Effects of Egg Yolk, Glycerol and the Freezing Rate on the Viability and Acrosomal Structures of Frozen Ram Spermatozoa

Watson, P. F.; Martin, Ian

doi:10.1071/bi9750153

Cited by 68 publications

(27 citation statements)

References 10 publications

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“…The incorporation of up to 18.0% v/v egg yolk in the pre-freezing diluent is known to improve the post-thawing motility of cryopreserved ram spermatozoa (Salamon and Visser, 1972;Watson and Martin, 1975). The active fraction of egg yolk is the low density lipoprotein (LDL); the protein component binds to the sperm surface and phosphotidylcholine appears to be instrumental in providing cryoprotection (Watson, 1981).…”

Section: Discussionmentioning

confidence: 99%

In vitro evaluation of zwitterion buffers in diluents for freezing ram spermatozoa

Molinia

Evans²,

Maxwell³

1994

Reprod. Nutr. Dev.

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Summary ― Diluents based on the zwitterion buffers Tes, Hepes and Pipes titrated to pH 7.0 with NaOH or Tris were compared with Tris-citrate diluents by assessment of post-thawing motility and acrosome integrity of frozen ram spermatozoa. Varying buffer osmolalities between 300 and 360 mosmol had no effect but there was a decrease (P < 0.001 ) in motility between 360 and 420 mosmol. There was a quadratic effect (P < 0.001 ) on motility of increasing glucose concentration in the diluent (30, 85, 140, 195 and 250 mM) with a maximum at 85 mM. At this sugar concentration, motility was higher (P < 0.001 ) for diluents containing glucose and fructose than lactose, sucrose or trehalose. Inclusion of egg yolk (optimum at 13.5% v/v; P < 0.05), centrifugation of the diluents (P < 0.01 use of low dilution rates (3 or 6-fold; P < 0.05) and freezing in pellets rather than minitubes or straws (P < 0.001 ) improved both the motility and acrosome integrity of spermatozoa in the zwitterion diluents. It is concluded that zwitterion buffers are superior to Tris-citrate in ram semen freezing diluents. zwitterion buffer / motility / acrosome integrity / frozen ram spermatozoa Résumé ― Effet de l'addition de tampons amphotères à des dilueurs sur la congélabilité des spermatozoïdes de bélier. Nous avons comparé l'efficacité de dilueurs à base de tampons amphotères TES, HEPES et PIPES ajustés à pH 7,0 avec NaoH ou tris, à celle de milieux à base de tris-citrate, en évaluant la motilité des spermatozoïdes de bélier et l'intégrité de leur acrosome après congélation. Entre 300 et 360 milliosmoles, l'osmolarité des tampons n'a aucun effet. Au-delà, et jusqu'à 420 milliosmoles, on observe une baisse significative (P < 0, 001) de la motilité. On obtient un effet biphasique sur la motilité en augmentant la concentration de glucose dans le dilueur (30, 85, 140, 195, 250 mM) avec un maximum pour 85 mM. À cette concentration, la motilité est plus élevée avec le milieu contenant du glucose et du fructose qu'avec ceux contenant du lactose, du sucrose ou du tréhalose (P < 0,001). L'addition de jaune d'œuf (optimum à 13,5% vlv; P < 0,05), la centrifugation des dilueurs (P < 0,01), l'utilisation d'un taux de dilution peu élevé (3 ou 6 fois ; P < 0,05) et la congélation en pastilles plutôt qu'en minitubes ou en minipaillettes (P < 0,001) améliorent la motilité et l'intégrité de l'acrosome des spermatozoïdes congelés dans des milieux contenant un tampon amphotère. Cette étude montre donc que les tampons amphotères sont plus efficaces que le tampon tris-citrate pour les dilueurs utilisés pour la congélation du sperme de bélier.bélier / mobilité l acrosome / sperme congelé / dilueur / tampon * Current address:

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Section: Discussionmentioning

confidence: 99%

In vitro evaluation of zwitterion buffers in diluents for freezing ram spermatozoa

Molinia

Evans²,

Maxwell³

1994

Reprod. Nutr. Dev.

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“…Sperm damage during preservation in liquid nitrogen at -196°C has been attributed by many authors to cold shock, ice crystal formation, oxidative stress, osmotic changes, cryoprotectant toxicity and reorganizations of lipid-protein and phospholipid layers within the cell membranes [6][7][8]. It is proposed that membrane is thought to be a primary target of cold shock or freezing damage in cells, which leads to a loss of integrity and selective permeability of the sperm plasma membrane [9,10].…”

Section: Introductionmentioning

confidence: 99%

Pure Egyptian Cattle Bulls Show both Individual Variation and Different Interaction with Extender in the Post-Thawing Sperm Parameters

