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| 副溶血弧菌(Vibrio parahaemolyticus)中溶源噬菌体与其宿主菌致病力的相关性 |
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王 娜1,2, 王海亮1, 白 楠1, 黄 倢1,2,3, 谢达祥4, 谢起浪5, 於俊琦5, 万夕和6
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1.中国水产科学研究院黄海水产研究所 农业部海洋渔业可持续发展重点实验室 青岛 266071;2.上海海洋大学水产与生命学院 上海 201306;3.青岛海洋科学与技术国家实验室 海洋渔业科学与食物产出过程功能实验室 青岛 266237;4.广西水产科学研究院 南宁 530021;5.浙江省海洋水产养殖研究所 温州 324005;6.江苏省海洋水产研究所 南通 226007
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| 摘要: |
| 急性肝胰腺坏死病(Acute Hepatopancreatic Necrosis Disease, AHPND)是由副溶血弧菌(Vibrio parahaemolyticus)引起的对虾病害，本研究从患AHPND的凡纳滨对虾样品中分离得到5株副溶血弧菌，采用致AHPND的副溶血弧菌(VPAHPND)的相关质粒的引物AP2进行PCR检测，表明这5株菌中均存在AHPND相关质粒。利用丝裂霉素C进行溶源性噬菌体筛选和噬菌体诱导发现，其中2株副溶血弧菌(20130629002S01和20130726001S01)可能存在溶源性噬菌体感染；从经0.5 μg/ml丝裂霉素C诱导的20130629002S01和20130726001S01中分别分离得到两种噬菌体phage1和phage2。透射电镜观测显示，phage1为有尾噬菌体，phage2为球形噬菌体。将上述5株副溶血弧菌进行卤虫无节幼体人工感染实验，结果显示，它们对卤虫无节幼体均有致病力，且各分离株的毒力表现出显著性差异；20130629002S01和20130726001S01两株带有溶源性噬菌体的副溶血弧菌的致病力显著低于无噬菌体的副溶血弧菌(20130721001S02)。本研究结果表明，5株VPAHPND分离株都含有AHPND相关的质粒，表现出显著的毒力差异，可能携带不同的溶源噬菌体，也可能不携带溶源噬菌体，溶源噬菌体与副溶血弧菌各分离株的毒力并无必然相关性。 |
| 关键词: 副溶血弧菌 溶源噬菌体 急性肝胰腺坏死病 |
| DOI：10.11758/yykxjz.20150417002 |
| 分类号: |
| 基金项目:公益性行业科研专项经费项目(201103034)、现代农业产业技术体系(CARS-47)、山东省泰山学者建设工程专项经费和农业部科研杰出人才和创新团队专项经费共同资助 |
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| Isolation of Lysogenic Phage in Vibrio parahaemolyticus and Its Relationship with the Pathogenicity of the Host Bacteria |
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WANG Na11,2, WANG Hailiang1, BAI Nan1, HUANG Jie1,2,3, XIE Daxiang4, XIE Qilang5, YU Junqi5, WAN Xihe6
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1.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;2.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306;3.Function Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200;4.Guangxi Academy of Fishery Science, Nanning 530021;5.Zhejiang Mariculture Research Institute, Wenzhou 324005;6.Jiangsu Institute of Oceanology and Marine Fisheries, Nantong 226007
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| Abstract: |
| Acute hepatopancreatic necrosis disease (AHPND) is caused by infection with Vibrio parahaemolyticus. In this study, five strains of V. parahaemolyticus were isolated from Litopenaeus vannamei with AHPND. The primer set AP2 targeting the plasmid harbored in AHPND-causing V. parahaemolyticus (VPAHPND) was used in the PCR amplification, and it was found that all five isolates carried the AHPND related plasmid. Mitomycin C was used to induce and screen the lysogenic phages in VPAHPND isolates. The growth curves showed that two isolates of V. parahaemolyticus, 20130629002S01 and 20130726001S01, contained lysogenic phages. Phage 1 and Phage 2 were released from 20130629002S01 and 20130726001S01 respectively after the induction with 0.5 μg/ml mitomycin C. Transmission electron microscopy revealed that Phage 1 had a tailed shape and Phage 2 had a spherical shape. The challenge test on Artimia nauplii suggested that the virulence was significantly different between the 5 strains of V. parahaemolyticus. The pathogenicity of two lysogenic phages-containing strains, 20130629002S01 and 20130726001S01, was significantly lower than that of phage-free 20130721001S02. Our study suggested that the presence of lysogenic phage might not necessarily correlate with the pathogenicity of the host bacterium V. parahaemolyticus. |
| Key words: Vibrio parahaemolyticus Lysogenic phage Acute Hepatopancreatic Necrosis Disease (AHPND) |