Immunohistochemical Evidence for the Localization of Bovine Viral Diarrhea Virus, a Single-stranded RNA Virus, in Ovarian Oocytes in the Cow

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Abstract
Bovine viral diarrhea virus (BVDV) is a single-stranded RNA virus responsible for enteric disease and reproductive failure in cattle. The virus can pass vertically from cow to fetus, causing abortion, birth of malformed calves, and calves born with persistent and life-long infections. In this study, we investigated the tropism of BVDV in ovarian tissue from persistently infected animals. Three heifers persistently infected with BVDV were euthanatized and their ovaries were recovered. A specimen of each ovary was taken ( n = 6) for virus isolation, and the remaining ovarian tissue was stored at −70 C. Cryosections (6 μm) cut from each ovary were analyzed for the presence of BVDV antigens by indirect immunofluorescence. The immunofluorescent analysis employed two monoclonal antibodies, WB103 and WB162, previously raised against the nonstructural protein NS3 and the envelop glycoprotein E2, respectively. High titers (6.97 ± 0.17 log10 tissue culture infective dose50/ml) of BVDV were recovered from 6/6 ovarian samples; NS3 and E2 were widely distributed within the ovarian stroma, the cumulus cell population, and the oocytes maturing in primordial, primary, and secondary follicles. Overall, 362/1,939 (18.7%) of the oocytes contained BVDV antigens, and there was no significant ( P > 0.05) difference in the proportion of BVDV-infected oocytes recorded within the primordial (227/1,247, 18.2%), primary (122/630, 19.4%), and secondary (13/62, 21.0%) follicle populations. Although the developmental potential of the infected oocytes could not be established in the present study, we conclude that bovine oocyte and the cumulus cells are susceptible to BVDV infection.