Development of primary shrimp hemocyte cultures of Penaeus chinensis to study white spot syndrome virus (WSSV) infection

Hemocytes were withdrawn from hearts of Penaeus chinensis, cultured with seven kinds of culture media, the hemocytes remained viable for about 20 days using Leibovitz's 2 × L-15 medium consisting of 20% fetal bovine serum, 20% GIT, and 2% glucose, adjusting osmolarity with sodium chloride, pH 7.2–7.4, and incubated at a temperature of 25 °C. Different concentrations of virus inoculum were inoculated into the primary cultured hemocytes, and diverse cytopathic effects were observed with an inverted phase-contrast microscope. WSSV infection was confirmed by immunocytochemical tests with monoclonal antibodies against the virus. These revealed that WSSV could infect hemocytes of P. chinensis and propagate in the nuclei of infected cells.