Isolation and preliminary characterization of an orbivirus of the Palyam serogroup from biting midge Culicoides oxystoma in Japan

An orbivirus of the Palyam serogroup was isolated from Culicoides oxystoma collected in a cowshed in Kagoshima, Southern Kyushu Island, Japan. This is the first isolation of an orbivirus of the Palyam serogroup in Japan. The virus was a spherical non-enveloped RNA virus, approximately 60 nm in diameter. The virus was resistant to ethyl ether, sodium deoxycholate and freezing-thawing, but readily inactivated by trypsin. The virus was not stabilized by 1 M MgCl2, was labile at pH 3.0 and was not precipitated by protamine sulfate. Indirect immunofluorescent staining of infected Vero cells indicated the virus to be antigenically related to D'Aguilar and Bunyip Creek viruses of the Palyam serogroup. Neutralization tests showed the virus to have no relationship with D'Aguilar virus, but to have a one-way cross-reaction with Bunyip Creek virus. The virus was tentatively designated as Kagoshima virus. A serological survey indicated dissemination of the virus in cattle populations in Kagoshima Prefecture.