Diversity of bacteria at healthy human conjunctiva

Ocular surface (OS) microbiota contributes to infectious and autoimmune diseases of the eye. Comprehensive analysis of microbial diversity at the OS has been impossible because of the limitations of conventional cultivation techniques. This pilot study aimed to explore true diversity of human OS mic...

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Bibliographic Details
Published in:Investigative ophthalmology & visual science 2011-07, Vol.52 (8), p.5408-5413
Main Authors: Dong, Qunfeng, Brulc, Jennifer M, Iovieno, Alfonso, Bates, Brandon, Garoutte, Aaron, Miller, Darlene, Revanna, Kashi V, Gao, Xiang, Antonopoulos, Dionysios A, Slepak, Vladlen Z, Shestopalov, Valery I
Format: Article
Language:English
Subjects:
Adult
Bacteria - genetics
Conjunctiva - microbiology
DNA, Bacterial - genetics
Gene Library
Genes, rRNA - genetics
High-Throughput Nucleotide Sequencing
Humans
Male
Metagenome - physiology
Middle Aged
Pilot Projects
Polymerase Chain Reaction
RNA, Ribosomal, 16S - genetics
Sequence Analysis, DNA
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Summary:Ocular surface (OS) microbiota contributes to infectious and autoimmune diseases of the eye. Comprehensive analysis of microbial diversity at the OS has been impossible because of the limitations of conventional cultivation techniques. This pilot study aimed to explore true diversity of human OS microbiota using DNA sequencing-based detection and identification of bacteria. Composition of the bacterial community was characterized using deep sequencing of the 16S rRNA gene amplicon libraries generated from total conjunctival swab DNA. The DNA sequences were classified and the diversity parameters measured using bioinformatics software ESPRIT and MOTHUR and tools available through the Ribosomal Database Project-II (RDP-II). Deep sequencing of conjunctival rDNA from four subjects yielded a total of 115,003 quality DNA reads, corresponding to 221 species-level phylotypes per subject. The combined bacterial community classified into 5 phyla and 59 distinct genera. However, 31% of all DNA reads belonged to unclassified or novel bacteria. The intersubject variability of individual OS microbiomes was very significant. Regardless, 12 genera-Pseudomonas, Propionibacterium, Bradyrhizobium, Corynebacterium, Acinetobacter, Brevundimonas, Staphylococci, Aquabacterium, Sphingomonas, Streptococcus, Streptophyta, and Methylobacterium-were ubiquitous among the analyzed cohort and represented the putative "core" of conjunctival microbiota. The other 47 genera accounted for
ISSN:1552-5783
0146-0404
1552-5783
DOI:10.1167/iovs.10-6939