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1
AD, Omur. "Antioxidant Potential of Ferulic Acid on the Freezability of Bull Semen." Open Access Journal of Veterinary Science & Research 4, no. 3 (2019): 1–4. http://dx.doi.org/10.23880/oajvsr-16000187.
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Ejaculates were collected twice a week from the bulls, via an artificial vagina, during two weeks. The suitable ejaculates obtained for sperm density (≥ 1.4 × 10 9 spermatoz oa / ml) and for motility (≥ 75%) were used for dilution and freezing of semen. A Tris - based extender (Tris 297.58mM, citric acid 96.32mM, fructose 82.66mM, egg yolk 15% (v/v), glycerol 5% (v/v), gentamicin 0.1 ml / 100ml, pH 6.8 - 7.0) was used as the base extender (cryopreservation diluent). Pooled ejaculate was split into 2 equal aliquots and diluted at 32 °C with base extender containing ferulic acid (100 μM) and no antioxidant (control), respectively. Each aliquot was diluted to a final semen concentrati on of approximately 1.2 × 10 8 sperm/ml (single step dilution), in 15 - ml polypropylene centrifuge tubes. After dilution, semen samples were kept at room temperature for 10 minutes then, the diluted semen samples were aspirated into 0.25 ml French straws, seal ed with polyvinyl alcohol powder and equilibrated at 5 °C for 3 h. After equilibration, the straws were frozen in liquid nitrogen vapour (4 cm above the liquid nitrogen, ~ - 100 o C ) for 10 min and then plunged into liquid nitrogen for storage, - 196 o C . In the study, sperm samples containing antioxidant and non - antioxidant were evaluated for spermatozoa motility and membrane integrity after freezing / thawing. In the present study, no statistically significant difference was found between the control and experim ental groups for motility and membrane integrity after freeze - thawing. The application consisted of 4 replications.
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Jovičić, Marija, Eva Chmelíková, and Markéta Sedmíková. "Cryopreservation of boar semen." Czech Journal of Animal Science 65, No. 4 (April 30, 2020): 115–23. http://dx.doi.org/10.17221/47/2020-cjas.
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Sperm cryopreservation is the best technology for long-term storage of the semen. However, the damage of boar spermatozoa by cryopreservation is more severe than in other animal species and a standardized freezing protocol for efficient cryopreservation has not been established yet. Semen quality and freezability vary greatly between breeds as well as between individual boars and even the season. Boar spermatozoa are sensitive to low temperatures; they sustain damage and a high rate of mortality and freezing/thawing the boar semen may strongly impair the sperm function and decrease the semen quality. The freezability of boar semen can be influenced by a cryopreservation procedure, and also by using various additives to freezing and thawing extenders such as antioxidants. In order to obtain acceptable results after thawing the boar semen, it is necessary to combine an optimal amount of additives (glycerol, egg yolk, sugars, antioxidants), cooling and warming velocities.
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Ardebili, R., A. Towhidi, S. Zeinodini, and M. Bahreini. "117. THE COMPARISON OF SPERM FREEZABILITY USING TWO EGG YOLK-FREE DILUENTS IN ZANDI RAM." Reproduction, Fertility and Development 21, no. 9 (2009): 36. http://dx.doi.org/10.1071/srb09abs117.
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The aim of this study was to investigate the effect of two kinds of commercial egg yolk-free extenders: Bioxcell and AndroMed on in vitro sperm function of sheep. A total of 10 ejaculates were collected from 5 adult Iranian Zandi rams using artificial vagina. Semen samples were mixed and diluted with Bioxcell or AndroMed. Freezing was conducted using imv semi-automatic equipment. The percentage of motility and progressive motility of sperm were evaluated before freezing (at 37°C), after refrigeration (at 5°C) and thawing, and recovery rate was calculated. Data was analyzed using proc GLM of SAS. The effect of extender on sperm qualitative characteristics after thawing was significant. Mean percentage of motility, progressive motility and recovery rate were higher (P≤0.01) in AndroMed than Bioxcell. Results suggested that AndroMed in comparison with Bioxcell had more ability to preserve sperm quality during freezing process.
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Acha, D., M. Hidalgo, I. Ortiz, M. J. Gálvez, J. J. Carrasco, V. Gómez-Arrones, and J. Dorado. "Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants." Reproduction, Fertility and Development 28, no. 12 (2016): 1990. http://dx.doi.org/10.1071/rd14449.
