Effects of prolonged exposure to alkaline pH on Enterococcus faecalis survival and specific gene transcripts

O. K. Appelbe, Christine Sedgley

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Introduction: The persistence of Enterococcus faecalis in treated root canals has been attributed to its resistance to the high pH of antimicrobial agents used during treatment, but the specific mechanisms are not clear. We investigated the survival and gene expression of E. faecalis maintained in alkaline media. Methods: E. faecalis JH2-2 was maintained in media at pH 7, 10, 11 and 12 at either 25°C or 37°C for 1 week (168 h). At 24, 48, 72, 120 and 168 h, cell viability was determined in parallel with real-time quantitative polymerase chain reaction analyses of stress response genes (dnaK, fba, ftsZ, GroEL, napA, pbp5, tsf and tuf). Results: After 1 week the E. faecalis showed survival levels of 100% in pH 7, 1% in pH 10, 0.001% in pH 11 and 0.00001% in pH 12 media. At 37°C increased levels of gene transcripts occurred between 72 and 120 h in pH 7 media for ftsZ and dnaK, and in pH 10 media for ftsZ, pbp5, dnaK, napA, tsf, fba and GroEL. No increase in transcripts was observed at 37°C in media at pH 11 or pH 12, nor at 25°C in any media. Conclusion: Transcripts of ftsZ, a gene involved in cell division, increased by 37-fold after 120 h at pH 10 at 37°C. Overall, the greatest increase in levels of gene transcripts occurred in cultures maintained in pH 10 media at 37°C. These data may assist in understanding the survival strategies of E. faecalis following prolonged exposure to alkaline pH levels.

Original languageEnglish (US)
Pages (from-to)169-174
Number of pages6
JournalOral Microbiology and Immunology
Volume22
Issue number3
DOIs
StatePublished - Jun 2007
Externally publishedYes

Fingerprint

Enterococcus faecalis
Genes
Dental Pulp Cavity
Anti-Infective Agents
Cell Division
Real-Time Polymerase Chain Reaction
Cell Survival

Keywords

  • Alkaline pH
  • Enterococcus faecalis
  • Gene transcripts
  • Prolonged exposure
  • Survival

ASJC Scopus subject areas

  • Immunology
  • Microbiology (medical)
  • Dentistry(all)

Cite this

Effects of prolonged exposure to alkaline pH on Enterococcus faecalis survival and specific gene transcripts. / Appelbe, O. K.; Sedgley, Christine.

In: Oral Microbiology and Immunology, Vol. 22, No. 3, 06.2007, p. 169-174.

Research output: Contribution to journalArticle

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AB - Introduction: The persistence of Enterococcus faecalis in treated root canals has been attributed to its resistance to the high pH of antimicrobial agents used during treatment, but the specific mechanisms are not clear. We investigated the survival and gene expression of E. faecalis maintained in alkaline media. Methods: E. faecalis JH2-2 was maintained in media at pH 7, 10, 11 and 12 at either 25°C or 37°C for 1 week (168 h). At 24, 48, 72, 120 and 168 h, cell viability was determined in parallel with real-time quantitative polymerase chain reaction analyses of stress response genes (dnaK, fba, ftsZ, GroEL, napA, pbp5, tsf and tuf). Results: After 1 week the E. faecalis showed survival levels of 100% in pH 7, 1% in pH 10, 0.001% in pH 11 and 0.00001% in pH 12 media. At 37°C increased levels of gene transcripts occurred between 72 and 120 h in pH 7 media for ftsZ and dnaK, and in pH 10 media for ftsZ, pbp5, dnaK, napA, tsf, fba and GroEL. No increase in transcripts was observed at 37°C in media at pH 11 or pH 12, nor at 25°C in any media. Conclusion: Transcripts of ftsZ, a gene involved in cell division, increased by 37-fold after 120 h at pH 10 at 37°C. Overall, the greatest increase in levels of gene transcripts occurred in cultures maintained in pH 10 media at 37°C. These data may assist in understanding the survival strategies of E. faecalis following prolonged exposure to alkaline pH levels.

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