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Applied and Environmental Microbiology, March 2009, p. 1388-1394, Vol. 75, No. 5
0099-2240/09/$08.00+0     doi:10.1128/AEM.02195-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Use of a Bacteriophage Lysin, PlyC, as an Enzyme Disinfectant against Streptococcus equi{triangledown}

J. Todd Hoopes,1 Caren J. Stark,1 Han Ah Kim,1,2 Daniel J. Sussman,3 David M. Donovan,4 and Daniel C. Nelson1*

Center for Advanced Research in Biotechnology, University of Maryland Biotechnology Institute, Rockville, Maryland,1 Center for Advanced Research in Biotechnology/Thomas S. Wootton High School Science Internship Program, Rockville, Maryland,2 9110 Red Branch Road, Columbia, Maryland,3 Animal Biosciences and Biotechnology Laboratory, United States Department of Agriculture, Beltsville, Maryland4

Received 22 September 2008/ Accepted 24 December 2008

Streptococcus equi is the causative agent of the purulent infection equine strangles. This disease is transmitted through shedding of live bacteria from nasal secretions and abscess drainage or by contact with surfaces contaminated by the bacteria. Disinfectants are effective against S. equi, but inactivation by environmental factors, damage to equipment, and toxicity are of great concern. Bacteriophage-encoded lysins (cell wall hydrolases) have been investigated as therapeutic agents due to their ability to lyse susceptible gram-positive organisms. Here, we investigate the use of one lysin, PlyC, as a narrow-spectrum disinfectant against S. equi. This enzyme was active against >20 clinical isolates of S. equi, including both S. equi subsp. equi and S. equi subsp. zooepidemicus. Significantly, PlyC was 1,000 times more active on a per weight basis than Virkon-S, a common disinfecting agent, with 1 µg of enzyme able to sterilize a 108 CFU/ml culture of S. equi in 30 min. PlyC was subjected to a standard battery of tests including the Use Dilution Method for Testing Disinfectants and the Germicidal Spray Products Test. Results indicate that aerosolized PlyC can eradicate or significantly reduce the S. equi load on a variety of materials found on common stable and horse-related equipment. Additionally, PlyC was shown to retain full activity under conditions that mimic a horse stable, i.e., in the presence of nonionic detergents, hard water, or organic materials. We propose PlyC as the first protein-based, narrow-spectrum disinfectant against S. equi, which may augment or supplement the use of broad-spectrum disinfectants in barns and stables where equine strangles is prevalent.

* Corresponding author. Mailing address: Center for Advanced Research in Biotechnology, University of Maryland Biotechnology Institute, 9600 Gudelsky Drive, Rockville, MD 20850. Phone: (240) 314-6249. Fax: (240) 314-6225. E-mail: nelsond{at}umbi.umd.edu

{triangledown} Published ahead of print on 9 January 2009.

Applied and Environmental Microbiology, March 2009, p. 1388-1394, Vol. 75, No. 5
0099-2240/09/$08.00+0     doi:10.1128/AEM.02195-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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