文章摘要
韩琳,王秀华,杨冰,万晓媛,武和英,张庆利,黄倢.一例日本囊对虾暴发性死亡的病原分析[J].水产学报,2018,42(3):431~441
一例日本囊对虾暴发性死亡的病原分析
Analysis of pathogen in an outbreak death of Marsupenaeus japonicus
投稿时间:2016-12-01  修订日期:2017-05-25
DOI:10.11964/jfc.20161210623
中文关键词: 日本囊对虾  暴发性死亡  病毒  细菌  病因分析
英文关键词: Marsupenaeus japonicus  sudden death  virus  bacteria  etiology analysis
基金项目:“九四八”项目(2016-X56);公益性行业(农业)科研专项(201103034)
作者单位E-mail
韩琳 上海海洋大学水产与生命学院, 上海 201306
中国水产科学研究院黄海水产研究所, 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, 农业部海水养殖病害防治重点实验室, 青岛市海水养殖流行病学与生物安保重点实验室, 山东 青岛 266071 
 
王秀华 中国水产科学研究院黄海水产研究所, 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, 农业部海水养殖病害防治重点实验室, 青岛市海水养殖流行病学与生物安保重点实验室, 山东 青岛 266071 wangxh@ysfri.ac.cn 
杨冰 中国水产科学研究院黄海水产研究所, 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, 农业部海水养殖病害防治重点实验室, 青岛市海水养殖流行病学与生物安保重点实验室, 山东 青岛 266071  
万晓媛 中国水产科学研究院黄海水产研究所, 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, 农业部海水养殖病害防治重点实验室, 青岛市海水养殖流行病学与生物安保重点实验室, 山东 青岛 266071  
武和英 中国水产科学研究院黄海水产研究所, 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, 农业部海水养殖病害防治重点实验室, 青岛市海水养殖流行病学与生物安保重点实验室, 山东 青岛 266071  
张庆利 中国水产科学研究院黄海水产研究所, 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, 农业部海水养殖病害防治重点实验室, 青岛市海水养殖流行病学与生物安保重点实验室, 山东 青岛 266071  
黄倢 中国水产科学研究院黄海水产研究所, 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, 农业部海水养殖病害防治重点实验室, 青岛市海水养殖流行病学与生物安保重点实验室, 山东 青岛 266071  
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中文摘要:
      为了阐明山东省潍坊市一对虾养殖场发生日本囊对虾暴发性死亡的原因,采用分子生物学检测方法,对发病对虾进行了白斑综合征病毒(white spot syndrome virus, WSSV)、桃拉综合征病毒(Taura syndrome virus, TSV)、黄头病毒(yellow head virus, YHV)、传染性皮下及造血组织坏死病毒(infectious hypodermal and hematopoietic necrosis virus, IHHNV)、传染性肌肉坏死病毒(infectious myonecrosis virus,IMNV)、偷死野田村病毒(covert mortality nodavirus,CMNV)及急性肝胰腺坏死病(acute hepatopancreatic necrosis disease, AHPND)7种病原的检测,且对发病对虾进行了常规组织病理学观察。同时采用16S rDNA细菌鉴定方法及浸泡回接感染实验对分离自发病对虾体内的可疑病原菌进行了分子鉴定及毒力测试。结果显示,发病对虾样品核酸检测呈现WSSV强阳性,IHHNV和CMNV为弱阳性,其他4种病原为阴性。组织病理学观察发现,在对虾的胃、鳃等上皮组织中存在WSSV包涵体,头部肌肉纤维出现离散。对分离编号为2901、2902、2903的3株优势可疑病原菌鉴定结果显示,3株菌分别与印度格里蒙菌、交替单胞菌及溶藻弧菌相似,相似度分别为99%、99%及100%。攻毒结果显示,3株可疑病原菌的LC50分别为9.8×107 、1.1×108与2.3×108 CFU/mL,各细菌毒力均较弱,非导致对虾出现暴发性死亡的病原。研究表明,导致本次日本囊对虾暴发性死亡的病因与混合感染病原WSSV、IHHNV、CMNV有关,其中WSSV感染是造成日本囊对虾暴发性死亡的主因,研究结果可为解析当前养殖日本囊对虾疾病暴发及其成因提供参考。
英文摘要:
      In order to clarify the cause of sudden death that occurred in a shrimp (Marsupenaeus japonicus) farm in Weifang from July to August 2015, seven main shrimp pathogens including White spot syndrome virus (WSSV), Taura syndrome virus (TSV), Yellow head virus (YHV), Infectious hypodermal and hematopoietic necrosis virus (IHHNV), Infectious myonecrosis virus (IMNV), Covert mortality nodavirus (CMNV) and Vibrio alginolyticus that causing acute hepatopancreatic necrosis disease (AHPND) were detected by Polymerase chain reaction (PCR) method. Meanwhile, histopathology of the diseased shrimp was observed. Furthermore, bacterial strains, isolated from the moribund shrimp and identified by the 16 SrDNA gene, were used for the evaluating of virulence by immersion challenge. The results showed that all the three shrimp samples gave strong positive PCR results (an amplicon of 941 bp) with primers WSSV 146 F2/R2, and two of three sample show weak positive results (an amplicon of 389 bp) with primers IHHNV 389 F/R and two of three sample show strong positive results (an amplicon of 167 bp) with primers CMNV-7 F2/R2. However the results of PCR amplification for the other four pathogens (TSV, YHV, IMNV and AHPND) show negative. Histopathology results show that there were inclusion body of WSSV in the epithelial tissue of stomach and gill, and the muscle fibres dispersed. Three dominant strains of bacterial microorganisms 2901, 2902, 2903 were isolated and 16 S rDNA sequencing indicated that they had higher similarity with Grimontia indica (99%)、Alteromonas sp. (99%) and Vibrio alginolyticus (100%) respectively. Challenge test show that LC50 of bacteria 2901, 2902, 2903 were 9.8×107 CFU/mL, 1.1×108 CFU/mL and 2.3×108 CFU/mL, which means the bacterial microorganisms had lower virulence and may not be the main pathogen of shrimp. Comprehensive analysis suggested that sudden death of M.japanicus might be caused by coinfection of WSSV, IHHNV and CMNV, in which WSSV infection might play a key role. The results of this study could provide strong support for clarifying the cause of outbreak death in shrimp M. japonicus.
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