Thun Rico, Hurtado Maria, Janett F
Department of Reproduction, University of Zürich, Switzerland.
Theriogenology. 2002 Feb;57(3):1087-94. doi: 10.1016/s0093-691x(01)00704-x.
For optimizing routine freezing of bull semen, we examined three different cryopreservation methods using either TRIS-egg yolk-citrate extender or Biociphos-Plus. Biociphos-Plus (IMV, France) has been marketed as an extender, in which egg yolk is replaced by a sterile soybean extract to reduce the contamination risk derived from animal borne substances. We used 78 bulls of various breeds (Brown Swiss, Holstein, Simmental) between 12 and 23 months of age, and we produced a total of 800-1000 straws (0.25 ml, 20 x 10(6) spermatozoa) from each bull using three different methods. In method A, we used TRIS-egg yolk as extender and packaged at 4 degrees C. In method B, we also used TRIS-egg yolk but packaged at room temperature (RT) between 18 and 22 degrees C. In method C, Biociphos-Plus served as extender and we packaged at RT. We compared methods A, B and C by using post-thaw motility, viability, morphology and osmotic resistance as semen quality parameters. In addition, we recorded 75-day nonreturn rates (NR75) to detect the effect of extenders on fertility. With the exception of primary defects, all laboratory parameters investigated were significantly (P < 0.05) better in methods A (TRIS-egg yolk, 4 degrees C) and B (TRIS-egg yolk, RT), compared to method C (Biociphos-Plus, RT). We recorded no significant difference between methods A and B. We could not verify the differing laboratory results by fertility data (NR75). However, when we analyzed NR75 for a single breed, significant (P < 0.05) differences existed between methods A and B compared to method C in Simmental and Holstein but not in Brown Swiss. We obtained best results in Simmental using method A (69%, n = 3384), while method C (61.4%, n = 763) was superior to methods A (57.6%, n = 698) and B (57.3%, n = 737) in Holstein. After considering various factors like preparation of extender, cost of materials and ambient working temperature, we concluded from our data that bull semen processing using TRIS-egg yolk extender and RT for packaging (method B) produced the best semen quality and field fertility.
为优化公牛精液的常规冷冻,我们使用TRIS - 蛋黄 - 柠檬酸盐稀释液或Biociphos - Plus研究了三种不同的冷冻保存方法。Biociphos - Plus(法国IMV公司)作为一种稀释液进行销售,其中蛋黄被无菌大豆提取物替代,以降低动物源性物质带来的污染风险。我们使用了78头12至23月龄的不同品种公牛(瑞士褐牛、荷斯坦牛、西门塔尔牛),并使用三种不同方法从每头公牛采集总共800 - 1000支细管精液(0.25毫升,含20×10⁶个精子)。方法A中,我们使用TRIS - 蛋黄作为稀释液,并在4℃下进行包装。方法B中,我们同样使用TRIS - 蛋黄,但在18至22℃的室温(RT)下进行包装。方法C中,Biociphos - Plus作为稀释液,并在室温下进行包装。我们以解冻后活力、生存力、形态和抗渗透性作为精液质量参数,比较了方法A、B和C。此外,我们记录了75天不返情率(NR75)以检测稀释液对生育力的影响。除了初级缺陷外,与方法C(Biociphos - Plus,室温)相比,方法A(TRIS - 蛋黄,4℃)和方法B(TRIS - 蛋黄,室温)中所有研究的实验室参数均显著更好(P < 0.05)。我们记录到方法A和方法B之间无显著差异。我们无法通过生育力数据(NR75)验证不同的实验室结果。然而,当我们分析单一品种的NR75时,西门塔尔牛和荷斯坦牛中,方法A和方法B与方法C相比存在显著(P < 0.05)差异,但瑞士褐牛中没有。西门塔尔牛使用方法A获得了最佳结果(69%,n = 3384),而在荷斯坦牛中,方法C(61.4%,n = 763)优于方法A(57.6%,n = 698)和方法B(57.3%,n = 737)。在考虑了稀释液制备、材料成本和环境工作温度等各种因素后,我们从数据中得出结论,使用TRIS - 蛋黄稀释液并在室温下进行包装的公牛精液处理方法(方法B)产生了最佳的精液质量和田间生育力。