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草鱼暴发性出血病病原分离、毒力基因检测与药敏分析
引用本文:徐先栋,付辉云,饶毅,盛银平,王海华,傅义龙,李彩刚,周永灿.草鱼暴发性出血病病原分离、毒力基因检测与药敏分析[J].上海海洋大学学报,2016,25(3):350-358.
作者姓名:徐先栋  付辉云  饶毅  盛银平  王海华  傅义龙  李彩刚  周永灿
作者单位:海南大学 海洋学院, 海南 海口 570228;江西省水产科学研究所, 江西 南昌 330039,江西省水产科学研究所, 江西 南昌 330039,江西省水产科学研究所, 江西 南昌 330039,江西省水产科学研究所, 江西 南昌 330039,江西省水产科学研究所, 江西 南昌 330039,江西省水产科学研究所, 江西 南昌 330039,江西省水产科学研究所, 江西 南昌 330039,海南大学 海洋学院, 海南 海口 570228
基金项目:国家科技支撑计划项目(2012BAD25B07);海南省重点科技计划项目(ZDXM20120005);江西省发明专利产业化技术示范项目(20143BBM26038);江西省水产科学研究所青年基金项目(SC201302)
摘    要:从草鱼体内分离到致病菌株Jxsks1,该菌株对草鱼的LD50值为2.3×103CFU/g鱼体质量,为强致病菌;经生理生化、16S r DNA和gyr B基因序列测序鉴定为嗜水气单胞菌。ERIC-PCR分型结果表明,该菌株为迄今未见报道的类型。选取丝氨酸蛋白酶ahp、热稳定性肠毒素ast、气溶素aer A、热不稳定性肠毒素alt、鞭毛基因fla、脂酶lip等6种毒力基因特异性引物对该菌株进行PCR检测,结果表明其毒力基因型为aer A~+、alt~+、ast~+、ahp~+、lip~+、fla~-。选取28种抗生素进行药敏检测结果表明,该菌株对喹诺酮类、氨基糖苷类、四环素类、氯霉素、第二代和第三代头孢类等药物敏感,对硝基呋喃类、大环内酯类、多肽类中度敏感,对第一代头孢类、青霉素类、复方新诺明和洁霉素耐药。

关 键 词:嗜水气单胞菌  ERIC-PCR分型  毒力因子  药敏试验
收稿时间:2015/6/18 0:00:00
修稿时间:2015/9/25 0:00:00

The virulence characteristics and drug susceptibility of outbreak hemorrhage disease pathogen isolated from grass carp Ctenopharyngodon idllus
XU Xiandong,FU Huiyun,RAO Yi,SHENG Yinping,WANG Haihu,FU Yilong,LI Caigang and ZHOU Yongcan.The virulence characteristics and drug susceptibility of outbreak hemorrhage disease pathogen isolated from grass carp Ctenopharyngodon idllus[J].Journal of Shanghai Ocean University,2016,25(3):350-358.
Authors:XU Xiandong  FU Huiyun  RAO Yi  SHENG Yinping  WANG Haihu  FU Yilong  LI Caigang and ZHOU Yongcan
Institution:College of Marine Science, Hainan University, Haikou 570228, Hainan, China;Jiangxi Institute for Fisheries Science, Nanchang 330039, Jiangxi, China,Jiangxi Institute for Fisheries Science, Nanchang 330039, Jiangxi, China,Jiangxi Institute for Fisheries Science, Nanchang 330039, Jiangxi, China,Jiangxi Institute for Fisheries Science, Nanchang 330039, Jiangxi, China,Jiangxi Institute for Fisheries Science, Nanchang 330039, Jiangxi, China,Jiangxi Institute for Fisheries Science, Nanchang 330039, Jiangxi, China,Jiangxi Institute for Fisheries Science, Nanchang 330039, Jiangxi, China and College of Marine Science, Hainan University, Haikou 570228, Hainan, China
Abstract:A strong pathogenic bacterial strain Jxsks1 was isolated from grass carp, Ctenopharyngodon idllus. The pathogen was identified as Aeromonas hydrophila based on the physiological and biochemical analysis and the sequencing of housekeeping gene 16S rDNA and gyrB. The LD50 of the bacteria infecting the grass carp via intraperitoneal injection was 2.3×103 CFU/g. The results of genetic typing by ERIC-PCR (enterobacterial repetitive intergenic consensus PCR)-fingerprinting showed that it was a type of A. hydrophila which has not been reported before. The distribution patterns of six virulence genes, including aerolysin (aerA), heat-labile cytotonic enterotoxin (alt), heat-stable cytotoxic enterotoxin (ast), elastase (ahpB), lipase (lip) and flagella (fla), were tested by PCR method and the results showed that its virulence-gene pattern was aerA+, alt+, ast+, ahp+, lip+, fla-. The drug susceptibility test of 28 antibiotics by paper extension method showed that the strain was susceptible to quinolones, aminoglycoside, tetracyclines, chloramphenicol, the second and the third generation cephalosporins, intermediately sensitive to nitrofurans, macrolides and polypeptides, but resistant to the first generation cephalosporins, penicillins, sulfamethoxazole compound and lincomycin.
Keywords:Aeromonas hydrophila  ERIC-PCR  virulence gene  drug susceptibility test
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