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养殖大菱鲆烂鳍病病原菌的鉴定及系统发育学研究 总被引：9，自引：0，他引：9
从山东半岛3个养殖场各分离到一株大菱鲆烂鳍病的病原菌,均为革兰氏阴性,短杆状,极生鞭毛,有运动能力,菌落半透明。经常规生理生化特征测试和16SrRNA基因序列对比,表明它们都是同一种细菌。理化特征的结果显示这3株菌均与V．anguillarum的表性特征非常相似。对其16SrRNA基因序列进行分析,并构建了系统发育树,结果表明这些菌株与V．anguillarum亲缘关系最近。综合上述研究结果,将该菌鉴定为V．anguillarum. 相似文献
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Chunmei Li Xubo Wang Quanqi Zhang Zhigang Wang Jie Qi Qilin Yi Zhipeng Liu Yanan Wang Haiyang Yu 《中国海洋大学学报(英文版)》2012,11(1):32-44
Major histocompatibility complex class II antigens are important in vertebrate immune system.In the present study,the full cDNA sequence of class II A gene was synthesized by RACE-PCR from half-smooth tongue sole(Cynoglossus semilaevis),and its open reading frame(ORF) polymorphism was studied.The whole cDNA sequence was 992 bp in length,including the ORF with 717 bp.Twenty-five alleles were identified and clustered into two distinct groups according to the specific nucleotides/amino acids in specific positions.Eleven alleles belonged to Cyse-DAA while fourteen alleles belonged to Cyse-DBA.Four Cyse-DAA alleles were observed in one individual,and three to five Cyse-DBA alleles were observed in each of the three detected individuals,which indicated that at least two loci existed in each gene.Moreover,in order to study the function of the alleles in resistance to infection,200 individuals were intraperitoneally injected with Vibrio anguillarum and the first 20 dead individuals and 20 surviving ones were selected for genotype analysis.Fifty-six alleles were identified among the 40 individuals.Twenty-nine alleles belonged to Cyse-DAA and the other 27 alleles belonged to Cyse-DBA.Eighteen alleles were selected for studying their function in resistance to infection.Alleles Cyse-DAA*0201,Cyse-DAA*1101,Cyse-DBA*0401,Cyse-DBA*1102,Cyse-DBA*1801 and Cyse-DBA*2201 were identi-fied only in surviving individuals,while alleles Cyse-DAA*0901,Cyse-DBA*1101 and Cyse-DBA*1401 occurred more frequently in dead individuals.This study confirmed the existence and polymorphism of two class II A genes as well as the relationship between alleles of class II A genes and disease susceptibility/resistance in half-smooth tongue sole. 相似文献
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Vibrio anguillarum is a common bacterial pathogen in fish.However,little is known about its pathogenic mechanism,in part,because the entire genome has not been completely sequenced.We constructed a fosmid library for V.anguillarum containing 960 clones with an average insert size of 37.7 kb and 8.6-fold genome coverage.We characterized the library by end-sequencing 50 randomly selected clones.This generated 93 sequences with a total length of 57 485 bp covering 1.4% of the whole genome.Of these sequences,58... 相似文献