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The aim of this study was to compare the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. First, 32 ejaculates were pooled, split and frozen in either Gent B or INRA 96 with egg yolk and glycerol. Second, 12 pooled semen samples were simultaneously frozen in Gent B (glycerol) or Gent A containing ethylene glycol (EG; 1 or 1.5%) or dimethyl sulfoxide (DMSO; 1.5 or 2%). Finally, nine pooled samples were simultaneously cryopreserved in Gent A containing 1% EG (as control), dimethylformamide (DMFA; 1 or 2.5%) or a combination of 1% EG and 1.5% DMFA. Gent B yielded a higher (P < 0.01) post-thaw sperm motility than modified INRA96. EG 1% increased the sperm membrane integrity (P < 0.001), whereas DMSO affected sperm motility and membrane integrity (P < 0.001). DMFA 2.5% yielded higher (P < 0.001) values for sperm motility and membrane integrity. We concluded that Gent B improves in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% EG or 2.5% DMFA increased sperm protection against cryodamage. The use of DMSO for freezing donkey semen was unsuccessful and a toxic effect is suspected. These extenders should be included in the pre-freeze test for each donkey.
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Swelum, A. A., I. M. Saadeldin, M. Bahadi, M. Afifi, M. Al-Mutary, and A. N. Alowaimer. "The effect of heterologous seminal plasma from ram, buck or camel on the freezability of ram semen." Veterinární Medicína 63, No. 11 (November 19, 2018): 500–512. http://dx.doi.org/10.17221/52/2018-vetmed.
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Here, we compared the effects of ram, buck and dromedary camel seminal plasma mixed with TRIS-egg yolk glycerol extender on the freezing preservation of ram semen. Awassi ram semen samples underwent primary evaluation and were then pooled and diluted with the following diluents: TRIS-egg yolk glycerol mixed with (1) whole ram semen as a control (T); (2) ram sperm after seminal plasma removal (W); or (3) ram, (4) buck or (5) camel seminal plasma (R, B and C, respectively). The diluted semen was frozen using liquid nitrogen vapor. Various sperm parameters were evaluated in the frozen semen. Total motility before and after freezing was significantly higher in R, B and C diluents than in T and W diluents. Progressive motility after freezing was significantly higher in R, B, C and T diluents than in W diluent. Vitality after freezing was significantly higher in B than in W diluent. DNA fragmentation before and after freezing was significantly lower in R, B, C and T diluents than in W diluent. Plasma membrane integrity before and after freezing was significantly higher in R, B and C diluents than in W diluent. Sperm abnormalities before freezing were significantly lower in R, B and C diluents than in W diluent. Malondialdehyde concentration was significantly higher in T and W diluents than other diluents. Reduced glutathione concentration was significantly higher in B diluent than other diluents. Moreover, reduced glutathione concentration was significantly higher in C, R and W diluents than in T diluent. Thus, the addition of ram, buck or camel seminal plasma to TRIS-egg yolk glycerol extender improved the quality of frozen ram semen, while seminal plasma removal adversely affected it. Ram, buck and camel seminal plasma had similar effects, with no significant differences between them on the evaluated parameters of frozen ram semen.
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Ardebili, R., A. Towhidi, S. Zeinodini, M. Bahreini, and E. Dirandeh. "85 THE COMPARISON OF SPERM FREEZABILITY USING TWO EGG YOLK-FREE DILUENTS IN ZANDI RAM." Reproduction, Fertility and Development 22, no. 1 (2010): 201. http://dx.doi.org/10.1071/rdv22n1ab85.
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The aim of this study was to investigate the effect of 2 different commercial egg yolk-free extenders, Bioxcell and AndroMed, on post-thaw ram sperm motility and recovery rate. A total of 10 ejaculates were collected from 5 adult Iranian Zandi rams using artificial vagina during 3 weeks, once per week. Preexamination was conducted on semen, and semen motility >70% were selected. Semen samples were pooled and diluted with Bioxcell or AndroMed. Pooled semen was split into 2 treatments and extended to a final concentration of 200 × 108 spermatozoa per mL with extenders. The extended semen was manually packed into 0.25-mL mini-straws and sealed with PVC powder. After 4 h of equilibration in a waterbath at 5°C, the straws were dried with paper tissue, placed horizontally on a rack, and transferred to an isothermal box to be frozen in vapor 5 cm above liquid nitrogen. After 10 min, the straws were plunged into liquid nitrogen. Freezing was conducted using IMV semi-automatic equipment. The percentage of motility and progressive motility of sperm were evaluated before freezing (at 37°C), after refrigeration (at 5°C) and thawing, and recovery rate was calculated. To evaluate indices of sperm motility including the percentage of motile sperm and the progressive forward motility, semen was diluted with 0.9% NaCl w/v (1:100). Ten μL of diluted semen was placed on the prewarmed (37°C) microscope slide (76.2 mm × 24.5 mm; Pearl, China) covered with a cover slip (24 × 24 mm; Menzel-Glaser, Germany) and examined 10 different fields under a phase contrast microscope (Nikon, Tokyo, Japan), at ×200. Dilution factor 1:5 was used. The microscope was equipped with warm plate set at 37°C. Data were analyzed using Proc GLM of SAS. The effect of extender on sperm qualitative characteristics after thawing was significant. Least squares mean percent motility and progressive motility of AndroMed (36.05% ± 0.75; 29.0% ± 0.84) was significantly higher (P < 0.01) than Bioxcell (8.02% ± 0.35; 4.0% ± 0.60); least squares mean recovery rate in AndroMed (34.23% ± 1.05) also was higher (P < 0.01) than Bioxcell (10.0% ± 0.92). Results suggested that AndroMed in comparison with Bioxcell had more ability to preserve sperm quality during the freezing and thawing process.
