Neutralization of white spot syndrome virus (WSSV) for Penaeus chinensis by antiserum raised against recombinant VP19.

This study was carried out to neutralize the WSSV one of the most virulent pathogen causing large economic damage in shrimp culture industry using the antiserum produced against recombinant WSSV envelope protein VP19 (rVP19) as a tool to evaluate WSSV infection mechanism. A fragment of VP19 was expressed in Sf21 insect cell using baculovirus expression system as fusion protein with 6 His-tag. Then, polyclonal antiserum against rVP19 was raised in white rabbit. A constant amount of WSSV (at 10(4) diluted stock) was incubated with various antiserum concentrations and injected into shrimp, Penaeus chinensis, for the neutralization challenge. At 9 days post injection, the shrimp in the positive control injected with WSSVshowed 100% mortality The shrimps injected with WSSV preincubated with preimmune serum showed 83.3% mortality at 15 days post injection. The shrimps injected with the WSSV preincubated with 1 microl, 5 microl or 10 microl r VP19 antiserum and shrimp mortalities showed 66.6%, 40.0% and 26.6% at 15 days post injection, respectively The high concentration of antiserum group showed lower mortality than those of the low concentration of antiserum group. This indicates that the WSSV can be neutralized by the rVP19 antiserum in a dose-dependent manner. The neutralization challenge result suggested that VP19 might play an important role in WSSV infection to shrimp.

Shrimps survive white spot syndrome virus challenge following treatment with Vibrio bacterin.

Taking an innovative approach, a vaccination study using five bacterial strains viz. Vibrio campbelli (B60), V. alginolyticus (B73), V. parahaemolyticus-like (B79), V. parahaemolyticus (R8) and V. harveyi (RG203) was conducted in Penaeus monodon against white spot syndrome virus (WSSV) infection, considered as one of the serious pathogens of shrimps. Oral challenge with shrimps infected with WSSV showed a relative percentage survival of 5 and 47% in the P. monodon juveniles vaccinated with V. parahaemolyticus and V. harveyi, respectively. Results showed that there is a possibility of specifically immunising the shrimps against WSSV using bacterin prepared out of Vibrio harveyi isolates taken from shrimps infected with WSSV. Also, there was a level of protection attained by the shrimps due to immunisation with Vibrio strains.

Recombinant expression of marine shrimp lysozyme in Escherichia coli

Shrimp Lysozyme (Lyz) is a key component of the antibacterial response as part of the innate defense in Crustacea; however, it has not been possible to purify this protein because of the very low amount present in the shrimp blood cells (hemocytes). In an effort to produce enough protein to study its function and biochemical properties we have overexpressed Lysozyme from marine shrimp (Penaeus vannamei) in E. coli. A bacterial protein expression system based on the T7 polymerase promoter was used. Although Lyz was produced as insoluble protein in inclusion bodies, its refolding led to an active protein with a yield of ~10 percent. Details of the protein recombinant expression techniques applied to this shrimp protein are presented.