Advanced

Effect of Cold Starvation, Acid Stress, and Nutrients on Metabolic Activity of Helicobacter pylori.

Nilsson, Hans-Olof LU ; Blom, Jens; Abu Al-Soud, Waleed LU ; Ljungh, Åsa LU ; Andersen, Leif P and Wadström, Torkel LU (2002) In Applied and Environmental Microbiology 68(1). p.11-19
Abstract
Helicobacter pylori can transform, in vivo as well as in vitro, from dividing spiral-shaped forms into nonculturable coccoids, with intermediate forms called U forms. The importance of nonculturable coccoid forms of H. pylori in disease transmission and antibiotic treatment failures is unclear. Metabolic activities of actively growing as well as nonculturable H. pylori were investigated by comparing the concentrations of cellular ATP and total RNA, gene expression, presence of cytoplasmic polyphosphate granules and iron inclusions, and cellular morphology during extended broth culture and nutritional cold starvation. In addition, the effect of exposing broth-cultured or cold-starved cells to a nutrient-rich or acidic environment on the... (More)
Helicobacter pylori can transform, in vivo as well as in vitro, from dividing spiral-shaped forms into nonculturable coccoids, with intermediate forms called U forms. The importance of nonculturable coccoid forms of H. pylori in disease transmission and antibiotic treatment failures is unclear. Metabolic activities of actively growing as well as nonculturable H. pylori were investigated by comparing the concentrations of cellular ATP and total RNA, gene expression, presence of cytoplasmic polyphosphate granules and iron inclusions, and cellular morphology during extended broth culture and nutritional cold starvation. In addition, the effect of exposing broth-cultured or cold-starved cells to a nutrient-rich or acidic environment on the metabolic activities was investigated. ATP was detectable up to 14 days and for at least 25 days after transformation from the spiral form to the coccoid form or U form in broth-cultured and cold-starved cells, respectively. mRNAs of VacA, a 26-kDa protein, and urease A were detected by using reverse transcription-PCR in cells cultured for 2 months in broth or cold starved for at least 28 months. The ATP concentration was not affected during exposure to fresh or acidified broth, while 4- to 12-h exposures of nonculturable cells to lysed human erythrocytes increased cellular ATP 12- to 150-fold. Incubation of nonculturable cold-starved cells with an erythrocyte lysate increased total RNA expression and ureA mRNA transcription as measured by quantitative real-time reverse transcription-PCR. Furthermore, the number of structurally intact starved coccoids containing polyphosphate granules increased almost fourfold (P = 0.0022) under the same conditions. In conclusion, a specific environmental stimulus can induce ATP, polyphosphate, and RNA metabolism in nonculturable H. pylori, indicating viability of such morphological forms. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Gene Expression Regulation Bacterial, Heat-Shock Response, Hydrogen-Ion Concentration, Helicobacter pylori : growth & development : metabolism : ultrastructure, Microscopy Electron, RNA Bacterial : metabolism, RNA Messenger : metabolism, Support Non-U.S. Gov't, Culture Media, Cold, Adenosine Triphosphate : metabolism
in
Applied and Environmental Microbiology
volume
68
issue
1
pages
11 - 19
publisher
American Society for Microbiology
external identifiers
  • pmid:11772603
  • wos:000173085000002
  • scopus:0036135877
ISSN
0099-2240
DOI
10.1128/AEM.68.1.11-19.2002
language
English
LU publication?
yes
id
0e711310-5dcb-45d4-b2be-789352718de6 (old id 106897)
date added to LUP
2007-07-10 08:49:09
date last changed
2017-10-29 03:36:40
@article{0e711310-5dcb-45d4-b2be-789352718de6,
  abstract     = {Helicobacter pylori can transform, in vivo as well as in vitro, from dividing spiral-shaped forms into nonculturable coccoids, with intermediate forms called U forms. The importance of nonculturable coccoid forms of H. pylori in disease transmission and antibiotic treatment failures is unclear. Metabolic activities of actively growing as well as nonculturable H. pylori were investigated by comparing the concentrations of cellular ATP and total RNA, gene expression, presence of cytoplasmic polyphosphate granules and iron inclusions, and cellular morphology during extended broth culture and nutritional cold starvation. In addition, the effect of exposing broth-cultured or cold-starved cells to a nutrient-rich or acidic environment on the metabolic activities was investigated. ATP was detectable up to 14 days and for at least 25 days after transformation from the spiral form to the coccoid form or U form in broth-cultured and cold-starved cells, respectively. mRNAs of VacA, a 26-kDa protein, and urease A were detected by using reverse transcription-PCR in cells cultured for 2 months in broth or cold starved for at least 28 months. The ATP concentration was not affected during exposure to fresh or acidified broth, while 4- to 12-h exposures of nonculturable cells to lysed human erythrocytes increased cellular ATP 12- to 150-fold. Incubation of nonculturable cold-starved cells with an erythrocyte lysate increased total RNA expression and ureA mRNA transcription as measured by quantitative real-time reverse transcription-PCR. Furthermore, the number of structurally intact starved coccoids containing polyphosphate granules increased almost fourfold (P = 0.0022) under the same conditions. In conclusion, a specific environmental stimulus can induce ATP, polyphosphate, and RNA metabolism in nonculturable H. pylori, indicating viability of such morphological forms.},
  author       = {Nilsson, Hans-Olof and Blom, Jens and Abu Al-Soud, Waleed and Ljungh, Åsa and Andersen, Leif P and Wadström, Torkel},
  issn         = {0099-2240},
  keyword      = {Gene Expression Regulation Bacterial,Heat-Shock Response,Hydrogen-Ion Concentration,Helicobacter pylori : growth & development : metabolism : ultrastructure,Microscopy Electron,RNA Bacterial : metabolism,RNA Messenger : metabolism,Support Non-U.S. Gov't,Culture Media,Cold,Adenosine Triphosphate : metabolism},
  language     = {eng},
  number       = {1},
  pages        = {11--19},
  publisher    = {American Society for Microbiology},
  series       = {Applied and Environmental Microbiology},
  title        = {Effect of Cold Starvation, Acid Stress, and Nutrients on Metabolic Activity of Helicobacter pylori.},
  url          = {http://dx.doi.org/10.1128/AEM.68.1.11-19.2002},
  volume       = {68},
  year         = {2002},
}