Rakha B A, Ansari M S, Akhter S, Zafar Z, Naseer A, Hussain I, Blesbois E, Santiago-Moreno J
Department of Wildlife Management, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, 46300, Pakistan.
Department of Zoology, University of Lahore, Sargodha Campus, Sargodha, 40100, Pakistan.
Theriogenology. 2018 Dec;122:61-67. doi: 10.1016/j.theriogenology.2018.09.003. Epub 2018 Sep 7.
Glycerol is a least toxic and most effective cryoprotectant for cryopreservation of poultry semen, but due to its contraceptive properties removal of glycerol is usually needed prior to artificial insemination. Dimethylsulfoxide (DMSO), a small amphiphilic molecule used as penetrating cryoprotectant for biological cells, has been recognized as an adequate alternative for cryopreservation of sperm from several species. This study was designed to evaluate the efficacy of different concentrations of DMSO as cryoprotectant for Indian red jungle fowl (Gallus gallus murghi) sperm. Semen was collected from Indian red jungle fowl cocks, pooled and divided into five aliquots. Different concentrations of DMSO (0%, 4%, 6%, 8% and 10%) were compared. Diluted semen was cooled from 37 °C to 4 °C (-0.275 °C min, 20% glycerol added to control and equilibrated for 10 min. After equilibration, semen was filled in 0.5 mL French straws, kept over liquid nitrogen vapors for 10 min and plunged into liquid nitrogen. Semen samples were thawed at 37 °C for 30 s. Cryo-survival of Indian red jungle fowl sperm was affected by cryopreservation stages and different concentrations of cryoprotectant used. Highest sperm motility (85.0 ± 2.9; 80.0 ± 3.5; 71.3 ± 4.3; 60.0 ± 1.3), plasma membrane integrity (79.5 ± 3.8; 75.3 ± 2.4; 72.8 ± 3.3; 60.3 ± 2.8), viability (80.8 ± 4.6; 75.5 ± 2.9; 71.0 ± 7.6; 58.8 ± 1.3) and acrosomal integrity (76.3 ± 2.4; 72.0 ± 6.0; 62.5 ± 4.3; 55.0 ± 3.2) were recorded in a diluent having 8% DMSO at post-dilution, cooling, equilibration and freeze-thawing. Highest fertility results were obtained after artificial insemination with 8% DMSO compared to 20% glycerol (73.0 ± 4.4 vs 53.1 ± 4.3, P < 0.05). It is concluded that 8% DMSO as a permeable cryoprotectant improves the post thaw semen quality and fertility in Indian red jungle fowl and can be used effectively to avoid the contraceptive effects of glycerol.
甘油是家禽精液冷冻保存中毒性最小且最有效的冷冻保护剂,但由于其具有避孕特性,通常在人工授精前需要去除甘油。二甲基亚砜(DMSO)是一种用作生物细胞穿透性冷冻保护剂的小分子两亲分子,已被认为是几种物种精子冷冻保存的合适替代品。本研究旨在评估不同浓度的DMSO作为印度红原鸡(Gallus gallus murghi)精子冷冻保护剂的效果。从印度红原鸡公鸡采集精液,混合并分成五等份。比较了不同浓度的DMSO(0%、4%、6%、8%和10%)。将稀释后的精液从37℃冷却至4℃(-0.275℃/分钟),向对照组添加20%甘油并平衡10分钟。平衡后,将精液装入0.5mL法式细管中,在液氮蒸汽上放置10分钟,然后投入液氮中。精液样本在37℃解冻30秒。印度红原鸡精子的冷冻存活率受冷冻保存阶段和所用冷冻保护剂不同浓度的影响。在稀释后、冷却、平衡和冻融后,在含有8%DMSO的稀释液中记录到最高的精子活力(85.0±2.9;80.0±3.5;71.3±4.3;60.0±1.3)、质膜完整性(79.5±3.8;75.3±2.4;72.8±3.3;60.3±2.8)、活力(80.8±4.6;75.5±2.9;71.0±7.6;58.8±1.3)和顶体完整性(76.3±2.4;72.0±6.0;62.5±4.