Comparison of different extenders for the cryopreservation of Atlantic salmon spermatozoa

Gallant, R. K.; Richardson, Gavin F.; McNiven, Mary A.

doi:10.1016/0093-691x(93)90401-p

Cited by 16 publications

(5 citation statements)

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“…In previous studies on cryopreservation of Atlantic salmon sperm, DMSO has been primarily used as the cryoprotectant (Truscott et al 1968;Truscott and Idler 1969;Mounib 1978;Stoss and Refstie 1983;Alderson and Macneil 1984;Gallant et al 1993). In this study, methanol provided a higher level of cryoprotection than did DMSO based on the fertilization results.…”

Section: Discussionmentioning

confidence: 77%

Methanol and Egg Yolk as Cryoprotectants for Atlantic Salmon Spermatozoa

et al. 2007

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The effects of four extenders on the fertilization rates of eggs fertilized with cryopreserved sperm of Atlantic salmon Salmo salar were tested. We used (1) glucose extender (54.04 g glucose/L and 1.7 g KCl/L) with 5% DMSO, (2) glucose extender with 5% DMSO supplemented with 13.3% egg yolk, (3) glucose extender with 10% methanol, and (4) glucose extender with 10% methanol supplemented with 13.3% egg yolk. Fertilization rates, expressed as the percentage of eyed embryos, ranged from 52.7% to 83.5%. Sperm cryopreserved with the glucose extender and 10% methanol supplemented with 13.3% egg yolk yielded significantly higher fertilization rates (83.5%) than did sperm cryopreserved with the other three extenders. Our fertilization rates compare favorably with those observed for eggs from the same year-class fertilized with fresh milt (81.4%) and reared at the White River National Fish Hatchery. The presence of egg yolk in extenders incorporating 10% methanol provided additional protection to salmonid sperm during the freezing and thawing processes and resulted in an increase in survival from 72.9% to 83.5%. However, the cryoprotective effect of egg yolk may be specific to the individual formulation of extenders. In our trials, glucose and 5% DMSO without egg yolk yielded a 66.9% fertilization rate, while glucose and 5% DMSO supplemented with 13.3% egg yolk produced only 52.7% fertilization after cryopreservation.

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Section: Discussionmentioning

confidence: 77%

Methanol and Egg Yolk as Cryoprotectants for Atlantic Salmon Spermatozoa

et al. 2007

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“…Diluent 2 was 0.3 m glucose with 0.011 m KCl added to inhibit sperm activation before fertilization ( Stoss & Holtz 1983). Diluent 3 was a modification of that described by Horton & Ott (1976) and has been used to cryopreserve Atlantic salmon Salmo salar L. and rainbow trout semen ( Gallant, Richardson & McNiven 1993; McNiven, Gallant & Richardson 1993). This diluent contained 0.137 m NaCl, 0.011 m KCl, 0.004 m Na 2 HP0 4 .7H 2 0 and 7.5 g L −1 l ‐α‐lecithin and was adjusted to pH 7.5.…”

Section: Methodsmentioning

confidence: 99%

Cryopreservation of Arctic charr,Salvelinus alpinus(L.), semen in various extenders and in three sizes of straw

Richardson

Miller

McNiven

2000

Aquaculture Research

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Cryopreservation of Arctic charr Salvelinus alpinus (L.) semen was investigated using three diluents, three cryoprotectants [10% dimethyl sulphoxide (DMSO), 10% dimethylacetamide (DMA) or 20% glycerol] and three sizes of straw. The three diluents and three cryoprotectants were combined, resulting in nine extenders. One part semen was added to three parts extender, and motility was evaluated to assess the toxicity of six of the extenders. Semen in nine extenders was frozen in 0.5‐mL straws using liquid nitrogen vapour. Semen extended in 0.3 m glucose and each of the cryoprotectants was also frozen in 0.5‐mL, 1.7‐mL (flat) or 2.5‐mL straws. The freezing rate in each size of straw was measured. Fertility trials were conducted to determine the post‐thaw viability of the frozen semen. The motility of activated spermatozoa was higher in the DMA and DMSO extenders than in the glycerol extender. For the trial using 0.5‐mL straws, post‐thaw fertility results were higher for all extenders containing DMSO, or 0.3 m glucose and DMA, than for all other combinations of diluent and cryoprotectant. For the straw size comparison, the highest fertility was obtained for the 1.7‐mL straw using either DMSO or DMA and for the 2.5‐mL straw using DMSO. For all cryopreservation trials, fertility was low for extenders containing glycerol.

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“…目前鱼类的种质细胞保存中, 精液保存是重要组成部分之一 (Chen, 2007)。自 20 世纪 50 年代初首次成功冷冻保存大西洋鲱(Clupea harengus)精巢以来 (Blaxter, 1953), 世界各国学者对鱼类配子的冷冻保存进行了大量研究, 从稀释液的选择 (Gallant et al, 1993)、保护剂的使用浓度 (Kopeika et al, 2003;Muchlisin, 2005)、不同冷冻保护剂的配合、平衡时间 (He & Woods, 2003)、降温速率、解冻方式及复温速率 , 到先进低温仪器设备的研制和利用等方面解决了不同生物材料的超低温冷冻保存问题 (Leung & Jamieson, 1991;Li et al, 1998)。目前, 已经报道了 200 余种鱼类精子冷冻保存的研究, 形成了 60 余种淡、海水鱼类精液成熟的冷冻保存技术, 并建设了首个淡水鲤科鱼类冷冻精子库 (Chen, 2007)。软鳍新光唇鱼(Neolissochilus benasi) (Pellegrin & Chevey, 1936), 旧称软鳍四须鲃 (Barbodes benasi), 隶属于鲤科(Cyprinidae)鲃亚科(Barbinae) 新光唇鱼属(Neolissochilus), 有关软鳍新光唇鱼的研究主要集中在其分类地位上 (Chen & Yang, 2003), 在精子冷冻方面未见有报道。随着中国境内红河水系水电的开发, 对河道内多种珍稀土著鱼类产生了诸多的不利影响 (Yang et al, 2011) (Yang et al, 2010;Yang et al, 2011) Fleming & Gross, 1993); 而很多的研究报道将造成这一结果的原因归结于环境的改变, 而这种环境改变对塘养环境下雌鱼所产鱼卵质量造成的影响更能引起人们的广泛关注 (Pan et al, 2009) (Chen et al, 1992)稀释成相应比例, 以一定体积装入 1.8 mL 冻存管中, 4 ℃平衡一段时间, 然后将冻存管在液氮面上 6 cm 放置 2 min, 液氮面上放置 2 min, 然后投入液氮中 (Chen, 2007)。24 h 后, 取出液氮中的样品, 一定温度水浴解冻 30 s。 …”

Section: 多样性已成为水产养殖业中亟待解决的重要科技问题。unclassified