Abstract
Shrimp viral diseases have caused severe production and economic losses in the past two decades. A complete understanding of shrimp viruses is dependent upon the development of laboratory techniques for the maintenance and culturing of these viruses and host cells. This investigation was done to characterize the cell line culture from specific pathogen free Litopenaeus vannamei and its susceptibility to revise the cytopathic effects of white spot syndrome virus. A cell culture was successfully developed from insect cell SF9. Cytopathic changes like enlarged cells, focal lesions, shrunken and clumped cells were observed in white spot syndrome viruses (WSSV) infected SF9 cell cultures from 24 to 120 h duration. In the present study, conditions for the successful primary culture of insect SF9 cells for WSSV infection have been established. The conformation of WSSV infection in SF9 cell line was by polymerase chain reaction using target gene of WSSV419 like proteins and followed by electrophoresis. The WSSV result was positively as 550 bp in SF9 cell line and in the control sample. This is the first report on the development of primary cell culture of WSSV, host species of L. vannamei using insect cell line SF9. Key words: White spot syndrome viruses (WSSV), SPF, L. vannamei, SF9 cell line, cytopathic effect, pathogen, economy.
Highlights
Crustacean cell culture has gained momentum due to viral diseases affecting commercially important species
white spot syndrome viruses (WSSV) infection in shrimp was confirmed by polymerase chain reaction (PCR) using PCR primers for target genes such as WSSV419 like protein which is similar to VP28 and their amplification size and by specific clinical symptoms like the presence of white spots, lethargy and reddish discoloration of the body
The cell culture was successfully developed from insect cell SF9 in serum-free medium (SFM)
Summary
Crustacean cell culture has gained momentum due to viral diseases affecting commercially important species. Cell culture techniques were developed: (a) to assist in understanding the mechanism of host pathogennesis interaction (Chen et al, 1989), (b) to produce large amount of viral material for their characterization and (c) to improve tools for diagnosis and cure of diseases. Attempts have been made to establish several cell culture systems of shrimps (Al-Mohanna and Nott 1987; Chen et al, 1986, 1995; Ke et al, 1990; Nadala et al., 1993; Hsu et al, 1995; Toullec et al, 1996; Mulford and Austin, 1998; Mulford et al, 2001; Uma et al, 2002) and other crustaceans (Peponnet and Quiol, 1971).
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