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Hernández, M., J. Roca, J. Ballester, J. M. Vázquez, E. A. Martínez, A. Johannisson, F. Saravia, and H. Rodríguez-Martínez. "97 DIFFERENCES IN SPERM CHROMATIN STRUCTURE AMONG GOOD AND BAD BOAR SPERM FREEZERS." Reproduction, Fertility and Development 18, no. 2 (2006): 157. http://dx.doi.org/10.1071/rdv18n2ab97.
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Inter- and intra-boar differences in sperm freezability are observed independent of the sperm quality before freezing, the breed, or the genetic line. This study aimed to determine whether boars with different post-thaw sperm quality also show differences in sperm DNA integrity. Sperm-rich fractions (3 to 10 ejaculates per boar) from 19 fertile mature boars were extended in Beltsville thawing solution (BTS) and cooled to 17�C for 16 h. Then, samples were centrifuged at 2400g for 3 min, extended in freezing extender (lactose/egg yolk/glycerol/Equex STM; Nova Chemical Sales, Inc., Scituate, MA, USA) to a final concentration of 1 � 109 spermatozoa/mL, dispensed into 0.5 mL straws, and frozen in a programmable cell freezer at a rate of -20�C min. Thawing was carried out in a water bath at 37�C for 20 s. Frozen-thawed spermatozoa were evaluated for progressive sperm motility (PSM) using a computer-assisted sperm analysis (CASA) system, and sperm viability (PMI) using flow cytometry. All data generated were used for a multivariate pattern analysis (PATN; CSIRO, Canberra, Australia) which objectively classified all boars into two groups, categorized as good (n = 10; >60% PSM and PMI) or bad (n = 9; <40% PSM and PMI) based on their sperm freezability. Post-thaw sperm quality was consistent for different ejaculates within boars (P < 0.05). The DNA-integrity of frozen-thawed spermatozoa was evaluated according to the sperm chromatin structure assay (SCSA; Evenson et al. 1980 Science 210, 1131-1133). All SCSA variables (X mean, DNA fragmentation index (DFI), and the standard deviation of the DFI), were significantly higher for bad freezers (P < 0.001). The percentage of spermatozoa with abnormal chromatin structure ranged from 1.06 to 3.42% for good and 3.06 to 6.04% for bad freezers. Although these differences exist between good and bad sperm freezers, and can only to some extent be the product of cryopreservation, the levels of affected spermatozoa can not explain the differences on post-thaw sperm survival seen in the two categories of sires. This work was supported by CICYT, AGL05-0471 (Spain), SLF and Formas (Sweden).
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Martínez-Fresneda, Lucía, Emma O'Brien, Rosario Velázquez, Adolfo Toledano-Díaz, Carlos M. Martínez-Cáceres, Dawit Tesfaye, Karl Schellander, Francisco A. García-Vázquez, and Julian Santiago-Moreno. "Seasonal variation in sperm freezability associated with changes in testicular germinal epithelium in domestic (Ovis aries) and wild (Ovis musimon) sheep." Reproduction, Fertility and Development 31, no. 10 (2019): 1545. http://dx.doi.org/10.1071/rd18511.
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The aim of this study was to examine ovine sperm cryoresistance during the rutting season (RS) and its association with sperm head area and seminiferous epithelium proliferation. Small ruminants show fluctuating testosterone levels throughout the year, which could interfere with spermatogenesis and sperm cryopreservation. Ejaculates, testicular biopsies and blood were collected during the middle and at the end of the RS (Middle-RS vs End-RS) during periods of high and low testosterone levels in Merino and Mouflon rams. Fresh and frozen–thawed sperm quality, sperm morphometry, seminiferous tubule morphometry and testicular proliferation markers (proliferating cell nuclear antigen, proliferation marker protein Ki-67 and transcription factor GATA-4) were evaluated. Post-thaw sperm viability was higher in the End-RS group in both Merino (69.9±8.2 vs 41.6±7.3%; P=0.020) and Mouflon rams (40.9±3.3 vs 24.2±5.0%; P=0.008). Mouflons had larger sperm head area at the End-RS (38.3±0.2 vs 34.3±0.1µm2; P=0.029), whereas there was no difference between Merino groups (35.7±0.5 vs 34.8±1.0µm2). Seminiferous tubule morphometry and proliferation markers showed higher levels of germinal epithelium proliferation in the Middle-RS of both species. In conclusion, sperm freezability is affected during the RS in domestic and wild rams, which could be correlated with changes that occur during spermatogenesis, since there is an effect of season on cell proliferation in the testis.
