分光光度法测定海洋河弧菌510脂多糖     

张运涛  顾谦群  方玉春  吕元萌  管华诗 《中国海洋大学学报(自然科学版)》2003,33(3):355-360

本文研究了直接测定海洋河弧菌 (vibrio fluvailis) 5 10脂多糖的分光光度法。利用 1,9-二甲基亚甲蓝 (DMB) ,在适当条件下同脂多糖发生染色反应 ,在 5 5 0 nm波长下 ,反应体系的吸光值同脂多糖含量呈线性相关。相关系数为 0 .9993,最低检出限为 10 μg/m L,相对标准偏差为 1.6 2 ,平均回收率为 (10 5± 2 .18) %。  相似文献   

鳗弧菌(Vibrio anguillarum)M3菌株生长条件及其对蛋白酶产量的影响   总被引：2，自引：5，他引：2  

陈师勇  张培军  莫照兰  邹玉霞  张振冬  徐永立 《海洋与湖沼》2004,35(2):159-166

采用体外测定细菌浓度、胞外产物(ECP)蛋白含量和蛋白酶活力的方法，进行了鳗弧菌M3菌株在2216E培养基中的培养条件研究。结果表明，该菌株用固体培养基培养至24h左右，可得到较高的菌体浓度、ECP蛋白含量和蛋白酶活力。采用响应面分析方法设计实验，用SAS统计软件分析数据，得到NaCl浓度、pH值和温度对菌体生长及蛋白酶产量影响的回归模型。在2216E培养基的基础上，添加不同氮源、碳源物质以及不同浓度蛋白胨进行生长研究。结果表明，胰大豆蛋白胨能促进菌体生长及ECP蛋白分泌；NH4Cl与酪蛋白水解物可抑制蛋白酶的产生；牙鲆肌肉匀浆对菌体、ECP蛋白产量和蛋白酶产生有不同程度的促进作用；培养基中蛋白胨浓度为4％时菌体量与ECP蛋白含量达最高值，在蛋白胨浓度为2％时蛋白酶分泌量已稳定；1％的葡萄糖、蔗糖、甘油均能显著地提高菌体及ECP蛋白产量，却抑制了蛋白酶的产生。  相似文献   

哈维氏弧菌的生物学特性、流行病学及检测技术   总被引：4，自引：0，他引：4  

张晓华  钟英斌  陈吉祥 《中国海洋大学学报(自然科学版)》2007,37(5):740-748

哈维氏弧菌是1种革兰氏阴性、发光的海洋细菌,广泛分布于海洋环境中。它是最近10多年才被认识到的水产养殖动物的重要致病菌。文中综述哈维氏弧菌的生物学特性、流行病学、致病机制、密度感应系统、检测技术、疾病的防治及其应用。  相似文献   

用16S rRNA基因的内切酶图谱快速鉴别几种对虾病原菌   总被引：4，自引：0，他引：4  

孔杰  刘萍  张岩  杨丛海  叶军  徐怀恕  郭华荣 《海洋与湖沼》1997,28(5):449-452

坎普氏弧菌是青岛海洋大学生物系微生物实验室于１９８９－１９９０年自对虾养殖场中国对虾红腿病心脏及血淋巴中分离并鉴定的菌株，副溶血菌和溶藻胶弧菌两菌株于１９９４年９月得自中国科学院微生物研究所，为建立快速、准确的中国对虾病原菌的诊断技术，根据几种细菌的１６ＳｒＲＮＡ基因的序列，设计并合成该基因的多聚酶反应的引物ＰＬ１和ＰＬ２。并用该对引物分别从坎普氏弧菌、副溶血弧菌和溶藻胶弧菌的ＤＮＡ要品中扩增出分  相似文献   

Rapid detection of virulent protease secreted by Vibrio anguillarum by dot enzyme-linked immunosorbent assay     

ZHANG Zhendong  ZHANG Peijun  MO Zhaolan  WANG Chunling  YU Yang 《海洋学报(英文版)》2005,24(4):155-161

Dot enzyme-linked immunosorbent assay （dot-ELISA）, indirect ELISA and Westem blot were performed to detect the virulent protease secreted by Vibrio anguillarum which was isolated from the diseased left-eyed flounder, Paralichthys olivaceous. Sensitivity results showed that dot-ELISA is a more sensitive, rapid and simple technique for the protease detection. The minimal detectable amount of protease is about 7 pg in the dot-ELISA test, while 7.8 ng in the indirect ELISA and 6.25 ng in the Westem blot respectively. Protease could be detected 2 h after incubation of V. anguillarum in the 2216E liquid medium but enzyme activity was very low at that period. From 6 to 12 h, the amount and enzyme activity of protease increased markedly and reached maximum at stationary phase. Analysis of serum samples periodically collected from the infected flounders showed that after 2 h of infection by V. anguillarum, the pathogenic bacteria could be detected in the blood of the infected flounders but no protease was found. It was 5-6 h after infection that the protease was detected in blood and then the amount increased as infection advanced. Quantitative detection of protease either incubation in the medium or from the blood of infected flounders could be accomplished in virtue of positive controls of quantificational protease standards （＂marker＂） so that the alterations ofprotease secretion both in vitro and in vivo could be understood generally. In addition, the indirect ELISA and dot-ELISA were also performed to detect V. anguillarum cells. Results indicated that the sensitivity of indirect ELISA to bacteria cells is higher than that of the dot-ELISA, and that the minimal detectable amount is approximately 10^4 cell/mL in the indirect ELISA, while 10^5 cell/mL in the dot-ELISA.  相似文献   

