致病性哈维氏弧菌:海洋寄生性纤毛虫的内共生细菌     

QIAO Ying  WANG Jun  MAO Yong  LIU Min  CHEN Ruanni  SU Yongquan  KE Qiaozhen  HAN Kunhuang  ZHENG Weiqiang 《海洋学报(英文版)》2017,36(10):115-119

哈维氏弧菌被观察到与海洋寄生性纤毛虫刺激隐核虫具有内共生现象,其作为一种致病性细菌,可导致感染刺激隐核虫的大黄鱼产生严重的继发性细菌感染。我们通过16s宏测序技术对刺激隐核虫的细菌群落进行鉴定,并通过标准的细菌培养方法分离鉴定出哈维氏弧菌。通过透射电镜和荧光原位杂交方法,在刺激隐核虫的细胞胞质中,均观察到内共生的哈维氏弧菌的存在;而在其细胞核中,则没有相关信号存在。哈维氏弧菌与刺激隐核虫的相关关系以及内共生的哈维氏弧菌对刺激隐核虫的影响作用,仍需要深入进行研究。  相似文献   

一株副溶血弧菌噬菌体VPp1 的分离鉴定及裂解性能     

彭勇  丁云娟  林洪  王静雪 《海洋科学》2013,37(1):96-101

为探究副溶血弧菌(Vibrio parahaemolyticus)的生物防治方法,从水产品市场处污水中分离出一株副溶血弧菌噬菌体 VPp1.并借助噬菌斑形态、电镜、酶切等技术对其进行了分类鉴定,同时测定了其裂解谱、最佳感染复数、一步生长曲线以研究其裂解性能.分类鉴定结果表明,其核酸是线型双链DNA,大小在15 kb 左右.具有一个正二十面体的头部,头部直径大约为44 nm,无尾,属盖噬菌体科(Tectivirus);裂解性能研究结果表明,其在双层平板上培养12 h 可形成中心清亮的噬菌斑,周围有大而明显的晕环.最佳感染复数为0.0001,潜伏期为10 min,裂解量为90.3,是符合条件的潜伏期短裂解量大的理想噬菌体,可用作进一步的应用.  相似文献   

海洋细菌脂肪酸的气相色谱分析     

徐敏  王静  柴子涵  牟海津 《海洋科学》2013,37(2):76-83

以副溶血性弧菌VP-X-3为试验对象,利用气相色谱技术分析了其细胞脂肪酸的组成和含量,以色谱峰的数量、色谱峰高及脂肪酸含量作为评价指标,探讨了不同的皂化和甲酯化条件对细菌细胞脂肪酸测定结果的影响。结果表明:皂化和甲酯化条件都对细胞脂肪酸的组成和含量产生一定的影响。较低温度皂化处理利于增加色谱峰数量和色谱峰高。甲酯化温度过高容易破坏脂肪酸,尤其是长链脂肪酸和不饱和脂肪酸的结构。高温甲酯化条件下,时间越长,甲酯化效果越差。试验得到较为理想的气相色谱前处理条件:2 mol/L NaOH-甲醇溶液2 mL,70℃水浴10 min;10%(V/V)H2SO4-甲醇溶液2 mL,70℃水浴15 min。此条件下的色谱峰的数量多达29种,色谱峰高明显高于其他处理条件下的峰高。  相似文献   

鳗弧菌(Vibrio anguillarum)对青蛤(Cyclina sinensis)的毒性及半致死浓度研究   总被引：1，自引：1，他引：0  

葛端阳  赵婷  潘宝平 《海洋与湖沼》2012,43(6):1192-1195

利用不同浓度的鳗弧菌对青蛤进行了急性毒性实验,观察青蛤在菌液胁迫下的存活情况,进一步统计得到鳗弧菌对青蛤的半致死浓度。结果表明,青蛤在鳗弧菌液浓度OD600为1.0—2.2范围内,胁迫96h以内死亡率呈逐渐上升趋势,并且在OD600=2.2时所有受试个体全部死亡。在相同的胁迫时间内,受试个体死亡率与胁迫浓度均呈正相关。经SPSS16.0软件分析后,最终得到鳗弧菌对青蛤的半致死浓度(LC50)为OD600=1.57,说明鳗弧菌对青蛤有明显的毒害作用。  相似文献   