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Anel, L., V. Garcia-Macias, F. Martinez-Pastor, M. Alvarez, S. Borragan, J. Bernardo, S. Alves, C. Chamorro, and P. Paz. "242 EFFECT OF SPERMATOZOA HEAD MORPHOMETRIC DIMENSIONS ON FREEZABILITY IN BROWN BEAR (URSUS ARCTOS)." Reproduction, Fertility and Development 19, no. 1 (2007): 237. http://dx.doi.org/10.1071/rdv19n1ab242.
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Having recently observed that survival of red deer spermatozoa after cryopreservation seemed to reflect the size of sperm heads, we hypothesized that cryoresistance of brown bear spermatozoa might also be dependent on head size, since in a preliminary study we had also observed significant differences in sperm head sizes among male brown bears (median values for area 22.2 �m2, perimeter 18.2, length 6.1 and width 4.4 �m). In the present report, we analyzed the post-thaw survival of spermatozoa of 6 brown bears that were assigned to 2 groups (3 bears/group) based on sperm size: Group A with large-size sperm heads; Group B with small-size heads. Ejaculates were obtained by electroejaculation of adult brown bears (semi-free ranging in Cabarceno Park, Cantabria, Spain) under general anesthesia (7 mg kg-1 tiletamine + zolazepan and 2 mg kg-1 ketamine). Semen was diluted (Tes-Tris-fructose, 8% glycerol, 20% egg yolk, EDTA, and Equex paste), loaded in 0.25-mL straws, and frozen in a biofreezer at 20�C min-1 to -100�C. After storage in liquid nitrogen, samples were thawed in water at 65�C for 6 s and survival was measured. Sperm motility (TM: total, and PM: progressive; %) was assessed microscopically, and sperm viability, acrosome integrity (PI/PNA-FITC), and mitochondrial status (JC-1) were assayed for fresh and thawed sperm by flow cytometry. Recovery rates (RR: thawed/fresh � 100) were calculated for all parameters. For measurement of head size, fresh sperm samples were fixed in glutaraldehyde and slides were air-dried for 2 h. The samples were then stained with Diff-Quik� staining at 37�C. The area (Ar), perimeter (P), length (L), and width (W) of the heads of >100 spermatozoa per slide were measured (Sperm Class Analyzer�; Microptic S.L., Barcelona, Spain). Data were analyzed with the SAS ver8 system, and the Wilcoxon test was applied. The respective morphometric dimensions of the 2 groups were practically identical (Ar = 22; P = 18; L = 6; W = 4). The post-thaw recovery rates of spermatozoa from Group A were: TM: 60.1 � 29.3; PM: 54.8 � 36.0; viability: 99.4 � 8.0; acrosomes: 96.2 � 3.1; mitochondria: 70.9 � 15.5. The recovery rates for Group B were: TM: 78.7 � 13.8; PM: 69.0 � 18.8; viability: 93.8 � 5.2; acrosomes: 98.2 � 9.8; mitochondria: 72.5 � 22.5. Because of the high variability of recovery rates between males within each group, there were no statistical differences between the 2 groups. The absence of differences can be explained by the small number of males examined and the high variability between them. More studies are necessary to determine whether large sperm cells of brown bears are more susceptible to damage during cryopreservation. This work was supported in part by CANTUR S.A. and CICYT (CGL 2004-0278/BOS).
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Cremades, T., G. Carvajal, M. Hernandez, J. M. Vazquez, E. A. Martinez, and J. Roca. "92THE ADDITION OF SEMINAL PLASMA FROM INDIVIDUAL BOARS TO FREEZING EXTENDER CAN IMPROVE THE POST-THAW SPERM SURVIVAL." Reproduction, Fertility and Development 16, no. 2 (2004): 167. http://dx.doi.org/10.1071/rdv16n1ab92.
