Effect of Composition of Tris-based Diluent on Survival of Boar Spermatozoa following Deep Freezing

1974 ◽  
Vol 27 (5) ◽  
pp. 485 ◽  
Author(s):  
D Visser ◽  
S Salamon
Keyword(s):  
Acetic Acid ◽  
Egg Yolk ◽  
Factorial Experiments ◽  
Deep Freezing ◽  
Boar Spermatozoa

Six factorial experiments were conducted to examine the effects of concentrations of 2-amino-2-hydroxymethylpropane-l,3-diol (tris), sugars, erythritol, catalase, ethylenediaminetetra-acetic acid (EDTA), glycerol and egg yolk in the freezing diluent on the survival of boar spermatozoa after freeze-thawing.

scholarly journals Deep Freezing of Ram Semen: Recovery of Spermatozoa After Pelleting and Comparison With Other Methods of Freezing

1968 ◽  
Vol 21 (2) ◽  
pp. 351 ◽  
Author(s):  
S Salamon
Keyword(s):  
Egg Yolk ◽  
Factorial Experiments ◽  
Deep Freezing

Four factorial experiments were conducted in which were examined and compared factors which affect revival of ram spermatozoa after pellet.freezing and other methods of freezing. Revival of spermatozoa after pellet.freezing was the best in egg yolk-lactose (333 mM), followed by egg yolk-raffinose (333 mM), egg yolk-glucose (355 mM), and egg yolk-fructose (355 mM) diluents. Within the glycerol range of 0-7%, lactose gave the best results with 3%, raffinose and glucose with 5%, and fructose with 7% (vJv) glycerol.

scholarly journals Deep Freezing of Boar Semen II. Effects of Method of Dilution, Glycerol Concentration, and Time of Semen-Glycerol Contact on Survival of Spermatozoa

1973 ◽  
Vol 26 (1) ◽  
pp. 231 ◽  
Author(s):  
I Wilmut ◽  
S Salamon ◽  
C Polge
Keyword(s):  
Glycerol Concentration ◽  
Factorial Experiments ◽  
Deep Freezing ◽  
Boar Semen ◽  
Boar Spermatozoa

Five factorial experiments were conducted to examine the effects of glycerol concentration and processing procedures prior to freezing on the revival of boar spermatozoa upon thawing.

Deep Freezing of Boar Semen III. Effects of Centrifugation, Diluent and Dilution Rate, Pellet Volume, and Method of Thawing on Survival of Spermatozoa

1973 ◽  
Vol 26 (1) ◽  
pp. 239 ◽  
Author(s):  
S Salamon
Keyword(s):  
Dilution Rate ◽  
Factorial Experiments ◽  
Deep Freezing ◽  
Boar Semen ◽  
Boar Spermatozoa

Five factorial experiments were conducted to examine the effects of centrifugation of semen, diluent and dilution rate, pellet volume, and method of thawing on the survival of boar spermatozoa after freezing by the pellet method.

scholarly journals Deep Freezing of Boar Semen I. Effects of Diluent Composition, Protective Agents, and Method of Thawing on Survival of Spermatozoa

1973 ◽  
Vol 26 (1) ◽  
pp. 219 ◽  
Author(s):  
S Salamon ◽  
I Wilmut ◽  
C Polge
Keyword(s):  
Molecular Weight ◽  
Low Molecular Weight ◽  
Factorial Experiments ◽  
Protective Agents ◽  
Deep Freezing ◽  
Boar Semen ◽  
Boar Spermatozoa

A series of factorial experiments was conducted to examine the effects of sugars and polyols (inositol, dulcitol), glycerol, low molecular weight polyols and cell "non-permeating" agents as cryoprotectives, and method of thawing on survival of boar spermatozoa after freezing by the pellet method.

