Cryopreservation of rabbit semen: Comparing the effects of different cryoprotectants, cryoprotectant-free vitrification, and the use of albumin plus osmoprotectants on sperm survival and fertility after standard vapor freezing and vitrification

Rosato, M. P.; Iaffaldano, Nicolaia

doi:10.1016/j.theriogenology.2012.11.008

Cited by 84 publications

(82 citation statements)

References 28 publications

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“…Other smallscale experiments using 1 and 2 M Me 2 SO (Hanada andNagase, 1980 andKashiwazaki et al, 2006, respectively) showed no differences between Me 2 SO and acetamide as cryoprotectants for rabbit sperm freezing. On the contrary, several authors have demonstrated the efficient sperm protection of 3.0 and 3.5 M Me 2 SO in rabbit sperm cryopreservation Viudes de Castro and Vicente, 1996;Mocé et al, 2003;Cortell and Viudes de Castro, 2008;Iaffaldano et al, 2012;Rosato and Iaffaldano, 2013). According to our results, when Me 2 SO was used v. acetamide extender, results on frozen-thawed sperm quality and reproductive parameters studied were improved.…”

Section: Discussionsupporting

confidence: 52%

“…It would be highly beneficial in routine artificial insemination (AI) programmes on commercial rabbit farms if semen could be cryopreserved without affecting fertility, although a multitude of factors affect the effectiveness of cryopreservation, including the initial quality of the semen samples, cryopreservation protocol, freezing extenders and environmental factors (Mocé and Vicente, 2009;Iaffaldano et al, 2012;Lavara et al 2013;Rosato and Iaffaldano, 2013). In addition, differences in reproductive parameters have been observed with fresh (Bolet et al, 2000;Vicente et al, 2000) and cryopreserved sperm belonging to different breeds or lines of rabbits.…”

Section: Introductionmentioning

confidence: 99%

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Effect of freezing extender composition and male line on semen traits and reproductive performance in rabbits

Viudes-de-Castro

Lavara

Safaa

et al. 2014

Animal

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This study was conducted to elucidate the effect of different freezing extenders on two lines selected for hyperprolificacy and longevity (H and LP, respectively). In extender A, dimethyl sulphoxide (Me 2 SO) and sucrose were used as cryoprotectants. In extenders B and C, the sucrose was replaced by 20% egg yolk, and in extender C the Me 2 SO was substituted by acetamide. Semen was packaged in 0.25 ml plastic straws and cooled at 5°C for 45 min, and then was frozen in liquid nitrogen vapour for 10 min before being plunged into the liquid nitrogen. Thawing was carried out by immersing the straws in a water bath at 50°C for 10 s. Frozen-thawed semen characteristics and reproductive parameters were affected by freezing. Extender C showed significantly lower post-thawing quality traits than any of the three extenders. Acrosome integrity was significantly improved when Me 2 SO was used as cryoprotectant. Sucrose replacement by 20% egg yolk had no effect on acrosome integrity but provided significantly lower sperm motility and viability. Freezing extender affected fertility rate, total born, number of implantation sites and gestational losses, obtaining better results when extender A was used. The acrosomal integrity after frozen-thawed process showed a significant correlation with fertility at 12 th day and also at birth, indicating that an increase in acrosomal integrity leads to an increase in both fertilities (12 th day and at birth). A positive correlation between motility of semen and implantation sites was found. The post-thawing quality traits of semen were not affected by the genetic line, although LP line showed higher total born and lower foetal and gestational losses. The findings of this study suggest that freezing extender composition has a significant effect on the success of rabbit sperm for preservation, and when Me 2 SO was used as permeable cryoprotectant sucrose provided better protection compared with egg yolk and improved reproductive traits, and, on the other hand, the male genotypes used in the present study had no effect on frozen-thawed sperm parameters but negatively affected some of the reproductive parameters.

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Section: Discussionsupporting

confidence: 52%

Section: Introductionmentioning

confidence: 99%

Effect of freezing extender composition and male line on semen traits and reproductive performance in rabbits

Viudes-de-Castro

Lavara

Safaa

et al. 2014

Animal

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“…Our results reveal a negative impact of crude glycerol on the spermatozoa kinetics. Rosato and Iaffaldano (2013) observed a beneficial effect of 5% and 10% glycerol on rabbit spermatozoa, where glycerol preserved 60-65% of spermatozoa motility for the first 45 minutes upon treatment. Then, however, the motility dropped to ~40% after 15 min incubation.…”

Section: Discussionmentioning

confidence: 91%

“…The effects of pure glycerol on animal and human spermatozoa have been previously well documented (Kashizawaki et al, 2006;Rosato and Iaffaldano, 2013). This study was designed to investigate the effects of crude glycerol on rabbit spermatozoa motility in vitro.…”