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Contradictory results have been reported about the effect of seminal plasma (SP) on the freezability of mammalian spermatozoa. In pigs, current methods for sperm cryopreservation involve removing seminal plasma. Therefore, no conclusive evidence of the potential effect of SP on the freezability of boar spermatozoa has been reported. In this study, we evaluate the effect of the addition of low concentrations of SP from individual boars to the freezing extender on post-thaw sperm survival. Sperm cryopreservation procedure included: dilution of sperm-rich fraction in Beltsville Thaw Solution extender (BTS), cooling to 17°C for 16h, centrifugation at 2400g for 3min, dilution in lactose/egg-yolk/glycerol/Equex Stem (freezing extender) to a final concentration of 1×109 spermmL−1, dispensing into 0.5-mL straws, and freezing in a programmable cell freezer at 20°Cmin−1. Thawing was carried out in a waterbath at 37°C for 20s. Post-thaw sperm survival was assessed by progressive sperm motility (PSM) using a CASA system (SCA); plasma membrane integrity (PMI) and acrosome membrane integrity (AMI) were assessed by flow cytometric procedures (SYBR-14/PI and FITC-PNA/PI, respectively) at 30 and 150min post-thawing in BTS-diluted thaw spermatozoa held in a waterbath at 37°C. Four individual seminal plasma donors (SP1 to SP4) were selected in a preliminary study in which 48 ejaculates from 12 boars (4 ejaculates/boar) were cryopreserved. Then the boars were classified into 3 groups (good, moderate and bad freezers) based on their post-thaw sperm survival. SP1 and SP2 were good freezers (>60% PSM and PMI), SP3 was a moderate freezer (40–60% PSM and PMI) and SP4 was a bad freezer (<40% PSM and PMI). In the main experiment, pooled sperm-rich fractions collected from 9 mature hybrid boars were divided into five aliquots and each was diluted with freezing extender supplemented with 0% (control) or 10% of SP (1–4). Data from eight replicates were analyzed as a split plot design using a PROMIXED model. The addition of SP to freezing extender had a significant effect (P<0.05) on post-thaw sperm survival compared to control. Moreover, there were significant differences (P<0.05) between SP donors. PSM, PMI and AMI were significantly (P<0.05) higher in SP1 (56.71±4.30; 57.16±4.01 and 57.22±4.01, respectively) and SP2 (59.48±4.30; 60.17±4.01 and 60.05±4.01, respectively) compared to control (50.39±4.30; 49.98±4.01 and 49.54±4.01, respectively). There were no differences (P>0.05) between SP3, SP4 and control. These results indicate that the addition of SP from particular boars (good freezers) to freezing extender may improve post-thaw sperm survival. Individual differences in the SP composition should explain the above results. Supported by INIA (RZ01-019) and MCYT (AGL2001-0471).
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KCHALIFA, TAA, M. M. WAHEED, and A. G. LYMBEROPOULOS (Α.Γ. ΛΥΜΠΕΡΟΠΟΥΛΟΣ). "An endeavor to improve longevity of cryopreserved equine sperm." Journal of the Hellenic Veterinary Medical Society 57, no. 3 (November 29, 2017): 195. http://dx.doi.org/10.12681/jhvms.15039.
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Exposure of sperm cells to the oxidative stress pending hypothermic storage of semen has been suggested to be responsible, in part, for the decline of their motility and fertility. This study was conducted to evaluate the in-vitro effects of antioxidants (AOs) and / or caffeine on longevity of cryopreserved stallion spermatozoa. Aliquots from the gel-free fraction of semen ejaculates (n=12), collected from 5 Arabian stallions (9-18 years old) of unknown sperm freezability, were mixed 1:1 with a Tris-egg yolk extender (TEYE), centrifuged at 500 χ g for 5 min and sperm cells were frozen in the form of 0.25-ml concentrated pellets after 2-step addition of TEYE supplemented with or without AOs (0.50 mg /ml Na pyruvate, 1 mg / ml Na thiosulfate, 5 mg / ml bovine serum albumin, 0.15 mg / ml zinc chloride and 0.50 mg / ml ferulic acid). The final pre-freeze concentrations of glycerol and sperm cells were 5% and 562-924 χ IO6 / ml, respectively. Frozen pellets from non-AOs and AOs-treated sperm were thawed in a Tris-citric acidglucose solution (40°C) containing 0, 0.49, 0.97 or 1.94 mg / ml caffeine and incubated (140-230 χ IO6 sperm / ml) at 30°C for 3 h. Sperm progressive motility (%) was assessed after centrifugation, before freezing and after 0, 1, 2 and 3 h of thawing. The results revealed significant (P<0.05) effects of sperm treatments only on post-thaw motility. Neither AOs alone nor caffeine alone could significantly ameliorate the maintenance of sperm motility. AOs plus 0.97 or 1.94 mg / ml caffeine were the superior supplements in improving the longevity of stallion spermatozoa.
