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Different strains of noncytopathic bovine viral diarrhea virus (BVDV) vary in their affinity for in vivo-derived bovine embryos

机译:不同菌株的非细胞病性牛病毒性腹泻病毒(BVDV)对体内来源的牛胚胎的亲和力有所不同

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Washing procedures (without trypsin treatment) recommended by the International Embryo Transfer Society (IETS) for use on in vivo-derived embryos effectively removed a cytopathic strain (NADL) of bovine viral diarrhea virus (BVDV) after artificial exposure. However, these washing procedures have not been evaluated using other isolates of BVDV, including representative non-cytopathic strains. Thus, the objective of this study was to evaluate the efficacy of the IETS procedures following artificial exposure of in vivo-derived bovine embryos to two different strains and biotypes of BVDV. One hundred and twenty-nine zona pellucida-intact (ZP-I) morulae and blastocysts (MB) and 56 non-fertile and degenerated (NFD) ova were collected 7 days following exposure to bulls from 32, BVDV-negative, superovulated cows. After collection, all MB and NFD ova were washed according to IETS standards. Subsequently, half of the MB and NFD ova were exposed for 1 h to approximately 10(6)-cell culture infective doses (50% endpoint) per milliliter of viral strain SD-1, and the other half were exposed to the same concentration of CD-87. After exposure, groups of greater than or equal to 3 and less than or equal to 10 MB or NFD ova were washed using methods that met or exceeded IETS standards. Then, the washed groups were sonicated, and sonicate fluids were assayed for presence of virus using virus isolation and a reverse transcription nested polymerase chain reaction. No virus was detected in any group of NIB or NFD ova that had been exposed to the CD-87 isolate. However, virus was detected in association with 50% of the groups of NIB and 33% of the groups of NFD ova that had been exposed to the SD-1 isolate. Therefore, standard embryo-washing procedures recommended by the IETS are more effective for removal of some isolates of BVDV than for others. It remains to be determined if the quantity of a high-affinity isolate of BVDV associated with individual washed embryos would infect recipients via the intrauterine route. Further, it should be determined if an alternative embryo processing procedure, washing and trypsin treatment, would be more effective for removal of high-affinity isolates.
机译:由国际胚胎移植协会(IETS)推荐的用于体内衍生胚胎的洗涤程序(未经胰蛋白酶处理)可在人工暴露后有效去除牛病毒性腹泻病毒(BVDV)的细胞病变株(NADL)。但是,尚未使用其他BVDV分离株(包括代表性非细胞毒株)评估这些洗涤程序。因此,本研究的目的是评估将体内来源的牛胚胎人工暴露于两种不同菌株和BVDV生物型后的IETS程序的功效。暴露于32头BVDV阴性,超排卵母牛的公牛暴露7天后,收集了129个完整透明带(ZP-1)桑ula和胚泡(MB)和56个非可育和变性(NFD)卵。收集后,根据IETS标准洗涤所有MB和NFD卵。随后,MB和NFD卵子的一半暴露于每毫升病毒菌株SD-1约10(6)个细胞培养感染剂量(终点为50%)1小时,另一半暴露于相同浓度的HBV。 CD-87。暴露后,使用达到或超过IETS标准的方法洗涤大于或等于3且小于或等于10 MB的组或NFD卵。然后,对清洗的组进行超声处理,并使用病毒分离和逆转录嵌套式聚合酶链反应分析超声处理液中病毒的存在。在已暴露于CD-87分离物的NIB或NFD卵的任何组中均未检测到病毒。但是,已检测到与暴露于SD-1分离物的NIB组的50%和NFD卵组的33%相关的病毒。因此,IETS推荐的标准胚胎清洗程序对于去除某些BVDV分离株比对其他分离株更有效。与单个清洗过的胚胎相关的BVDV高亲和力分离株的数量是否会通过子宫内途径感染受体还有待确定。此外,应该确定替代性的胚胎加工程序(洗涤和胰蛋白酶处理)对于去除高亲和力分离株是否更有效。

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