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为研究海洋附着细菌的群落结构及动态变化,在厦门近岸海区进行挂板实验.将无菌玻璃板浸没于海水中,连续放置14 d.分别于放置1 h和7、14 d后取玻璃板上附着生物样品.用细菌通用引物构建16S rRNA基因克隆文库,每个克隆文库随机挑选约40个克隆子测序,序列同源性分析和系统进化分析结果表明,所有的克隆子可分为六大类群:γ-变形菌纲(γ-Proteobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、α-变形菌纲(α-Proteobacteria)、蓝细菌门(Cyanobacteria)和真核硅藻类叶绿体,各类群分别占42.0%、4.5%、2.2%、2.2%、1.1%和45.0%.γ-变形菌纲变形斑沙雷氏菌(Serratia proteamaculans)为优势附着细菌,占测序克隆子的31.5%.这类细菌在1 h样品中的比例超过一半,说明变形斑沙雷氏菌在生物膜形成初期发挥着重要作用.随着挂板时间延长,检测到的细菌类群有所增加:附着7 d后检测到拟杆菌门细菌,附着14 d后检测到厚壁菌门细菌.γ-变形菌纲细菌所占比例随挂板时间的延长而逐渐降低,从挂板1 h的81%降至7 d的21%,14 d的18%.另外,在各阶段的附着样品中,都检测到较多的真核克隆子序列,约占16%~64%.本研究为进一步阐明海洋附着细菌的附着动态及其在生物膜形成过程中的作用奠定了基础. 相似文献
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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
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
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大黄鱼病原副溶血弧菌单克隆抗体制备及其应用 总被引:5,自引:0,他引:5
用甲醛灭活副溶血弧菌(Vibrio parahaemolyticus)和溶藻弧菌(V.alginolyticus)制成免疫原免疫BALB/c小鼠,利用淋巴细胞杂交瘤技术,获得1株特异的针对副溶血弧菌的单克隆抗体,命名为D6F3H5。腹水及培养上清液抗体的ELISA效价分别为:1∶5 120和1∶1 280,该单克隆抗体与其它细菌没有明显的交叉反应。利用该单克隆抗体和兔抗副溶血弧菌多克隆抗体,建立了检测副溶血弧菌的双抗体夹心ELISA方法。该方法对副溶血弧菌的最小检出量为5×104个/mL。用双抗体夹心ELISA检测大黄鱼(Pseudosciaena crocea)样品,14尾患病大黄鱼中有11尾检出副溶血弧菌,而10尾健康大黄鱼都没有检出副溶血弧菌。由此可见,本试验制备的高特异性的抗副溶血弧菌单克隆抗体,可以用于患病大黄鱼副溶血弧菌的快速诊断。 相似文献
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用平板涂布法从集美海边分离到381株细菌,用平板划线法或点种法检验这些海洋细菌对两株病原弧菌(副溶血弧菌(Vibrio Parahaemolyticus)和溶藻弧菌(Vibrio alginolyticus)的拮抗作用。从中挑选2株对海水养殖动物病原弧菌有较好拮抗作用的细菌,编号为GX75和GZ46。对拮抗菌(GX75和GZ46进行了抗菌谱测定,结果表明,GX75对9株指示菌中的副溶血弧菌、河流弧菌和坎普氏弧菌有较好的拮抗作用,GZ46对副溶血弧菌、溶藻弧菌和坎普氏弧菌有较好的桔抗作用。在营养肉汤中,测定了GX75和GZ46的抗菌作用,表明(GX75和GZ46对副溶血弧菌拮抗作用很明显。 相似文献
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应用本实验室建立的趋化模型对海水鱼类寄生虫刺激隐核虫(Cryptocaryon irritans)的趋化特性进行研究。结果显示, 刺激隐核虫幼虫在30min时趋化达到饱和, 在温度28℃时趋化率达到最高; 多种硬骨鱼的血清与粘液对刺激隐核虫的幼虫都有着强烈的吸引作用, 刺激隐核虫幼虫对碳水化合物、氨基酸等小分子物质没有趋化作用; 在组织匀浆液中, 刺激隐核虫幼虫对肌肉匀浆液有一定的趋化作用, 对肝、肠、胃的匀浆液都没有趋化作用; 感染后鱼类血清对刺激隐核虫的幼虫有着更加强烈的吸引作用(P<0.01); 18种中草药中的五倍子、大黄、槟榔、苦楝、虎杖、贯众等6种中药草对刺激隐核虫的幼虫有一定的驱虫作用。本文的研究结果有助于深化对刺激隐核虫致病性的认识并对“白点病”的防治有一定的参考价值。 相似文献
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Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups. Each bullfrog in bacterium-injected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension at a density of 5.2 × 10^6 CFU/ml, while each one in control group injected i.p. with 0.2 ml sterile saline solution (0.85%, w/v). Three bullfrogs in both groups were sampled at 0, 1, 3, 7, 11, 15 and 20 days