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EnteropathogenicEscherichia coli (EPEC) is an important causal agent of diarrheal illness throughout the world. Nevertheless, researchers have only recently begun to explore its capacity to form biofilms. Strain O55:H7 (DMS9) is a clinical isolate belonging to the atypical EPEC (aEPEC) group, which displays a high degree of genetic relatedness to enterohemorrhagicE. coli . Strain DMS9 formed a robust biofilm on an abiotic surface at 26°C, but not at 37°C. It also formed a dense pellicle at the air-liquid interface and developed a red, rough, and dry (RDAR) morphotype on Congo red agar. Unlike a previously describedE. coli O157:H7 strain, the aEPEC strain seems to express cellulose. Transposon mutagenesis was used to identify biofilm-deficient mutants. One of the mutants was inactivated in thecsgFG genes, required for assembly and secretion of curli fimbriae, while a second mutant had a mutation incrl , a thermosensitive global regulator that modulates σS activity and downstream expression of curli and cellulose. The two mutants were deficient in their biofilm formation capabilities and did not form a pellicle at the air-liquid interface. Unlike inSalmonella , thecsgFG mutant in aEPEC completely lost the RDAR phenotype, while thecrl mutant displayed a unique RDAR “pizza”-like morphotype. Genetic complementation of the two mutants resulted in restoration of the wild-type phenotype. This report is the first to describe and analyze a multicellular behavior in aEPEC and support a major role for curli and thecrl regulator in biofilm development at low temperatures corresponding to the nonmammalian host environment.

Biofilm Formation by and Multicellular Behavior of Escherichia coli O55:H7, an Atypical Enteropathogenic Strain