifers and a targeted group of cows, with conventional semen in the remainder of cows for the first AI and conventional beef semen used for the second AI (SS-BEEF); or (5) sexed semen in heifers and a targeted group of cows, with conventional semen in the remainder of cows for the first AI and short gestation length semen used for the second AI (SS-SGL). Each AI protocol was assessed under 3 scenarios of sexed semen conception rate (SS-CR): 100, 94, and 87% relative to that of conventional semen. Artificial insemination was used on heifers for the first 3 wk and on cows for the first 6 wk of the 12-wk breeding season. The initial herd size was 100 cows, and all available replacement heifers were retained to facilitate herd expansion, up to a maximum herd size of 300 cows. Once maximum herd size was reached, all excess heifer calves were sold at 1 mo old. All capital expenditure associated with expansion was financed with a 15-yr loan. Each AI protocol was evaluated in terms of annual farm profit, annual cash flow, and total discounted net profit. The SS-CONV protocol generated more replacement heifers than all other AI protocols, facilitating faster expansion, and reached maximum herd size in yr 9, 9, and 10 for 100, 94, and 87% SS-CR, respectively. All AI protocols, except SS-BEEF and SS-SGL at 87% SS-CR, reached maximum herd size within the 15-yr period

  15. 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.

    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.

  16. Quality of bull spermatozoa after preparation by single-layer centrifugation.

    PubMed

    Goodla, Lavanya; Morrell, Jane M; Yusnizar, Yulnawati; Stålhammar, Hans; Johannisson, Anders

    2014-01-01

    The present study aimed to evaluate the effect of single-layer centrifugation (SLC) through a species-specific colloid (Androcoll-B; patent pending, J. M. Morrell) on bull sperm quality. Computer-assisted sperm analysis of motility and flow cytometric analysis of sperm viability (SYBR-14/propidium iodide staining), chromatin integrity (acridine orange staining), reactive oxygen species production [Hoechst 33258-hydroethidine-2',7'-dichlorodihydrofluorescein diacetate (HO-HE-DCFDA) staining], mitochondrial membrane potential (staining with JC-1 probe), and protein tyrosine phosphorylation (specific antibody staining) were performed on unselected and SLC-selected sperm samples. Single-layer centrifugation of bull spermatozoa resulted in the selection of a sperm population that had high mitochondrial membrane potential, a higher content of phosphorylated protein, and more reactive oxygen species than control samples. Sperm chromatin damage was lower in the SLC samples although sperm viability and motility did not differ between SLC samples and controls. These observations suggest that SLC of bull semen in a soybean-containing extender improved some, but not all, parameters of sperm quality. PMID:24534497

  17. A multilaboratory study on the variability of bovine semen analysis.

    PubMed

    Brito, Leonardo F C

    2016-01-15

    To evaluate the variability of semen analysis, five replicates of 10 different bovine frozen semen batches were coded with different identification numbers and submitted to various laboratories for evaluation. Three studies were conducted: study I included eight laboratories in semen processing centers in the United States; study II included one laboratory in one semen processing center and five veterinary university laboratories in the United States; and study III included five veterinary university laboratories in Brazil. Evaluation methodology, sample classification criteria, and reporting format varied considerably among laboratories. There were laboratory effects (P < 0.05) on sperm concentration, motility, and morphology results in all studies. When Bland-Altman plots were evaluated, differences in sperm concentration were approximately between -5 and +5 × 10(6) sperm/mL in study I, when the same method of evaluation was used by all laboratories but ranged between -30 and +30 × 10(6) sperm/mL in studies II and III. Differences in the proportions of motile sperm were approximately -30% to +30%, and differences in the proportion of normal sperm were -15% to +15% in studies I and II; these differences were -15% to +15% and -10% to +10%, respectively, in study III. Mean absolute (one tail) proportional differences in estimates across all laboratories ranged from 9% to 31%, 16% to 37%, and 9% to 14% for sperm concentration, motile sperm, and normal sperm across studies; much larger (48%-86%) differences were observed for sperm abnormality categories. Intralaboratory and interlaboratory precision varied considerably across laboratories and seemed to be at least in part related to methods used for evaluation; precision was better when the NucleoCounter was used for evaluation of sperm concentration, whereas the use of computer-assisted sperm analysis for evaluation of sperm motility resulted in greater precision in some but not all laboratories. None of the

  18. 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. PMID:26097016

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

    PubMed Central

    PAGLIOSA, Ronaldo C.; DEROSSI, Rafael; COSTA, Deiler S.; FARIA, Fabio J.C.

