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Applied and Environmental Microbiology, February 2000, p. 632-637, Vol. 66, No. 2
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Role of rpoS in Acid Resistance and Fecal Shedding of Escherichia coli O157:H7

Stuart B. Price,1,* Chorng-Ming Cheng,2 Charles W. Kaspar,2 James C. Wright,1 Fred J. DeGraves,3 Thomas A. Penfound,4,dagger Marie-Pierre Castanie-Cornet,4 and John W. Foster4

Departments of Pathobiology1 and Large Animal Surgery and Medicine,3 College of Veterinary Medicine, Auburn University, Auburn, Alabama 36849; Department of Food Microbiology and Toxicology, Food Research Institute, University of Wisconsin, Madison, Wisconsin 537062; and Department of Microbiology and Immunology, University of South Alabama College of Medicine, Mobile, Alabama 366884

Received 9 August 1999/Accepted 18 November 1999

Acid resistance (AR) is important to survival of Escherichia coli O157:H7 in acidic foods and may play a role during passage through the bovine host. In this study, we examined the role in AR of the rpoS-encoded global stress response regulator sigma S and its effect on shedding of E. coli O157:H7 in mice and calves. When assayed for each of the three AR systems identified in E. coli, an rpoS mutant (rpoS::pRR10) of E. coli O157:H7 lacked the glucose-repressed system and possessed reduced levels of both the arginine- and glutamate-dependent AR systems. After administration of the rpoS mutant and the wild-type strain (ATCC 43895) to ICR mice at doses ranging from 101 to 104 CFU, we found the wild-type strain in feces of mice given lower doses (102 versus 103 CFU) and at a greater frequency (80% versus 13%) than the mutant strain. The reduction in passage of the rpoS mutant was due to decreased AR, as administration of the mutant in 0.05 M phosphate buffer facilitated passage and increased the frequency of recovery in feces from 27 to 67% at a dose of 104 CFU. Enumeration of E. coli O157:H7 in feces from calves inoculated with an equal mixture of the wild-type strain and the rpoS mutant demonstrated shedding of the mutant to be 10- to 100-fold lower than wild-type numbers. This difference in shedding between the wild-type strain and the rpoS mutant was statistically significant (P <=  0.05). Thus, sigma S appears to play a role in E. coli O157:H7 passage in mice and shedding from calves, possibly by inducing expression of the glucose-repressed RpoS-dependent AR determinant and thus increasing resistance to gastrointestinal stress. These findings may provide clues for future efforts aimed at reducing or eliminating this pathogen from cattle herds.

* Corresponding author. Mailing address: Department of Pathobiology, College of Veterinary Medicine, 264 Greene Hall, Auburn University, Auburn, AL 36849. Phone: (334) 844-2673. Fax: (334) 844-2652. E-mail: pricesb{at}vetmed.auburn.edu

dagger Present address: Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, TN 38105.

Applied and Environmental Microbiology, February 2000, p. 632-637, Vol. 66, No. 2
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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