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Applied and Environmental Microbiology, May 2003, p. 2563-2567, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2563-2567.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Enhanced Killing of Acanthamoeba Cysts with a Plant Peroxidase-Hydrogen Peroxide-Halide Antimicrobial System

Reanne Hughes, Peter W. Andrew, and Simon Kilvington*

Department of Microbiology and Immunology, University of Leicester, Leicester LE1 9HN, United Kingdom

Received 30 August 2002/ Accepted 31 January 2003

The activity of H2O2 against the resistant cyst stage of the pathogenic free-living amoeba Acanthamoeba was enhanced by the addition of KI and either horseradish peroxidase or soybean peroxidase or, to a lesser degree, lactoperoxidase. This resulted in an increase in the cysticidal activity of 3% (wt/vol) H2O2, and there was >3-log killing in 2 h, compared with the 6 h required for comparable results with the peroxide solution alone (P < 0.05). With 2% H2O2, enhancement was observed at all time points (P < 0.05), and total killing of the cyst inoculum occurred at 4 h, compared with 6 h for the peroxide alone. The activity of sublethal 1% H2O2 was enhanced to give 3-log killing after 8 h of exposure (P < 0.05). No enhancement was obtained when KCl or catalase was used as a substitute in the reaction mixtures. The H2O2 was not neutralized in the enhanced system during the experiments. However, in the presence of a platinum disk used to neutralize H2O2 in contact lens care systems, the enhanced 2% H2O2 system gave 2.8-log killing after 6 h or total cyst killing by 8 h, and total neutralization of the H2O2 occurred by 4 h. In contrast, 2% H2O2 alone resulted in <0.8-log killing of cysts in the presence of the platinum disk due to rapid (<1 h) neutralization of the peroxide. Our observations could result in significant improvement in the efficacy of H2O2 contact lens disinfection systems against Acanthamoeba cysts and prevention of acanthamoeba keratitis.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Leicester, Medical Sciences Building, P.O. Box 138, University Rd., Leicester LE1 9HN, United Kingdom. Phone: 44 (0) 116 252 2950. Fax: 44 (0) 116 252 5030. E-mail: sk46{at}le.ac.uk.

Applied and Environmental Microbiology, May 2003, p. 2563-2567, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2563-2567.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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