    2015-01-01

    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. PMID:26097016

  20. 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. PMID:26794460

  1. Frozen-intensity test research of frozen coal with steel

    NASA Astrophysics Data System (ADS)

    Zhang, Xiaopeng; Huang, Cheng; Liu, Weibo

    2002-05-01

    As a sort of multiple component, and dispersed state granule aggregation, frozen coal behaves similar to frozen soil. On the basis of its unique ice-cementation effect and not-frozen water along with dynamical balance state between the frameworks of mineral granule, the mechanical behavior of frozen coal is more complex than usual in compact medium, restrictedly with force amount, process time period and temperature. In all factors which impact on frozen intensity of frozen coal frozen with steel plate, water content is relatively easy to control. From results of this test research, values of frozen intensity is changeable under different water content. Up to the critical water content, the value of frozen intensity increase rapidly till a certain steady value. Under a certain temperature and water content condition, the granule component of frozen coal has somewhat effect on the frozen intensity. Usually, the frozen intensity of large granule coal is greater than the small granule's However, the distributing of coal granule size present a steady probability rule. So the effect from granule size is tiny.

  2. 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.…

  3. 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

  4. 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

  5. Histamine-2 Receptor Antagonists and Semen Quality.

    PubMed

    Banihani, Saleem A

    2016-01-01

    Histamine-2 receptor antagonists are a class of drugs used to treat the acid-related gastrointestinal diseases such as ulcer and gastro-oesophageal reflux disease. Although such drugs, especially ranitidine and famotidine, are still widely used, their effects on semen quality, and hence on male infertility, is still unclear. This MiniReview systematically addresses and summarizes the effect of histamine-2 receptor antagonists (cimetidine, ranitidine, nizatidine and famotidine) on semen quality, particularly, on sperm function. Cimetidine appears to have adverse effects on semen quality. While the effects of ranitidine and nizatidine on semen quality are still controversial, famotidine does not appear to change semen quality. Therefore, additional studies will be required to clarify whether histamine-2 receptor-independent effects of these drugs play a role in semen quality as well as further clinical studies including direct comparison of the histamine-2 receptor antagonists. PMID:26176290

  6. Expanding the dairy herd in pasture-based systems: the role for sexed semen use on virgin heifers.

    PubMed

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

    2013-02-01

    A model was developed to examine the effects of sexed semen use on replacement heifer numbers and rate of herd expansion in a seasonal dairy production system. Three separate herds were established according to the type of semen used on virgin heifers: conventional frozen-thawed (Conv), sexed fresh (SFre), or sexed frozen-thawed (SFro). In the model, sexed semen was used for the first and second inseminations in heifers only. Pregnancy rates achieved with sexed fresh and sexed frozen-thawed semen were assumed to be 94% and 75% of those achieved with conventional frozen-thawed semen, respectively. Initial herd size was 100 cows, which was maintained for the first 2 yr of the 15-yr simulation, after which all available replacement heifers were retained to facilitate herd expansion. Two different scenarios of land availability (S1 and S2) were examined for each of the 3 herds using different semen types: land available allowed expansion to a maximum herd size of 150 cows (S1) or 300 cows (S2). Once maximum herd size was reached, sexed semen use was discontinued and all excess heifer calves were sold at 1 mo of age. All capital expenditure associated with expansion was financed with a 15-yr loan. Each of the different options was evaluated in terms of annual farm profit, annual cash flow, and total discounted net profit. The analysis was completed at a milk price of € 0.27/L, and sensitivity around milk price was carried out at € 0.22/L and € 0.32/L. The use of SFre generated more replacement heifers and thus faster herd expansion compared with SFro and Conv semen. Maximum herd size was reached in yr 5, 6, and 7 under S1, and in yr 10, 12, and 14 under S2 for SFre, SFro, and Conv herds, respectively. Total discounted net profit under S1 for the SFre herd was € 19,929 greater than that of the SFro herd and € 41,852 greater than that of the Conv herd. Under S2, discounted net profit for the SFre herd was € 138,587 greater than that of the SFro herd and

  7. 9 CFR 98.34 - Import permits for poultry semen and animal semen.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... will be denied for semen from ruminants or swine from any region where it has been declared, under... rinderpest or foot-and-mouth disease exists. Importation of semen of ruminants or swine, originating in any...-mouth disease is determined to exist, is prohibited, except that semen from ruminants or...

  8. 9 CFR 98.34 - Import permits for poultry semen and animal semen.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... will be denied for semen from ruminants or swine from any region where it has been declared, under... rinderpest or foot-and-mouth disease exists. Importation of semen of ruminants or swine, originating in any...-mouth disease is determined to exist, is prohibited, except that semen from ruminants or...

  9. 9 CFR 98.34 - Import permits for poultry semen and animal semen.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... will be denied for semen from ruminants or swine from any region where it has been declared, under... rinderpest or foot-and-mouth disease exists. Importation of semen of ruminants or swine, originating in any...-mouth disease is determined to exist, is prohibited, except that semen from ruminants or...

  10. 9 CFR 98.34 - Import permits for poultry semen and animal semen.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... will be denied for semen from ruminants or swine from any region where it has been declared, under... rinderpest or foot-and-mouth disease exists. Importation of semen of ruminants or swine, originating in any...-mouth disease is determined to exist, is prohibited, except that semen from ruminants or...