African Journal of
Microbiology Research

  • Abbreviation: Afr. J. Microbiol. Res.
  • Language: English
  • ISSN: 1996-0808
  • DOI: 10.5897/AJMR
  • Start Year: 2007
  • Published Articles: 5177

Full Length Research Paper

Development of a cell line from the gill of koi and molecular characterization of KHV isolated in Taiwan

Chien Tu1,2, Chia-Yu Hsieh3, Shao-Kuang Chang2, Je-Fu Su1 and Meei-Mei Chen2*
1Animal Health Research Institute, Council of Agriculture, Executive Yuan, No. 376, Chung-Cheng Road, Tansui, New Taipei City 25158, Taiwan. 2School of Veterinary Medicine, National Taiwan University, No. 1, Sec. 4, Roosevelt Road, Taipei 10617, Taiwan. 3Animal Hospital, National Pingtung University of Science and Technology, Pingtung 912, Taiwan.
Email: [email protected]

  •  Accepted: 05 July 2013
  •  Published: 12 July 2013

Abstract

A new cell line (KoG) was developed from the gill of koi, Cyprinus carpio. KoG cells were composed of epithelial-like cells and fibroblast-like cells. The cells were maintained well at 25°C in Leibovitz L15 medium supplemented with 10% fetal calf serum. Chromosome analysis showed that the KoG cells had 43 diploid chromosomes at passage 40. The KoG cells were used to test the susceptibility of a koi herpesvirus isolate using a Taiwanese koi herpesvirus isolate (strain TW BL98). The KHV-infected KoG cells revealed vacuolation in the cytoplasm and marginal hyperchromation in the nucleus, which was confirmed by an indirect immunofluorescence assay and transmission electron microscopy. A multistep growth curve showed the virus reaching the highest titer of 106.2 TCID50/ml at 5 days post inoculation. Sequence analysis targeting the alleles of the two molecular markers indicated that the Taiwanese isolate displayed the I++IIallele and of the Asian lineage, which was identical to a Japanese strain (Asian genotype variant 1). Collectively, our results demonstrate that the KoG cells may provide a tool for the replication of KHV andthe Taiwanese isolate belongs to a typical Asian genotype.

 

Key words: Taiwan, KoG cell line, koi herpesvirus, phylogenetic analysis.

Abbreviation

Abbreviations: KHV, Koi herpesvirus; CyHV-3, cyprinid herpesvirus-3; IFA, indirect immunofluorescence assay; KoG, koi gill cell; CPE, cytopathic effect.