Galik Patricia K, Givens M Daniel, Stringfellow David A, Crichton Elizabeth G, Bishop Michael D, Eilertsen Ken J
College of Veterinary Medicine, Auburn University, AL 36849, USA.
Theriogenology. 2002 Mar 1;57(4):1219-27. doi: 10.1016/s0093-691x(02)00633-7.
Bovine viral diarrhea virus (BVDV) can be found in cells and fluids from ovaries collected at the abattoir. On the other hand, immunoglobulins are also found in the fluid of ovarian follicles. Anti-BVDV antibodies in follicular fluid might reduce cross-contamination of COCs at the time of collection or hinder the use of virus isolation to test for the presence of virus. One objective of this study was to determine the frequency with which BVDV could be found in pooled follicular fluid collected during the periodic aspiration of COCs from abattoir-origin ovaries. A second objective was to determine the prevalence and neutralizing activity of anti-BVDV antibodies in these blended samples. We collected samples of pooled follicular fluid (n = 55) over a 20-month period as part of our routine oocyte collection activities. We assayed each sample for BVDV using virus isolation as well as reverse transcription nested polymerase chain reaction (RT-nPCR) procedures. We also tested follicular fluid for antibody that would neutralize four representative strains of BVDV (SD-1, a genotype 1a strain; NY-1, a genotype lb strain; CD-87, a genotype 2 strain, and PA-131, a divergent genotype 2 strain). We detected no BVDV by virus isolation, but we did identify the virus by RT-nPCR in one of the 55 samples of follicular fluid. Automated dye terminator nucleotide sequencing of the amplified portion of the viral genome indicated a genotype 1 strain that was distinct from any of our laboratory strains. In addition, each of the samples of follicular fluid contained sufficient antibody to neutralize large quantities of each of the four laboratory strains that were used. Finding BVDV in just 1 of 55 samples was consistent with reports of similar studies in which the occurrence of BVDV in abattoir-origin materials ranged from 0.9 to 12%. We presumed that failure to isolate the virus was due to neutralizing antibody in the sample. Thus, the incidence of BVDV contamination of our IVF system at the level of pooling of follicular fluid was low for the 20-month period. The presence of anti-BVDV antibody in pooled follicular fluid provided a coincidental means of neutralizing BVDV when it was introduced in fluid aspirated from infected ovaries.
在屠宰场收集的卵巢细胞和液体中可发现牛病毒性腹泻病毒（BVDV）。另一方面，在卵泡液中也能发现免疫球蛋白。卵泡液中的抗BVDV抗体可能会在采集时减少卵母细胞- cumulus细胞复合体（COCs）的交叉污染，或阻碍使用病毒分离法检测病毒的存在。本研究的一个目的是确定在从屠宰场来源的卵巢定期抽吸COCs过程中收集的混合卵泡液中发现BVDV的频率。第二个目的是确定这些混合样本中抗BVDV抗体的流行率和中和活性。在为期20个月的时间里，作为我们常规卵母细胞采集活动的一部分，我们收集了混合卵泡液样本（n = 55）。我们使用病毒分离以及逆转录巢式聚合酶链反应（RT-nPCR）程序对每个样本进行BVDV检测。我们还检测卵泡液中是否存在能中和四种代表性BVDV毒株（SD-1，1a基因型毒株；NY-1，1b基因型毒株；CD-87，2基因型毒株；以及PA-131，一种不同的2基因型毒株）的抗体。通过病毒分离未检测到BVDV，但在55份卵泡液样本中的1份样本中，我们通过RT-nPCR鉴定出了该病毒。对病毒基因组扩增部分进行的自动染料终止子核苷酸测序表明是一种1基因型毒株，与我们实验室的任何毒株都不同。此外，每份卵泡液样本都含有足够的抗体来中和所使用的四种实验室毒株中的每一种。仅在55份样本中的1份样本中发现BVDV，这与类似研究的报告一致，在这些研究中，屠宰场来源材料中BVDV的发生率在0.9%至12%之间。我们推测未能分离出病毒是由于样本中的中和抗体。因此，在为期20个月的时间里，在卵泡液混合水平上，我们的体外受精（IVF）系统中BVDV污染的发生率较低。当从受感染的卵巢抽吸的液体中引入BVDV时，混合卵泡液中抗BVDV抗体的存在提供了一种偶然的中和BVDV的方式。
