变形假单胞菌免疫刺激复合物对大黄鱼口服免疫效果的评价  

Evaluation of the Oral Immune Effects of Pseudomonas plecoglossicida Immune Stimulating Complexes on Large Yellow Croaker(Larimichthys crocea)

作　　者：许斌福[1] 陈秀霞[1] 池洪树[1] 李素一[1] 郑在予[1] 徐梦婷 江秋欢 郑炜强 林能锋[1] XU Bin-Fu;CHEN Xiu-Xia;CHI Hong-Shu;LI Su-Yi;ZHENG Zai-Yu;XU Meng-Ting;JIANG Qiu-Huan;ZHENG Wei-Qiang;LIN Neng-Feng(Fujian Provincial Aquatic Disease Prevention and Control Engineering Technology Research Center,Institute of Biotechnology,Fujian Academy of Agricultural Sciences,Fuzhou 350003,China;State Key Laboratory of Mariculture Breeding/Ningde FuFa Fisheries Company Limited,Ningde 352103,China)

机构地区：[1]福建省农业科学院生物技术研究所福建省水产病害防治工程技术研究中心,福州350003 [2]海水养殖生物育种全国重点实验室/宁德市富发水产有限公司,宁德352103

出　　处：《农业生物技术学报》2024年第9期2181-2189,共9页Journal of Agricultural Biotechnology

基　　金：国家重点研发计划(2022YFD2401002);福建省重大专项专题(2021NZ033025);福建省公益专项专题(2022R1027005,2023R1025006);宁德市揭榜挂帅项目(ND2023J005)。

摘　　要：变形假单胞菌(Pseudomonas plecoglossicida)是大黄鱼(Larimichthys crocea)内脏白点病的主要病原,可导致巨大经济损失。前期研究表明免疫刺激复合物(immune stimulating complexes, ISCOMs)具有较好的口服免疫效果。为了评价口服变形假单胞菌ISCOMs免疫大黄鱼的效果,本研究制备具有笼格状纳米颗粒结构的变形假单胞菌ISCOMs疫苗(Pp-ISCOMs),检测口服免疫后21 d大黄鱼头肾的各免疫因子的基因表达水平,通过菌体血清凝集方法测定免疫后大黄鱼血清效价,并测定其免疫保护力。结果显示,与灭活变形假单胞菌(Pp)和空白对照(CK)相比,口服免疫Pp-ISCOMs的大黄鱼头肾的髓样分化因子88 (myeloid differentiation factor 88, MyD88)(P<0.05)和白介素2 (interleukin 2, IL-2)(P<0.001)等免疫相关因子表达显著提高;受免21 d的大黄鱼血清凝集效价为2~2;用5.0×10^(2) CFU/g剂量的变形假单胞菌腹腔注射攻毒后,Pp-ISCOMs和Pp受免鱼的相对免疫保护率分别为73.3%和40%。研究表明变形假单胞菌ISCOMs口服免疫可提高大黄鱼抗变形假单胞菌感染能力,具有良好的应用前景。本研究为大黄鱼内脏白点病的免疫防控提供了技术支撑。Pseudomonas plecoglossicida is the main pathogen which causes the visceral white spot disease in large yellow croaker(Larimichthys crocea),leading to serious economic loss.Previous studies have shown that immune stimulating complexes(ISCOMs)have good oral immune effects.To evaluate the oral immune effects of P.plecoglossicida immune stimulating complexes(Pp-ISCOMs)on L.crocea,Pp-ISCOMs vaccine was prepared,which presented a cage-like nanoparticle structure.At 21 d post oral vaccination,the expression of immune-related genes in the head-kidney of immunized L.crocea were detected,and the immune serum titer was measured using agglutination tests.The immune protective effects of Pp-ISCOMs were then determined by challenge assay with P.plecoglossicida.The results showed that the expression of immune or anti-inflammatory factors such as myeloid differentiation factor 88(MyD88)(P<0.05),interleukin 2(IL-2)(P<0.001)in the head kidney of Pp-ISCOMs-immunized L.crocea were significantly higher than those of the inactived-P.plecoglossicida-immunized group(Pp)or the blank control group(CK);and the agglutination titer of large yellow croaker serum(21 d post immunization)was 22.After a P.plecoglossicida attack with a dose of 5×10^(2) CFU/g by intraperitoneal injection,the relative percent survival(RPS)of L.crocea vaccinated by PpISCOMs and Pp were estimated to be 73.3%and 40%respectively.The results indicate that the Pp-ISCOMs could improve the resistance of L.crocea to P.plecoglossicida,and suggested a promising prospect for application.This study provides technical support for the prevention and control by immune to visceral white spot disease in L.crocea.

关 键 词：变形假单胞菌 免疫刺激复合物(ISCOMs) 口服 大黄鱼 免疫效果 

分 类 号：S943[农业科学—水产养殖]