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Mata-Campuzano, M., M. Alvarez, S. Borragan, F. Martinez-Pastor, M. Nicolas, J. Tamayo, L. Anel, and P. de Paz. "160 FREEZABILITY OF ELECTROEJACULATED AND EPIDIDYMAL SPERMATOZOA FROM BLUE WILDEBEEST (CONNOCHAETES TAURINUS)." Reproduction, Fertility and Development 21, no. 1 (2009): 179. http://dx.doi.org/10.1071/rdv21n1ab160.
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Maintenance of population viability, especially endangered species, requires preservation of as much genetic variability as possible, therefore genetic resource banks are very important. For male gametes, preservation of all available sources (ejaculates and epididymal) are useful. Information regarding sperm characteristics of most wild ruminant species is limited compared to that from domestic species. The objective of this work was to characterize the freezability of electroejaculated and epididymal spermatozoa from a wildebeest (5 year old; housed in Cabárceno Nature Park, Cantabria, Spain) that was castrated because of behavioral problems. After general anesthesia (ethorfine + xilazine, 1.8 mL + 0.5 mg kg–1) with dart, semen was collected by electroejaculation (3 V and 75 mA). Sperm concentration was 250 × 106 mL–1 (total spermatozoa: 1128.6 × 106). After castration, epididymides were disected and spermatozoa were collected by making several incisions in the caudal epididymis. Concentration was 12 441 × 106 spermatozoa mL–1 (total spermatozoa: 24 882 × 106). Samples were diluted to 200 × 106 spermatozoa mL–1 (TesT-Fructose-Egg yolk-Glycerol-Antibiotics) and chilled to 5°C during 2 h. Diluted semen was packaged in 0.25-mL straws and frozen from 5°C to –100°C (–20°C min–1) in a programmable cell freezer (Kryo 10, Planer). Straws were plunged into liquid nitrogen until analysis and thawed in a water bath (65°C, 6 s). Fresh, pre-freezing and post-thawed samples were analysed for motility (total motility TM, %; progressive motility PM, %; path velocity VAP, μm s–1; track speed VCL, μm s–1; progressive velocity VSL, μm s–1) using a CASA (ISAS, Proiser, Valencia, Spain). Viability (VIAB %) (SYBR-14 and propidium iodide) and mitochondrial membrane potential (MIT %) (JC1) were assessed by flow cytometry. Post-thawing results for electroejaculated v. epididymal samples were, respectively: TM: 87.0 v. 64.6%; PM: 68.7 v. 33.4%; VAP: 95.9 v. 49.8 μm s–1; VCL: 108.3 v. 71.6 μm s–1; VSL: 86.7 v. 40.2 μm s–1; VIAB: 57.0 v. 73.9%; MIT: 59.5 v. 77.5%. Motility parameters were higher for the electroejaculated sample; however, viability was higher for the epididymal sample. Recovery rates (post-thawed value/pre-freezing value × 100) for electroejaculated v. epididymal samples were: TM: 97.2 v. 93.4%; PM: 113.3 v. 103.2%; VAP: 88.9 v. 122.0 μm s–1; VCL: 87.5 v. 126.0 μm s–1; VSL: 93.4 v. 125.3 μm s–1; VIAB: 75.0 v. 97.7%; MIT: 69.2 v. 95.9%). These rates suggest a good freezability of electroejaculated and epididymal spermatozoa in blue wildebeest. This work was supported in part by Cantur. 3 Supported by Juan de la Cierva program (MICINN, Spain).
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Cabrera, F., F. Gonzalez, M. Batista, P. Calero, A. Medrano, and A. Gracia. "The Effect of Removal of Seminal Plasma, Egg Yolk Level and Season on Sperm Freezability of Canary Buck (Capra hircus)." Reproduction in Domestic Animals 40, no. 3 (June 2005): 191–95. http://dx.doi.org/10.1111/j.1439-0531.2005.00544.x.
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Deori, S., B. C. Deka, R. K. Biswas, N. Nahardeka, A. Arangasamy, D. C. Bhuyan, D. J. Kalita, R. S. Borah, and Arundhati Phookan. "Characteristics and freezability of Assam Hill goat semen." Indian Journal of Animal Research, OF (May 19, 2017). http://dx.doi.org/10.18805/ijar.v0iof.7821.
