河流弧菌(Vibrio fluvialis)感染后牙鲆(Paralichthys olivaceus)血清抗菌物质的研究  

ANTIBACTERIAL SUBSTANCE IN PARALICHTHYS OLIVACEUS SERUM INFECTED BY VIBRIO FLUVIALIS

作　　者：邹文政[1] 纪荣兴[1] 鄢庆枇[1] 覃映雪[1] 苏永全[2] 徐晓津[1] 邢颜丽[1] 

机构地区：[1]集美大学水产学院福建省高校水产科学与食品安全重点实验室,厦门361021 [2]厦门大学海洋学系,厦门361005

出　　处：《海洋与湖沼》2010年第4期477-482,共6页Oceanologia Et Limnologia Sinica

基　　金：国家"863"高技术研究发展计划;2007AA09Z115号;厦门市科技项目;3502Z20073019号;3502Z20073020号;集美大学水产科学技术与食品安全省高校重点实验室开放基金项目;2008J103号

摘　　要：采用1×107cells/ml的河流弧菌悬液背部肌肉注射感染牙鲆,注射后24h、48h试验组的血清抗菌活力分别极显著性(P<0.01)和显著性(P<0.05)高于对照组。用Sephadex G-25凝胶柱对第二次人工注射感染后24h牙鲆血清进行分离,其中第7—15收集管对河流弧菌具有抗菌活性,对大肠杆菌、枯草芽孢杆菌等指示菌也具有明显的抗菌作用,在4—100℃范围内,大部分活性分离组分随着温度的升高其抗菌活力有所增强。具有抗河流弧菌活性收集管(第9管)经弱阳离子交换柱分离后,OD280显示出2个蛋白吸收峰,具有抗河流弧菌活性的物质主要集中在第1吸收峰。SDS-PAGE分析显示经Sephadex G-25凝胶柱和弱阳离子交换柱分离的抗菌蛋白分子量较大。以上结果表明,牙鲆被河流弧菌感染后能很快产生大量抗菌物质释放到血清中;主要抗菌物质的分子量较大,且热稳定性较好;这些活性物质的抗菌活性具有较强的特异性。To investigate the antibacterial substance in Paralichthys olivaceus serum upon infection,P.olivaceus were injected intramuscularly at the dorsal region with Vibrio fluvialis（1×107cells/ml）.The antibacterial activity of the serum of infected P.olivaceus increased significantly in comparison to the control group at 24h and 48h post-injection.Sera of another group of P.olivaceus infected by V.fluvialis were collected at 24h post-injection.Three protein peaks were obtained by fractionating the sera with a Sephadex G-25 column;and the component in the first peak（No.7—15） exhibited strong anti-V.fluvialis activity.The antibacterial range of the antibacterial fractions were determined by six strains of indicator bacteria,among which Escherichia coli and Bacillus subtilis were inhibited.The antibacterial activity of the antibacterial fractions increased with the temperature from 4℃ to 100℃.The antibacterial fractions（No.9） were separated by ion-exchange chromatography,and two main peaks were obtained.The extractions in the first peak exhibited antibacterial activities against V.fluvialis.SDS-PAGE showed the antibacterial products were of high-molecular-weight.These results indicate that P.olivaceus produces multi-antibacterial-component and releases them into serum soon after being infected by V.fluvialis;the major antibacterial proteins are of high molecular weight and rather stable from 4℃ to 100℃;and the antibacterial substances have specific antibacterial activity.

关 键 词：河流弧菌 牙鲆 抗菌物质 抗菌活性 

分 类 号：S917[农业科学—水产科学]