Escherichia coli O157:H7 Curli Fimbriae Promotes Biofilm Formation, Epithelial Cell Invasion, and Persistence in Cattle

被引:17
作者
Sheng, Haiqing [1 ]
Xue, Yansong [1 ]
Zhao, Wei [1 ]
Hovde, Carolyn J. [1 ]
Minnich, Scott A. [1 ]
机构
[1] Univ Idaho, Bistate Sch Food Sci, Moscow, ID 83844 USA
基金
美国国家卫生研究院;
关键词
curli; E; coli O157; biofilm; invasion; cattle; SALMONELLA-TYPHIMURIUM; GENOME SEQUENCE; BINDING CURLI; O157-H7; STRAIN; COLONIZATION; PREVALENCE; EXPRESSION; PROTEIN; K-12;
D O I
10.3390/microorganisms8040580
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Escherichia coli O157:H7 (O157) is noninvasive and a weak biofilm producer; however, a subset of O157 are exceptions. O157 ATCC 43895 forms biofilms and invades epithelial cells. Tn5 mutagenesis identified a mutation responsible for both phenotypes. The insertion mapped within the curli csgB fimbriae locus. Screening of O157 strains for biofilm formation and cell invasion identified a bovine and a clinical isolate with those characteristics. A single base pair A to T transversion, intergenic to the curli divergent operons csgDEFG and csgBAC, was present only in biofilm-producing and invasive strains. Using site-directed mutagenesis, this single base change was introduced into two curli-negative/noninvasive O157 strains and modified strains to form biofilms, produce curli, and gain invasive capability. Transmission electron microscopy (EM) and immuno-EM confirmed curli fibers. EM of bovine epithelial cells (MAC-T) co-cultured with curli-expressing O157 showed intracellular bacteria. The role of curli in O157 persistence in cattle was examined by challenging cattle with curli-positive and -negative O157 and comparing carriage. The duration of bovine colonization with the O157 curli-negative mutant was shorter than its curli-positive isogenic parent. These findings definitively demonstrate that a single base pair stably confers biofilm formation, epithelial cell invasion, and persistence in cattle.
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页数:17
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