PDF(Pubmed)
Abstract：
Semen cryopreservation is an important technique for preserving the genetic material of numerous species. However, frozen semen is highly susceptible to sperm DNA damage and reduced motility, resulting in decreased fertility. The standard method for cryopreservation and several approaches have not been elucidated. This study aimed to determine the effects of supplementing rooster semen extender with a combination of phosphorus and vitamin B12 on cryopreserved semen quality. Semen was collected weekly via dorso-abdominal massage from 57 Burmese × Vietnam-crossbred Thai native roosters aged 1-3 years. In total, 139 semen samples were collected, pooled, and diluted to 200 million sperm per dose. The pooled sample was divided into six experimental groups: a control group (0.00%) diluted with modified Beltville Poultry Semen Extender (BPSE) and five treatment groups diluted with modified BPSE supplemented with phosphorus and vitamin B12 at concentrations 0.02, 0.04, 0.06, 0.08, and 0.10%, respectively. The semen samples were frozen and evaluated at 0, 15, and 30 min after thawing. Sperm kinematic parameters were determined using a computer-assisted sperm analysis system. Sperm quality was evaluated by measuring sperm viability, mitochondrial activity, acrosome integrity, and plasma membrane integrity. Statistical analyses were performed using a general linear mixed model (MIXED) in SAS. Factors in the statistical model were experimental groups, time after thawing, and interaction between experimental groups and time after thawing. Total and progressive motilities were greater in semen supplemented with 0.04% phosphorus and vitamin B12 compared with those in the control (p < 0.05). At 15 min post-thawing, VCL, VAP, and HPA in the 0.04% phosphorus and vitamin B12 supplementation group was greater than that in the control (p < 0.05). Phosphorus and vitamin B12 supplementation did not affect sperm kinematics at 0 and 30 min after thawing (p > 0.05). All the sperm parameters that were tested for the 0.04% phosphorus and vitamin B12 supplementation group in modified BPSE were the highest at all the timepoints after thawing. Thus, supplementing frozen semen extender with 0.04% phosphorus and vitamin B12 increased sperm motility, sperm kinematic parameters, and sperm quality.
摘要：
精液冷冻保存是保存许多物种遗传物质的重要技术。然而,冷冻精液极易受到精子DNA损伤和运动能力降低的影响，导致生育率下降。冷冻保存的标准方法和几种方法尚未阐明。本研究旨在确定添加磷和维生素B12组合的公鸡精液补充剂对冷冻保存的精液质量的影响。每周通过背腹部按摩从57只1-3岁的缅甸×越南杂交泰国本地公鸡中收集精液。总的来说,收集了139份精液样本，池化,稀释到每剂量2亿个精子。将合并的样品分为六个实验组：对照组（0.00％），用改良的Beltville家禽精液增量剂（BPSE）稀释，五个处理组，用改良的BPSE稀释，并补充了浓度为0.02、0.04、0.06、0.08和0.10％的磷和维生素B12，分别。将精液样品冷冻并在解冻后0、15和30分钟进行评估。使用计算机辅助精子分析系统确定精子运动学参数。通过测量精子活力来评估精子质量，线粒体活性，顶体完整性,和质膜完整性。使用SAS中的一般线性混合模型(MIXED)进行统计分析。统计模型中的因素是实验组，解冻后的时间，以及实验组之间的相互作用和解冻后的时间。与对照组相比，补充0.04％磷和维生素B12的精液的总运动和进行性运动更大（p<0.05）。解冻后15分钟，VCL,VAP,0.04%磷和维生素B12补充组的HPA高于对照组(p<0.05)。在解冻后0和30min时，补充磷和维生素B12对精子运动学没有影响（p>0.05）。改良BPSE中0.04％磷和维生素B12补充剂组测试的所有精子参数在解冻后的所有时间点都是最高的。因此，补充冷冻精液补充剂0.04%磷和维生素B12可增加精子活力，精子运动学参数，和精子质量。
