April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Diversity And Prevalence Of Microorganisms At Healthy Ocular Surface
Author Affiliations & Notes
  • Valery I. Shestopalov
    Bascom Palmer Eye Institute, Department of Ophthalmology,
    Univ of Miami Miller Sch of Med, Miami, Florida
  • Qunfeng Dong
    Departments of Biological Sciences, University of North Texas, Denton, Texas
  • Lakshmi Akileswaran
    Bascom Palmer Eye Institute, Department of Ophthalmology,
    University of Washington, Seattle, Washington
  • Jay Shendure
    Department of Genome Science, University of Washington, Miami, Washington
  • Choli Lee
    Molecular Pharmacology, Department of Genome Science,
    University of Washington, Seattle, Washington
  • Jennifer M. Brulc
    Institute for Genomics and Systems Biology, Argonne National Laboratory, Argonne, Illinois
  • Darlene Miller
    Bascom Palmer Eye Institute, Department of Ophthalmology,
    Univ of Miami Miller Sch of Med, Miami, Florida
  • Dionysios A. Antonopoulos
    Institute for Genomics and Systems Biology, Argonne National Laboratory, Argonne, Illinois
  • Vladlen Z. Slepak
    Molecular Pharmacology, Department of Genome Science,
    Univ of Miami Miller Sch of Med, Miami, Florida
  • Russell N. Van Gelder
    Ophthalmology,
    University of Washington, Seattle, Washington
  • Footnotes
    Commercial Relationships  Valery I. Shestopalov, None; Qunfeng Dong, None; Lakshmi Akileswaran, None; Jay Shendure, None; Choli Lee, None; Jennifer M. Brulc, None; Darlene Miller, None; Dionysios A. Antonopoulos, None; Vladlen Z. Slepak, None; Russell N. Van Gelder, None
  • Footnotes
    Support  1R21EY019974 (V.I.S.), RO1EY018666 (V.S.); P30 EY014801, Unrestricted RPB to Dept. Ophthalmol., Burroughs-Wellcome Foundation grant (RVG)
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 1488. doi:https://doi.org/
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      Valery I. Shestopalov, Qunfeng Dong, Lakshmi Akileswaran, Jay Shendure, Choli Lee, Jennifer M. Brulc, Darlene Miller, Dionysios A. Antonopoulos, Vladlen Z. Slepak, Russell N. Van Gelder; Diversity And Prevalence Of Microorganisms At Healthy Ocular Surface. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1488. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Ocular surface (OS) microbiota contribute to infectious diseases of the eye. Comprehensive analysis of microbial diversity at the OS has been impossible because of the limitations of conventional microbial culture techniques. This study aimed to explore true diversity of human OS microbiota using DNA sequencing-based detection and identification of microbes at healthy OS and on contact lenses.

Methods: : We utilized 454 Roche deep sequencing of 16S rRNA gene amplicon libraries and Biome Representational in Silico Karyotyping (BRiSK) to obtain microbial DNA reads. These were classified using ESPRIT, MOTHUR Ribosomal Database Project-II software. Endonuclease-generated fragments were matched to known microbial genomes.

Results: : Deep sequencing of conjunctival rDNA from four subjects yielded 115,003 quality DNA reads corresponding to an average of 221 bacterial species per subject. BRiSK was applied to three conjunctival samples and yielded an average of 434 microbial DNA reads, corresponding to 17 known microbial (bacterial, fungal and viral) species. Significantly, BRiSK analysis of microbiota on used contact lenses detected 28 known microbial species. The combined bacterial community was classified into 5 phyla and 64 distinct genera. In addition, >40% of 16S rRNA-detected bacteria and >90% of BRiSK-detected microbes belonged to unclassified or novel species. Despite high inter-subject variability, 12 known genera were ubiquitous among the analyzed cohort and represented putative "core" of the OS microbiome. The other 52 genera accounted for less than 4% and likely belonged to transient species. Unexpectedly, healthy conjunctiva contained many genera that are commonly identified as ocular surface pathogens.

Conclusions: : Our first DNA sequencing-based survey of healthy OS have revealed unexpectedly diverse microbial community. All analyzed samples contained ubiquitous "core" genera that included commensal, environmental and opportunistic pathogenic bacteria, fungi and viruses.

Keywords: conjunctiva • cornea: epithelium • microbial pathogenesis: experimental studies 
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