Glycerol, Methyl-Formamide and Dimethyl-Formamide in Canine Semen Cryopreservation

Futino, D.O.; Mendes, M C B; Matos, W N L; Mondadori, Rafael Gianella; Lucci, Carolina Madeira

doi:10.1111/j.1439-0531.2008.01208.x

Cited by 47 publications

(36 citation statements)

References 43 publications

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“…These results are supported by the findings of Futino et al (2010) who reported that lecithin may have played a protective role during cryopreservation due to its low viscosity, lower presence of debris, improves the kinematics of sperm cells and rearrangements structure of the plasma membrane. The reduction in motility parameters in extenders containing higher (1.5% SL and 2 % SL) levels of lecithin compared to 1% SL may be related to high viscosity (Van Wagtendonk-de Leeuw et al, 2000).…”

Section: Resultssupporting

confidence: 84%

Preservation of Black Bengal buck semen in soybean lecithin based chemically defined extender

Konyak

Mandal

Mondal

et al. 2018

IJAR

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This experiment was conducted with the aim to study the effect of replacing egg yolk with soybean lecithin (SL) for cryopreservation of Black Bengal buck semen. Sexually matured Black Bengal buck (n = 5) were used and the ejaculates were obtained using an artificial vagina method. The semen samples were pooled and diluted in Tris extender with 5% Glycerol containing either 15% egg yolk (control group) or SL at different concentrations (1% SL, 1.5% SL and 2% SL). The semen samples were filled in straws and cooled gradually to 5 °C. Semen straws were equilibrated for 3 hours at 5°C and were frozen in static liquid nitrogen vapor and stored in liquid nitrogen. Semen samples were evaluated after initial dilution, after completion of equilibration and after freeze thawing for in vitro sperm characters such as sperm motility, functional membrane integrity and malondioldehyde (MDA) concentration. Semen samples preserved in extender containing 1% SL was able to maintain in vitro sperm characters similar to the extender containing egg yolk. However, significant (P<0.05) reduction in all semen parameters was observed as the concentration of soybean lecithin increased above 1% level. It is concluded that an extender containing soybean lecithin @ 1% with 5% Glycerol can be used for replacing egg yolk for cryopreservation of Black Bengal buck semen.

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Section: Resultssupporting

confidence: 84%

Preservation of Black Bengal buck semen in soybean lecithin based chemically defined extender

Konyak

Mandal

Mondal

et al. 2018

IJAR

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“…Contraceptive effects of glycerol in mares has also been published (Demick et al 1976). There are a number of works dealing with the search for alternative cryoprotectants not only for stallion ejaculate, but also for the cryopreservation of sperm in other animal species and humans (Futino et al 2010;Swelum et al 2011). Investigated substances are specifically compounds of glycol: ethylene glycol, propylene glycol, diethylene glycol, dimethyl sulphoxide (Chenier et al 1998;Moore et al 2006;Hoffmann et al 2010) or various amides: formamide, methylformamide, dimethylformamide, acetamide or methylacetamide (Gomes et al 2002;Vidament et al 2002;Squires et al 2004;Alvarenga et al 2005;Ferreira et al 2008).…”

mentioning

confidence: 99%

Effects of glycerol concentration on the motility of equine spermatozoa after thawing

et al. 2017

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The aim of this study was to evaluate the effect of different glycerol concentrations on stallion sperm motility after thawing. For statistical analysis 228 ejaculates were used. The semen was filtrated to remove gel fraction; macroscopic and microscopic evaluation was done. After evaluation the ejaculates were centrifuged, the supernatant was removed and the spermatozoa were re-suspended in French diluent with different concentrations of glycerol (2.0; 2.5; 4.0 and 6.0%). The choice of concentration of glycerol for a particular ejaculate was completely random. The spermatozoa were packed into 0.5 ml straws and placed for 2 h in a fridge (4 °C). Then the straws were placed in liquid nitrogen vapor (-80 to -100 °C) and after 10 min plunged into liquid nitrogen and stored at -196 °C for at least 48 h. The selected straws were individually thawed in a 38 °C water bath for 30 s prior to post-freezing analysis. Two progressive motilities using phase contrast microscopy (magnification × 400) were recorded: motility II immediately after thawing and motility III after 2 h incubation in a 38 °C water bath. The Spearmen/Kendall rang correlation test was selected to prove whether there is a correlation between the selected indices (glycerol concentration and motility II and motility III). Nonparametric multiple group analysis (Steel-Dwass test) was applied for finding the differences between groups. The Spearman/Kendall rang correlation proved a relationship between motility II and glycerol concentration. It can be stated that in this study the best glycerol concentration for freezing equine spermatozoa is with a concentration of 4.0% glycerol.

