Molecular epidemiology of bluetongue virus serotype 4 isolated in the Mediterranean Basin between 1979 and 2004

The nucleotide sequences of genome segments 2, 7, 8, 9 and 10, coding for viral proteins (VP) and non-structural proteins (NS)--VP2, VP7, NS2, VP6 and NS3/NS3A, respectively, were determined and compared for 10 strains of bluetongue virus (BTV) serotype 4 isolated in the Mediterranean Basin between 1979 and 2004, and the South African attenuated BTV 4 vaccine strain. The sequence data generated for the BTV 4 strains isolated in Greece in 1979, 1999 and 2000 showed that they had a common origin but were distinct from the lineage of the BTV 4 strains isolated from 2003 onward in the western Mediterranean Basin (Italy, Morocco, Spain and Corsica). The nucleotide and deduced amino acid (aa) sequences of the BTV 4 strains within each lineage were identical to each other, irrespective of the year of isolation or the geographical location. Although the sequence of VP2 from the Turkish and Greek strains were highly similar, there were sufficient differences in the VP6, VP7 and NS2 proteins to suggest that the Turkish BTV 4 belongs to a third lineage. Alignment of the NS3 sequences from the attenuated BTV 4 vaccine strain and the field strains showed 13 aa substitutions, which may, either singularly or together, be responsible for attenuation and hence determining the virulence of the virus.