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当前位置: 首页 > 文章 > 1株大肠杆菌O157鸭源噬菌体的分离、纯化及其特性鉴定 畜牧与兽医 2011,43 (7) 36-38
Position: Home > Articles > Isolation,purification and characterization of an E.coli O157 bacteriophage from duck faeces Animal Husbandry & Veterinary Medicine 2011,43 (7) 36-38

1株大肠杆菌O157鸭源噬菌体的分离、纯化及其特性鉴定

作  者：
王雪松;李晓;杨光远;刘鹏刚;汤芳;张炜;陆承平
单  位：
南京农业大学动物医学院
关键词：
大肠杆菌O157;噬菌体;分离;特性
摘  要：
将安徽某鸭场的400份鸭粪样品离心,微孔滤器过滤后提取上清液,将上清液与均匀涂布于LB固体培养基上的大肠杆菌O157ATCC43889滴定,6 h后初步鉴定噬菌体。将疑似存在噬菌体的上清液采用双层琼脂平板法进一步鉴定,结果分离鉴定了1株噬菌体,命名为EC43889-30。3次纯化后得到直径约为2 mm的噬菌斑,富集得到纯培养物。电镜观察,噬菌体粒子无尾,为直径50 nm左右的正六边形,形态与复层病毒科(Tectiviridae)的成员相似。将纯培养物稀释至107 pfu/mL,每隔10 min(至50 min)分别测其在50℃,60℃和70℃时噬菌体存活数,热稳定曲线显示,50℃时存活率均在70%左右,60℃时存活率明显下降,70℃时完全失活。宿主谱检测结果显示,其宿主范围较窄,对测试的42株鸭源大肠杆菌无裂解作用。
译  名：
Isolation,purification and characterization of an E.coli O157 bacteriophage from duck faeces
作  者：
WANG Xue-song,LI Xiao,YANG Guang-yuan,LIU Peng-gang, TANG Fang,ZHANG Wei,LU Cheng-ping(College of Veterinary Medicine,Nanjing Agriculture University,Nanjing 210095,China)
关键词：
E.coli O157;bacteriophage;isolation;characterization
摘  要：
Four hundreds of faecal samples from ducks in Anhui Province were centrifugated and filtered by 0.22um filters.The supernatants were titrated on LB solid medium pooled with E.coli O157 ATCC43889 and the plaques were picked up after 6 hours.By using the double-deck agar,a strain of bacteriophage named EC43889-30 was isolated.After purification for three times,the plaques in 2mm diameter were enriched.The electron microscopic observation revealed that the bacteriophage had an autho-sexangle head with approximately 50nm diameter,but no tail,which is similar in morphology to a member of Tectiviridae.After the bacteriophages were diluted to 107PFU/mL and the plaques were tested at 50℃,60℃ and 70℃ every 10 minutes in 5 times,respectively,the thermal stability curve was determined.It showed that the survival rate at 50℃ was about 70 percent,declined apparently at 60℃ and died completely at 70℃.The host spectrum of EC43889-30 was narrow and no lysis happened in other 42 E.coli isolates from duck faeces.

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