共培养沙门菌对粪肠球菌N41株生长特性及毒力基因的影响  

Influence of the growth traits and virulence genes of Enterococcus faecalis isolate N41 by cocultivation with Salmonella

作　　者：武二丽[1] 和春昊 金钺[1] 刘玲玲[1] 胡慧[1,2] 曹晗[1] 王亚宾[1,2] 陈丽颖[1,2] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]河南省动物性食品安全重点实验室,河南郑州450002

出　　处：《中国兽医学报》2017年第8期1528-1533,共6页Chinese Journal of Veterinary Science

基　　金：河南省重点科技攻关资助项目(152102310081)

摘　　要：为探讨共培养条件下沙门菌对粪肠球菌N41分离株生长特性及其不同生长时期26种毒力基因转录的影响,将N41菌株、沙门菌菌株单独/混合培养后,分别进行菌落计数并绘制生长曲线,观察其生长状况及影响,同时提取其对数中期、对数后期、稳定前期和稳定期的总RNA,SYBR GreenⅠ实时荧光定量PCR方法分析N41菌株26种毒力基因的转录情况。结果显示,共培养后沙门菌和N41生长均受到影响,其中,在N41菌体浓度降低约3.9倍,而沙门菌菌体浓度降低约110倍。在N41菌株26种毒力基因中,沙门菌促进了ebpA、ebpC、rnjB、ace、ebpR、psr等12种毒力基因在4个不同生长时期的转录,但同时也降低了毒力基因SlyA和sprE在4个生长期中转录水平;另外,除了CylL-S、CylL-L、efaA和AS在4个生长时期变化不明显外,其余的大部分毒力基因都在对数后期转录水平有明显上升,但在其他生长时期则表现不同。这说明,两菌共培养后,N41菌株明显抑制了沙门菌的生长,同时沙门菌也整体促进了N41菌株毒力基因的转录。本试验为研究细菌间相互作用机制以及肠球菌的致病机制奠定基础。To explore the impact of Salmonella on Enterococcusfaecalis isolate N41 under the coex-isting,the growth traits and the transcription of 26 virulence genes of N41 at various growth pha- ses were detected. Salmonella and/or N41 were inoculated and done plate counting,then growth curves were drawn and bacterial total RNA were extracted at given time points, quantitive real- time PCR was used to analyze the RNA transcription levels of 26 virulence genes of N41. The result showed that comparison with single incubation, the bacteria concentration of N41 dropped a-bout 3.9 times,and Salmonella dropped about ll0-times. Among 26 virulence genes of N41, the RNA transcription levels of 12 virulence genes such as ebpA,ebpC,rnjB,ace,ebpR, psr and so on were promoted at the four growth phases, but the RNA transcription levels of SlyA and sprE were dropped. Except that the RNA transcription levels of CylL-S ,CylL-L ,efaA and AS had no signifi- cant changes at the four growth phases, the mostly rest genes increased dramatically at post-log phase. This indicated that when incubated together, N41 inhibited the growth of Salmonella signif- icantly,and Salmonella promoted the transcription levels of virulence genes of N41 as a whole.The results lay a foundation for further research on the interaction between bacteria species and the pathogenicity mechanism of Enterococci.

关 键 词：粪肠球菌N41 沙门菌 生长特性 毒力基因 荧光定量PCR 

分 类 号：S852.61[农业科学—基础兽医学]