| 哈氏弧菌胞外产物对红鳍东方鲀的致病性 |
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| 引用本文： | 王斌,于兰萍,袁甜,姜志强.哈氏弧菌胞外产物对红鳍东方鲀的致病性[J].中国水产科学,2010,17(1). |
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| 作者姓名： | 王斌 于兰萍 袁甜 姜志强 |
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| 作者单位： | 大连水产学院,农业部海洋水产增养殖学重点开放实验室,辽宁,大连,116023 |
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| 基金项目： | 辽宁省科学技术计划重大项目 |
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| 摘 要： | 采用平板玻璃纸覆盖技术提取哈氏弧菌(Vibrio harveyi)H-06091菌株的胞外产物(ECP),通过肌肉和腹腔注射2种方式研究其对红鳍东方鲀(Fugu obscurus)的致病性。采用双层纸片法对其主要酶成份进行初步研究;测定其对绵羊红细胞和红鳍东方鲀红细胞的溶血活性;将哈氏弧菌在不同温度下(15℃、20℃、25℃、30℃、35℃、40℃)培养36h和25℃下培养不同时间(12h、18h、24h、36h、48h、60h、72h)后提取其胞外产物并对其蛋白质含量进行测定;经SDS-PAGE分离粗提胞外产物中蛋白成分,并用Dot-ELISA和Western-blot方法对其成分进行分析。实验结果显示,该胞外产物对红鳍东方鲀具有致病性;具有淀粉酶和酪蛋白酶活性,不具有卵磷脂酶、脂肪酶和明胶酶活性;对红鳍东方鲀红细胞有溶血活性,对绵羊红细胞无溶血活性;胞外产物蛋白质总含量在35℃培养36h后出现高值,与温度变化无明显相关性;25℃下培养24h、36h和48h的粗提胞外产物经SDS-PAGE分离,有3条蛋白质条带最明显,分子量分别约为31kD、38kD、60kD;其中分子量约为60kD蛋白条带,经Dot-ELISA和Western-blot方法证实其为半胱氨酸蛋白酶-1。根据以上结果认为哈氏弧菌H-06091菌株胞外产物含有多种毒力因子,与该菌的致病性密切相关。
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| 关 键 词： | 红鳍东方纯 哈氏弧菌 胞外产物 |
Pathogenicity of extracellular products of Vibrio harveyi to Fugu obscurus |
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| WANG Bin,YU Lanping,YUAN Tian,JIANG Zhiqiang.Pathogenicity of extracellular products of Vibrio harveyi to Fugu obscurus[J].Journal of Fishery Sciences of China,2010,17(1). |
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| Authors: | WANG Bin YU Lanping YUAN Tian JIANG Zhiqiang |
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| Institution: | WANG Bin,YU Lanping,YUAN Tian,JIANG Zhiqiang(Key Laboriatory of Mariculture , Biotechnology of Ministry of Agriculture,Dalian Fisheries University,Dalian 116023,China) |
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| Abstract: | Extracellular products(ECPs)were extracted from pathogenic strain H-06091 of Vibrio harveyi with cellophane overlay method.The pathogenesis of ECPs were investigated by intramuscular injection and intraperitoneal injection.The method of double slips was used for detecting the main enzyme ingredients.Moreover,haemolytic activity of ECP to sheep erythrocytes and Fugu obscurus erythrocytes(Fish erythrocytes were collected freshly through venepuncture from healthy fish maintained in aquarium)was determined.ECPs... |
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| Keywords: | Fugu obscurus Vibrio harveyi extracellular products |
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