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Assam Hill goat (AHG) is an important goat germplasm found in Assam and its adjoining areas of India. The study was designed with an objective to study the semen characteristics and freezability of AHG buck semen using Tris -Egg yolk-Citrate-Fructose diluent. The mean values of fresh semen characteristics in AHG bucks viz., ejaculate volume (ml), initial sperm motility (%), sperm concentration (x106/ml), live sperm (%), sperm abnormality (%), HOST-reacted sperm (%) and intact acrosome (%) recorded were 0.39 ± 0.01, 77.97 ± 0.73, 3201.00 ± 143.78, 83.02 ± 0.65, 7.66 ± 0.73, 66.95 ± 0.74 and 93.34 ± 0.51, respectively. Mean values for post-thaw semen characteristics i.e., sperm motility (%), live sperm (%), HOST-reacted sperm (%) and intact acrosome (%) were 55.39 ± 0.97, 71.01 ± 0.78, 54.77 ± 0.55 and 82.16 ± 0.43, respectively. It can be concluded that AHG bucks donate acceptable quality of semen which can be frozen successfully in Tris-Egg yolk-Citrate-Fructose diluents for using in Artificial Insemination.
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Kumar, Narendra, B. Rai, Chetna Gangwar, S. A. Lone, Anshuman Kumar, S. K. Gupta, and Vipin Maurya. "Effect of different levels of egg-yolk on freezability of Jakhrana buck semen." Indian Journal of Animal Research, OF (August 31, 2016). http://dx.doi.org/10.18805/ijar.11326.
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The present study was designed to determine the effect of different levels of egg-yolk on freezability of Jakhrana buck semen. Six healthy Jakhrana bucks (BW=30 ± 2kg, age=12 ± 0.5 month) were used for semen collection. These bucks were maintained under semi-intensive system at Jakhrana Unit of C.I.R.G. Makhdoom, Mathura. A total of 48 ejaculates (6 bucks × 8 replicates) were collected twice a week using artificial vagina. Each ejaculate was divided into 4 groups (G1, G2, G3 and G4). The G1, G2, G3 and G4 were extended with Tris-egg yolk-citric acid- fructose-glycerol (TEYCFG) extenders containing 5, 10, 15 and 20% egg yolk level, respectively. Each ejaculate was evaluated for sperm motility, viability, abnormality, and hypo-osmotic swelling (HOS) response and acrosome integrity before and after freezing. At pre-freeze stage no significant (P>0.05) difference in sperm motility and viability was found among all groups. Sperm abnormality was significantly (P<0.05) higher in G4 as compared to other groups (G1, G2, G3). The HOS response and acrosomal integrity was significantly (P<0.05) higher in G1, G2 and G3 as compared to G4. However, no significant (P>0.05) difference was observed in HOS response and acrosomal integrity among G1, G2 and G3. At post thaw stage, sperm motility, viability and HOS response was significantly (P<0.05) higher in G1 and G2 as compared to G3 and G4. Sperm abnormality was significantly (P<0.05) lower in G2 as compared to other groups. The acrosomal integrity was significantly (P<0.01) higher in G1 and G2 as compared to G3 and G4. It is concluded that 10% egg yolk in Tris based extender may be the best for successful cryopreservation of Jakhrana buck semen.
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Chaudhari, D. V., J. A. Patel, K. K. Hadiya, and A. J. Dhami. "Influence of Seasons and Extenders on Quality and Freezability of Gir Bull Semen under Middle Gujarat Climate." INDIAN JOURNAL OF VETERINARY SCIENCES AND BIOTECHNOLOGY 13, no. 03 (January 7, 2018). http://dx.doi.org/10.21887/ijvsbt.v13i03.10632.
Full textAbstract:
The study was conducted to evaluate the seasonal influence (peak winter and summer) and the efficacy of three extenders (egg yolk based TFYG extender and egg yolk free soya bean based commercial extenders Optixcell and Andromed) on quality and freezability of Gir bull semen in Middle Gujarat. Semen ejaculates (6/bull/season, total36) revealed mean ejaculate volume 6.49±0.30 ml, sperm concentration1212.36±58.10 million/ml, progressive motility 74.17±0.78 %, live sperm 81.39±0.80 %, abnormal sperm 7.36±0.31 %, and sperm with intact plasma membrane 81.31±0.98 % and intact acrosome 94.81±0.24 %. Only the progressive sperm motility was significantly (P<0.05) higher(76.39±0.97 % vs. 71.94±1.00 %) with lesser sperm abnormality(6.17±0.37 % vs. 8.56±0.30 %) during winter than in summer. Semen samples split diluted with TFYG, Optixcell and Andromed extenders recorded the overall mean values of progressive sperm motility, livability, abnormality, plasma membrane integrity and acrosomal integrity during winter season as 77.87±0.51, 77.50±0.45, 5.56±0.20,76.02±0.81 and 94.35±0.29 on dilution; 72.41±0.51, 70.50±0.64, 5.96±0.26, 71.20±0.79 and 93.09±0.32 at pre-freeze stage; 41.30 ±0.94, 50.28±1.03, 9.15±0.31, 29.89±0.40 and 90.65±0.40 at post-thaw stage, respectively. The respective values in summer season were 72.13±0.60, 75.50±0.60, 7.48±0.25, 75.61 ±0.55 and 94.09±0.30 on dilution; 65.46±0.66, 69.41±1.05, 8.89±0.28, 69.70±0.66 and 92.63 ±0.33 at pre-freeze stage; 31.48±0.52, 45.09±0.85, 13.48±0.33, 26.85±0.71 and91.26±0.38 at post-thaw stage. The overall mean sperm post-thaw motility/longevity at 0, 30, 60 and 120 min of incubation at 37°C was 41.20±1.51, 35.19±1.47, 28.80±1.75 and 17.50±1.47 % during winter season and 31.57±0.89, 26.20±0.77, 20.37±0.83 and 13.80±0.77% in summer season, respectively. The initial quality as well as freezability of semen in terms of motile, live, normal and HOS reactive sperm including post thaw longevity were better in winter season than in summer season. Further, the values of all the five semen quality parameters studied were comparatively better in Optixcell than TFYG and Andromed extenders with significant differences only in sperm progressive motility in both the seasons.The season x extender interaction was not significant for any of the sperm quality parameters studied.
