September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Rose bengal-mediated photodynamic antimicrobial therapy to inhibit Pseudomonas aeruginosa isolates in agar plates and contaminated contact lens cases
Author Affiliations & Notes
  • Heather Ann Durkee
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Nidhi Relhan
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Alejandro Arboleda
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Mariela C Aguilar
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Karam AlRahman Alawa
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Francisco Halili
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Cornelis Rowaan
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Guillermo Amescua
    Anne Bates Leach Eye Hospital, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Harry W Flynn
    Anne Bates Leach Eye Hospital, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Darlene Miller
    Ocular Microbiology Laboratory, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Jean-Marie A Parel
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
    CHU Sart-Tillman, Department of Ophthalmology, University of Liege, Liege, Belgium
  • Footnotes
    Commercial Relationships   Heather Durkee, None; Nidhi Relhan, None; Alejandro Arboleda, None; Mariela Aguilar, None; Karam Alawa, None; Francisco Halili, None; Cornelis Rowaan, None; Guillermo Amescua, None; Harry Flynn, None; Darlene Miller, None; Jean-Marie Parel, None
  • Footnotes
    Support  This work was supported in part by the Florida Lions Eye Bank, the Edward D. and Janet K. Robson Foundation. Drs. KR Olsen and ME Hildebrandt, Drs. Raksha Urs and Aaron Furtado, NIH center grant P30EY14801, an unrestricted grant from Research to Prevent Blindness to the Department of Ophthalmology, Brien Holden Vision Research Institute, and the Henri and Flore Lesieur Foundation (JMP). The authors are grateful to Alex Gonzalez, Carolina de Freitas, Juan Silgado, Victor Hernandez and Adriana Henao-Pink for their technical contributions.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, No Pagination Specified. doi:
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      Heather Ann Durkee, Nidhi Relhan, Alejandro Arboleda, Mariela C Aguilar, Karam AlRahman Alawa, Francisco Halili, Cornelis Rowaan, Guillermo Amescua, Harry W Flynn, Darlene Miller, Jean-Marie A Parel; Rose bengal-mediated photodynamic antimicrobial therapy to inhibit Pseudomonas aeruginosa isolates in agar plates and contaminated contact lens cases. Invest. Ophthalmol. Vis. Sci. 201657(12):.

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

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Abstract

Purpose : Pseudomonas aeruginosa is often present in contact lens cases and can adhere to the contact lens and cause keratitis in patients. Keratitis secondary to Pseudomonas aeruginosa is difficult to treat medically and, in severe cases, can cause corneal ulceration and perforation. In this study we evaluate an alternative antimicrobial treatment, rose bengal-mediated photodynamic antimicrobial therapy (PDAT), for the inhibition of Pseudomonas aeruginosa on agar plates and in patients’ contaminated contact lens cases.

Methods : Pseudomonas aeruginosa isolates were prepared into inoculum of concentration 104 cfu/mL. Isolates were mixed with one of three concentrations of rose bengal (0.1%, 0.05% & 0.01%) or water (control) and plated onto agar plates. Plates were kept in the dark or underwent irradiation with a custom-built green LED light source for a total energy density of 5.4 J/cm2. At 72 hours, agar plates were photographed and growth was measured with LabVIEW software.
Contaminated contact lens cases were collected from six patients with suspected contact lens-induced keratitis of mixed microbial communities. Cases were rehydrated with balanced salt solution for 24 hours and cultured onto sheep’s blood agar. Rose bengal-mediated PDAT was performed on contact lens cases, as above. The cases were then decanted, 2 mL of tryptic soy broth was added, and cases were cultured again. Photographs of agar plates were taken for analysis.

Results : PDAT using all concentrations of rose bengal was able to inhibit Pseudomonas aeruginosa growth within central 47mm diameter, corresponding to the irradiation source diameter, and extended outwards towards the edge of the plate. Rose bengal without irradiation showed no inhibition. Contact lens case experiments showed no difference in growth between pre- and post-PDAT cultures.

Conclusions : Pseudomonas aeruginosa can be inhibited by rose bengal-mediated PDAT with 0.1%, 0.05%, and 0.01% concentrations on agar plates. In the contact lens cases, PDAT could not inhibit Pseudomonas aeruginosa, likely because the rose bengal could not penetrate the biofilm present. Further studies are needed to understand microbial biofilm properties and how this influences PDAT and other treatments.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

 

Rose bengal-mediated PDAT of Pseudomonas aeruginosa compared to controls on agar plates.

Rose bengal-mediated PDAT of Pseudomonas aeruginosa compared to controls on agar plates.

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