Complement fixation test of Nebraska calf diarrhea virus with calf serum

A complement fixation (CF) test for neonatal calf diarrhea has been developed with Nebraska calf diarrhea virus (NCDV) as antigen. The CF antigenicity of NCDV appeared in BK cell cultures for the first time 3 days after inoculation. Usually, the unconcentrated BK cell culture fluid infected with NCDV possessed poor CF antigenicity. The fluid concentrated by ultracentrifugation and ultrafiltration and with polyethylene glycol 6000 revealed a high-titered CF antigenicity to antiserum against NCDV. The antigen was not sensitive to lipid solvents, but was relatively resistant to heating. It was recognized that the concentrated fluid of cell culture infected with NCDV was available as antigen in the CF test for the serological demonstration of infection with a calf reovirus-like agent. The wide prevalence of diarrhea among Japanese calves with the calf reovirus-like agent was revealed from the results of CF and neutralization tests of NCDV.