Community Analysis of Chronic Wound Bacteria Using 16S rRNA Gene-Based Pyrosequencing: Impact of Diabetes and Antibiotics on Chronic Wound Microbiota

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

Comparison of orange oil and chloroform as gutta- percha solvents in endodontic retreatment

Objective: To compare the efficacy of gutta-percha removal achieved with simple mechanical technique using two different solvents.MATERIALS AND Methods: It was an in vitro experimental study conducted at the Aga Khan University Hospital, Karachi, Pakistan on 27 extracted human molars divided into two groups by alternate sampling technique. The teeth were prepared by manual filing and obturated with gutta-percha and calcium hydroxide sealer. Two drops of the assigned solvent were placed on the orifice of the obturated canal and Gates Glidden drills #1, 2 and 3 were used for removal of coronal gutta-percha. Manual filing was done for removal of remaining gutta-percha and the solvent was used in drops until needed. Postoperative periapical radiographs were taken to visualize the remaining gutta-percha left in the canals. Descriptive statistics (mean and SD) of the variables such as tooth type, obturation length and canal curvatures were determined. Chi-square and independent sample t-tests were applied and level of significance was set at 0.05.Results: There was 5.19 (3.8) mm of remaining gutta-percha in the orange oil group and 5.37 (4.2) mm in the chloroform group (p=0.90).CONCLUSION: There is no statistically significant difference between the orange oil and chloroform when used as solvent for removing gutta-percha