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2011 Abstract List | < Previous | Next > 2011 OMIG Abstract 21 Use of metagenomics to document Acanthamoeba microbial communities 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 >
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