The Cooper isolate of bovine herpesvirus 1 (BHV1) was used to produce a thymidine kinase-negative (TK-) recombinant by insertion of a beta-galactosidase (bgal) expression cassette into the TK coding region. The recombinant virus (TK- bgal+) was tested for abortifacient activity in cattle by inoculation of 5 pregnant heifers at 25 to 29 weeks gestation. Five additional heifers were inoculated with the Cooper TK-positive (TK+) virus to serve as controls. After inoculation, both groups of heifers developed similar febrile responses and neutralizing antibody titers. Virus was isolated from blood of all heifers during the first postinoculation (PI) week, and isolation frequencies were similar for both groups. In contrast, whereas virus was isolated from many of the nasal and vaginal swab specimens of heifers inoculated with TK+ virus, only rare virus isolations were made from the heifers given TK- bgal+ virus. All heifers inoculated with TK+ virus aborted between PI days 19 and 35. The finding of characteristic microscopic lesions and viral antigen in fetal tissues indicated that the abortions were caused by BHV-1 infection. Virus was isolated from 3 fetuses, and all isolates were TK+. Two heifers inoculated with TK- bgal+ virus aborted at PI days 25 and 39. Fetal tissues had typical BHV-1 microscopic lesions and viral antigen. Virus was isolated from blood of both fetuses, and the isolates were TK- bgal+. Results of this study indicate that inactivation of the TK gene reduces, but does not eliminate, the abortifacient activity of BHV-1.

Bovine immunodeficiency virus (BIV), a lentivirus, is prevalent in dairy and beef cattle in southeastern United States and may be associated with a lymphoproliferative disease. The mode(s) of BIV transmission are undefined. Because artificial insemination is a common practice in dairy production, contaminated stud semen could serve as an important source of infection if the virus is harbored in seminal fluids. To evaluate this possibility, we procured 11 cryopreserved semen specimens from a stud semen repository. Leukocytes were purified from the specimens, and the leukocyte DNA was used as template in a polymerase chain reaction procedure that targeted a 235-base pair, highly conserved domain of the BIV pol gene. The target sequence was amplified from the seminal leukocyte DNA of 9 of the specimens (82%), and nucleotide sequencing confirmed the BIV specificity of the fragment. This finding provides evidence that stud bull semen may serve as an important reservoir of BIV, suggesting the possibility that artificial insemination of dairy cows may have a major role in transmission and wide-spread dissemination of this bovine lentivirus.