Rakha Bushra Allah, Ansari Muhammad Sajjad, Akhter Shamim, Zafar Zartasha, Hussain Iftikhar, Santiago-Moreno Julian, Blesbois Elisabeth
Department of Wildlife Management, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi 46300, Pakistan.
Department of Zoology, University of Sargodha, Lyallpur Campus, Faisalabad 38000, Pakistan.
Cryobiology. 2017 Oct;78:27-33. doi: 10.1016/j.cryobiol.2017.07.006. Epub 2017 Jul 27.
The Indian red jungle fowl is a sub-species of the genus Gallus native to South Asia; facing high risk of extinction in its native habitat. During cryopreservation, permeable cryoprotectants like glycerol are usually employed and we previously showed encouraging results with 20% glycerol. Because bird spermatozoa contain very little intracellular water, the possibility of replacing an internal cryoprotectant by an external one is opened. In the present study, we tested the replacement of internal cryoprotectant glycerol by the external cryoprotectant Polyvinylpyrrolidone (PVP). PVP is a non-permeable cryoprotectant and keeps the sperm in glassy state both in cooling and warming stages without making ice crystallization within the sperm cell. We evaluated the effect of various levels of polyvinylpyrrolidone (PVP) on Indian red jungle fowl semen quality and fertility outcomes. The qualifying semen ejaculates collected from eight mature cocks were pooled, divided into five aliquots, diluted (37 °C) with red fowl semen extender having PVP [0% (control) 4% (w/v), 6% (w/v), 8% (w/v) and 10% (w/v)]. Diluted semen was cryopreserved and stored in liquid nitrogen. The whole experiment was repeated/replicated for five times independently. Sperm motility, plasma membrane integrity, viability and acrosome integrity were recorded highest (P < 0.05) with 6% PVP at post-dilution, cooling, equilibration and freeze-thawing. Higher (P < 0.05) no. of fertile eggs, fertility, no. of hatched chicks, percent hatch and hatchability was recorded with 6% PVP compared to control. It is concluded that 6% PVP maintained better post-taw quality and fertility of Indian red jungle fowl spermatozoa than glycerol and can be used in routine practice avoiding the contraceptive effects of glycerol.
印度红原鸡是鸡属的一个亚种，原产于南亚，在其原生栖息地面临着极高的灭绝风险。在冷冻保存过程中，通常会使用像甘油这样的渗透性冷冻保护剂，我们之前使用20%甘油取得了令人鼓舞的结果。由于鸟类精子细胞内含水量极少，因此开启了用外部冷冻保护剂替代内部冷冻保护剂的可能性。在本研究中，我们测试了用外部冷冻保护剂聚乙烯吡咯烷酮（PVP）替代内部冷冻保护剂甘油的效果。PVP是一种非渗透性冷冻保护剂，能使精子在冷却和升温阶段都保持玻璃态，且不会在精子细胞内形成冰晶。我们评估了不同浓度的聚乙烯吡咯烷酮（PVP）对印度红原鸡精液质量和受精结果的影响。从八只成熟公鸡采集的合格精液射精样本混合后，分成五等份，用含有PVP[0%（对照）、4%（w/v）、6%（w/v）、8%（w/v）和10%（w/v）]的红原鸡精液稀释液在37℃下进行稀释。稀释后的精液进行冷冻保存并储存在液氮中。整个实验独立重复进行了五次。在稀释后、冷却、平衡和冻融后，6% PVP组的精子活力、质膜完整性、存活率和顶体完整性记录最高（P < 0.05）。与对照组相比，6% PVP组的受精卵数量、受精率、孵化出的雏鸡数量、孵化率百分比和孵化率均更高（P < 0.05）。结论是，6% PVP比甘油能更好地维持印度红原鸡精子解冻后的质量和受精能力，并且可用于常规操作，避免甘油的不良影响。
