渔业科学进展2018，Vol.39Issue(3)：103-111,9.DOI:10.19663/j.issn2095-9869.20170222001
凡纳滨对虾急性肝胰腺坏死病(AHPND)病原分离鉴定及其致病性分析
Identification and Pathogenicity Analysis of Bacterial Pathogen Associated with Acute Hepatopancreatic Necrosis Disease (AHPND) in the Pacific Shrimp Litopenaeus vannamei
摘要
Abstract
Acute hepatopancreatic necrosis disease (AHPND) is an emerging shrimp disease causing great losses for the shrimp culture industry worldwide since 2010. In the present study, a bacterial strain 20160303005-1 was isolated from the hepatopancreas tissue of Litopenaeus vannamei with early mortality syndrome (EMS), and was identified as Vibrio parahaemolyticus based on its physiological and biochemical characteristics; and the analysis of both 16S rRNA and groEL gene sequence. The serotype of the bacterium is O1:KUT (K untypeable). It revealed positive amplification of the genes pirAVP and pirBVP which is related to cause AHPND in a virulence plasmid harboring this strain. However, the isolate examined showed negative amplification results for the virulent clinical V. parahaemolyticus strain markers—thermostable direct hemolysin gene tdh and TDH-related hemolysin gene trh. The immersion challenge test with L. vannamei was also employed to study pathogenicity and histopathology. The results showed that the isolate was highly virulent, with a median lethal dose (LD50) value of 7.96×103 CFU/ml. The empty gut in shrimp was observed at 6 h post-challenged. The hepatopancreas appeared pale and atrophic at 9 h. More than half of the shrimps died at 12 h, and up to 100％ died at 24 h. Subsequent histological analyses showed that the hepatopancreas tubules collapsed with massive sloughing of hepatopancreas epithelial cells, which was the typical pathological characteristics of AHPND. Among 21 antibiotics tested, the isolate was resistant to amoxicillin, cefalotin, ticarcillin, cefuroxime, and cotrimoxazol; however, it was sensitive to gentamicin, ciprofloxacin, and other 14 antibiotics tested. These results provide basic data for epidemiology and drug control research on V. parahaemolyticus in aquaculture.关键词
凡纳滨对虾/急性肝胰腺坏死病/副溶血弧菌/细菌鉴定Key words
Litopenaeus vannamei/AHPND/Vibrio parahaemolyticus/Bacterial identification分类
农业科技引用本文复制引用
贾丹,谢国驷,史成银,黄倢,张庆利,万晓媛,许华,刘冉阳,王海波,郭程程..凡纳滨对虾急性肝胰腺坏死病(AHPND)病原分离鉴定及其致病性分析[J].渔业科学进展,2018,39(3):103-111,9.基金项目
中国水产科学研究院基本科研业务费专项(2017HY-ZD0502 （）
2017HY-ZD1002)、青岛海洋科学与技术国家实验室鳌山科技创新计划项目(2015ASKJ02)、现代农业产业技术体系(CARS-48)、山东省泰山学者建设工程专项(S41201203)共同资助[This work was supported by Central Public-Interest Scientific Institution Basal Research Fund, CAFS (2017HY-ZD0502 （2015ASKJ02）
2017HY-ZD1002), the Scientific and Technological Innovation Project Financially supported by Qingdao National Laboratory for Marine Science and Technology (2015ASKJ02), China Agriculture Research System (CARS-48), and Construction Programme for "Taishan Scholarship" of Shandong Province of China (S41201203)]. （2015ASKJ02）
