Community analysis of chronic wound bacteria using 16S rRNA gene-based pyrosequencing

Impact of diabetes and antibiotics on chronic wound microbiota

Lance B. Price, Cindy M. Liu, Johan H. Melendez, Yelena M. Frankel, David Engelthaler, Maliha Aziz, Jolene Bowers, Rogan Rattray, Jacques Ravel, Chris Kingsley, Paul S Keim, Gerald S. Lazarus, Jonathan M. Zenilman

Research output: Contribution to journalArticle

121 Citations (Scopus)

Abstract

Background: Bacterial colonization is hypothesized to play a pathogenic role in the non-healing state of chronic wounds. We characterized wound bacteria from a cohort of chronic wound patients using a 16S rRNA gene-based pyrosequencing approach and assessed the impact of diabetes and antibiotics on chronic wound microbiota. Methodology/Principal Findings: We prospectively enrolled 24 patients at a referral wound center in Baltimore, MD; sampled patients' wounds by curette; cultured samples under aerobic and anaerobic conditions; and pyrosequenced the 16S rRNA V3 hypervariable region. The 16S rRNA gene-based analyses revealed an average of 10 different bacterial families in wounds - approximately 4 times more than estimated by culture-based analyses. Fastidious anaerobic bacteria belonging to the Clostridiales family XI were among the most prevalent bacteria identified exclusively by 16S rRNA gene-based analyses. Community-scale analyses showed that wound microbiota from antibiotic treated patients were significantly different from untreated patients (p = 0.007) and were characterized by increased Pseudomonadaceae abundance. These analyses also revealed that antibiotic use was associated with decreased Streptococcaceae among diabetics and that Streptococcaceae was more abundant among diabetics as compared to non-diabetics. Conclusions/Significance: The 16S rRNA gene-based analyses revealed complex bacterial communities including anaerobic bacteria that may play causative roles in the non-healing state of some chronic wounds. Our data suggest that antimicrobial therapy alters community structure - reducing some bacteria while selecting for others.

Original languageEnglish (US)
Article numbere6462
JournalPLoS One
Volume4
Issue number7
DOIs
StatePublished - Jul 31 2009

Fingerprint

Microbiota
plant damage
Medical problems
rRNA Genes
diabetes
Bacteria
Genes
antibiotics
ribosomal RNA
Anti-Bacterial Agents
bacteria
Wounds and Injuries
Streptococcaceae
genes
Anaerobic Bacteria
Pseudomonadaceae
Clostridiales
microbiome
bacterial colonization
Baltimore

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Community analysis of chronic wound bacteria using 16S rRNA gene-based pyrosequencing : Impact of diabetes and antibiotics on chronic wound microbiota. / Price, Lance B.; Liu, Cindy M.; Melendez, Johan H.; Frankel, Yelena M.; Engelthaler, David; Aziz, Maliha; Bowers, Jolene; Rattray, Rogan; Ravel, Jacques; Kingsley, Chris; Keim, Paul S; Lazarus, Gerald S.; Zenilman, Jonathan M.

In: PLoS One, Vol. 4, No. 7, e6462, 31.07.2009.

Research output: Contribution to journalArticle

Price, LB, Liu, CM, Melendez, JH, Frankel, YM, Engelthaler, D, Aziz, M, Bowers, J, Rattray, R, Ravel, J, Kingsley, C, Keim, PS, Lazarus, GS & Zenilman, JM 2009, 'Community analysis of chronic wound bacteria using 16S rRNA gene-based pyrosequencing: Impact of diabetes and antibiotics on chronic wound microbiota', PLoS One, vol. 4, no. 7, e6462. https://doi.org/10.1371/journal.pone.0006462
Price, Lance B. ; Liu, Cindy M. ; Melendez, Johan H. ; Frankel, Yelena M. ; Engelthaler, David ; Aziz, Maliha ; Bowers, Jolene ; Rattray, Rogan ; Ravel, Jacques ; Kingsley, Chris ; Keim, Paul S ; Lazarus, Gerald S. ; Zenilman, Jonathan M. / Community analysis of chronic wound bacteria using 16S rRNA gene-based pyrosequencing : Impact of diabetes and antibiotics on chronic wound microbiota. In: PLoS One. 2009 ; Vol. 4, No. 7.
@article{849ce9677d4b4873ba5c420a5986aebb,
title = "Community analysis of chronic wound bacteria using 16S rRNA gene-based pyrosequencing: Impact of diabetes and antibiotics on chronic wound microbiota",
abstract = "Background: Bacterial colonization is hypothesized to play a pathogenic role in the non-healing state of chronic wounds. We characterized wound bacteria from a cohort of chronic wound patients using a 16S rRNA gene-based pyrosequencing approach and assessed the impact of diabetes and antibiotics on chronic wound microbiota. Methodology/Principal Findings: We prospectively enrolled 24 patients at a referral wound center in Baltimore, MD; sampled patients' wounds by curette; cultured samples under aerobic and anaerobic conditions; and pyrosequenced the 16S rRNA V3 hypervariable region. The 16S rRNA gene-based analyses revealed an average of 10 different bacterial families in wounds - approximately 4 times more than estimated by culture-based analyses. Fastidious anaerobic bacteria belonging to the Clostridiales family XI were among the most prevalent bacteria identified exclusively by 16S rRNA gene-based analyses. Community-scale analyses showed that wound microbiota from antibiotic treated patients were significantly different from untreated patients (p = 0.007) and were characterized by increased Pseudomonadaceae abundance. These analyses also revealed that antibiotic use was associated with decreased Streptococcaceae among diabetics and that Streptococcaceae was more abundant among diabetics as compared to non-diabetics. Conclusions/Significance: The 16S rRNA gene-based analyses revealed complex bacterial communities including anaerobic bacteria that may play causative roles in the non-healing state of some chronic wounds. Our data suggest that antimicrobial therapy alters community structure - reducing some bacteria while selecting for others.",
author = "Price, {Lance B.} and Liu, {Cindy M.} and Melendez, {Johan H.} and Frankel, {Yelena M.} and David Engelthaler and Maliha Aziz and Jolene Bowers and Rogan Rattray and Jacques Ravel and Chris Kingsley and Keim, {Paul S} and Lazarus, {Gerald S.} and Zenilman, {Jonathan M.}",
year = "2009",
month = "7",
day = "31",
doi = "10.1371/journal.pone.0006462",
language = "English (US)",
volume = "4",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "7",

