ITRA-ULE, INDEGSAL, University of León, Spain.
Anim Reprod Sci. 2012 Nov;135(1-4):113-21. doi: 10.1016/j.anireprosci.2012.09.009. Epub 2012 Sep 27.
Cryopreservation of brown bear (Ursus arctos) semen requires centrifugation to increase concentration and/or remove urine contamination. However, a percentage of the spermatozoa are lost in the process. This percentage varies considerably between males and ejaculates, and we have studied the effect of sperm quality and seminal plasma characteristics on the spermatozoa recovery rate after centrifugation. One hundred and thirty one sperm samples obtained from fifteen brown bear males by electroejaculation under general anaesthesia were used. The ejaculates were centrifuged 600 × g for 6 min. Motility was assessed by CASA, and acrosomal status (PNA-FITC) and viability (SYBR-14/propidium iodide) were determined by flow cytometry. Seminal plasma characteristics (albumin, alkaline phosphatase, calcium, cholesterol, creatine, glucose, glutamic oxaloacetic transaminase (GOT), lactate, lipase, magnesium, phosphate and total protein) were determined by a biochemical and gas analysis. Total motility (r = 0.26; P=0.005) and cell viability (r = 0.20; P = 0.033) were positively correlated with the percentage of recovered spermatozoa. Sperm recovery was correlated with the concentration of several components of seminal plasma: negatively with glucose concentration (r = -0.47; P = 0.005) and positively with the enzymes GOT (r = 0.36; P = 0.040) and lactate dehydrogenase (r = 0.36; P = 0.041). After sorting the data into classes according to sperm recovery (Low: 0-39, Medium: 40-69, High: 70-100), we observed that the samples with a lower recovery rate derived from ejaculates with lower values for TM, VAP and viability (P<0.05). Multiple regression analysis rendered two models to define the post-centrifugation spermatozoa recovery which included total motility and damaged acrosome or glucose, GOT and lactate dehydrogenase. We discuss these relationships and their implications in the electroejaculation procedure and the handling of the sample during centrifugation.
棕熊(Ursus arctos)精液的冷冻保存需要离心以增加浓度和/或去除尿液污染。然而,在此过程中会有一定比例的精子丢失。这种损失在不同的雄性和精液之间差异很大,我们研究了精子质量和精浆特性对离心后精子回收率的影响。我们使用了 131 个来自 15 只棕熊雄性电刺激射精获得的精子样本。将精液在 600×g 下离心 6 分钟。通过 CASA 评估精子活力,通过流式细胞术评估顶体状态(PNA-FITC)和活力(SYBR-14/碘化丙啶)。通过生化和气体分析测定精浆特性(白蛋白、碱性磷酸酶、钙、胆固醇、肌酸、葡萄糖、谷草转氨酶(GOT)、乳酸、脂肪酶、镁、磷酸盐和总蛋白)。总活力(r=0.26;P=0.005)和细胞活力(r=0.20;P=0.033)与回收精子的百分比呈正相关。精子回收率与精浆中几种成分的浓度相关:与葡萄糖浓度呈负相关(r=-0.47;P=0.005),与 GOT(r=0.36;P=0.040)和乳酸脱氢酶(r=0.36;P=0.041)呈正相关。根据精子回收率(低:0-39、中:40-69、高:70-100)对数据进行分类后,我们观察到回收率较低的样本来源于总活力、曲线速度和活力较低的精液(P<0.05)。多元回归分析得出了两个模型来定义离心后精子回收率,包括总活力和顶体损伤或葡萄糖、GOT 和乳酸脱氢酶。我们讨论了这些关系及其在电刺激射精过程和离心过程中样本处理中的意义。