Journal of Microbiology

Establishment and Characterization of the Epithelioma Papulosum Cyprini (EPC) Cell Line Persistently Infected with Infectious Pancreatic Necrosis Virus (IPNV), an Aquabirnavirus: Hyoung Jun Kim , Jae-Kwon Cho , Hyung-Kyu Hwang , Myung-Joo Oh , Toyohiko Nishizawa; J. Microbiol. 2012;50(5):821-826. Published online November 4, 2012; DOI: https://doi.org/10.1007/s12275-012-2364-2

Abstract: Infectious pancreatic necrosis virus (IPNV), a type species of aquabirnaviruses in the family Birnaviridae, is an etiological agent of infectious pancreatic necrosis and has been isolated from epizootics of cultured salmonids. In the present study, an epithelioma papulosum cyprini (EPC) cell line persistently infected with IPNV (PI-EPC) was experimentally established by subculturing EPC cells surviving IPNV infection, and was characterized. PI-EPC cells were morphologically indistinguishable from EPC, but continued to grow and yield IPNV. PI-EPC cells showed no cytopathic effect due to IPNV inoculation, and susceptibility of PI-EPC cells against heterologous viruses was not different from that of EPC cells. Only one cell of 103.5 PI-EPC cells produced IPNV at approximately 100.5 50% tissue culture infectious dose (TCID50)/cell/day, which was approximately 1,000 times lower than that of normal EPC cells. PI-EPC cells that did not yield IPNV (N-PI-EPC) were screened. The IPNV genome was detected from both PI-EPC and N-PI-EPC cells, and the IPNV VP2 structural protein was detected from both cell lines, but no other IPNV proteins were observed by Western blot analysis with anti-IPNV serum. Thus, multiplication of IPNV in PI-EPC cells was regulated by some host cell factors, except interferon.

Distribution of Marine Birnavirus (MABV) in Marine Organisms from Okinawa, Japan, and a Unique Sequence Variation of the VP2/NS Region: Manami Inaba , Satoru Suzuki , Shin-Ichi Kitamura , Norichika Kumazawa , Hiroshi Kodama; J. Microbiol. 2009;47(1):76-84. Published online February 20, 2009; DOI: https://doi.org/10.1007/s12275-008-0250-8

Abstract: Distribution of marine type of Aquabirnavirus (MABV) was examined in shellfish and fish from Okinawa and Ishigaki Islands, Japan, where water temperature is higher than 25°C through the year. Genome detection and virus isolation were performed for shellfish and fish samples, and the results revealed the prevalent distribution of MABV in diverse species in the area, although isolation was not frequently. Detection rate of MABV genome in bivalves was higher than gastropods, which was similar result to former report in mainland of Japan. Furthermore, the unique five-nucleotide deletion was found with a high rate of occurrence in the MABV genome from shellfish and fish. This study showed distribution status of MABV in organisms in subtropical waters by wide monitoring, and discovered new genome variation in VP2/NS region of this virus.

Distribution of Marine Birnavirus in Cultured Olive Flounder Paralichthys olivaceus in Korea: Sung-Ju Jung , Seok-Ryel Kim , Il-Yong Joung , Shin-Ichi Kitamura , Hee-Taek Ceong , Myung-Joo Oh; J. Microbiol. 2008;46(3):265-273. Published online July 5, 2008; DOI: https://doi.org/10.1007/s12275-008-0004-7

Abstract: Surveys of marine birnavirus (MABV) were undertaken in cultured olive flounder Paralichthys olivaceus from the south and west coastal areas and Jeju in Korea during the period January 1999 to April 2007. MABV was detected in all seasons from the fry, juveniles and adult fish from the areas examined. Evident cytopathic effects of the virus including rounding and cell lysis were observed in chinook salmon embryo (CHSE-214) and rainbow trout gonad (RTG-2) cells, but not in fathead minnow (FHM) and epithelial papilloma of carp (EPC) cells. Nucleotide sequences of the VP2/NS junction region of the Korean isolates showed 97.8%~100% similarity, and they belonged to the same genogroup. Cross neutralization tests with serotype-specific rabbit antisera against MABV strains exhibited a close antigenic relationships between strains, and were distinct from infectious pancreatic necrosis virus (IPNV) strains. Coinfection of MABV with bacteria (Streptococcus iniae, Vibrio spp.) and viruses (nervous necrosis virus, lymphocystis disease virus, viral hemorrhagic septicemia virus) was observed.

Genetic Analysis of the VP2/NS Junction Region on Segment A of Marine Birnavirus Isolated from Cultured Flounder Paralichthys olivaceous: Seong-Joon Joh , Jeong-Ho Kim , Gang-Joon Heo; J. Microbiol. 2000;38(1):44-47.

Abstract: The cDNA of VP2/NS junction region on segment A of the marine birnavirus (MBV) isolated from flounder (DS strain) was amplified using the reverse transcription (RT)-polymerase chain reaction (PCR). Its cDNA nucleotide and deduced amino acid sequences were analyzed, and compared with the reference strains of MBV and infectious pancreatic necrosis virus (IPNV). Analyses of the nucleotide and deduced amino acid sequences revealed that the DS strain is very similar to the reference strains of MBV, distant from the IPNV.

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