河流弧菌(Vibrio fluvialis)对大黄鱼( Pseudosciaena crocea ) 鳃粘液粘附特性研究   总被引：1，自引：0，他引：1  

朱苏琴  纪荣兴  苏永全  王军  覃映雪  马英  邹文政  李海平  鄢庆枇 《海洋与湖沼》2012,43(2):389-393

采用间接ELISA法(最低检测值约104cfu)研究不同培养条件、不同培养阶段、抗体处理、营养饥饿等对河流弧菌粘附作用的影响。结果表明,经TSB培养的菌体的粘附作用极显著强于TSA培养的菌体的粘附作用(P<0.01);河流弧菌能很好地粘附于大黄鱼粘液,粘附量随菌浓度升高而增大并符合饱和粘附动力学:y=0.1782ln(x)1.6923(R2=0.9810);不同生长阶段河流弧菌的粘附能力不同,在培养初期阶段细菌的粘附量先是随着培养时间的延长而增大,并在培养24h后粘附量达到最大,而后随着培养时间的延长其粘附量急剧下降;抗体处理后河流弧菌的粘附量显著低于对照组(P<0.05);营养饥饿菌体的粘附量明显降低。以上结果表明:海水中的河流弧菌能很好地粘附于大黄鱼鳃粘液,其粘附作用受细菌培养条件、营养状况等自身因素的显著影响。本研究结果有助于了解河流弧菌的流行病学和致病机理。  相似文献   

创伤弧菌和哈氏弧菌双重PCR检测方法的建立及初步应用     

陈福艳  程光平  欧阳贤华  陈晓汉  梁万文  韦友传  黄婷  杨学明  陈明  王瑞  李丽萍  韦明利 《广东海洋大学学报》2013,(3)

根据创伤弧菌(Vibrio vulnificus)的溶细胞毒素基因序列和哈氏弧菌(Vibrio harveyi)的toxR基因序列,分别设计并合成两对特异性引物,通过PCR反应条件优化,测试两种菌的特异性和敏感性,建立双重PCR方法,同时快速检测V.vulnificus和V.harveyi。结果表明:纯培养V.vulnificus和V.harveyi的检测灵敏度分别是12 cfu/mL和18 cfu/mL,与无乳链球菌、海豚链球菌、副溶血弧菌及美人发光杆菌无交叉反应;此PCR检测方法具有良好的特异性、敏感性,具快速、高效等优点,对细菌V.vulnificus和V.harveyi诊断与防治具有较好的临床应用性。  相似文献   

Effects of dietary β-glucan and glycyrrhizin on non-specific immunity and disease resistance of the sea cucumber (<Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka) challenged with <Emphasis Type="Italic">Vibrio splendidus</Emphasis>     

Jie Chang  Wenbing Zhang  Kangsen Mai  Hongming Ma  Wei Xu 《中国海洋大学学报(英文版)》2010,9(4):389-394

Sea cucumbers, Apostichopus japonicus Selenka, were fed diets containing non-immunostimulant (basal diet), 0.2% β-glucan and 0.02% glycyrrhizin in a recirculatory water system for 45 days, and subsequently challenged with Vibrio splendidus by injection at 1.0×10⁸ cfu / sea cucumber for 15 days. Phagocytic capacity (PC), intracellular superoxide anion production (ISAP), lysozyme (LSZ) activity and superoxide dismutase (SOD) activity in the coelomic fluid were analyzed on the 0^th, 5^th, 10^th and 15^th days after injection. Results showed that after the 45-day feeding period, PC, ISAP, LSZ activity and SOD activity in sea cucumbers fed with dietary β-glucan or glycyrrhizin were significantly higher than in those fed with the basal diet. On the 5th day after infection, all the immune parameters examined in the sea cucumbers injected with V. splendidus decreased in value significantly. On the 15^th day, PC, ISAP and LSZ activity returned to levels similar to those on the 0^th day. For the sea cucumbers injected with saline, there were no significant differences in all the immune parameters examined and in the cumulative morbidity during the 15-day challenging trial. After injecting with V. splendidus, the cumulative morbidity of sea cucumbers fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin when challenged with V. splendidus challenged sea cucumber fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin. There was no significant difference in cumulative morbidity between the dietary β-glucan and glycyrrhizin treatments over time.  相似文献   

