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Lu, Yongzhong; Lm, Tapay; Pc, Loh; Rb, Gose; Ja, Brock

doi:10.1023/a:1018399720619

Cited by 14 publications

(1 citation statement)

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“…Cells derived from the ovary and lymphoid tissues have been used and some are susceptible to virus infection, for example, primary cell cultures of the lymphoid organs that are susceptible to WSSV are from Penaeus monodon (Kasornchandra et al, 1999;Wang et al, 2000), from the blue shrimp, Litopenaeus stylirostris (Tapay et al, 1995(Tapay et al, , 1997 and from the kuruma shrimp, Marsupenaeus japonicus (Itami et al, 1999) as well as ovarian primary cultures from kuruma shrimp (Maeda et al, 2003(Maeda et al, , 2004. Primary cell cultures of lymphoid organ were also reported to be susceptible to YHV (Lu et al, 1995;Assavalapsakul et al, 2003). Only one report (West et al, 1999) used a mixture of haematopoietic tissues (hpt) and lymphoid cells from P. monodon to develop primary cell cultures, but these were not used for viral studies.…”

Section: Introductionmentioning

confidence: 99%

Characterization of white spot syndrome virus replication in in vitro-cultured haematopoietic stem cells of freshwater crayfish, Pacifastacus leniusculus

Jiravanichpaisal

Söderhäll

2006

Journal of General Virology

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Replication of White spot syndrome virus (WSSV) was investigated in haematopoietic cells (hpt cells) derived from haematopoietic tissue (hpt) of freshwater crayfish, Pacifastacus leniusculus. Temperature and type of inoculum for virus replication were studied. The cell culture remained viable at a wide range of temperatures ranging from 4 to 25 6C. WSSV replicated in cells, as evidenced by in situ hybridization, RT-PCR and by the presence of virions visualized with an electron microscope. Moreover, the results showed that the infectivity of WSSV to hpt cells is dependent on temperature and a supplemented growth factor (cytokine) astakine. WSSV replicated more rapidly at higher temperatures than at lower temperatures. No virus replication was observed at 4 6C. Detectable WSSV-infected cells were present as early as 36 h post-inoculation, demonstrated by in situ hybridization or RT-PCR of VP28 expression at 25 6C. Hpt cells can survive a few weeks at 25 or 16 6C and longer than several months at 4 6C. INTRODUCTIONAmong cultured crustaceans, penaeid shrimp are of considerable economic importance. Rapid development and expansion of shrimp farming have occurred throughout the world especially in South East Asia. However, diseases, mainly viral, have resulted in the collapse of the shrimp aquaculture worldwide and hence caused considerable economic losses. Over the past decade, several viral diseases have been discovered and they have had large economic impacts and still continue to threaten shrimp production (Flegel, 1997;Spann & Lester, 1997). Among these causative viruses, Yellow head virus (YHV) and White spot syndrome virus (WSSV) have been the two most serious causes of major production losses (Kasornchandra et al., 1995;Wongteerasupaya et al., 1995). Although reliable techniques for diagnosis have been established for these two viruses by using PCR, the majority of studies on viral agents have been limited to light or electron microscopic observations. Cell culture is a basic tool for the study of pathogenic infections, especially for those pathogens that replicate intracellularly, such as viruses. A complete understanding of these viruses is dependent upon the development of laboratory techniques for maintenance and culturing of these viruses and their host cells. There are no continuous crustacean cell lines established so far for replication of viruses, although over the past decade primary cultures obtained from various organ sources have been used and are reported with increasing frequency in the literature. Explants or dissociation techniques are often used to obtain cells from various tissues and organs of penaeid shrimp, including the lymphoid (Oka) organ (Chen & Kou, 1989;Chen & Wang, 1999;Hsu et al., 1995; Kasornchandra et al., 1999;Itami et al., 1999;Owens & Smith, 1999; West et al., 1999;Wang et al., 2000), the heart (Chen et al., 1986;Chen & Wang, 1999;Owens & Smith, 1999), nerve cord (Nadala et al., 1993;Owens & Smith, 1999), gut (Nadala et al., 1993, hepatopancreas (Owens & Smith, 1999) and gonad...

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