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Martínez-Fresneda, Lucía, Cristina Castaño, Paula Bóveda, Dawit Tesfaye, Karl Schellander, Julián Santiago-Moreno, and Francisco A. García-Vázquez. "Epididymal and ejaculated sperm differ on their response to the cryopreservation and capacitation processes in mouflon (Ovis musimon)." Scientific Reports 9, no. 1 (October 30, 2019). http://dx.doi.org/10.1038/s41598-019-52057-0.
Full textAbstract:
Abstract Spermatozoa must undergo the process of capacitation to fertilize the egg which involves a cell destabilizing process. Capacitation-like changes such as protein tyrosine phosphorylation (PTP) are associated with cryopreservation. The aim of this study was to compare the cryoresistance and capacitation response of epididymal and ejaculated sperm of European mouflon (Ovis musimon). Post-thaw sperm parameters were analysed from epididymal and ejaculated samples cryopreserved by slow-freezing or ultrarapid-freezing for comparison. Sperm capacitation status was assessed by the semiquantification of PTP levels, cell localization of PTP and kinematic clustering. Epididymal sperm had higher cryoresistance than ejaculated sperm in both freezing techniques, and slow-freezing rendered better results than ultrarapid-freezing in both sperm samples. Ejaculated sperm had higher PTP levels than epididymal sperm and, additionally, ejaculated sperm showed higher phosphorylation in capacitating (CA) than in non-capacitating (NCA) conditions while there was no effect of medium in epididymal sperm. There was a higher tail PTP in CA than in NCA conditions in both types of sperm. Kinematic analysis revealed that the cluster associated with hyperactivated movement increased in ejaculated sperm incubated in CA whereas no effect of medium was observed in epididymal sperm clusters. In conclusion, epididymal sperm showed better freezability and lower capacitation status compared to ejaculated sperm.
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Monaco, Davide, Miguel Batista, Olga Amann, Barbara Padalino, Wouter Pieters, Mariacristina Morelli, Gianluca Accogli, Salvatore Desantis, and Giovanni Michele Lacalandra. "Retrograde flushing collection and freezing of dromedary camel epididymal spermatozoa with seminal plasma." Acta Veterinaria Hungarica, November 4, 2020. http://dx.doi.org/10.1556/004.2020.00042.
Full textAbstract:
AbstractThe objectives of this study were to describe the parameters of dromedary camel epididymal spermatozoa collected by retrograde flushing (RF) technique and to evaluate the freezability of the collected sperm, diluted with and without the supplementation of seminal plasma (SP). Two experiments were conducted: in Experiment 1, ES were recovered within 6–8 h after castration; selected samples were diluted with a Tris-citrate egg-yolk glycerolated buffer and frozen. In Experiment 2, epididymides were stored for 24 h at 4 °C before RF and semen samples were frozen after dilution with a Tris-lactose egg-yolk glycerolated extender with and without 15% SP. In Experiment 1, eight semen samples were obtained from ten epididymides with a mean of 500 × 106 total spermatozoa recovered, per flushed epididymis. Mean post-thaw motility and progressive motility were 75 and 17%, respectively. In Experiment 2, 15 samples were collected, out of the 18 epididymides (mean number of collected spermatozoa: 700 × 106), and 13 of these samples were of excellent quality. Post-thaw parameters were not satisfactory but the supplementation of the freezing medium with 15% SP improved the progressive motility and kinematic parameters of the spermatozoa.