鮰爱德华菌外膜微孔蛋白ompN1、ompN2和ompN3基因生物信息学分析及截段基因的原核表达与其抗原性检测     

潘延乐  汪开毓  王均  肖孟玮  李岚敏  陈德芳 《中国兽医学报》2015,35(6):900-908

根据GenBank中鮰爱德华菌(Edwardsiella ictaluri)外膜微孔蛋白N(outer membrane porin protein,OMP)基因序列(NC_012779.2)设计引物,以鮰爱德华菌Ms12基因组DNA为模板,经PCR扩增后克隆至pMD19-T载体,将阳性克隆菌送公司测序。测序结果通过生物信息学软件对ompN1、ompN2和ompN3序列的氨基酸序列相对分子质量、分子式、信号肽、跨膜区、亲/疏水性、B细胞表位进行预测,并构建系统发育树;根据去信号肽的B细胞表位相对集中区域设计特异性引物,经PCR扩增后,将截段序列克隆至pMD19-T载体,鉴定成功后克隆至原核表达载体pET32a(+)并转化至BL21(DE3)/Rosseta(DE3)进行IPTG诱导表达,最后用兔抗鮰爱德华菌血清对重组蛋白OmpN1、OmpN2和OmpN3进行免疫原性检测。结果显示扩增得到ompN1(1 143bp)、ompN2(1 056bp)和ompN3(1 053bp)的序列,获得的基因登录号分别为KJ831564、KJ831565和KJ831566。经生物信息学分析显示,OmpN1、OmpN2和OmpN3蛋白分子式分别为C1940H2993N543O622S7、C1817H2723N497O579S5和C1896H2843N519O563S7,相对分子质量大小分别为44 100、40 900和41 800,均为含有1个信号肽和跨膜区的疏水性蛋白,具有1个革兰阴性细菌OM_channels超家族结构域,是1种嵌入型的膜外蛋白;SDS-PAGE电泳检测发现,诱导表达的融合重组蛋白均以包涵体的形式出现在沉淀中,大小分别约为61 800、58 700和59 100;Western-blot分析表明,重组蛋白OmpN1、OmpN2和OmpN3均能与兔抗鮰爱德华菌全菌血清结合,表明鮰爱德华菌外膜微孔蛋白N具有亚单位疫苗开发的潜力。  相似文献   

鳗鲡爱德华氏病病原菌及一新种   总被引：16，自引：2，他引：16       下载免费PDF全文

王国良  徐兴林  路正 《水产学报》1993,17(3):224-229

本文报道在我国浙江省发现的鳗鲡爱德华氏病病原菌特征。从患病鳗鲡内脏分离到9株菌,用其中的E895205等4株菌进行人工感染,死亡率均为100％,与自然发病症状相似。这些菌株的特征一致。均为G~-短杆菌,单个,周鞭毛,兼性厌氧,发酵葡萄糖产酸产气。氧化酶阴性,接触酶阳性,产H_(?)S。不能利用柠檬酸盐和丙二酸盐作为唯一碳源,鸟氨酸和赖氨酶脱羧酶阳性。属爱德华氏菌属细菌。比较已报道的四个种,认为E895205等菌株为一新种,定名为浙江爱德华氏菌(Edwardsiella zhe jiangensis sp. nov.)。  相似文献   

Development of a multiplex PCR assay to detect Edwardsiella tarda,Streptococcus parauberis,and Streptococcus iniae in olive flounder (Paralichthys olivaceus)     

Seong Bin Park  Kyoung Kwon  In Seok Cha  Ho Bin Jang  Seong Won Nho  Fernand F. Fagutao  Young Kyu Kim  Jong Earn Yu  Tae Sung Jung 《Journal of veterinary science (Suw?n-si, Korea)》2014,15(1):163-166