Mohammed¹,

Ahmed²

2017

Andrology

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Objectives: As very few studies were done on the freezability of pure Egyptian cattle bull sperm. So, we designed this study to evaluate the individual variations in freezability of native bull semen extended in Tris based diluent and sodium citrate-based diluent. Methods:Semen was collected by artificial vagina, examined at once in farm laboratory. Only semen samples fit the minimum parameters are extended in two extenders first the universal one (TRIS) and second modified Sodium citrate extender by adding glycerol (CU-16) which created at Cornell University to evaluate the individual bull variation and interaction between extender and bull.Results: Post-Thawing individual sperm motility, live, abnormal, acrosome integrity percentage was evaluated in addition to Hypo-Osmotic Swelling test (HOS). The highest value for motility, live abnormal and acrosome percentage were 44.00 ± 1.12, 52.25 ± 1.60, 21.25 ± 0.81 and 62.80 ± 2.58 from bull 2, 1, 3 and 2, respectively for semen extended in TRIS, and 42.25 ± 1.61, 52.00 ± 1.76, 22.15 ± 0.85 and 57.40 ± 3.07, from bull 1, 1, 4, 3, respectively for semen extended in CU-16. The results of HOS were 59.25 ± 1.76 and 55.95 ± 2.13 from bull 1 extended in TRIS and CU-16, respectively. Conclusion:A significant variation (p<0.05) in tested parameter was clear when semen extended in TRIS extender with the absence of such variation when semen extended in CU-16 except in live sperm percentage. With ignoring the extender effect a clear significant variations were detected between bulls in all tested parameters.

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“…Preparation of uniform semen extenders containing egg yolk is difficult because individual egg yolk quality may vary depending on the number of days after laying and the storage period. Furthermore, addition of egg yolk reduces the acrosome integrity of goat spermatozoa [2], and high egg yolk concentrations reduce the postthawing viability of ejaculated spermatozoa in several species, such as goats [3], rams [4] and water buffaloes [5]. Finally, development of a chemically defined semen extender would be required in order to eliminate possible infectious origins from egg yolk added to the semen extender.…”

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confidence: 99%

Fertility of Ewes Inseminated Intrauterinally with Frozen Semen Using Extender Containing Bovine Serum Albumin

et al. 2007

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Abstract. The present study was conducted to examine the fertility of ewes inseminated intrauterinally with frozen semen using semen extender containing either egg yolk or bovine serum albumin (BSA). Sixty Suffolk and cross-bred ewes were treated with controlled internal drug release (CIDR) devices during the non-breeding season (July 2006). A CIDR was inserted into the vagina for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin was administered one day before its removal. Ejaculates from a suffolk ram were diluted with a Tris-based extender containing either 15% (v/v) egg yolk or 10% (w/v) BSA, and the diluted semen was frozen in 0.25 ml straws. A fixed-time intrauterine artificial insemination (AI) was performed 43-47 h after CIDR removal, regardless of incidence of estrus. There was no significant difference in pregnancy rates at 60 days after AI between the extenders containing egg yolk (66.7%, 20/30 animals) or BSA (65.5%, 19/29 animals). Furthermore, there were no significant difference in the lambing rates (66.7% and 62.1%) and prolificacy (1.25 and 1.56) between the two semen extenders. The present study indicates that a semi-defined semen extender containing 10% BSA produces fertility after intrauterine AI that is similar to that achieved with semen extender containing egg yolk. o s t s e m e n e x t e n d e r s f o r f r e e z i n g o f spermatozoa, including ram semen contain egg yolk, milk, or a combination of the two as a basic ingredient. Although egg yolk is beneficial for sperm cryopreservation because it protects a g a i n s t c o l d s h o c k [ 1 ] , t h e r e a r e s e v e r a l disadvantages of addition of egg yolk to semen extender. Preparation of uniform semen extenders containing egg yolk is difficult because individual egg yolk quality may vary depending on the number of days after laying and the storage period. Furthermore, addition of egg yolk reduces the acrosome integrity of goat spermatozoa [2], and high egg yolk concentrations reduce the postthawing viability of ejaculated spermatozoa in several species, such as goats [3], rams [4] and water buffaloes [5]. Finally, development of a chemically defined semen extender would be required in order to eliminate possible infectious origins from egg yolk added to the semen extender. Several investigations have been conducted for development of semen extenders that do not contain egg yolk, such as a defined extender containing soybean lecithin for bovine and wild gazelle semen [6][7][8][9][10], and bovine serum albumin (BSA) has also been used as a substitute of egg yolk for rainbow trout and turkey spermatozoa [11,12].

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Effects of Egg Yolk, Glycerol and the Freezing Rate on the Viability and Acrosomal Structures of Frozen Ram Spermatozoa

Cited by 68 publications

References 10 publications

In vitro evaluation of zwitterion buffers in diluents for freezing ram spermatozoa

In vitro evaluation of zwitterion buffers in diluents for freezing ram spermatozoa

Pure Egyptian Cattle Bulls Show both Individual Variation and Different Interaction with Extender in the Post-Thawing Sperm Parameters

Fertility of Ewes Inseminated Intrauterinally with Frozen Semen Using Extender Containing Bovine Serum Albumin

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