Influence of Equilibration, Freezing Rate, Method of Dilution, and Diluent on the Survival of Deep-Frozen Bull Spermatozoa

1965 ◽  
Vol 18 (4) ◽  
pp. 901 ◽  
Author(s):  
ICA Martin
Keyword(s):  
Freezing Rate ◽  
Slow Freezing ◽  
Factorial Experiments ◽  
Deep Freezing ◽  
Rate Method ◽  
Fast Freezing

From the results of four factorial experiments on the deep-freezing of bull spermatozoa: (1) Revival rates of semen treated with lecithin followed by cooling to 5�C with dilution just before freezing did not differ significantly from samples diluted at 30�C soon after collection of the ejaculate. Aging the spermatozoa at 5�C for 6 hr before freezing was beneficial and a slow freezing rate of O� 5 to 1 degC fall per minute to -15�C followed by 3 degC fall below this temperature gave better results than faster rates. Time of storage at 5�C and freezing rate interacted, as fast freezing was much better tolerated by spermatozoa which had been aged at 5�C for 6 or 18 hr.

75 GLUCOSE CONCENTRATION OF FREEZING EXTENDER MODULATES THE TYROSINE PHOSPHORYLATION PATTERN OF FROZEN-THAWED BOAR SPERMATOZOA

2009 ◽  
Vol 21 (1) ◽  
pp. 138
Author(s):  
J. E. Rodríguez-Gil ◽  
M. Hernández ◽  
M. M. Rivera ◽  
L. Ramió-Lluch ◽  
J. Ballester ◽  
...  
Keyword(s):  
Protein Phosphorylation ◽  
Tyrosine Phosphorylation ◽  
Glucose Concentration ◽  
Egg Yolk ◽  
Freezing And Thawing ◽  
Precise Control ◽  
Boar Sperm ◽  
Phosphorylation Pattern ◽  
Thawing Process ◽  
Boar Spermatozoa

The optimization of freezing extenders is an essential issue for enhancing boar sperm cryosurvival. The aim of the present study was to disclose the role of glucose concentration of freezing extender on the metabolic activity of frozen–thawed spermatozoa. To achieve it, pooled sperm-rich ejaculate fractions from 5 mature and fertile boars (3 ejaculates per boar) were collected using the gloved-hand method. After centrifugation (2400g for 3 min), the sperm pellet was split into 7 aliquots. The aliquots were diluted to a final concentration of 1 × 109 sperm mL–1, in a Tris-citric extender supplemented with 20% egg-yolk, 3% glycerol, and 0, 0.05, 2, 4, 10, 55, or 185 mm glucose. All the extenders were adjusted to a pH of 6.8 and 310 mOsm kg–1 to avoid osmolarity effects. Extended semen samples were dispensed into 0.5-mL straws, and frozen in a programmable cell freezer at 20°C min–1. Thawing was carried out in a water bath at 37°C for 20 s. Afterward, an analysis of protein phosphorylation in tyrosine residues was carried out through bi-dimensional electrophoresis followed by a Western blot analysis. This analysis indicated that sperm samples frozen in extenders without glucose showed specific changes in the tyrosine phosphorylation pattern compared with fresh sperm. Furthermore, the addition of glucose in increasing concentrations to the freezing extender was accompanied by a concentration-dependent decrease in the overall tyrosine phosphorylation pattern, especially in proteins with a molecular weight ranging from 150 to 200 kDa and an acidic isoelectric point (pI). The maximal decrease was observed in spermatozoa frozen in the extender containing 185 mm glucose, in which an additional decrease in the tyrosine phosphorylation of proteins ranging from 60 to 80 kDa, and a basic pI was also observed. These results suggest that glucose is a modulator in the resistance of boar sperm to support freezing and thawing process, because the precise protein phosphorylation pattern of spermatozoa is directly linked to their functional status. In this way, a precise control of the glucose concentration of the freezing extender would be required to improve boar sperm cryoresistance. Supported by CICYT (AGL2005-00760 and AGL2004-04756-C02-02/GAN), Madrid and GERM (04543/07), Murcia, Spain.

scholarly journals Cryopreservation of boar semen

2020 ◽  
Vol 65 (No. 4) ◽  
pp. 115-123
Author(s):  
Marija Jovičić ◽  
Eva Chmelíková ◽  
Markéta Sedmíková
Keyword(s):  
Animal Species ◽  
Semen Quality ◽  
Egg Yolk ◽  
High Rate ◽  
Freezing And Thawing ◽  
Long Term Storage ◽  
Boar Semen ◽  
Boar Spermatozoa ◽  
Term Storage

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.

