Isolation of Bluetongue Virus Serotype 1 from Culicoides vector Captured in Livestock Farms and Sequence Analysis of the Viral Genome Segment-2

• Published in: Transboundary and emerging diseases Vol. 59; no. 4; pp. 361 - 368
• Main Authors: Dadawala, A. I., Biswas, S. K., Rehman, W., Chand, K., De, A., Mathapati, B. S., Kumar, P., Chauhan, H. C., Chandel, B. S., Mondal, B.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.08.2012; Hindawi Limited
• Subjects: Agriculture; Animals; Bluetongue virus; Bluetongue virus - classification; Bluetongue virus - isolation & purification; bluetongue virus serotype-1; Cell Line; Ceratopogonidae - virology; Cricetinae; Culicoides; Culicoides oxystoma; Farms; Genetics; Genome, Viral; India; Insects; Livestock; PAGE; Phylogeny; RNA; RNA, Viral - genetics; Serotyping; Viruses; VP2 gene; India; Gujarat India
• ISSN: 1865-1674; 1865-1682
• DOI: 10.1111/j.1865-1682.2011.01279.x

Summary Bluetongue virus serotype‐1 (BTV‐1) was isolated from Culicoides oxystoma vectors captured on livestock farms in two places of Gujarat, India. The viruses were isolated on BHK‐21 cells, which produced characteristic BTV‐related cytopathic effects between 24 and 48 h post‐infection. Virus antigen was demonstrated in infected cells at different passage by a BTV‐specific sandwich ELISA. Further, polyacrylamide gel electrophoresis and silver staining of viral genomic RNA revealed ten double‐stranded RNA segments characteristic of BTV. Serotype of the isolates was identified by virus neutralization and PCR coupled with sequencing. The isolates were designated as SKN‐7 and SKN‐8 and their genome segment‐2 (VP2) were sequenced. Phylogenetic analyses revealed very close relationship between them although they are not identical. SKN‐8 showed closer relationship with a recently isolated BTV‐1 from goat. Bluetongue virus was earlier isolated from Culicoides in adjacent state more than 20 years ago, although the serotype of the virus was not determined.