The Charles T. Campbell Eye Microbiology Lab
UPMCUniversity of Pittsburgh Schools of the Health Sciences
HomeContact InformationLab Diagnostic TestingAntibiotic SusceptibilityAntimicrobial TherapyCurrent ResearchPhotos

Ocular Microbiology and Immunology Group
Back to OMIG Main Page

2011 Abstract List | < Previous | Next >

2011 OMIG Abstract 21

Use of metagenomics to document Acanthamoeba microbial communities
D. Miller1, J. Maestre1, M. Diaz1, E. Perez1, V. Shestopalov1, R. Van Gelder2, E. Alfonso1
1Bascom Palmer Eye Institute, University of Miami, Miami, FL, 2University of Washington, Seattle, WA

Purpose: The pathology of Acanthamoeba keratitis is still being deciphered. Acanthamoeba may harbor unique microbiomes and, similar to Wobachia, these may contribute to the pathology of the disease. Metagenomics is a culture- independent, genomic approach to studying mixed microbial communities.   We implemented a small pilot study using metagenomics (454 pyrosequencing, Biome representation in silico karyotyping [BRISK]) to document the presence and diversity of amebic microbiomes (endosymbionts) recovered from clinical samples isolates and compared these with culture-dependent recovery.

Methods: DNA from Acanthamoeba hosts recovered  from patients with amebic keratitis (N=9) were extracted, purified, aliquoted and forwarded to the respective laboratories (454 pyrosoequencing Argonne National Laboratories and BRISK-Van Gelder laboratory at University of Washington) for processing and analysis.

Results: Abundant mixed microbial communities were documented with both metagenomics techniques. More than 12 phyla and 23 different bacteria species were documented in the 7 samples evaluated by 454 based pyrosequencing vs 13 phyla and greater than 290 total microbial species detected among the 4 samples evaluated with BRISK. Despite the difference in species diversity and richness, a “core” microbiome composed of Delftia acidovrans, Stenotrophomonas maltophilia, and Pseudomonas aeruginosa was observed. Culture results revealed 7 phyla and 12 species, including all three members of the “core” amebic microbiome. Severity and disease duration correlated with increasing community mix. 

Conclusions: Use of metagenomics can serve as a tool to help decipher the epidemiology and pathology of amebic keratitis. Acanthamoeba hosts harbor a diverse microbiota that might play a role in the immunology and pathology of the disease.

Disclosures: N

2011 Abstract List | < Previous | Next >