Howard T H, Bowen R A, Pickett B W
Prog Clin Biol Res. 1985;178:127-34.
The efficacy of inoculation of Vero cell cultures or intravenous inoculation of chicken embryos in the isolation and titration of seminal bluetongue virus (BTV) was studied, as was the toxicity of bull semen for these 2 isolation systems. Frozen and thawed BTV-contaminated ejaculates collected during periods of viremia from 2 bulls experimentally infected with cell culture-adapted BTV serotype 17 were used in isolation, titration and fractionation studies. Blood collected from the 2 bulls concurrently with the semen was titrated in chicken embryos. Bull semen was toxic for both isolation systems. Toxicity was associated with both the spermatozoa and seminal plasma. Dilution of the semen at least 1:25, addition of peptone or tryptose broth to the diluent, limitation of adsorption time and postinoculation washing of cell culture monolayers all reduced the destructive effects of semen. Isolation of BTV was successful from 11 ejaculates and was titratable in 9 of these. Blind passage of surviving embryos or cell cultures at the endpoints of the titrations produced BTV isolations in 4 instances. The virus was never isolated from semen in the absence of concurrent viremia. Peak seminal BTV titers of 10(5.5) CEIVLD50/ml and 10(5.7) TCID50/ml were observed.(ABSTRACT TRUNCATED AT 250 WORDS)
研究了接种Vero细胞培养物或静脉接种鸡胚在精液蓝舌病毒（BTV）分离和滴定中的效果，以及公牛精液对这两种分离系统的毒性。从2头经细胞培养适应的BTV 17型实验感染公牛的病毒血症期采集的冻融BTV污染射精物用于分离、滴定和分级研究。与精液同时采集的2头公牛的血液在鸡胚中进行滴定。公牛精液对两种分离系统均有毒性。毒性与精子和精浆有关。精液至少1:25稀释、在稀释剂中添加蛋白胨或胰蛋白胨肉汤、限制吸附时间以及接种后对细胞培养单层进行洗涤，均降低了精液的破坏作用。从11份射精物中成功分离出BTV，其中9份可滴定。在滴定终点对存活胚胎或细胞培养物进行盲传，有4次获得了BTV分离株。在无并发病毒血症的情况下，从未从精液中分离出病毒。观察到精液中BTV的峰值滴度为10(5.5) CEIVLD50/ml和10(5.7) TCID50/ml。（摘要截短于250字）