最小弧菌热不稳定型溶血素基因克隆与序列分析     

李涛  吴后波 《台湾海峡》2006,25(3):324-329

根据已发表的最小弧菌热不稳定型溶血素（heat-labile hemolysin）基因核苷酸序列，设计和合成一对特异性引物，以最小弧菌96-6-3的基因组DNA为模板，PCR扩增得到热不稳定型溶血素基因片段，克隆到质粒载体pMD-18T中进行测序和分析．其结果表明扩增的热不稳定型溶血素基因片段与已发表的最小弧菌热不稳定型溶血素的同源性高达98％，扩增片段编码由446个氨基酸组成的多肽，推测分子量为50200．软件分析表明编码的氨基酸有较高的抗原性，可作为基因工程疫苗的候选基因片段．  相似文献   

人工养殖点带石斑鱼弧菌病病原菌的分离及鉴定   总被引：8，自引：1，他引：8  

陈晓燕  胡超群  陈偿  张吕平  任春华 《海洋科学》2003,27(6):68-72

广东、海南两省多个海水网箱养殖场养殖的点带石斑鱼(Epinephelus malabaricus)发生严重的弧菌病,从海南三亚和广东深圳的发病鱼病灶上分离到2株致病性细菌VAL02和VAL00,经人工回接感染实验证明这2株菌引起点带石斑鱼弧菌病,VAL02对点带石斑鱼苗的半致死量是2.25×10~5 cfu/g。经形态及生理生化特征分析,鉴定出VAL02及VAL00均为溶藻弧菌。应用药敏纸片法研究了22种化学疗剂对VAL02及溶藻弧菌标准株的生长抑制作用,发现VAL02对氯霉素、氟哌酸、卡那霉素和乙酰螺旋霉素等产生耐药性,对复方新诺明和环丙沙星等敏感。  相似文献   

褐藻胶裂解酶产生菌Vibro sp.QY102的发酵条件优化     

傅晓妍  李京宝  韩峰  路新枝  于文功 《中国海洋大学学报(自然科学版)》2007,37(3)

对海洋来源的Vibro sp.QY102的产褐藻胶裂解酶的发酵条件进行研究。结果表明,该菌株最适液体培养基成分为(w/v):0.5%褐藻酸钠;0.4%蛋白胨;0.3%KH2PO4;0.7%K2HPO4.3H2O;2%NaCl;0.01%MgSO4.7H2O,pH=6.0。按3%的接种量接入培养基,30℃150 r/min振荡培养120 h,产酶达到10.2 U/mL,为优化前的4.5倍。Mg2 是该菌株产酶所必需的,这在其他褐藻胶裂解酶生产菌株中未见报道。该菌株产酶发酵条件的研究,为褐藻胶裂解酶的大规模制备及应用奠定了基础。  相似文献   

Pathogenicity of vibrios in fish: An overview   总被引：1，自引：0，他引：1  

Norman Y.S.Woo 《中国海洋大学学报(英文版)》2003,2(2):117-128

Bacteria of the genus Vibrio are ubiquitously distributed in the marine environment. Due to the rapid expansion of intensive mariculture and the consequent deterioration of culture conditions, more and more Vibrio spp. have been recognized as pathogenic agents in outbreaks of vibriosis, a serious epizootic disease affecting most wild and farmed fish species worldwide, which has become the most important limiting factor for the development of intensive mariculture industry. Attempts have been made to understand the pathogenicity of vibrios in host fish with the ultimate aim of elucidating the best means for disease control. After an extensive literature survey of the recent advances in the field of fish vibriosis, the pathological changes, virulence factors and associated potential pathogenic mechanisms, transmission routes and related environmental factors involved in outbreak of vibriosis, as well as the controlling strategies are reviewed in the present paper.  相似文献   

不同条件下哈维氏弧菌(Vibrio harvey)对墨吉明对虾(Fenneropenaeus merguiensis)致病性研究     

赵吉臣  王刚  邓中宁  陈兆明  孙成波 《海洋与湖沼》2015,46(6):1413-1419

为研究墨吉明对虾弧菌病的防控方法, 本文通过人工注射感染的方式, 研究不同数量哈维 氏弧菌及在不同温度、盐度、pH、不同硝酸氮、亚硝酸氮和氨氮浓度条件下哈维氏弧菌对墨吉明对 虾的致病性的影响。结果表明: 当人工感染哈维氏弧菌数量为3.16×10⁸ CFU 时, 24 h 内对虾累计死 亡率为96.67%; 哈维氏弧菌数量为3.16×10⁶ CFU 以上, 实验结束时, 对虾累计死亡率都超过了 50.00%.在20-32℃ 范围内, 随着温度升高哈维氏弧菌致病力增强, 并且低温组(20℃ 实验组与 26℃ 实验组)对虾累计死亡率显著低于高温组(32℃ 实验组)(P<0.05).在温度为(24±1)℃ 时, 盐度为 20 时对虾累计死亡率处于较低水平; 在温度为(24±1)℃, 盐度为(22±1)时, 96 h, pH 为8.0 时对虾 累计死亡率最低, 并且与另外两个实验组差异显著(P<0.05); 温度为(24±1)℃, 盐度为(22±1), 硝 酸氮对感染哈维氏弧菌的墨吉明对虾有胁迫作用; 在亚硝酸氮浓度(1.0-10.0 mg/L)、氨氮浓度 (0.5-1.5 mg/L)范围内, 哈维氏弧菌对墨吉明对虾的致病性, 随着浓度升高而增强。本实验为墨吉明 对虾弧菌病的防控提供了理论基础。  相似文献