Ocular Microbiology and Immunology Group
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2013 OMIG Abstract 10

The Ocular Surface Microbiome: Indigenous Communities Colonize the Healthy Cornea and Conjunctiva
Valery I. Shestopalov^1A,B, Nabeel Shalabi^1A, Alexander Tuzhikov^1A, Anat Galor^1A, Onsiri Thanahantee^1A, Qunfeng Dong², Evelyn Toh³, David Nelson³, Lakshmi Akileswaran⁴, Elena Linerdopoulou⁴; Darlene Miller^1A, Russell N. Van Gelder⁴, Terrence O’Brien^1A
1ABascom Palmer Eye Institute,^1BCell Biology and Anatomy Univ. of Miami School of Medicine, Miami, FL;
²Department of Biological Sciences, University of North Texas, Denton, TX; ³Department of Biology, Indiana University, Bloomington, IN; ⁴Department of Ophthalmology, University of Washington, Seattle, WA

Purpose. Infections of the ocular surface (OS) are a major cause of visual disability in the US and worldwide, however the composition of indigenous microbiota colonizing OS remains poorly characterized.

Methods. This study utilized two next generation sequencing strategies to assess microbial composition at the conjunctiva and cornea. We compared the results obtained by 16S rRNA gene sequencing and a Biome Representational in Silico Karyotyping (BRiSK). Conjunctivae were sampled by swabbing; corneal epithelia were removed by Microkeratome (Debauer) and collected during surgery. Only eyes with healthy OS were sampled from patient cohort of both genders with age average 56±21.

Results. Using 454 Roche pyrosequencing technology to analyze bacteria, we detected 59 bacterial genera belonging to 5 phyla in conjunctiva, and 56 genera from the same phyla in the cornea. BRiSK analysis added 18 species of viruses and retroviruses, 6 fungal and 4 protozoans species within the same cohort. Genders differed significantly in bacterial alpha-diversity, which was up to three-fold higher in female vs. male corneas. In addition to known bacteria, approximately 30% of 16S rRNA-detected bacteria and up to 90% of BRiSK-detected sequences belonged to unclassified or novel species. The application of TUIT algorithm, an  alternative to the mainstream RDP-II Classifier, have increased the efficiency of DNA-based bacterial identification.

Conclusions. Our results indicate that all conjunctival and the majority of corneal epithelia are colonized by diverse microbiota. Despite technical complexity, next generation sequencing provides a powerful tool for the analysis of the OS microbiome.

Support: EY019974 (V.I.S); EY022038 (V.I.S and RVG); RC2HG005806-01 (Q.D. & D.N.), Burroughs-Wellcome Foundation (R.V.G); P30 EY014801 and an unrestricted RPB grants (Departments of Ophthalmology)

2013 Agenda and Abstracts | < Previous | Next >