In vitro propagation of hepatopancreatic parvo-like virus (HPV) of shrimp in C6/36 (Aedes albopictus) cell line.

Abstract Hepatopancreatic parvovirus (HPV) which causes infection in many species of penaeid shrimp is a serious viral pathogen in the young life stages of shrimp. An attempt was made to develop an in vitro system using C6/36 subclone of Aedes albopictus cell line for propagation of HPV. The results revealed that C6/36 cells were susceptible to this virus and the infected cells showed CPE in the form of vacuole formation. The results of PCR, immunocytochemistry and Western blot revealed the HPV-infection in C6/36 cell line. The RT-PCR analysis confirmed the replication of HPV in C6/36 cell line. The HPV load was quantified at different time intervals by ELISA and real time PCR, and the results showed the increase of viral load in C6/36 cell line in time course of infection. HPV propagated in C6/36 cell line was used to infect post-larvae of shrimp and the results showed that the twentieth passage of HPV propagated in C6/36 cell line caused 100% mortality in post-larvae after 6 weeks post infection (d.p.i.). The infected post-larvae showed clinical signs of reduced growth, reduced preening, muscle opacity and atrophy of hepatopancreas. The HPV-infection was confirmed by PCR. The results of the present study showed that C6/36 cell line can be used as an in vitro model for HPV replication instead of whole animal.