Response to alkaline stress by root canal bacteria in biofilms.

AIM To determine whether bacteria isolated from infected root canals survive alkaline shifts better in biofilms than in planktonic cultures. METHODOLOGY Clinical isolates of Enterococcus faecalis, Lactobacillus paracasei, Olsenella uli, Streptococcus anginosus, S. gordonii, S. oralis and Fusobacterium nucleatum in biofilm and planktonic cultures were stressed at pH 10.5 for 4 h, and cell viability determined using the fluorescent staining LIVE/DEAD BacLight bacterial viability kit. In addition, proteins released into extracellular culture fluids were identified by Western blotting. RESULTS Enterococcus faecalis, L. paracasei, O. uli and S. gordonii survived in high numbers in both planktonic cultures and in biofilms after alkaline challenge. S. anginosus, S. oralis and F. nucleatum showed increased viability in biofilms compared with planktonic cultures. Alkaline exposure caused all planktonic cultures to aggregate into clusters and resulted in a greater extrusion of cellular proteins compared with cells in biofilms. Increased levels of DnaK, HPr and fructose-1,6-bisphosphate aldolase were observed in culture fluids, especially amongst streptococci. CONCLUSIONS In general, bacteria isolated from infected roots canals resisted alkaline stress better in biofilms than in planktonic cultures, however, planktonic cells appeared to use aggregation and the extracellular transport of specific proteins as survival mechanisms.

[1]  L. E. Chávez de Paz,et al.  Streptococci from root canals in teeth with apical periodontitis receiving endodontic treatment. , 2005, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[2]  P. Stewart,et al.  Adaptive responses to antimicrobial agents in biofilms. , 2005, Environmental microbiology.

[3]  H. Ceri,et al.  Persister cells, the biofilm matrix and tolerance to metal cations in biofilm and planktonic Pseudomonas aeruginosa. , 2005, Environmental microbiology.

[4]  K. Lewis.  Persister cells and the riddle of biofilm survival , 2005, Biochemistry (Moscow).

[5]  D. Cvitkovitch,et al.  Quorum sensing in streptococcal biofilm formation. , 2005, Trends in microbiology.

[6]  E. Greenberg,et al.  Sociomicrobiology: the connections between quorum sensing and biofilms. , 2005, Trends in microbiology.

[7]  P Stoodley,et al.  Survival strategies of infectious biofilms. , 2005, Trends in microbiology.

[8]  G. Svensäter,et al.  Biofilms in endodontic infections , 2004 .

[9]  S. Leibler,et al.  Bacterial Persistence as a Phenotypic Switch , 2004, Science.

[10]  L. E. Chávez de Paz,et al.  Gram-positive rods prevailing in teeth with apical periodontitis undergoing root canal treatment. , 2004, International endodontic journal.

[11]  G. Svensäter,et al.  Protein Expression by Streptococcus mutans during Initial Stage of Biofilm Formation , 2004, Applied and Environmental Microbiology.

[12]  G. Trueba,et al.  Cell aggregation: a mechanism of pathogenic Leptospira to survive in fresh water. , 2004, International microbiology : the official journal of the Spanish Society for Microbiology.

[13]  K. Lewis,et al.  Persister cells and tolerance to antimicrobials. , 2004, FEMS microbiology letters.

[14]  G. Sundqvist,et al.  Life as an endodontic pathogen , 2003 .

[15]  F. Vogensen,et al.  Identification of proteins induced at low pH in Lactococcus lactis. , 2003, International journal of food microbiology.

[16]  Hiroshi Kobayashi,et al.  Bacterial Responses to Alkaline Stress , 2003, Science progress.

[17]  U. Endal,et al.  Persistent, recurrent, and acquired infection of the root canal system , 2003 .

[18]  S. Fey,et al.  Effect of acid shock on protein expression by biofilm cells of Streptococcus mutans. , 2003, FEMS microbiology letters.

[19]  D. Beighton,et al.  Effect of Acidic pH on Expression of Surface-Associated Proteins of Streptococcus oralis , 2003, Applied and Environmental Microbiology.

[20]  C. Hill,et al.  Surviving the Acid Test: Responses of Gram-Positive Bacteria to Low pH , 2003, Microbiology and Molecular Biology Reviews.

[21]  L. E. Chávez de Paz,et al.  Bacteria recovered from teeth with apical periodontitis after antimicrobial endodontic treatment. , 2003, International endodontic journal.

[22]  I. R. Hamilton,et al.  Acid tolerance response of biofilm cells of Streptococcus mutans. , 2003, FEMS microbiology letters.

[23]  J. McPherson,et al.  In vitro antimicrobial activity of various medication preparations on E. faecalis in root canal dentin. , 2003, Journal of endodontics.

[24]  T. Larsen.  Susceptibility of Porphyromonas gingivalis in biofilms to amoxicillin, doxycycline and metronidazole. , 2002, Oral microbiology and immunology.

[25]  G. Robson,et al.  Comparative susceptibility of resident and transient hand bacteria to para‐chloro‐meta‐xylenol and triclosan , 2002, Journal of applied microbiology.

[26]  Yuliya N. Yoncheva,et al.  pH-Dependent Expression of Periplasmic Proteins and Amino Acid Catabolism in Escherichia coli , 2002, Journal of bacteriology.

[27]  M. Evans,et al.  Mechanisms involved in the resistance of Enterococcus faecalis to calcium hydroxide. , 2002, International endodontic journal.

