WSSV inoculum

Description

The ICDSA can provide WSSV inoculum prepared from the WSSV-TW strain (GenBank No. AF440570) This strain was originally isolated from a batch of WSSV-infected moribund farmed Penaeus monodon collected in southern of Taiwan in 1994. Inoculun Stock must be hept at −80 °C until used.

Brief protocol

WSSV inoculum preparation and experimental challenge methodology

WSSV inoculum was prepared as follows. Briefly, 0.5 g of the frozen infected specimens of P. monodon were minced and then homogenized in 4.5 ml of sterile phosphate-buffered saline (PBS) (137 mM NaCl, 2.7 mM KCl, 10 mM Na₂HPO₄, 2 mM KH₂PO₄). After centrifugation at 400 ×g for 10 min at 4 °C, the supernatant was filtered through 0.45 μm membrane. After a further 10× dilution, the virus suspension was used to infect adult specific pathogen free (SPF) Penaeus (Litopenaeus) vannamei (body weight ~45 g; High Health Aquaculture Inc., Hawaii) by injection into each shrimp (5 μl/g of body weight) at the dorsal lateral area of the fourth abdominal segment, between the tergal plates forward into the third abdominal segment. Hemolymph from moribund challenged shrimp was collected, removed the hemocytes by centrifugation and filtration, diluted 4× with PBS, and stored at −80 °C for viral stock. For challenge by intra muscular injection, the viral stock use in penaeid shrimp and crayfish should be further diluted 25× and 2.5×, respectively and with a dosage of 5 μl/g of body weight.

Some of our publications that have used this platform

1. Chang Y-S, Liu W-J, Chou T-L, Lee Y-T, Lee T-L, et al. (2008) Characterization of white spot syndrome virus envelope protein VP51A and its interaction with viral tegument protein VP26. J Virol 82: 12555-12564.
2. C.H. Wang, C.F. Lo, J.H. Leu, C.M. Chou, P.Y. Yeh, H.Y. Chou, M.C. Tung, C.F. Chang, M.S. Su and G.H. Kou, Purification and genomic analysis of baculovirus associated with white spot syndrome (WSBV) of Penaeus monodon, Dis Aquat Organ 23 (1995), pp. 239–242.
3. Lo, C.-F., H.-C. Hsu, M.-F. Tsai, C.-H. Ho, S.-E. Peng, G.-H. Kou, and D. V. Lightner., Specific genomic DNA fragment analysis of different geographical clinical samples of shrimp white spot syndrome virus, Dis Aquat Organ 35 (1999), pp. 175–185.
4. H.C. Wang, H.C. Wang, G.H. Kou, C.F. Lo and W.P. Huang, Identification of icp11, the most highly expressed gene of shrimp white spot syndrome virus (WSSV), Dis Aquat Organ 74 (2007), pp. 179–189.
5. H.C. Wang, H.C. Wang, J.H. Leu, G.H. Kou, A.H. Wang and C.F. Lo, Protein expression profiling of the shrimp cellular response to white spot syndrome virus infection, Dev Comp Immunol 31 (2007), pp. 672–686.
6. M.F. Tsai, G.H. Kou, H.C. Liu, K.F. Liu, C.F. Chang and S.E. Peng et al., Long-term presence of white spot syndrome virus (WSSV) in a cultivated shrimp population without disease outbreaks, Dis Aquat Organ 38 (1999), pp. 107–114.