INFLUENCE OF GLYCEROL CONCENTRATION AND FREEZING TO −79 C ON OXYGEN UPTAKE AND LIVABILITY OF BOAR AND BULL SPERMATOZOA

1972 ◽  
Vol 52 (1) ◽  
pp. 65-72 ◽  
Author(s):  
L. M. SANFORD ◽  
G. J. KING ◽  
J. W. MACPHERSON
Keyword(s):  
Oxygen Uptake ◽  
Incubation Period ◽  
Skim Milk ◽  
Egg Yolk ◽  
Freezing Process ◽  
Glycerol Concentration ◽  
Motile Cells ◽  
Boar Semen ◽  
Boar Spermatozoa

Boar and bull spermatozoa were diluted in a skim milk–egg yolk–glucose extender containing 0, 7.5, or 15% glycerol (v/v) and incubated aerobically for 6 hr at 37 C. Other partially diluted boar semen samples were cooled to 5 C. Glycerol was added to a final concentration of 0, 7.5, and 15%. Samples were frozen to −79 C, rewarmed, and incubated for 3 hr at 37 C. The presence of glycerol in the extender depressed (P < 0.01) the oxygen uptake by nonfrozen boar and bull spermatozoa during the 6-hr incubation period. The reduction of oxygen uptake by semen samples increased as the level of glycerol in the extender increased. There was a corresponding decrease (P < 0.01) in the number of motile cells at the conclusion of the incubation period. Glycerol appeared to have more of a detrimental effect on boar spermatozoan oxygen uptake. The rate of oxygen uptake by boar semen samples postfreezing was extremely depressed, suggesting that spermatozoa surviving the freezing process metabolize at a much lower rate than normal. Active progressive motility of most of the surviving boar spermatozoa ceased within 1–2 hr of incubation under the in vitro conditions of this experiment.

The use of butylated hydroxytoluene in cryopreservation of boar semen

2016 ◽  
Vol 12 (2) ◽  
pp. 21-28
Author(s):  
Monika Trzcińska ◽  
Magdalena Baryła
Keyword(s):  
Dna Fragmentation ◽  
Sperm Quality ◽  
Annexin V ◽  
Egg Yolk ◽  
Butylated Hydroxytoluene ◽  
Preimplantation Embryos ◽  
Progressive Motility ◽  
Boar Semen ◽  
Boar Spermatozoa

The objective of the study was to determine the effect of butylated hydroxytoluene (BHT) on the quality and fertilizing capacity of frozen-thawed (FT) boar semen. Semen from five boars (36 ejaculates) was resuspended in lactose-egg yolk-glycerol extender supplemented with 0 (control), 1.0 (R1), 1.5 (R2) or 2.0 mM BHT (R3). Sperm quality was assessed based on motility (CASA; TM: total motility; PM: progressive motility), phosphatidylserine (PS) translocation across the plasma membrane (Annexin-V-FLuos Staining Kit) and DNA fragmentation (TUNEL Assay). The FT semen was also used for intrauterine artificial insemination (AI) of synchronized gilts. The fertilizing capacity of the FT semen was assessed on the basis of the gilt insemination rate and the number of morphologically normal embryos. The quality of the preimplantation embryos was determined by observing a TUNEL-positive reaction. The highest percentage of progressive motile and viable spermatozoa was noted in extender R3 (74.8 ±4.4% and 63.7 ±5.8%), as compared with the control (38.3 ±2.8% and 36.1 ±2.6%). The addition of BHT to the extender did not increase early apoptotic changes in the frozen-thawed spermatozoa with respect to the control. Irrespective of the variant of the extender, cryopreservation and thawing did not induce fragmentation in the boar spermatozoa. The highest number of morphologically normal embryos from inseminated gilts was observed in the case of semen cryopreserved in extender supplemented with 1.5 mM BHT. No significant differences were observed in DNA fragmentation in the expanded blastocysts from gilts inseminated with FT semen cryopreserved in the extenders analysed.

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