March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Non-Cultivatable Bacterial Biofilm Communities in Used Contact Lens Cases
Author Affiliations & Notes
  • Judith L. Flanagan
    Brien Holden Vision Institute, Sydney, Australia
  • Martin Allgeier
    Joint Genome Institute, Walnut Creek, California
  • Mark D. Willcox
    Brien Holden Vision Institute, Univ of New South Wales, Sydney, Australia
  • Philip Hugenholtz
    Australian Centre for Ecogenomics, School of Chemistry and Molecular Biosciences & Institute for Mol, University of Queensland, Queensland, Australia
  • Footnotes
    Commercial Relationships  Judith L. Flanagan, None; Martin Allgeier, None; Mark D. Willcox, None; Philip Hugenholtz, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 6087. doi:
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      Judith L. Flanagan, Martin Allgeier, Mark D. Willcox, Philip Hugenholtz; Non-Cultivatable Bacterial Biofilm Communities in Used Contact Lens Cases. Invest. Ophthalmol. Vis. Sci. 2012;53(14):6087.

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

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Abstract

Purpose: : Using phylogentic modeling, we sought to document the microbial ecology of contaminating biofilms in used contact lens cases as the first step in an effort to determine how these communities contribute to, or help to protect against, adverse ocular events such as corneal infiltrative events (CIEs).

Methods: : Lens cases from subjects (and control cases) were swabbed with PBS (100 µl). DNA was extracted using Wizard Genomic DNA Isolation System (Promega, CA, USA). SSU rRNA was amplified (35 cycles PCR) using primers 926F and 1392R with barcodes and 454 linker. Pyrosquencing was performed using 454 titanium technology. Raw sequence reads were analysed using software pyrotaggers followed by custom analysis.

Results: : 6000 independent reads were generated from each lens case and indicated a high microbial diversity including the presence of phyla previously implicated in MK such as Propionibacterium, Staphylococcus, Streptococcus, Gammaproteobacteria (including Stenotrophomonads, Pseudomonads), and Tsukamurella. Shannon’s diversity index (SDI) which typically ranges from 1.5 (low species richness and evenness) to 3.5 (high species evenness and richness) indicated high diversity in the used contact lens cases (3.853 and 2.840), similar to that found on human skin in which most sequences are assigned to 4 phyla: Actinobacter, Firmicutes, Proteobacteria and Bacteroidetes. Most abundant phyla in the lens cases were α- and γ-Proteobacteria, Firmicutes and Actinobacter. These commensal profiles differ markedly from bacteria detected by culture from asymptomatic subjects.

Conclusions: : This pilot study provided evidence of very high microbial diversity in used contact lens cases of healthy wearers. Whether these commensal bacteria offer some protection against infection needs to be addressed through a larger temporal study in which bacterial community dynamics can be monitored for both healthy wearers and wearers who suffer a CIE.

Keywords: microbial pathogenesis: experimental studies • contact lens • bacterial disease 
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