| 对虾养殖场底泥中副溶血性弧菌的接合型质粒多样性及其与毒力之间的关系 |
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| 引用本文： | 郝婧薇,华昕彤,傅松哲,周灿,刘鹰,胡惠秩.对虾养殖场底泥中副溶血性弧菌的接合型质粒多样性及其与毒力之间的关系[J].水产学报,2023,47(3):039416-039416. |
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| 作者姓名： | 郝婧薇 华昕彤 傅松哲 周灿 刘鹰 胡惠秩 |
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| 作者单位： | 大连海洋大学海洋科技与环境学院，辽宁 大连 116023;大连海洋大学，设施渔业教育部重点实验室，辽宁 大连 116023;湖北大学环境与资源学院，湖北 武汉 430062 |
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| 基金项目： | 国家重点研发计划 (2019YFD0900500)；辽宁省教育厅科研经费(QL202005)；广东省重点研发计划 (2019B020215001，GML2019ZD0402)；国家自然科学基金 (81903372) |
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| 摘 要： | 为了阐明引起急性肝胰腺坏死病(acute hepatopancreatic necrosis disease, AHPND)副溶血性弧菌的接合型质粒在对虾养殖环境中的遗传多样性，实验从中国5个沿海省份的虾场收集了100个底泥样品，以质粒上编码接合转移蛋白的保守基因为目标，利用PCR法检测相关质粒的存在情况，并对质粒进行测序。结果显示，100个样品中有39个样品含有质粒的接合转移蛋白片段。从100个底泥样品中分离出15株副溶血性弧菌，其中13株含有1～2个质粒。质粒序列测序结果显示，这些质粒可分为8种类型/谱型，其中7种不携带pirAB，但均含有编码接合转移的基因簇。根据分离副溶血性弧菌携带质粒的8种谱型，分别选择8株副溶血性弧菌进行凡纳滨对虾攻毒实验，发现这些菌株对凡纳滨对虾的毒性有显著差异，实验虾死亡率为15%～100%。只有pirAB阳性菌株会对实验虾产生AHPND症状，死亡率为100%。对质粒组成进行分析表明，质粒之间遗传物质交换频繁，大部分质粒的遗传组成都来自一个183 kb的超大质粒pVP2HP。综上，本实验通过探究对虾养殖场底泥中结合性质粒的多样性，增强了人们对副溶血性弧菌...
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| 关 键 词： | 对虾 养殖 急性肝胰腺坏死病(AHPND) 副溶血性弧菌 质粒多样性 |
| 收稿时间： | 2021/3/8 0:00:00 |
| 修稿时间： | 2021/12/31 0:00:00 |
Conjugative plasmid diversity of Vibrio parahaemolyticus in the sediment of shrimp farm and its association with bacterial virulence |
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| HAO Jingwei,HUA Xintong,FU Songzhe,ZHOU Can,LIU Ying,HU Huizhi.Conjugative plasmid diversity of Vibrio parahaemolyticus in the sediment of shrimp farm and its association with bacterial virulence[J].Journal of Fisheries of China,2023,47(3):039416-039416. |
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| Authors: | HAO Jingwei HUA Xintong FU Songzhe ZHOU Can LIU Ying HU Huizhi |
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| Institution: | College of Marine Technology and Environment, Dalian Ocean University, Dalian 116023, China;Key Laboratory of Environment Controlled Aquaculture (KLECA), Ministry of Education, Dalian Ocean University, Dalian 116023, China; College of Environment and Resources, Hubei University, Wuhan 430062, China |
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| Abstract: | Acute hepatopancreatic necrosis disease (AHPND) caused by Vibrio parahaemolyticus is an emerging bacterial disease in cultured shrimp. The disease has caused the huge economic losses to the global shrimp aquaculture industry, with annual losses of over $1 billion. All AHPND related V. parahaemolyticus contain a 70 kb plasmid with genes encoding homologues of the Photorhabdus insect-related (Pir) binary toxin PirABvp, which is key virulence factor for shrimp. In order to clarify the genetic diversity of the conjugative plasmid of the V. parahaemolyticus strain that causes AHPND in the shrimp culture environment and understand the prevalence of APPND-associated plasmid, this study was conducted in five coastal provinces in China. A total of 100 sediment samples were collected from the shrimp farms in the five coastal provinces. PCR was then conducted to amplify the pirAB and conserved genes encoding the conjugative transfer protein on the plasmids. V. parahaemolyticus strains were then isolated from the pirAB-positive sediment samples to obtain the plasmid. Sanger sequencing was then conducted to obtain the sequence of the plasmid. Finally comparative sequence analysis was performed to assess the plasmid diversity. The results showed that 39 out of 100 samples contained plasmid conjugative transfer protein fragments. 15 strains of V. parahaemolyticus were isolated from 100 sediment samples, 13 of which contained 1-2 plasmids. The results of plasmid sequencing showed that these plasmids can be divided into 8 types/profiles, of which 7 do not carry pirAB gene, but all contain gene clusters encoding conjugation transfer. According to the isolation of 8 types of plasmids carried by V. parahaemolyticus, 8 strains of V. parahaemolyticus were selected for prawn challenge experiments. It was found that the toxicity of these strains to prawns was significantly different, and the mortality rate of prawns was between 15%-100%. Only pirAB positive strains will produce AHPND symptoms, and the mortality rate of shrimp is 100%. The analysis of the plasmid composition shows that the genetic material exchange between plasmids is frequent, and the genetic composition of most of the plasmids comes from a 183 kb super-large plasmid pVP2HP. In this study, we found that the conjugative plasmids isolated from the sediments of shrimp farms had rich genetic diversity. Most of the plasmids are genetically composed of plasmid pVP2HP, and only pirAB positive strains will produce AHPND symptoms. This study improved understanding of plasmid genetic diversity of V. parahaemolyticus and its relationship with AHPND, and provided theoretical guidance for the prevention and control of AHPND. |
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| Keywords: | shrimp farming acute hepatopancreatic necrosis disease (AHPND) Vibrio parahaemolyticus plasmid diversity |
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