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Bacteria-infected turbots Scophthalmus maximus with septicaemia were examined between 2001 and 2004 in aspects of the conditions of disease occurrence, clinical syndromes and pathological changes. The phenotypic information of pathogenic bacteria was studied, including morphology, physiological and biochemical characteristics, and the mol% G C of the DNA. In addition, representative strains (S010623-1, LH031120-1) were selected for molecular identification by partial 16S rRNA gene sequencing. The results show that the isolates (LH031120-1 to LH031120-6, HT040308-1 to HT040308-6, HT040620-1 to HT040620-6) from three farms were identified as Edwardsiella tarda. The isolates (S010610-1 to S010610-10, S010623-1 to S010623-20) from one farm were identified as Listonella anguillarum. We conducted studies on the pathogenicity of isolates by artificial infection, and revealed all infected groups in morbidity and mortality. The septicaemia infected turbot showed a syndrome similar to that of the naturally infected fish. Antibiotic sensitivity showed that of 37 antimicrobial agents, E. tarda was sensitive to 27 agents, and L. anguillarum was sensitive to 21 agents. 相似文献
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江汉平原江陵地区9ka以来古气候演化的沉积记录 总被引：1，自引：0，他引：1
从一起自然发病的中华绒螯蟹(Eriocheir sinensis H．Milne-Edwards)肝胰腺中,分离做纯培养的6株(HQ010516A-1至HQ010516A-6)鳗利斯顿氏菌(Listonella anguillarum),进行了形态特征、主要理化特性等方面的检验．同时择代表菌株(HQ010516A-1)测定了16S rRNA基因序列、构建了系统发育树．另外,以代表菌株(HQ010516A-1)对健康蟹进行了人工感染试验,结果表明了该菌株在所检病例的病原学意义;用37种抗菌类药物对3株菌所做的药敏试验,结果均表现对头孢噻肟等29种药物呈现敏感或高度敏感(抑菌圈直径在16-40 mm)、对头孢唑啉等3种药物呈低度敏感(抑菌圈直径在10-12 mm)、对青霉素G等5种药物耐药(无抑菌圈形成)． 相似文献
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为了研究大菱鲆(Scophthalmus maximus)中趋化因子(Chenokine)的特征及其在大菱鲆免疫过程中发挥的作用,设计了本文中的实验。本实验从大菱鲆基因组和转录组数据库中鉴定了一个硬骨鱼特有的趋化因子CC亚家族成员—CCL34,并选取大菱鲆的8个健康组织以及2种细菌感染后的肠道和皮肤组织,采用实时荧光定量PCR(qRT-PCR)对其表达特征进行研究。序列分析结果显示该趋化因子全长mRNA包含1个327 bp的5’非编码区(UTR),1个246 bp的3’非编码区,和1个编码103个氨基酸残基长度为312 bp的开放阅读框(ORF)。此CCL34基因含有4个外显子和3个内含子。通过系统发育分析、共线性分析,将该大菱鲆趋化因子命名为CCL34。实时荧光定量PCR表明CCL34在大菱鲆健康组织中普遍表达,尤其在肾脏、肝脏、皮肤中有高水平表达。鳗弧菌(Vibrio anguillarum)和无乳链球菌(Streptococcus agalactiae)感染后,肠道组织CCL34基因表达量较对照组显著性上调(p<0.05),这表明CCL34可能在大菱鲆的肠道粘膜免疫中起重要作用。本研究为趋化因子家族功能的研究奠定了基础,为增强鱼类免疫力和抗病能力提供了理论依据。 相似文献
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本研究将新发现的鳗弧菌毒力相关基因mltD(膜绑定溶胞壁质转糖酶基因)克隆于表达载体pET-32a(+),在大肠杆菌BL21(DE3)中诱导表达,并以镍琼脂糖亲和层析柱纯化表达的融合蛋白;经SDS-PAGE分析,纯化的融合蛋白为单一条带,分子量约为70.2 kDa,与理论预测值相符。生物信息学分析表明,鳗弧菌MltD由523个氨基酸组成,与其他弧菌的MltD氨基酸序列有较高的相似性;二级结构以α螺旋和无规则卷曲为主;功能区包括N端的信号肽区域、1个糖基转移酶结构域和C端的3个溶解酶结构域;蛋白的N端具有一段脂蛋白信号肽,信号肽外的其他部分为非细胞质蛋白。鳗弧菌MltD蛋白为亲水性;不稳定系数为27.40,属于稳定性蛋白;整个蛋白分子共有24个可能的抗原决定簇,具有较强的抗原性。推断MltD蛋白为1种外周膜脂蛋白,在弧菌中相对保守,抗原性强,可应用于抗弧菌病疫苗的开发研制。 相似文献
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TANG Xiaoqian ZHOU Li ZHAN Wenbin 《青岛海洋大学学报(英文版)》2008,(3):343-351
Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among 14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD50 value of 〈1.0×10^5 colony forming units （CFU）mL^-1, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillantm with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay （IFA） was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature. 相似文献