Viral hemorrhagic septicemia virus (VHSV) is the etiological agent of viral hemorrhagic septicemia (VHS), one of the most severe viral diseases affecting cultured olive flounder (Paralichthys olivaceus) in Far East Asia. VHS occurs during the winter or spring season when the water temperature is low (9–15 °C). In our previous study found that VHSV infection had controlled by using water temperature (above 17 °C). By using water temperature, we demonstrated optimal live VHSV immersion vaccine treatment concentration, also live VHSV immersion vaccine treatment method. We confirmed that the effective VHSV immersion treatment was 10^5.5 TCID₅₀/mL at 17 °C. It was no need pretreatment before live VHSV immersion vaccination. The VHSV titer of vaccinated fish organs was under the estimated limit (<1.8 log TCID₅₀/mL) within 3 days in 10^5.5 TCID₅₀/mL live VHSV immersion at 17 °C. High survival rates were observed in live VHSV immersion with 10^5.5 and 10^7.5 TCID₅₀/mL at 17 °C and then infected VHSV at 10 °C. VHSV specific antibody was not detected from in the surviving flounder under VHSV infection after immersion treatment with live VHSV. In addition, the potentiality of natural immunization against VHS in olive flounder was suggested by live VHSV immersion vaccine at temperature controlled fish culture condition.

病毒性出血性败血症病毒 (VHSV) 是病毒性出血性败血症 (VHS) 的病原体，VHS 是影响远东地区养殖的橄榄比目鱼 ( Paralichthys olivaceus )的最严重的病毒性疾病之一。VHS 发生在水温较低 (9–15 °C) 的冬季或春季。在我们之前的研究中发现 VHSV 感染已通过使用水温（17 °C 以上）得到控制。通过使用水温，我们展示了最佳的活 VHSV 浸渍疫苗处理浓度，以及活 VHSV 浸渍疫苗处理方法。我们确认有效的 VHSV 浸泡处理为 10 ^5.5 TCID ₅₀/mL 在 17 °C。VHSV活体浸泡接种前无需预处理。在 10 ^5.5 TCID ₅₀ /mL 活 VHSV 浸入 17 °C 下，3 天内接种疫苗的鱼器官的 VHSV 滴度低于估计限度（<1.8 log TCID ₅₀ /mL）。在 10 ^5.5和 10 ^7.5 TCID ₅₀ /mL 在 17 °C 下浸入活 VHSV 中观察到高存活率，然后在 10 °C 下感染 VHSV。在用活的 VHSV 浸泡处理后，在 VHSV 感染下的存活比目鱼中未检测到 VHSV 特异性抗体。此外，在温度控制的鱼类培养条件下，活 VHSV 浸入疫苗表明了对橄榄比目鱼中 VHS 进行自然免疫的潜力。