Section: Discussionmentioning

confidence: 99%

Crude glycerol negatively affects rabbit spermatozoa motility in vitro

Paál

Strejcek

Slanina

et al. 2018

J Hellenic Vet Med Soc

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In recent years the conventional diet for farmed animals has been increasingly substituted by alternative energy sources such as crude glycerol. There is an urgent need for investigation of the impact of crude glycerol on the male reproductive tract. Rabbit semen was cultured in a TRIS/NaCl-based medium containing 0% (control), 5%, 10%, 15% and 20% crude glycerol. Sperm kinetic characteristics were assessed immediately and 30 min, 60 min, 120 min and 180 min upon treatment by computer assisted analyzer. In general, a decrease in motility was recorded for the spermatozoa treated with 5% glycerol (P<0.001). A substantial reduction in spermatozoa motility was observed in the samples containing 10% and 15% crude glycerol (P<0.05). The lowest motility was observed for spermatozoa incubated with 20% crude glycerol. Presented data suggest toxic effects of crude glycerol on the rabbit spermatozoa kinetics in vitro.

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“…Similarly, vitrification (ultra-rapid freezing) of spermatozoa without the use of cryoprotectants has achieved post-thaw recovery of DNA integrity comparable to fresh spermatozoa in the rabbit (Rosato and Iaffaldano 2013) and human (Isachenko et al 2004ab). This observation has practical implications for the assessment of SDF in the field.…”

Section: Discussionmentioning

confidence: 99%

Amphibian sperm DNA fragmentation

Pollock¹

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There are extremely limited studies that have investigated DNA fragmentation in amphibians and even less information exists pertaining to DNA fragmentation in the spermatozoa.A DNA fragmentation assay applied and validated for amphibian spermatozoa would not only provide the first evidence of this phenomenon in a new major taxon, but also prove a valuable tool for developing improvements in ART used in the captive propagation of threatened and endangered species. Consequently, the fundamental aim of this project was the development and validation of the sperm chromatin dispersion (SCD) test for amphibian spermatozoa with the purpose of investigating the dynamic behaviour of amphibian sperm DNA in response to cryopreservation.The SCD test was first applied to African clawed frog (Xenopus laevis) as an amphibian sperm model using a species-specific modified lysing solution for protein depletion. Sperm DNA fragmentation (SDF) was assessed immediately following activation of sperm motility (T0) and again after one hour and 24 hours of incubation in order to produce a range of spermatozoa with differing levels of DNA damage. The SCD procedure resulted in the production of three nuclear morphotypes; amphibian sperm morphotype 1 (ASM-1) and ASM-2 showed no evidence of DNA damage, whereas ASM-3 spermatozoa were highly fragmented with large halos of dispersed DNA fragments and a reduced nuclear core. Levels of SDF revealed by the SCD test were highly correlated with results produced using in situ nick hybridisation with DNA-specific molecular probes (ISNT) and the double comet assay (r = 0.9613). The alkaline step of the double-comet assay revealed the extensive presence of structural single-stranded DNA breaks (SSB) and provided grounds for the suggestion that alkali labile sites (ALS) are a constitutive feature of African clawed frog sperm chromatin.SDF is a dynamic process and has been shown to increase with incubation at room temperature as well as following cryopreservation. However, the mechanisms of DNA cryoinjury and the resulting effect on the potential reproductive output of ART remain open to interpretation.Consequently, dynamic changes to the basal level of DNA fragmentation were examined in fresh African clawed frog spermatozoa in comparison with the post-thaw integrity of cryopreserved (-80 °C) and frozen (-20 °C) sperm DNA. SDF was examined initially at T0 and over incubation of up to 60 minutes. The difference in SDF was only marginal for fresh, frozen (-20 °C) and cryopreserved (-80 °C) spermatozoa examined at the onset of the experiment immediately upon thawing, however, a significant increase (p = 0.004) in SDF, albeit of a low magnitude, was observed in activated ii cryopreserved (-80 °C) spermatozoa following incubation. Although African clawed frog spermatozoa appeared to be highly resistant to cryopreservation-induced DNA damage, cryopreserved-thawed spermatozoa yielded a significantly lower (p = 0.0065) fertilisation rate than fresh spermatozoa.In order to provide further insight into cryo...

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Cryopreservation of rabbit semen: Comparing the effects of different cryoprotectants, cryoprotectant-free vitrification, and the use of albumin plus osmoprotectants on sperm survival and fertility after standard vapor freezing and vitrification

Cited by 84 publications

References 28 publications

Effect of freezing extender composition and male line on semen traits and reproductive performance in rabbits

Effect of freezing extender composition and male line on semen traits and reproductive performance in rabbits

Crude glycerol negatively affects rabbit spermatozoa motility in vitro

Amphibian sperm DNA fragmentation

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