[28]  K. Otto,et al.  Surface sensing and adhesion of Escherichia coli controlled by the Cpx-signaling pathway , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[29]  T. Srisuwan,et al.  A comparison of the antimicrobial efficacy of three calcium hydroxide formulations on human dentin infected with Enterococcus faecalis. , 2002, Journal of endodontics.

[30]  K. Lewis,et al.  Biofilms and Planktonic Cells of Pseudomonas aeruginosa Have Similar Resistance to Killing by Antimicrobials , 2001, Journal of bacteriology.

[31]  G. Svensäter,et al.  Protein expression by planktonic and biofilm cells of Streptococcus mutans. , 2001, FEMS microbiology letters.

[32]  M. E. Vianna,et al.  In vitro antimicrobial activity of several concentrations of sodium hypochlorite and chlorhexidine gluconate in the elimination of Enterococcus faecalis. , 2001, International endodontic journal.

[33]  J. Costerton,et al.  Antibiotic resistance of bacteria in biofilms , 2001, The Lancet.

[34]  K. Lewis,et al.  Riddle of Biofilm Resistance , 2001, Antimicrobial Agents and Chemotherapy.

[35]  J. Potempa,et al.  pH-regulated Secretion of a Glyceraldehyde-3-Phosphate Dehydrogenase from Streptococcus gordonii FSS2: Purification, Characterization, and Cloning of the Gene Encoding this Enzyme , 2001, Journal of dental research.

[36]  D. Steinberg,et al.  The Anticariogenic Effect of Amine Fluorides on Streptococcus sobrinus and Glucosyltransferase in Biofilms , 2000, Caries Research.

[37]  C. Prigent-Combaret,et al.  Abiotic Surface Sensing and Biofilm-Dependent Regulation of Gene Expression in Escherichia coli , 1999, Journal of bacteriology.

[38]  J. Costerton,et al.  Bacterial biofilms: a common cause of persistent infections. , 1999, Science.

[39]  G A Leonard,et al.  The crystal structure of Escherichia coli class II fructose-1, 6-bisphosphate aldolase in complex with phosphoglycolohydroxamate reveals details of mechanism and specificity. , 1999, Journal of molecular biology.

[40]  B. Henderson,et al.  Are molecular chaperones microbial virulence factors? , 1998, Trends in microbiology.

[41]  P. Gilbert,et al.  Biofilm Susceptibility to Antimicrobials , 1997, Advances in dental research.

[42]  J. R. Scott,et al.  Iron starvation causes release from the group A streptococcus of the ADP-ribosylating protein called plasmin receptor or surface glyceraldehyde-3-phosphate-dehydrogenase , 1996, Infection and immunity.

[43]  B. Pişkin,et al.  Observation of bacteria and fungi in infected root canals and dentinal tubules by SEM. , 1995, Endodontics & dental traumatology.

[44]  K. Marshall,et al.  Microbial Biofilms: Genetic Responses of Bacteria at Surfaces , 1995 .

[45]  C. Vadeboncoeur.  HPr: heteromorphous protein. , 1995, Research in microbiology.

[46]  J. Foster,et al.  Molecular responses of microbes to environmental pH stress. , 1995, Advances in microbial physiology.

[47]  T. A. Krulwich,et al.  Proton-coupled bioenergetic processes in extremely alkaliphilic bacteria , 1992, Journal of bioenergetics and biomembranes.

[48]  M. Pavlík,et al.  The characterization of two new low molecular weight proteins (LMPs) from Streptococcus pyogenes. , 1992, Zentralblatt fur Bakteriologie : international journal of medical microbiology.

[49]  I. Olsen,et al.  Scanning electron microscopy of bacteria in the apical part of root canals in permanent teeth with periapical lesions. , 1991, Endodontics & dental traumatology.

[50]  M. Fletcher.  The physiological activity of bacteria attached to solid surfaces. , 1991, Advances in microbial physiology.

[51]  P. Nair.  Light and electron microscopic studies of root canal flora and periapical lesions. , 1987 .

[52]  J. Costerton,et al.  Bacterial biofilms in nature and disease. , 1987, Annual review of microbiology.

[53]  I. Booth,et al.  Regulation of cytoplasmic pH in bacteria. , 1985, Microbiological reviews.

[54]  R. Claesson,et al.  The antibacterial effect of camphorated paramonochlorophenol, camphorated phenol and calcium hydroxide in the treatment of infected root canals. , 1985, Endodontics & dental traumatology.

[55]  B. Boyan,et al.  The effects of calcium hydroxide on bovine pulp tissue: variations in pH and calcium concentration. , 1985, Journal of endodontics.

[56]  H. Moyed,et al.  hipA, a newly recognized gene of Escherichia coli K-12 that affects frequency of persistence after inhibition of murein synthesis , 1983, Journal of bacteriology.

[57]  R. G. Kroll,et al.  Regulation of cytoplasmic pH (pH1) in bacteria and its relationship to metabolism. , 1983, Biochemical Society transactions.

[58]  G. Dahlén,et al.  A retrospective study of microbiologic samples from oral mucosal lesions. , 1982, Oral surgery, oral medicine, and oral pathology.

[59]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[60]  G. Bergenholtz.  Micro-organisms from necrotic pulp of traumatized teeth. , 1974, Odontologisk revy.

[61]  J. Bigger.  TREATMENT OF STAPHYLOCOCCAL INFECTIONS WITH PENICILLIN BY INTERMITTENT STERILISATION , 1944 .