An experimental inactivated virus vaccine against bovine ephemeral fever 1. Studies of the virus

Abstract

Studies were carried out to find methods for obtaining optimum yields of bovine ephemeral fever (BEF) virus and for concentrating the virus in order to develop inactivated virus vaccines. Cells from the SVP cell line, which was derived from the pig kidney PS cell line, were most satisfactory for growing and assaying BEF virus. BHK 21 and Vero cells also gave similar yields of virus but were not as useful for virus assay. A plaque assay in SVP cells, in which there was 0.1 μg of actinomycin D per ml of overlay, produced reproducible clear plaques and was slightly more sensitive than assays in BHK 21 cell roller tubes. High multiplicities of infection (MOI), around 1 PFU/cell, produced low yields of infectious virus, whereas decreasing the MOI approximately 100-fold led to an increase in virus yield of up to four logs. BEF virus could be concentrated using zinc acetate or ammonium sulphate but not polyethylene glycol 6000. Ammonium sulphate proved most suitable and produced an easily handled precipitate with up to 100% recovery of virus infectively, and 100-fold concentration was possible. This concentrated virus could be rapidly desalted by gel filtration through Sephadex G-75. The virus could be further purified by sucrose density gradient ultracentrifugation provided the gradient contained a protein stabilizer of 0.1% bovine serum albumin. Inactivation kinetics with 0.025% β-propiolactone was similar to that reported for other rhabdoviruses.