Detection of bovine herpesvirus 1 in the semen of experimentally infected bulls by dot-blot hybridisation, polymerase chain reaction and virus isolation

Two 18-month-old bovine herpesvirus 1 (BHV1)-seronegative bulls were inoculated experimentally with BHV1 via their prepuces. Semen collected at intervals was examined by optimised virus isolation, dot-blot hybridisation and the polymerase chain reaction (PCR) for detection of BHV1, and the infection was monitored serologically by using a virus neutralisation test. Antibodies were first detected 10 days after inoculation and were still present 40 days after inoculation. Semen collected from four to 40 days after inoculation was positive by PCR with Southern blot hybridisation whereas only the semen collected on day 4 was positive by dot-blot hybridisation, virus isolation and PCR with ethidium bromide staining. These results indicate that the bulls started to shed the virus in semen before they developed any detectable antibody. PCR with Southern blot hybridisation was the most sensitive of the three methods and detected virus for the longest period.