University of São Paulo State, UNESP, Laboratory of Animal Reproduction, College of Veterinary Medicine, São Paulo State, 16050-680, Brazil.
Anim Reprod Sci. 2011 Mar;124(1-2):55-60. doi: 10.1016/j.anireprosci.2011.01.004. Epub 2011 Feb 12.
The influence of Bovine Herpesvirus type 5 (BoHV-5) infection on semen variables and sperm morphology collected from healthy bulls with no reproductive disorder was evaluated in ten ejaculates distributed into two experimental groups: group I, bull semen exposed to 10(2.3) (tissue culture infectious dose) TCID(50)/50 μl of a Brazilian strain of BoHV-5 (US9/BR/2007; GU9457818) and group II, unexposed bull control semen. After experimental infection, the semen was frozen-thawed prior to computerized analysis (CASA) of sperm motility and movement. Also analyzed were sperm phosphatidylserine transposition, acrosomal integrity, mitochondrial function, plasma membrane integrity and Annexin V expression. Viable BoHV-5 particles and their DNA were detected in infected semen after virus isolation and in situ hybridization (ISH) assay. The ISH revealed the BoHV-5 US9 gene in the acrosome and tail of infected spermatozoa. The only remarkable differences between groups I and II were the sperm kinetic variables, whereby infected sperm had a lesser mean velocity (VAP) and curvilinear velocity (VCL) values as compared to controls (P≤0.05). However, the straightness coefficient (STR) and beat cross frequency (BCF) values were higher in infected sperm. These results indicate that BoHV-5 can be found in infected sperm but induces no functional and morphological damage even after freeze-thawing, and, importantly, BoHV-5 can be spread via in vitro and in vivo reproductive biotechnology procedures.
评估了无生殖障碍健康公牛的精液变量和精子形态，这些公牛的精液分别来自于两个实验组的 10 个射精：实验组 I，暴露于 10（2.3）（组织培养感染剂量）TCID（50）/50μl 的巴西 BoHV-5 株（US9/BR/2007；GU9457818）的公牛精液；实验组 II，未暴露的公牛对照精液。实验感染后，对精液进行冷冻-解冻处理，然后进行计算机分析（CASA）精子活力和运动。还分析了精子磷脂酰丝氨酸易位、顶体完整性、线粒体功能、质膜完整性和 Annexin V 表达。通过病毒分离和原位杂交（ISH）检测到感染精液中存在有感染性的 BoHV-5 颗粒及其 DNA。ISH 显示 BoHV-5 US9 基因存在于感染精子的顶体和尾部。组 I 和组 II 之间唯一明显的差异是精子动力学变量，即感染精子的平均速度（VAP）和曲线速度（VCL）值低于对照组（P≤0.05）。然而，感染精子的直线性系数（STR）和鞭打频率（BCF）值较高。这些结果表明，BoHV-5 可以在感染的精子中找到，但即使在冷冻-解冻后也不会引起功能和形态损伤，而且重要的是，BoHV-5 可以通过体外和体内生殖生物技术程序传播。
