The pestiviruses are classified into four established species: Bovine viral diarrhea virus 1 (BVDV-1) and BVDV-2 of cattle, Border disease virus (BDV) of sheep and goats, and Classical swine fever virus (CSFV) of swine, and a tentative pestivirus species of giraffe in the genus Pestivirus of the family Flaviviridae (8). Although these viral species are identified traditionally according to the animal host species from which they have been isolated, there is an extensive antigenic cross reactivity among them, and they can cross the host species barrier and infect different species of cloven-hoofed animals (7,22,23,27). Serological data indicate that the host range of pestiviruses includes most even-toed ungulates belonging to the order Artiodactyla. Pestiviruses have been isolated not only from domestic animals but also from wildlife. It is reasonable to suppose that many species of wild ungulates harbor pestiviruses (1) because interspecies transmission is easily achieved and it is prudent to believe that it will occur readily in domestic and free-living wild ruminants when permitted to do so by new husbandry practices or changes in population dynamics. Taxonomic study of isolates from wild ungulates will provide useful information on the ecology and evolution of pestiviruses. In the present paper we first describe pestivirus infection in the free-living Japanese serow, Capricornis crispus (synonym for Nemorhaedus crispus), of the family Bovidae.Serum samples collected from 16 free-living Japanese serows during 4 years from 2001 to 2004 in Morioka were examined for the presence of pestivirus by the reverse transcription (RT)-nested PCR procedure described in 1993 (15). Ribonucleic acids were isolated from serum samples by the single-step guanidinium isothiocyanate-phenol-chloroform method (2) by using the RNAzol B extraction kit (Biotecx Laboratories, Inc., Houston, Tex., U.S.A.). Briefly, 200 µl of the serums were mixed vigorously with 800 µl of a solution containing guanidinium isothiocyanate-phenol in a 1.5-ml Eppendorf tube. One hundred microliters of chloroform were added and the tube was vigorously shaken for 30 sec by hand. The tube was chilled on ice for 5
Evidence for Pestivirus Infection in
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