Freezing canine sperm: Comparison of semen extenders containing Equex® and LDL (Low Density Lipoproteins)

Bencharif, D.; Amirat-Briand, L.; Garand, Annabelle; Anton, Marc; Schmitt, Éric; Desherces, S.; Delhomme, Guy; Langlois, Marie-Laure; Barrière, P.; Destrumelle, S.; Vera-Munoz, O.; Tainturier, D.

doi:10.1016/j.anireprosci.2010.01.009

Cited by 37 publications

(19 citation statements)

References 31 publications

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“…After freezingthawing, the total sperm motility (approximately 60-70%) of the semen chilled for up to 48 h did not show any differences from the samples frozen by the conventional cryopreservation method (63.2%); even the progressive fast spermatozoa percentages were higher in the chilled R1 (p = 0.079) and R6 (p < 0.05) aliquots than in the control group. These results were comparable to the sperm motility percentages described in the frozenthawed canine semen frozen with the usual freezing protocol (without previous chilling), which have shown mean values between 45% and 70% (Hay et al 1997;Stro¨m et al 1997;Rota et al 1999;Yildiz et al 2000;Iguer-Ouada and Verstegen 2001b;Martins-Bessa et al 2006;Bencharif et al 2010). Therefore, the results for sperm motility may indicate that cooling the semen at 4°C for up to 48 h before freezing is not disadvantageous as compared to direct cryopreservation.…”

Section: Discussionsupporting

confidence: 83%

Influence of Cool Storage before Freezing on the Quality of Frozen–Thawed Semen Samples in Dogs

Santana

Batista

Alamo

et al. 2012

Reprod Domestic Animals

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

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Section: Discussionsupporting

confidence: 83%

Influence of Cool Storage before Freezing on the Quality of Frozen–Thawed Semen Samples in Dogs

Santana

Batista

Alamo

et al. 2012

Reprod Domestic Animals

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“…Moreover, some researchers (e.g., Amirat et al 2005) have demonstrated that extenders based on egg yolk can have negative effects on sperm respiration and motility due to other specific substances they contain. Pace & Graham (1974) purified egg yolk by ultracentrifugation and found that a fraction of egg yolk known as »low-density lipoprotein« (LDL) has a cryoprotective effect on the integrity of the plasma membrane, as well as on the percentage of normal spermatozoa and sperm motility (Bencharif et al 2010) and preserves bull semen and maintains its fertility during freezing, storage, and thawing (Amirat et al 2004). The motility of sperm was almost twice as high in LDL (54.4 %) compared to Optidyl (30.2 %, P<0.05).…”

Section: Introductionmentioning

confidence: 99%

Effect of sire and extender on sperm motility and share of live or dead sperm in bulls’ fresh ejaculate and in AI doses after thawing

et al. 2012

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The objectives were to evaluate the effects of sire and content of fresh or ionised egg yolk in extenders on sperm motility and share of live and dead sperm in collected ejaculate, in thawed artificial insemination (AI) doses, and during thermodynamic testing compared to extenders not containing egg yolk. Ejaculates were collected once a week from 4 Holstein bulls. Each of the 20 ejaculate samples from each bull was diluted with 4 different extenders. AndroMed and Bioxcell (no egg yolk) and Triladyl and Optidyl (fresh, ionised egg yolk) were used. A total of 640 AI doses were analysed. The volume of samples, sperm concentration, and percentage of motile spermatozoa were evaluated after collection, as was sperm motility after thawing of AI doses and during thermodynamic testing. Percentages of live and dead sperm were also evaluated. The data set was analysed using SAS/STAT 9.1 (SAS Institute Inc., Cary, NC, USA). The results confirmed significant (P<0.05-0.01) between-sire differences in the volume, density, and activity of sperm as well as in share of live and dead sperm after collection; in decline of sperm motility in the fresh ejaculate, after thawing, and during the entire thermodynamic test, as well as in the share of live and dead sperm after thawing. The extenders ranked by sperm motility are: Optidyl, Triladyl, AndroMed, and Bioxcell, demonstrating the higher quality of AI doses produced using egg yolk extenders. Differences in sperm motility were significant (P<0.05-0.01) during the entirety of thermodynamic testing. Egg yolk extenders had a significantly (P<0.05-0.01) higher percentages of live sperm after thawing.

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“…Many factors that affect canine sperm quality during freezing and thawing have been studied. In particular, the presence of vital factors that potentiate the resistance of dog ejaculates to freezing and thawing damage are important (Bencharif et al, , ; Holt, ; Peña et al, ; Ponglowhapan & Chatdarong, ; Thurston, Watson, & Holt, ). Therefore, many compounds have been used to protect spermatozoa during freezing and thawing processes.…”

Section: Introductionmentioning

confidence: 99%

Use of polyvinyl alcohol as a chemically defined compound in egg yolk‐free extender for dog sperm cryopreservation

Nabeel

Jeon

2019

Reprod Domestic Animals

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The objectives of this study were to investigate the effects of polyvinyl alcohol (PVA) as a chemically defined compound in egg yolk (EY)‐free extender by determining the appropriate concentration of PVA and the effect of pH adjustment in EY‐free PVA extenders on dog spermatozoa. Spermatozoa (1 × 108 cells/ml) were frozen with EY‐free extenders supplemented with 0 (control), 0.025, 0.05, 0.1, 0.2 or 0.3 g/100 ml PVA. Sperm progressive motility (PM) was assessed immediately after thawing (IAT) and post‐thaw incubation (PTI), while viability, acrosome integrity and reactive oxygen species (ROS) levels were evaluated after PTI. Additionally, spermatozoa were frozen using EY‐free PVA extenders before pH adjustment (6.45) and after adjustment of pH (6.85). Viability, PM, ROS and gene expression (BCL2 and SMCP) were assessed. Supplementation with 0.05 g/100 ml or more PVA significantly increased PM compared to the control group in the IAT and PTI. Post‐thaw incubation significantly increased sperm motility in all groups. The acrosome integrity in all PVA groups was higher (p < .05) than the control without an effect on ROS and viability. Adjustment of the pH to 6.85 improved (p < .05) sperm PM compared to the non‐adjusted groups without affecting viability, ROS or expression of BCL2 and SMCP. We suggest that PVA supplementation in EY‐free Tris extenders can effectively protect dog spermatozoa during freezing and can maintain higher motility and acrosome integrity. Adjustment of pH in EY‐free PVA extenders can improve post‐thaw sperm motility. Therefore, PVA can be used as a compound in EY‐free extender for the cryopreservation of dog spermatozoa.

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Freezing canine sperm: Comparison of semen extenders containing Equex® and LDL (Low Density Lipoproteins)

Cited by 37 publications

References 31 publications

Influence of Cool Storage before Freezing on the Quality of Frozen–Thawed Semen Samples in Dogs

Influence of Cool Storage before Freezing on the Quality of Frozen–Thawed Semen Samples in Dogs

Effect of sire and extender on sperm motility and share of live or dead sperm in bulls’ fresh ejaculate and in AI doses after thawing

Use of polyvinyl alcohol as a chemically defined compound in egg yolk‐free extender for dog sperm cryopreservation

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