}

TY - JOUR

T1 - Community analysis of chronic wound bacteria using 16S rRNA gene-based pyrosequencing

T2 - Impact of diabetes and antibiotics on chronic wound microbiota

AU - Price, Lance B.

AU - Liu, Cindy M.

AU - Melendez, Johan H.

AU - Frankel, Yelena M.

AU - Engelthaler, David

AU - Aziz, Maliha

AU - Bowers, Jolene

AU - Rattray, Rogan

AU - Ravel, Jacques

AU - Kingsley, Chris

AU - Keim, Paul S

AU - Lazarus, Gerald S.

AU - Zenilman, Jonathan M.

PY - 2009/7/31

Y1 - 2009/7/31

N2 - Background: Bacterial colonization is hypothesized to play a pathogenic role in the non-healing state of chronic wounds. We characterized wound bacteria from a cohort of chronic wound patients using a 16S rRNA gene-based pyrosequencing approach and assessed the impact of diabetes and antibiotics on chronic wound microbiota. Methodology/Principal Findings: We prospectively enrolled 24 patients at a referral wound center in Baltimore, MD; sampled patients' wounds by curette; cultured samples under aerobic and anaerobic conditions; and pyrosequenced the 16S rRNA V3 hypervariable region. The 16S rRNA gene-based analyses revealed an average of 10 different bacterial families in wounds - approximately 4 times more than estimated by culture-based analyses. Fastidious anaerobic bacteria belonging to the Clostridiales family XI were among the most prevalent bacteria identified exclusively by 16S rRNA gene-based analyses. Community-scale analyses showed that wound microbiota from antibiotic treated patients were significantly different from untreated patients (p = 0.007) and were characterized by increased Pseudomonadaceae abundance. These analyses also revealed that antibiotic use was associated with decreased Streptococcaceae among diabetics and that Streptococcaceae was more abundant among diabetics as compared to non-diabetics. Conclusions/Significance: The 16S rRNA gene-based analyses revealed complex bacterial communities including anaerobic bacteria that may play causative roles in the non-healing state of some chronic wounds. Our data suggest that antimicrobial therapy alters community structure - reducing some bacteria while selecting for others.

AB - Background: Bacterial colonization is hypothesized to play a pathogenic role in the non-healing state of chronic wounds. We characterized wound bacteria from a cohort of chronic wound patients using a 16S rRNA gene-based pyrosequencing approach and assessed the impact of diabetes and antibiotics on chronic wound microbiota. Methodology/Principal Findings: We prospectively enrolled 24 patients at a referral wound center in Baltimore, MD; sampled patients' wounds by curette; cultured samples under aerobic and anaerobic conditions; and pyrosequenced the 16S rRNA V3 hypervariable region. The 16S rRNA gene-based analyses revealed an average of 10 different bacterial families in wounds - approximately 4 times more than estimated by culture-based analyses. Fastidious anaerobic bacteria belonging to the Clostridiales family XI were among the most prevalent bacteria identified exclusively by 16S rRNA gene-based analyses. Community-scale analyses showed that wound microbiota from antibiotic treated patients were significantly different from untreated patients (p = 0.007) and were characterized by increased Pseudomonadaceae abundance. These analyses also revealed that antibiotic use was associated with decreased Streptococcaceae among diabetics and that Streptococcaceae was more abundant among diabetics as compared to non-diabetics. Conclusions/Significance: The 16S rRNA gene-based analyses revealed complex bacterial communities including anaerobic bacteria that may play causative roles in the non-healing state of some chronic wounds. Our data suggest that antimicrobial therapy alters community structure - reducing some bacteria while selecting for others.

UR - http://www.scopus.com/inward/record.url?scp=68149089830&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=68149089830&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0006462

DO - 10.1371/journal.pone.0006462

M3 - Article

VL - 4

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 7

M1 - e6462

ER -