A multiplex PCR protocol was established to simultaneously detect major bacterial pathogens in olive flounder (Paralichthys olivaceus) including Edwardsiella (E.) tarda, Streptococcus (S.) parauberis, and S. iniae. The PCR assay was able to detect 0.01 ng of E. tarda, 0.1 ng of S. parauberis, and 1 ng of S. iniae genomic DNA. Furthermore, this technique was found to have high specificity when tested with related bacterial species. This method represents a cheaper, faster, and reliable alternative for identifying major bacterial pathogens in olive flounder, the most important farmed fish in Korea.  相似文献   

A novel 'skinny pot-belly' disease in Asian seabass fry, Lates calcarifer (Bloch)     

Gibson-Kueh S  Crumlish M  Ferguson HW 《Journal of fish diseases》2004,27(12):731-735

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Blood parameters in Nile tilapia (Oreochromis niloticus L.) spontaneously infected with Edwardsiella tarda   总被引：6，自引：0，他引：6  

A Ca&#;lan Karasu Benli  & Hijran Yavuzcan Yildiz 《Aquaculture Research》2004,35(14):1388-1390

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A multi-epitope chimeric protein elicited a strong antibody response and partial protection against Edwardsiella ictaluri in Nile tilapia     

Vimbai Irene Machimbirike  Natapol Pornputtapong  Saengchan Senapin  Eakapol Wangkahart  Prapansak Srisapoome  Pongsak Khunrae  Triwit Rattanarojpong 《Journal of fish diseases》2022,45(1):1-18

Edwardsiella ictaluri infects several fish species and protection of the all the susceptible fish hosts from the pathogen using a monovalent vaccine is impossible because the species is composed of host-based genotypes that are genetic, serological and antigenic heterogenous. Here, immunoinformatic approach was employed to design a cross-immunogenic chimeric EiCh protein containing multi-epitopes. The chimeric EiCh protein is composed of 11 B-cell epitopes and 7 major histocompatibility complex class II epitopes identified from E. ictaluri immunogenic proteins previously reported. The 49.32 kDa recombinant EiCh protein was expressed in vitro in Escherichia coli BL-21 (DE3) after which inclusion bodies were successfully solubilized and refolded. Ab initio protein modelling revealed secondary and tertiary structures. Secondary structure was confirmed by circular dichroism spectroscopy. Antigenicity of the chimeric EiCh protein was exhibited by strong reactivity with serum from striped catfish and Nile tilapia experimentally infected with E. ictaluri. Furthermore, immunogenicity of the chimeric EiCh protein was investigated in vivo in Nile tilapia juveniles and it was found that the protein could strongly induce production of specific antibodies conferring agglutination activity and partially protected Nile tilapia juveniles with a relative survival percentage (RPS) of 42%. This study explored immunoinformatics as reverse vaccinology approach in vaccine design for aquaculture to manage E. ictaluri infections.  相似文献   

爱德华氏菌疫苗的研究进展     

胡林玲  郭松林  关瑞章  冯建军 《安徽农业科学》2013,(25):10353-10355

爱德华民菌为重要的致病菌之一,能感染多种鱼类并造成大量死亡.该病原菌的暴发已经给美国、欧洲、亚洲（包括日本、中国等）国家带来了严重的社会和经济损失.长期以来,用于鱼类细菌性疾病防治的西药类药物,可导致病原菌产生耐药性,鱼体的药物残留会危害人类健康等,因而被限制使用.针对该菌的鱼类疫苗的研究又再度兴起,包括天然亚单位疫苗、活体重组疫苗、基因缺失苗和基因工程亚单位疫苗等.综述了爱德华氏菌的基本特征和各种疫苗的研究进展,以期为鱼类爱德华氏菌病的防治提供重要的参考资料.  相似文献   

Emergence of Edwardsiella piscicida in Farmed Channel ♀, Ictalurus punctatus × Blue ♂, Ictalurus furcatus,Hybrid Catfish Cultured in Mississippi     

Matt J. Griffin  Stephen R. Reichley  Wes A. Baumgartner  Suja Aarattuthodiyil  Cynthia Ware  James M. Steadman  Marsha Lewis  Patricia S. Gaunt  Lester H. Khoo  David J. Wise 《Journal of the World Aquaculture Society》2019,50(2):420-432