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“…In dogs, there is a shortage of studies regarding the periods used for cooling to 5 C [22, 23]. To minimize the cold shock process, the majority of the protocols for dog semen cryopreservation include long cooling periods from 23 C to 5 C. The most frequently employed period is from 60 to 120 min, corresponding to approximate mean cooling rates within the range of 0.15–0.3 C/min [5, 13,14,15,16, 24]. Our results clearly demonstrated that the use of rapid cooling at a rate of 2.25 C/min prior to freezing provides the same post-thaw sperm quality, in terms of motility and viability, as the values obtained with the use of a control rate of 0.2 C/min.…”

Section: Discussionmentioning

confidence: 99%

“…Specifically, during cryopreservation, spermatozoa are very sensitive to the rapid reduction from room temperature to 5 C [7]. Although dog spermatozoa are considered relatively resistant to cooling at 5 C [9], slow cooling rates prior to freezing are generally used for this species [5, 10,11,12,13]. For this reason, our hypothesis was that a rapid cooling rate prior freezing could be used efficiently for dog semen cryopreservation.…”

mentioning

confidence: 99%

Effects of Rapid Cooling Prior to Freezing on the Quality of Canine Cryopreserved Spermatozoa

et al. 2014

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The aim of this study was to evaluate the effects of rapid cooling prior to freezing on frozen-thawed canine sperm quality. In experiment 1, centrifuged ejaculates from 6 dogs were pooled, split into 4 aliquots and cryopreserved by the Uppsala procedure using different cooling rates (control, cooling speed 18 C/90 min and average cooling rate 0.2 C/min; rapid, cooling speed 18 C/8 min and average cooling rate 2.25 C/min) in combination with 2 glycerol addition protocols (fractionated or unfractionated). In experiment 2, centrifuged ejaculates from 4 dogs were processed individually using the same cooling rates described in experiment 1 in combination with an unfractionated glycerol addition protocol. Each of the experiments was replicated 5 times. Sperm quality was evaluated after 30 and 150 min of post-thawing incubation at 38 C. Total motility (TM), progressive motility (PM) and quality of movement parameters were assessed using a computerized system, and sperm viability (spermatozoa with intact plasma and acrosome membranes) was assessed using flow cytometry (H-42/PI/FITC-PNA). Values for TM, PM, viable spermatozoa and the quality of movement parameters after thawing were not significantly affected by the cooling rate. The interaction between the cooling rate and the added glycerol protocol was not significant. There were significant differences among the males (P<0.01) in the sperm quality parameters evaluated after thawing. The interaction between the males and the cooling rate was not significant. In conclusion, canine spermatozoa can be cryopreserved using the Uppsala method at an average cooling rate of 2.25 C/min prior to freezing together with addition of fractionated or unfractionated glycerol.

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Glycerol, Methyl-Formamide and Dimethyl-Formamide in Canine Semen Cryopreservation

Cited by 47 publications

References 43 publications

Preservation of Black Bengal buck semen in soybean lecithin based chemically defined extender

Preservation of Black Bengal buck semen in soybean lecithin based chemically defined extender

Effects of glycerol concentration on the motility of equine spermatozoa after thawing

Effects of Rapid Cooling Prior to Freezing on the Quality of Canine Cryopreserved Spermatozoa

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