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Immunoaffinity-purified bovine respiratory syncytial virus (brsv) fusion (F) protein elicited anti-brsv-specific antibody responses in brsv-seronegative calves. After primary vaccination, all calves seroconverted to brsv as determined by the virus neutralization (vn) test and developed anti-F protein antibodies detectable by protein immunoblot analyses. Subsequent vaccinations induced > twofold increase in vn titer in 3 of 9 (33%) calves, and 1 calf became vn-negative, but still had nonneutralizing antibody detectable by protein immunoblot analysis. This calf remained seronegative after challenge exposure.

Two groups of calves were vaccinated im with immunoaffinity-purified brsv F protein. Each dose was 2 ml containing 20 μg of purified F protein. Freund's adjuvants were used for all vaccinations, with Freund's complete adjuvant used for the primary vaccination and Freund's incomplete adjuvant for subsequent vaccinations. The vaccine was administered to both groups at weeks 0 and 3; the first group received a third vaccination at week 21. Group-1 and -2 vaccinated calves and non-vaccinated contact controls were intranasally aerosol challenge-exposed with low cell culture-passage brsv on weeks 22 and 9, respectively.

Eight of 9 vaccinated calves did not develop a humoral anamnestic response following challenge exposure, as demonstrated by vn test and protein immunoblot analyses. Calf 14 from group 1 which had a 1:2 vn antibody titer prior to vaccination, was the only calf that developed an anamnestic response. This suggests that vaccine-induced antibodies interfered with the immune response or that the challenge virus (and the virus that calf 14 was infected with before challenge exposure) contained different F protein epitopes, compared with the purified F protein immunogen.

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in American Journal of Veterinary Research


Objective—To examine the role of bovine viral diarrhea virus (BVDV) biotype on the establishment of fetal infection in cattle.

Animals—30 mixed-breed pregnant cows.

Procedure—Pregnant cows were inoculated oronasally with either i-VVNADL, originating from an infectious BVDV cDNA clone of the National Animal Disease Laboratory (NADL) isolate, or the parental virus stock, termed NADL-A.

Results—All cows developed neutralizing antibodies to BVDV, and virus was commonly isolated from peripheral blood mononuclear cells or nasal swab specimens of NADL-A inoculated cows; however, virus was rarely isolated from specimens of i-VVNADL inoculated cows. i-VVNADL did not cause fetal infection, whereas all fetuses harvested from NADL-A inoculated cows at 6 weeks after inoculation had evidence of infection. Immunoblot analysis of fetal virus isolates revealed the absence of NS3, confirming a noncytopathic (NCP) biotype BVDV in the NADL-A stock. The sequence of the NCP contaminant (termed NADL-1102) and the i-VVNADL genome were virtually identical, with the exception of a 270 nucleotide-long insert in the i-VVNADL genome. Phylogenetic analyses revealed that NADL-1102 forms a monophyletic group with 6 other NADL genomes.

Conclusions and Clinical Relevance—These data suggest that the contaminating NCP virus in the NADL-A stock was the ancestral NADL virus, which originally infected a bovine fetus and recombined to produce a cytopathic (CP) variant. Following oronasal infection of pregnant cows, viremia and transplacental transmission of CP BVDV to the fetus is rare, compared with the high occurrence of maternal viremia and fetal infection observed with NCP BVDV. (Am J Vet Res 2002;63:1455–1463)

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in American Journal of Veterinary Research