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Assessment of microbial diversity in biofilms recovered from endotracheal tubes using culture dependent and independent approaches

Ilse Vandecandelaere UGent, Nele Matthijs UGent, Filip Van Nieuwerburgh UGent, Dieter Deforce UGent, PETER VOSTERS UGent, Liesbet De Bus UGent, Hans Nelis UGent, Pieter Depuydt UGent and Tom Coenye UGent (2012) PLOS ONE. 7(6).
abstract
Ventilator-associated pneumonia (VAP) is a common nosocomial infection in mechanically ventilated patients. Biofilm formation is one of the mechanisms through which the endotracheal tube (ET) facilitates bacterial contamination of the lower airways. In the present study, we analyzed the composition of the ET biofilm flora by means of culture dependent and culture independent (16 S rRNA gene clone libraries and pyrosequencing) approaches. Overall, the microbial diversity was high and members of different phylogenetic lineages were detected (Actinobacteria, beta-Proteobacteria, Candida spp., Clostridia, epsilon-Proteobacteria, Firmicutes, Fusobacteria and gamma-Proteobacteria). Culture dependent analysis, based on the use of selective growth media and conventional microbiological tests, resulted in the identification of typical aerobic nosocomial pathogens which are known to play a role in the development of VAP, e. g. Staphylococcus aureus and Pseudomonas aeruginosa. Other opportunistic pathogens were also identified, including Staphylococcus epidermidis and Kocuria varians. In general, there was little correlation between the results obtained by sequencing 16 S rRNA gene clone libraries and by cultivation. Pyrosequencing of PCR amplified 16 S rRNA genes of four selected samples resulted in the identification of a much wider variety of bacteria. The results from the pyrosequencing analysis suggest that these four samples were dominated by members of the normal oral flora such as Prevotella spp., Peptostreptococcus spp. and lactic acid bacteria. A combination of methods is recommended to obtain a complete picture of the microbial diversity of the ET biofilm.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
VENTILATOR-ASSOCIATED PNEUMONIA, SURVEILLANCE CULTURES, INTENSIVE-CARE-UNIT, NOSOCOMIAL PNEUMONIA, CYSTIC-FIBROSIS, ORAL CAVITY, BACTERIA, IMPACT, INFECTIONS, PREVENTION
journal title
PLOS ONE
PLoS One
volume
7
issue
6
article_number
e38401
pages
9 pages
Web of Science type
Article
Web of Science id
000305343900035
JCR category
MULTIDISCIPLINARY SCIENCES
JCR impact factor
3.73 (2012)
JCR rank
7/56 (2012)
JCR quartile
1 (2012)
ISSN
1932-6203
DOI
10.1371/journal.pone.0038401
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
2988511
handle
http://hdl.handle.net/1854/LU-2988511
date created
2012-09-17 15:37:43
date last changed
2012-09-19 10:17:21
@article{2988511,
  abstract     = {Ventilator-associated pneumonia (VAP) is a common nosocomial infection in mechanically ventilated patients. Biofilm formation is one of the mechanisms through which the endotracheal tube (ET) facilitates bacterial contamination of the lower airways. In the present study, we analyzed the composition of the ET biofilm flora by means of culture dependent and culture independent (16 S rRNA gene clone libraries and pyrosequencing) approaches. Overall, the microbial diversity was high and members of different phylogenetic lineages were detected (Actinobacteria, beta-Proteobacteria, Candida spp., Clostridia, epsilon-Proteobacteria, Firmicutes, Fusobacteria and gamma-Proteobacteria). Culture dependent analysis, based on the use of selective growth media and conventional microbiological tests, resulted in the identification of typical aerobic nosocomial pathogens which are known to play a role in the development of VAP, e. g. Staphylococcus aureus and Pseudomonas aeruginosa. Other opportunistic pathogens were also identified, including Staphylococcus epidermidis and Kocuria varians. In general, there was little correlation between the results obtained by sequencing 16 S rRNA gene clone libraries and by cultivation. Pyrosequencing of PCR amplified 16 S rRNA genes of four selected samples resulted in the identification of a much wider variety of bacteria. The results from the pyrosequencing analysis suggest that these four samples were dominated by members of the normal oral flora such as Prevotella spp., Peptostreptococcus spp. and lactic acid bacteria. A combination of methods is recommended to obtain a complete picture of the microbial diversity of the ET biofilm.},
  articleno    = {e38401},
  author       = {Vandecandelaere, Ilse and Matthijs, Nele and Van Nieuwerburgh, Filip and Deforce, Dieter and VOSTERS, PETER and De Bus, Liesbet and Nelis, Hans and Depuydt, Pieter and Coenye, Tom},
  issn         = {1932-6203},
  journal      = {PLOS ONE},
  keyword      = {VENTILATOR-ASSOCIATED PNEUMONIA,SURVEILLANCE CULTURES,INTENSIVE-CARE-UNIT,NOSOCOMIAL PNEUMONIA,CYSTIC-FIBROSIS,ORAL CAVITY,BACTERIA,IMPACT,INFECTIONS,PREVENTION},
  language     = {eng},
  number       = {6},
  pages        = {9},
  title        = {Assessment of microbial diversity in biofilms recovered from endotracheal tubes using culture dependent and independent approaches},
  url          = {http://dx.doi.org/10.1371/journal.pone.0038401},
  volume       = {7},
  year         = {2012},
}

Chicago
Vandecandelaere, Ilse, Nele Matthijs, Filip Van Nieuwerburgh, Dieter Deforce, PETER VOSTERS, Liesbet De Bus, Hans Nelis, Pieter Depuydt, and Tom Coenye. 2012. “Assessment of Microbial Diversity in Biofilms Recovered from Endotracheal Tubes Using Culture Dependent and Independent Approaches.” Plos One 7 (6).
APA
Vandecandelaere, I., Matthijs, N., Van Nieuwerburgh, F., Deforce, D., VOSTERS, P., De Bus, L., Nelis, H., et al. (2012). Assessment of microbial diversity in biofilms recovered from endotracheal tubes using culture dependent and independent approaches. PLOS ONE, 7(6).
Vancouver
1.
Vandecandelaere I, Matthijs N, Van Nieuwerburgh F, Deforce D, VOSTERS P, De Bus L, et al. Assessment of microbial diversity in biofilms recovered from endotracheal tubes using culture dependent and independent approaches. PLOS ONE. 2012;7(6).
MLA
Vandecandelaere, Ilse, Nele Matthijs, Filip Van Nieuwerburgh, et al. “Assessment of Microbial Diversity in Biofilms Recovered from Endotracheal Tubes Using Culture Dependent and Independent Approaches.” PLOS ONE 7.6 (2012): n. pag. Print.