National Veterinary Institute, SVA, Uppsala, Sweden.
Anim Reprod Sci. 2011 Jun;126(1-2):108-14. doi: 10.1016/j.anireprosci.2011.04.014. Epub 2011 Apr 29.
The virus porcine circovirus type 2 (PCV2) is associated with different disease entities, including reproductive failure. The objective of this study was to investigate the use of a semen processing technique for the elimination of infectious PCV2 in semen. PCV2 was chosen as a model virus because of its small size, high resistance to inactivation and as a known risk factor for boar semen contamination. Aliquots of ejaculates were spiked with PCV2 and processed by a double processing technique, consisting of Single Layer Centrifugation on Androcoll™-P followed by a "swim-up" procedure. Samples were collected from the resulting fractions during the selection process and analyzed for the presence of infectious PCV2. Virus titres were determined by performing a 50% tissue culture infective dose assay (TCID(50)) by end point dilution and with the use of an indirect peroxidise monolayer assay technique. With an initial infectious virus titre of 3.25-3.82 (TCID(50))/50μL the two-step sperm selection method eliminated 2.92±0.23 logs of infectious PCV2, corresponding to more than 99% reduction. Sperm quality was not affected by the selection procedure.
猪圆环病毒 2 型(PCV2)与不同的疾病实体有关,包括生殖失败。本研究的目的是研究一种精液处理技术,以消除精液中的传染性 PCV2。选择 PCV2 作为模型病毒,是因为其体积小、抗失活能力强,并且是公猪精液污染的已知危险因素。将精液等分试样与 PCV2 混合,并通过双处理技术进行处理,该技术由在 Androcoll™-P 上进行单层离心,然后进行“游泳”程序。在选择过程中从所得级分中收集样品,并分析存在的传染性 PCV2。通过终点稀释进行 50%组织培养感染剂量测定(TCID(50)),并使用间接过氧化物酶单层测定技术来确定病毒滴度。初始传染性病毒滴度为 3.25-3.82(TCID(50))/50μL,两步精子选择方法消除了 2.92±0.23 个对数的传染性 PCV2,对应于超过 99%的减少。精子质量不受选择过程的影响。