Replication of white spot syndrome virus in ovarian primary cultures from the kuruma shrimp, Marsupenaeus japonicus.
Propagation of white spot syndrome virus (WSSV) was investigated in primary ovarian cultures from the kuruma shrimp Marsupenaeus japonicus. A WSSV strain, purified by sucrose density gradient centrifugation, was inoculated into 10-day-old primary ovarian cultures. WSSV infection induced marked cytopathic effect (CPE) on primary ovarian cells. Initially, virus-infected cells began to shrink 72 h post-inoculation, followed by the rounding of most cells which detached finally from flask surface. Electron microscopic observations clearly showed that the replication of WSSV occurred in nuclei of ovarian cells. Immunoblot analysis with antibodies against the WSSV envelope protein VP28 provided the evidence that the levels of WSSV antigens in culture supernatant gradually increased during the period between 24 and 120 h after virus inoculation. The results suggest that the use of primary ovarian cultures of the kuruma shrimp will facilitate characterization of the WSSV infection.
Maeda
Minoru
M
Kyushu Medical Co Ltd, Kurume Research Center Building, 2432-3 Aikawa, Kurume, Fukuoka 839-0861, Japan. maeda-bio@ceres.ocn.ne.jp
Saitoh
Hiroyuki
H
Mizuki
Eiichi
E
Itami
Toshiaki
T
Ohba
Michio
M
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis / Q000276:immunology; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques; D002467:Cell Nucleus / Q000648:ultrastructure / Q000821:virology; D048429:Cell Size / Q000502:physiology; D002478:Cells, Cultured; D003588:Cytopathogenic Effect, Viral; D004267:DNA Viruses / Q000276:immunology / Q000502:physiology*; D005260:Female; D010053:Ovary / Q000166:cytology / Q000821:virology; D033561:Penaeidae / Q000821:virology*; D014759:Viral Envelope Proteins / Q000032:analysis / Q000276:immunology; D014764:Viral Proteins / Q000032:analysis / Q000276:immunology; D014779:Virus Replication / Q000502:physiology*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D014759:Viral Envelope Proteins; D014764:Viral Proteins