In vitro white spot syndrome virus (WSSV) replication in explants of the heart of freshwater crab, Paratelphusa hydrodomous.

Explants from different organs of freshwater crab, Paratelphusa hydrodomous were prepared to establish an in vitro system for replication of white spot syndrome virus (WSSV) of shrimp. Heart explants were maintained for 53 days without any morphological changes in EX-CELL™ 405 medium with and without serum whereas the explants of eye muscle, gill, shell membrane and appendage muscle died within 15 days of culture period. The heart explants on different days of culture were exposed to WSSV for 10 days to study the viral replication. The infection of WSSV in explants of the heart was confirmed by PCR, RT-PCR, Western blot, histology, immunohistochemistry, bioassay and transmission electron microscopy. The WSSV was quantified by real-time PCR and indirect ELISA. The WSSV inoculum prepared from the heart explants of crab caused significant mortality in Penaeus monodon in challenge experiments and the results indicate that the WSSV which replicated in the heart explants of freshwater crab maintains its infectivity as in marine shrimp. The results indicate that the heart explants of P. hydrodomous would be a good alternative to whole animals for production of WSSV.