绵羊大肠埃希菌生物被膜形成及相关基因的检测  

Detection of Biofilm Formation and Biofilm-associated Genes in E. coli XJ10 from Infected Sheep

作　　者：柴迎锦 邬琴[1] 顾晓晓 郭强强[1] 周霞[1] 韩猛立[2] 张星星[2] 黄新[2] CHAI Ying-jin;WU Qin;GU Xiao-xiao;GUO Qiang-qiang;ZHOU Xia;HAN Meng-li;ZHANG Xing-xing;HUANG Xin(College of Animal Science and Technology,Shihezi University,Shihezi,Xinjiang,832003,China;State Key Laboratory for Sheep Genetic Improvement and Healthy Production,Xinjiang Academy of Agricultural and Reclamation Science,Shihezi,Xinjiang,832000,China)

机构地区：[1]石河子大学动物科技学院,新疆石河子832003 [2]新疆农垦科学院,省部共建绵羊遗传改良与健康养殖国家重点实验室,新疆石河子832000

出　　处：《动物医学进展》2019年第11期36-39,共4页Progress In Veterinary Medicine

基　　金：省部共建绵羊遗传改良与健康养殖国家重点实验室优秀中青年人才培养引导计划专项项目(SKLSGIHP2016A02);兵团重大科技攻关项目(2014AA001-3);兵团现代农业科技攻关与成果转化计划项目(2016AC018)

摘　　要：检测绵羊大肠埃希菌中的XJ10生物被膜(biofilm,BF)形成情况并确定与BF形成相关基因的携带情况。采用96孔聚苯乙烯微孔板培养XJ10观察BF的形态和测定OD值分析XJ10BF在不同的时间段形成的情况,采用PCR方法检测BF 12种形成相关基因的分布情况。结果表明,在37℃,BF的OD值从2 h开始逐渐升高,24 h左右接近最高峰,维持至36 h后,OD值开始缓慢下降,直至72 h仍维持较高水平,在整个检测时间段内XJ10BF OD值显著高于对照组,且差异显著(P<0.05);6 h时BF形成初具形态,8 h^10 h呈现不完整的网格结构,12 h时BF形成较明显的网状结构,24 h^36 h形成有序的较为密集的网格结构,48 h后网状结构开始解离和脱落。同时在XJ10中检测到了其中8种与BF形成相关的基因,测序结果与GenBank公布的相应序列同源性为97.2%以上。试验表明XJ10具有形成BF的能力,但形成能力较弱,且携带8个与BF形成相关的基因。This experiment was performed to detect the biofilm(BF)formation of E.coli XJ10 from infected sheep and to determine the biofilm-associated genes.96-well polystyrene microplate culture and OD value were used to observe the BF formation of XJ10 in different period of time and detect distribution of biofilm-associated genes by molecular biological ways.The results showed that the OD values of BF gradually increased at 2 h,reached a peak at 24 h and continued until 36 h.Then the OD values began to decline slowly until 72 h,the XJ10 BF OD value was significantly higher than that of the control group during the whole test period,and the difference was significant(P<0.05);The BF began to form at 6h,an incomplete lattice structure was formed within 8-10 hours,BF had formed a more obvious network structure at 12 h,an ordered and dense grid structure had been observed at 24 h-36 h,the network structure began to dissociate and fall off after 48 h.At the same time,8 biofilm-associated genes were detected in XJ10 with PCR.The homologous of sequence of PCR amplified fragment was more than 97.2%compared to that of Genbank.XJ10 has the ability to form BF very weak,and it carries 8 of the 12 genes associated with BF formation.

关 键 词：绵羊大肠埃希菌 生物被膜 生物被膜形成相关基因 

分 类 号：S852.612[农业科学—基础兽医学]