| 摘要: |
| 鲫造血器官坏死病是由鲤疱疹病毒2型(CyHV-2)引起的危害我国养殖鲫鱼(Carassius auratus)最严重的病毒性疾病。为制备有效预防鲫造血器官坏死病的灭活疫苗,以CyHV-2代表流行株CNDF-TB2015为疫苗株,以CSC细胞系进行病毒增殖,并采用不同浓度的病毒对不同规格鲫鱼进行攻毒,建立攻毒模型。结果表明,培养的病毒滴度达1010 TCID50/mL,以4×107TCID50/尾的剂量腹腔注射感染体重300 g的鲫鱼可造成100%的死亡率。对病毒灭活剂甲醛、β-丙内酯和BEI灭活条件进行筛选优化,对疫苗免疫佐剂763A、IMS1312、铝胶和M402进行筛选,结果表明,浓度为108.3 TCID50/mL的病毒液最佳灭活条件为终浓度10 mmol/L的BEI 37 ℃灭活72 h,疫苗制备最佳佐剂为水佐剂IMS1312。对灭活疫苗的最小免疫剂量、最佳免疫剂量和有效免疫产生期进行测定,结果表明,要获得70%以上保护率的最小免疫剂量为8×106 TCID50/尾,最佳免疫剂量为4×107 TCID50/尾,在免疫4周后保护率可达80%, 5周后保护率达到100%。制备的灭活疫苗为鲫造血器官坏死病的免疫预防提供了有力手段,也为其他水产动物病毒灭活疫苗的制备提供了借鉴。 |
| 关键词: 鲤疱疹病毒2型 鲫造血器官坏死病 灭活疫苗 免疫效力 |
| DOI：10.11693/hyhz20211200329 |
| 分类号:S942.5 |
| 基金项目:浙江省重点研发计划,2019C02058号;浙江省科研院所扶持专项,2021YSZX001号。 |
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| PREPARATION AND EFFICACY OF INACTIVATED VACCINE AGAINST CYPRINID HERPESVIRUS-2 |
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PAN Xiao-Yi1, LIN Ling-Yun1, ZHU Yue2, YAO Jia-Yun1, YIN Wen-Lin1, LIU Yi-Han1, CHEN Fan3, SHEN Jin-Yu1,2
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1.Ministry of Agriculture and rural areas Key Laboratory of Healthy Freshwater Aquaculture, Key Laboratory of Fish Health and Nutrition of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China;2.School of Fishery, Zhejiang Ocean University, Zhoushan 316022, China;3.Hangzhou Centre for Agricultural Technology Extension, Hangzhou 310001, China
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| Abstract: |
| Herpesviral haematopoietic necrosis (HVHN) is a viral disease of crucian carp caused by Cyprinid herpesvirus-2(CyHV-2),which causes huge losses on aquaculture in China.To prevent from the outbreak of HVHN,the inactivated vaccine was developed using vaccine strain CNDF-TB2015 and the immune efficacy was assessed.The CyHV-2 culture showed that the titer of virus cultured with CSC cells reached 1010 TCID50/mL,and the challenge model test showed the dose of 4×107TCID50/ind could cause 100% mortality in crucian carp within 14 d post injection with the CyHV-2 CNDF-TB2015.The inactivated vaccine preparation test showed that the optimal inactivation condition for CyHV-2 at 108.3 TCID50/mL was 10 mmol/L BEI at 37℃ for 72 h and the best adjuvant was IMS1312.The minimum immune dose was 8×106 TCID50/ind,and the optimal immune dose was 4×107 TCID50/ind.The protection rate reached 80% in 4 weeks and 100% in 5 weeks after immunization with dose of 4×107 TCID50/ind.This study presents that the inactivated CNDF-TB2015 strain of CyHV-2 could serve as a potential vaccine candidate to prevent the prevalence of HVHN,and provides a reference for the preparation of other inactivated viral vaccines for fish in aquaculture. |
| Key words: CyHV-2 HVHN inactivated vaccine immune efficacy |
