Identification of microbial communities in healthy adult human conjunctiva using high throughput sequencing technology

Session Details

Session Title: Endophthalmitis/Ocular Infections

Session Date/Time: Wednesday 09/09/2015 | 08:00-09:30

Paper Time: 08:06

Venue: Room 8

First Author: : Y.Huang CHINA

Co Author(s): :    l. Xie                    

Abstract Details

Purpose:

To determine the microbiota in healthy adult human conjunctiva using DNA sequencing-based detection and identification of bacteria.

Setting:

Qingdao Eye Hospital, Shandong Eye Institute, Qingdao, China.

Methods:

Illumina high-throughout sequencing technology was used to sequence 16S rDNA V3-V4 hypervariable region of all microbes in conjunctival swab samples. The sequences were clustered using the CD-hit-est based clustering method. Software PyNAST(http:/qiime.org/pynast/) was used to analyze and calculate the numbers of sequence and operational taxonomic units (OUTs) for each sample, and then the species abundance and distribution were analyzed and followed by cluster analysis. The Ribosomal Database Project (RDP) classifier was used to classify the 16S rDNA into distinct taxonomic category. Sequences greater than 97% identity were used to associate a group of OUTs to specific species.

Results:

A total of 2220141 high-quality sequencing reads were generated from 31 sputum samples. The rarefaction curves showed that adequate sequencing depth was achieved. The species OTUs number ranged from 100 to 2027, indicating high microbial diversity. The combined bacterial community was classified into 27 phyla and 748 distinct genera. Proteobacteria, Actinobacteria, Firmicutes, Bacteroidetes, and Deinococcus-Thermus were the five major bacterial phyla recovered. Pseudomonas, Corynebacterium, Acinetobacter, Staphylococcus, Streptococcus, Achromobacter, Simonsiella, Millisia, and Caulobacter were the most predominant genera. The intersubject variability of individual microbiomes was very significant.

Conclusions:

The composition and diversity of microbiota in healthy adult human conjunctiva was characterized for the first time by using high-throughput sequencing. It lays a framework for examination of potential roles played by the diverse microbiota in disease related with ocular surface.

Financial Interest:

NONE