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. 2009 Jul 31;4(7):e6462.
doi: 10.1371/journal.pone.0006462.

"V体育平台登录" Community analysis of chronic wound bacteria using 16S rRNA gene-based pyrosequencing: impact of diabetes and antibiotics on chronic wound microbiota

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Community analysis of chronic wound bacteria using 16S rRNA gene-based pyrosequencing: impact of diabetes and antibiotics on chronic wound microbiota (V体育官网)

Lance B Price et al. PLoS One. .

Abstract

Background: Bacterial colonization is hypothesized to play a pathogenic role in the non-healing state of chronic wounds VSports手机版. 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 V体育安卓版. 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. V体育ios版.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Rarefaction and Shannon Weaver index analyses were performed for each wound specimen.
(A) Rarefaction curves were used to estimate richness (i.e., number of unique bacterial taxa) among samples. (B) Shannon Weaver Index curves were used estimate diversity (i.e., a combined assessment of the number of unique bacterial taxa and their abundance) among samples.
Figure 2
Figure 2. Heat map analysis of the 44 bacterial families detected using 16S rRNA gene-based pyrosequencing among chronic wound samples.
The families in red are those that were successfully cultured at least once during the study. The presence/absence plot on the left shows the bacteria present in each of the wound samples. The abundance plot on the right shows the number of 16S rRNA gene pyrosequences (300 maximum) in each of the wound samples. The average copy number per positive sample for each detected bacterial family is shown on the far right. Many rare bacterial families are only visible on the presence/absence plot on the left.
Figure 3
Figure 3. The nMDS ordination plot comparing wound bacterial communities from antibiotic treated participants and untreated participants.
Each data point in nMDS plot represent the bacterial community identified from a single wound specimen. Comparison using MRPP found that the antibiotic treated and untreated wound microbiota are significantly different (p = 0.0069).

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