摘要
建立嗜水气单胞菌(Aeromonashydrophila,Ah)的生物被膜(BF)体外形成模型,用银染法及扫描电镜观察检测Ah致病株J1、N1及非致病株W1、4332株产生BF的情况,同时研究温度和消毒剂对AhJ1株的游离(FC)细菌(以下简称FC细菌)和具有BF的细菌(以下简称BF细菌)的影响。实验表明:致病株AhJ1、N1株均可形成BF,而非致病株AhW1、4332株均不形成BF。AhJ1株的BF细菌经85℃30min、-20℃80d、0.01%新洁尔灭处理后均有存活,而FC细菌无存活细菌。根据已发表的AhA1株的转录调节蛋白基因(AhyR)核苷酸序列,设计并合成了一对引物,对AhJ1、N1、W1、4332株的基因组进行PCR扩增和测序分析。发现致病性AhJ1、N1株可扩增出912bp的核苷酸片段,该片段与已发表的AhAhyR基因的同源性达99%,而非致病性AhW1、4332株无该基因片段。
Aeromonas hydrophila （Ah） biofilm （BF） model in vitro was established. The BF formings of Ah J-1, N-l, W-1 and 4332 strains were detected by AgNO3 solution staining and scanning electron microscopy （SEM）. The influence of temperature and disinfectant on free cells （FC） and BF ofAh J-1 was tested. The pathogenic strain Ah J-1 and N-1 could form BF, while the non-pathogenic strains Ah W-1 and 4332 couldn＇t. Ah J-1 strain BF bacteria could survive from being treated respectively with 85 ℃ for 30 min, - 20 ℃ for 80 day, and 0. 01% benzalkonium bromide solution while Ah J-1 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 （AhyR） of Aeromonas species reported. One target fragment about 912 bp was amplified from strain Ah J-1 and N-1 genomic DNA via PCR, but not from strain Ah W-1 and 4332. Sequencing and analysis of PCR products were performed. The nucleotide sequence showed 99% homology of Ah J-1 to the sequence of Ah A1 AhyR gene registered in GenBank. It suggested that the AhyR gene was not only connected with virulence, but also with BF formation.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2005年第3期75-78,共4页
Journal of Nanjing Agricultural University
基金
江苏省科技攻关项目(三药)(BE2004609)
