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Cryopreservation of Ghagus chicken semen: effect of cryoprotectants diluents and thawing temperature
bioRxiv - Physiology Pub Date : 2020-05-16 , DOI: 10.1101/2020.05.16.099333
Shanmugam Murugesan , Ramkrishna Mahapatra

The present study evaluated the effects of cryoprotectants, semen diluents and thawing temperature during Ghagus chicken semen cryopreservation. Four different experiments were conducted; Experiment 1- semen was cryopreserved using 6% dimethylacetamide (DMA) and 2% dimethylsulfoxide (DMSO) in Sasaki diluent (SD) and Lake and Ravie diluent (LRD), Experiment 2 and 3- semen was cryopreserved using 8% Ethylene Glycol (EG) in SD, LRD and Red Fowl Extender (RFE), Experiment 4- semen was cryopreserved using 6% dimethylformamide (DMF) in SD, LRD and Beltsville Poultry Semen Extender (BPSE). Semen was cryopreserved in 0.5 ml French straws. Thawing was done at 5C for 100 sec in ice water in Experiments 1, 2 and 4, whereas in Experiment 3 thawing was done at 37C for 30 sec. The post-thaw sperm motility, live sperm and percent acrosome intact sperm were significantly (P<0.05) lower in cryopreserved samples in all the experiments. No fertile eggs were obtained from cryopreserved samples in Experiments 1 and 2, except for 8% EG RFE treatment where the fertility was 0.83%. In Experiments 3 and 4, highest fertility was obtained in LRD treatment 48.12 and 30.89% respectively. In conclusion, using cryoprotectant EG (8%) and thawing at 37C for 30 sec, and DMF (6%) resulted in acceptable level of fertility in Ghagus chicken. Though the diluents influenced post-thaw in vitro semen parameters the fertility was not affected. In addition, results indicated that thawing temperature may be a critical stage in the cryopreservation protocol.

中文翻译:

冻食加格斯鸡精液:冷冻保护剂稀释剂和解冻温度的影响

本研究评估了加格斯鸡精液冷冻保存过程中冷冻保护剂,精液稀释剂和解冻温度的影响。进行了四个不同的实验。使用Sasaki稀释剂(SD)和Lake和Ravie稀释剂(LRD)在6%的二甲基乙酰胺(DMA)和2%的二甲基亚砜(DMSO)中冷冻保存实验1精液,在8%的乙二醇(EG)中冷冻保存实验2和3的精液。 )在SD,LRD和Red Fowl延长剂(RFE)中,使用6%的二甲基甲酰胺(DMF)在SD,LRD和Beltsville家禽精液补充剂(BPSE)中冷冻实验4精液。将精液冷冻保存在0.5毫升的法国吸管中。在实验1、2和4中,在冰水中在5℃下融化100秒,而在实验3中,在37℃下融化30秒。解冻后的精子运动 在所有实验中,冷冻保存的样品中的活精子和顶体完整精子百分比均显着降低(P <0.05)。在实验1和2中,从冷冻保存的样品中未获得受精卵,除了8%EG RFE处理(受精率为0.83%)外。在实验3和4中,LRD处理的受精率最高,分别为48.12和30.89%。总之,使用冷冻保护剂EG(8%)并在37°C融化30秒钟,而DMF(6%)可使加格斯鸡的生育水平达到可接受的水平。尽管稀释剂影响解冻后的体外精液参数,但生育能力并未受到影响。另外,结果表明解冻温度可能是冷冻保存方案中的关键阶段。在实验1和2中,从冷冻保存的样品中未获得受精卵,除了8%EG RFE处理(受精率为0.83%)外。在实验3和4中,LRD处理的最高生育率分别为48.12和30.89%。总之,使用冷冻保护剂EG(8%)并在37°C融化30秒钟,而DMF(6%)可使加格斯鸡的生育水平达到可接受的水平。尽管稀释剂影响解冻后的体外精液参数,但生育能力并未受到影响。另外,结果表明解冻温度可能是冷冻保存方案中的关键阶段。在实验1和2中,从冷冻保存的样品中未获得受精卵,除了8%EG RFE处理(受精率为0.83%)外。在实验3和4中,LRD处理的受精率最高,分别为48.12和30.89%。总之,使用冷冻保护剂EG(8%)并在37°C融化30秒钟,而DMF(6%)可使加格斯鸡的生育水平达到可接受的水平。尽管稀释剂影响解冻后的体外精液参数,但生育能力并未受到影响。另外,结果表明解冻温度可能是冷冻保存方案中的关键阶段。使用冷冻保护剂EG(8%)并在37°C解冻30秒钟,而DMF(6%)可使加格斯鸡的生育水平达到可接受的水平。尽管稀释剂影响解冻后的体外精液参数,但生育能力并未受到影响。另外,结果表明解冻温度可能是冷冻保存方案中的关键阶段。使用冷冻保护剂EG(8%)并在37°C解冻30秒钟,而DMF(6%)可使加格斯鸡的生育水平达到可接受的水平。尽管稀释剂影响解冻后的体外精液参数,但生育能力并未受到影响。另外,结果表明解冻温度可能是冷冻保存方案中的关键阶段。
更新日期:2020-05-16
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