There is a trend toward the increased incidence and prevalence of Edwardsiella piscicida septicemia in US catfish aquaculture, particularly in channel ♀, Ictalurus punctatus, × blue ♂, I. furcatus, hybrid catfish. From 2013 to 2017, a total of 3242 disease case submissions were made to the Aquatic Research and Diagnostic Laboratory (ARDL) at the Thad Cochran National Warmwater Aquaculture Center in Stoneville, MS. Of these, 1400 (43.2%) were hybrids. E. piscicida was suspected in 138 (4.3%) of cases, the majority of which (89.1%) were from hybrid catfish. A molecular survey of these isolates confirmed the majority (92.0%) to be E. piscicida. Furthermore, cases of E. piscicida from hybrids submitted to the ARDL and the Aquatic Diagnostic Laboratory of the Mississippi State University College of Veterinary Medicine in Starkville, MS, were documented for gross lesions and histological analysis. Grossly, E. piscicida presents with small dermal ulcerations, a raised fluid‐filled cranial midline lesion that is frequently ulcerated, hemorrhage in the gills, exophthalmia, and abdominal distension. Internally, lesions include splenomegaly, straw‐colored ascites, renomegaly, and occasionally hemorrhagic intestines. Histopathological examination is in agreement with gross observations, and infected fish repeatedly demonstrate a mononuclear meningoencephalitis, hemorrhagic branchitis, splenitis, ulcerative dermatitis, granulomatous interstitial nephritis, and hepatitis coupled with a hemorrhagic enteritis.  相似文献   

迟缓爱德华氏菌检验程序的研究   总被引：10，自引：0，他引：10  

葛艳  陈怀青  陆承平 《中国动物检疫》1999,16(3):2-4

本试验研究了迟缓德华氏菌（Edwardsiellatrada,Et）的细菌学及致病因子检测,确定了相应检测指标,制定了检验程序。对不同来源的Et作分离及生化鉴定,确定10项生化鉴定指标。用三种方法即平板法（PlateAssay,PA）、接触法（ContactHemolysis,CH）和上清法（SunernatantAs－say,SA）检测Et溶血素,阳性率依次为75％、75％、57．14％。28株Et中,18／26的胞外产物（Extra－cellularProducts,ECP）对HEP－2细胞有细胞毒性,17／28菌株有侵袭力。用ATCC15947株ECP的抗血清进行Det－ELISA,检测阳性率为19／26（73．08％）。  相似文献   

Edwardsiella tarda infection in Korean catfish,Silurus asotus,in a Korean fish farm     

Jin‐Ha Yu  Jung Jo Han  Kwon Sam Park  Kwan Ha Park  Sung Woo Park 《Aquaculture Research》2009,41(1):19-26

Mass mortality of Korean catfish, Silurus asotus, occurred in a culture farm situated in Jeollabukdo Province, Korea. The cumulative mortality rates reached up to 5% of the total fish in the farm per day. In clinical signs, the affected fish showed abdominal distension, vent protrusion, enteritis, liver congestion and abscess‐like lesions in enlarged spleen and kidney. Histopathologically, in the liver, hepatocytes lost fat and underwent atrophy or necrosis. The spleen showed necrotized splenocytes and a haemorrhagic pulp. In the kidney, glomerular destruction, degeneration of renal tubular epithelial cell and haemorrhage were observed. However, necrotic muscular lesions were not observed. A pure bacterial isolate was obtained from the liver, spleen and kidney lesions of affected fish. Experimental infection of normal catfish with the isolate resulted in the development of clinical signs similar to those seen on the farm. The isolates were identified as Edwardsiella tarda through biochemical tests (99.4%) and analysis of bacterial genes (16S rDNA) sequences (98%). The bacteria possessed two virulent genes: sodB and katB genes. These results suggest that E. tarda can act as a pathogen of farmed catfish. This is the first report showing that E. tarda caused mortality in cultured Korean catfish.  相似文献