【作者】 王蓓； 吴灶和； 简纪常； 鲁义善； 蔡双虎； 汤菊芬；

【Author】 WANG Bei,WU Zao-he,JIAN Ji-chang,LU Yi-shan,CAI Shuang-hu, TANG Ju-fen (Fisheries College of Guangdong Ocean University,Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals & Key Laboratory of Diseases Controingl for Aquatic Economic Animals of GuangDong Higher Education Institutions,Zhanjiang 524025,China)

【机构】 广东海洋大学水产学院,广东省水产经济动物病原生物学及流行病学重点试验室暨广东省高等学校水产经济动物病害控制重点实验室；

【摘要】 建立溶藻弧菌(Vibrio alginolyticus)的生物被膜(BF)体外形成模型,用银染法观察检测溶藻弧菌HY9901生物被膜的形成;同时研究温度和消毒剂对HY9901的游离(FC)细菌和具有生物被膜(BF)的细菌的影响。实验结果表明,溶藻弧菌HY9901株可以形成BF,而非致病株ZJ017不形成BF。HY9901株的BF细菌经85℃30min、-20℃80d、0.01%新洁尔灭处理后均有存活,而游离(FC)细菌则无存活细菌。根据已发表的哈氏弧菌LuxR基因核酸序列,设计并合成了一对引物,对溶藻弧菌HY9901株的基因组进行PCR扩增和测序分析,扩增出了520bp的核苷酸片段,该片段与已发表的哈氏弧菌Lux L基因的同源性达95%,而非致病性菌株ZJ017株则扩增不出该基因片段。更多还原

【Abstract】 Vibrio alginolyticus biofilm（BF） model in vitro was established.The BF formings of HY9901 and ZJ017 strains were detected by AgNO₃ solution staining.The influence of temperature and disinfectant on free cells（FC） and BF of HY9901 was tested.The pathogenic strain HY9901 could form BF,while the non-pathogenic strains ZJ017 couldn’t.HY9901 strain BF bacteria could survive from being treated respectively with 85℃for 30 min,-20℃for 80 day,and 0.01%benzalkonium bromide silution while HY9901 FC bacteria hardly survived under the above conditions.A pair of primers were designed on the basis of the nucleotide sequences of transcriptional regular gene（Lux R） of Vibrio alginolyticus species reported.One target fragment about 520 bp was amplified from atrain HY9901 and ZJ017 genomin DNA via PCR.Sequencing and analysis of PCR products were performed.The nucleotide sequence showed 95%homology of Vibrio harveyi to the sequence of Lux R gene registered in GenBank.It suggested that the Lux R gene was not only connected with virulence,but also with BF formation.更多还原