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Journal of Bacteriology, May 2005, p. 3319-3328, Vol. 187, No. 10
0021-9193/05/$08.00+0     doi:10.1128/JB.187.10.3319-3328.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Persistence of Streptococcus pyogenes in Stationary-Phase Cultures

Daniel N. Wood,¹ Michelle A. Chaussee,² Michael S. Chaussee,² and Bettina A. Buttaro^1*

Department of Microbiology and Immunology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140,¹ Division of Basic Biomedical Science, University of South Dakota College of Medicine, Vermillion, South Dakota 57069²

Received 4 August 2004/ Accepted 1 February 2005

In addition to causing fulminant disease, Streptococcus pyogenes may be asymptomatically carried between recurrent episodes of pharyngitis. To better understand streptococcal carriage, we characterized in vitro long-term stationary-phase survival (>4 weeks) of S. pyogenes. When grown in sugar-limited Todd-Hewitt broth, S. pyogenes cells remained culturable for more than 1 year. Both Todd-Hewitt supplemented with excess glucose and chemically defined medium allowed survival for less than 1 week. After 4 weeks of survival in sugar-limited Todd-Hewitt broth, at least 10³ CFU per ml remained. When stained with fluorescent live-dead viability stain, there were a number of cells with intact membranes that were nonculturable. Under conditions that did not support persistence, these cells disappeared 2 weeks after loss of culturability. In persistent cultures, these may be cells that are dying during cell turnover. After more than 4 weeks in stationary phase, the culturable cells formed two alternative colony phenotypes: atypical large colonies and microcolonies. Protein expression in two independently isolated microcolony strains, from 14-week cultures, was examined by use of two-dimensional electrophoresis. The proteomes of these two strains exhibited extensive changes compared to the parental strain. While some of these changes were common to the two strains, many of the changes were unique to a single strain. Some of the common changes were in metabolic pathways, suggesting a possible alternate metabolism for the persisters. Overall, these data suggest that under certain in vitro conditions, S. pyogenes cells can persist for greater than 1 year as a dynamic population.

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* Corresponding author. Mailing address: Department of Microbiology and Immunology, Temple University School of Medicine, 3400 North Broad Street, Philadelphia, PA 19140. Phone: (215) 707-3212. Fax: (215) 707-7788. E-mail: bbuttaro{at}temple.edu.

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Journal of Bacteriology, May 2005, p. 3319-3328, Vol. 187, No. 10
0021-9193/05/$08.00+0     doi:10.1128/JB.187.10.3319-3328.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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