In vitro replication of Macrobrachium rosenbergii nodavirus and extra small virus in C6/36 mosquito cell line.
White tail disease (WTD) is a serious problem in hatcheries and nursery ponds of Macrobrachium rosenbergii in India and many parts of the world. The pathogenic agents have been identified as M. rosenbergii nodavirus (MrNV) associated with extra small virus (XSV), which is 27nm and 15nm in diameter, respectively. Replication of MrNV and XSV was investigated in apparently healthy C6/36 subclone of Aedes albopictus cell line. The results revealed that C6/36 cells were susceptible to these viruses. The replication of these viruses in C6/36 cells was confirmed by RT-PCR, acridine orange staining, infectivity study and electron microscopy. Cytoplasmic ribonucleic acid (RNA) stained by acridine orange increased by 48h, and by 72h larger proportion of cells which indicated alterations in quantity and localization of RNA in the infected cells. Post-larvae, challenged by immersion method using inoculum prepared from infected cells, exhibited lethargy, anorexia and opaqueness of abdominal muscle and 100% mortality was observed at 6 days post-infection. Experimentally infected C6/36 cells and post-larvae showed positive by RT-PCR, whereas control cells and healthy post-larvae showed negative. This is the first study to report the multiplication of MrNV and XSV in C6/36 cell line.
Sudhakaran
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Aquaculture Biotechnology Division, Department of Zoology, C. Abdul Hakeem College, Melvisharam 632509, Vellore Dist, Tamil Nadu, India.
Parameswaran
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V
Sahul Hameed
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D000330:Aedes / Q000166:cytology / Q000648:ultrastructure / Q000821:virology*; D000818:Animals; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral; D046529:Microscopy, Electron, Transmission; D030381:Nodaviridae / Q000302:isolation & purification / Q000502:physiology* / Q000648:ultrastructure; D033541:Palaemonidae / Q000821:virology; D012328:RNA Viruses / Q000302:isolation & purification / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014776:Virus Cultivation; D014779:Virus Replication