In Vitro propagation of Infectious Hypodermal Hematopoietic Necrosis Virus [IHHNV] / Penaeus stylirostris penstyldensovirus [PstDV1] in PmLyO-Sf9

Abstract

Abstract Isolation and in vitro propagation of Infectious Hypodermal Hematopoietic Necrosis Virus (IHHNV/ PstDVI) in PmLyO-Sf9 could be successfully performed. After few hours of post inoculation with the virus, cytopathic changes such as (a) clustering (b) enlargement (c) syncytium formation (d) granulation (e) vacuole formation (f) tapering (g) irregular plasma membrane with extended tails (h) detachment (i) cell death and cellular debris formation were observed. Expression of viral genes, presence of virions and cytological changes demonstrated through TEM suggested replication of the virus in the shrimp - insect hybrid cell line. The virus could be purified by ultracentrifugation, negatively stained, and demonstrated under electron microscope. The same was found to be infective both in vitro and in vivo. This development opens avenues for the study of basic molecular mechanism of IHHNV infection, pathogenesis and replication kinetics much required for developing antiviral strategy in aquaculture.