一种佐剂新材料在嗜水气单胞菌灭活疫苗浸泡免疫异育银鲫中的应用效果  

Efficacy of inactivated Aeromonas hydrophila vaccine with a new biodegradable material adjuvant in Carassius auratus gibelio by immersion immunization

作　　者：任燕 时云朵 石存斌 吴淑勤 

机构地区：[1]中国水产科学研究院珠江水产研究所广东省水产动物免疫技术重点实验室农业部渔用药物创制重点实验室,广东广州510380 [2]上海海洋大学水产与生命学院,上海201306 [3]四川省水产学校,四川成都611730

出　　处：《水产学报》2018年第1期112-119,共8页Journal of Fisheries of China

基　　金：广东省海洋渔业科技与产业发展专项(A201501C12);现代农业产业技术体系建设专项资金(CARS-46);国家科技支撑计划(2012BAD25B02)~~

摘　　要：为研究蓖麻油聚乙二醇单酯葡萄糖苷(CPMG)在细菌灭活疫苗浸泡免疫中的佐剂作用,将CPMG和嗜水气单胞菌灭活疫苗联合施用,二次浸泡免疫异育银鲫后饲养在室内玻璃缸中,用实时荧光定量PCR(RT-PCR)检测加强免疫后2、4、7、11、14和21 d脾脏中Ig M、IL-1β、C3、C-凝集素和溶菌酶的m RNA表达量,首次免疫30 d后用同源菌株活菌攻击实验鱼;同时,用含CPMG、山莨菪碱复合佐剂的灭活疫苗浸泡免疫异育银鲫一次后饲养在池塘网箱中,定期抽取血液检测血清抗体效价并用同源菌株人工感染实验鱼。结果显示,CPMG佐剂组和无佐剂组的5个免疫相关基因的相对表达量显著高于对照组,CPMG组Ig M基因较无佐剂组更早表达,补体C3、C-凝集素以及溶菌酶的基因表达量更高或表达持续时间更长。血清抗体效价在免疫后2周达到160,6周达到峰值640,12周降低为40。室内玻璃缸中二次免疫30 d后,CPMG组的平均相对保护率达到70.6%,复合佐剂组达到88.2%,均显著高于无佐剂组的56.0%。池塘网箱中一次浸泡免疫30、60和120 d后,平均相对保护率分别为54.6%、44.0%和12.5%。研究表明,CPMG与嗜水气单胞菌灭活疫苗共同浸泡免疫异育银鲫,增强了脾脏中Ig M、IL-1β、C3、C-凝集素以及溶菌酶基因表达,提高了异育银鲫抗嗜水气单胞菌感染的能力。Castor oil polyethylene glycol monoester glucoside（CPMG） is a kind of biodegradable and artificial material. Formalin-inactivated Aeromonas hydrophila bacteria with or without CPMG, CPMG ＋ anisodamine mixture were administered on days 1 and 7 to Carassius auratus gibelio through immersion. Spleens were sampled on days 2, 4, 7, 11, 14 and 21 post-second vaccination, and the expression quantities of Ig M, IL-1β, C3, C-lectin and lysozyme m RNAs in spleen were detected through the real-time fluorescence quantitative PCR（RT-PCR）. Formalin-inactivated A. hydrophila vaccine with CPMG ＋ anisodamine mixture was immersion administered just once,then, specific antibodies were detected and the fish were challenged intraperitoneally with lethal A. hydrophila at different time post-vaccination. The results of vaccination twice showed that fish immunized with inactivated A. hydrophila vaccine and CPMG exhibited enhanced expression of genes that are involved in both innate and adaptive immune responses（Ig M, IL-1β, C3, C-lectin and lysozyme） compared to no adjuvant control or saline control. The relative percent survival（RPS） was found to be 70.6% on day 30 post-vaccination, while immunization with the antigen alone followed by an experimental challenge gave an RPS of 56.0%, moreover, the CPMG ＋ anisodamine coupled vaccine showed an RPS of 88.2%.The results of vaccination just once showed that the CPMG ＋anisodamine coupled vaccine could augment the production of specific serum antibodies, which reached the highest level 6 weeks post-vaccination. The resulting RPS calculated was 54.6%, 44.0% and 12.5% on days 30, 60,120 post-vaccination, respectively. These results indicate that formalin-inactivated A. hydrophila vaccine with CPMG immersion administered to gibel carps can significantly promote the gene expression of Ig M, IL-1β, C3, Clectin and lysozyme in spleen and enhance specific immune responses against A. hydrophila infection.

关 键 词：异育银鲫 蓖麻油聚乙二醇单酯葡萄糖苷 山莨菪碱 灭活疫苗 浸泡免疫 

分 类 号：S942[农业科学—水产养殖]