post-injection (dpi) for the evaluation of non-specific immune parameters. It was observed that intraperitoneal injection of A. hydrophila significantly increased the number of leucocytes and that of NBT-positive cells in peripheral blood. Significant increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control group. However, a significant reduction was detected in vitro in phagocytosis activity of peripheral blood phagocytes. No significant difference in changes in the number of peripheral erythrocytes, serum superoxide dismutase (SOD) activity, and lysozyme activity was detected between the two groups. It is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the A. hydrophila infection. 相似文献
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用点种法从大黄鱼肠道优势菌群中筛选出104株病原性副溶血弧菌的拮抗菌,进一步研究拮抗效果最好的11株拮抗菌的抗菌谱和生长曲线,其中NAC平板上筛选的4株拮抗菌的抗菌范围较广,锰营养琼脂培养基上筛选的3株拮抗菌具有生长优势。用BIOLOG板测定了抗菌谱较广、有一定生长优势3株拮抗菌(R22,Y58和J312)的碳源利用情况,结果表明,3株拮抗菌的碳源谱都比较广,都能利用糖类、有机酸和氨基酸等,而且与鱼类营养的竞争较小。16S rRNA基因同源性分析的结果显示与R22,Y58,J312同源性达99%以上的菌株分别来自弧菌属、芽孢杆菌属和假单胞菌属。进一步对R22,J312和Y58三株拮抗菌进行生化鉴定,结果显示R22为溶藻弧菌,J312为荧光假单胞菌,但Y58的种属未能得到确认。所得到的3株拮抗菌R22,Y58,J312在拮抗病原菌、抗菌谱、生长特性和营养利用等方面都初步满足益生菌的筛选条件,有待进一步检测其安全性和实用效果以便能够成为能实用的益生菌。 相似文献
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Vibrio fluvialis incubated in trypticase soy broth (TSB) showed stronger adhesion to the skin mucus of Epinephelus awoara than V. fluvialis grown on trypticase soy agar (TSA), and this bacterial adhesion was assessed in terms of saturation kinetics. Treating bacteria
with antibody against O-antigens resulted in significantly reduced bacterial adhesion. In the early growth stage, the adhering
bacteria numbers increased with incubation time, peaked at 24 h, and then dropped sharply. Prior heat treatment of the mucus
at various temperatures resulted in different effects on bacterial adhesion. No remarkable effect on bacterial adhesion was
detected after treating mucus with protease K and trypsin, whereas periodic acid treatment significantly enhanced bacterial
adhesion. Extracts of V. fluvialis surface structures at high concentrations significantly inhibited bacterial adhesion, while relatively low concentrations
enhanced adhesion. V. fluvialis had strong affinity for a high molecular weight mucus component. These results indicate that V. fluvialis adhered strongly to E. awoara mucus by specific adhesion and that this adhesion was influenced by many factors, including bacterial conditions, mucus,
and the environment. 相似文献