An extracellular oligopeptide permease may be a potential virulence factor of Vibrio harveyi     

Qingfang He  Jixiang Chen  Caifeng Li 《中国海洋大学学报(英文版)》2011,10(4):343-350

An oligopeptide permease A(OppA)was purified from the extracellular product of Vibrio harveyi SF-1.The molecular weight of the purified protein was estimated to be 58 kDa on SDS-PAGE.The purified protein showed phospholipase C activity at the optimal values of temperature 50℃ and pH 8.0.The enzymatic activity decreased when the temperature increased to 40℃.The N-terminal sequence of the purified protein was determined as ADVPAGTKLA,which is similar to that of OppA.The OppA pre-cursor gene was cloned from th...  相似文献   

Characterization of EmpA protease in Vibrio anguillarum M3     

Yifan Han  Zhaolan Mo  Peng Xiao  Bin Hao  Jie Li  Guanpin Yang 《中国海洋大学学报(英文版)》2011,10(4):379-384

EmpA is an extracellular metalloprotease secreted by Vibrio anguillarum.For better understanding its role in the patho-genicity of V.anguillarum strain M3,empA insertion mutant was constructed.In the mutant it decreased in extracellular proteolytic activity,swarming motility,hemolytic activity and virulence on turbot(Scophthalmus maximus).Significant decline(by 5-fold)of extracellular proteolytic activity and similar growth curve between mutant and wild type strains indicated that EmpA was the major extracellular protease of M3.LD50 of mutant increased by 38-fold compared with wild type.No pro-EmpA was detected in the su-pernatant of culture,indicating that EmpA autolyzed to mature protein after 24 h.Secretion of EmpA in M3 was similar to that in NB10 strain.Attenuated virulence of mutant was similar to that of M93Sm strain.It was demonstrated that specific operation of EmpA was different from that in previous studies and EmpA contributed to the swarming motility and hemolytic activity in V.an-guillarum strain M3.The results provides insight into understanding the function of EmpA and its potential application in vaccine development.  相似文献   

Influence of dietary probiotic Bacillus TC22 and Prebiotic fructooligosaccharide on growth, immune responses and disease resistance against Vibrio splendidus infection in sea cucumber Apostichopus japonicus     

Yancui Zhao  Kangsen Mai  Wei Xu  Wenbing Zhang  Qinghui Ai  Yanjiao Zhang  Xiaojie Wang  Zhiguo Liufu 《中国海洋大学学报(英文版)》2011,10(3):293-300

The effects of probiotic Bacillus TC22 (isolated from intestine of infected sea cucumber) and prebiotic fructooligosaccharide (FOS) on growth, immunity and disease resistance in sea cucumber Apostichopus japonicus were studied. Six experimental diets were formulated with combinations of three levels of TC22 (0, 107 and 109 CFU g-1 diet) and two levels of FOS (0 and 0.5%) in a 3 × 2 factorial experiment. At the end of the 8-week feeding trial,animals were challenged by injecting Vibrio splendidus. The results revealed that the specific growth rates (SGR) of sea cucumbers were not affected by TC22 and FOS, or the interaction between TC22 and FOS (P > 0.05). However,there were significant interactions between TC22 and FOS for immune response and disease resistance in sea cucumbers (P < 0.05). When sea cucumbers were fed with TC22 at 109 CFU g?1 feed and 0.5% FOS alone or in combination, the phagocytosis, respiratory burst and phenoloxidase activity of sea cucumber coelomocytes were significantly enhanced;the disease resistance against V. splendidus infection was also increased significantly (P < 0.05).However, dietary combination of TC22 at 109 CFU g-1 diet and 0.5% FOS did not yield significantly higher efficiency than either TC22 at 109 CFU g-1 diet or 0.5% FOS used alone (P > 0.05). Therefore, further studies should examine the effects of combinations of other levels of FOS(> 0.5% or < 0.5%) and TC22 on the immunity and disease resistance of sea cucumbers.  相似文献