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

Author:

Hoopes J. Todd1,Stark Caren J.1,Kim Han Ah12,Sussman Daniel J.3,Donovan David M.4,Nelson Daniel C.1

Affiliation:

1. Center for Advanced Research in Biotechnology, University of Maryland Biotechnology Institute, Rockville, Maryland

2. Center for Advanced Research in Biotechnology/Thomas S. Wootton High School Science Internship Program, Rockville, Maryland

3. 9110 Red Branch Road, Columbia, Maryland

4. Animal Biosciences and Biotechnology Laboratory, United States Department of Agriculture, Beltsville, Maryland

Abstract

ABSTRACT 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 10 8 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.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Reference29 articles.

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2. AOAC International. 2005. AOAC germicidal spray products test, official method 961.02. In G. W. Latimer and W. Horwitz (ed.), Official methods of analysis of the AOAC International, 18th ed. Association of Official Analytical Chemists, Gaithersburg, MD.

3. AOAC International. 2005. Use dilution methods for testing disinfectants, official methods 955.15 and 964.02. In G. W. Latimer and W. Horwitz (ed.), Official methods of analysis of the AOAC International, 18th ed. Association of Official Analytical Chemists, Gaithersburg, MD.

4. Barnham, M., G. Cole, A. Efstratiou, J. R. Tagg, and S. A. Skjold. 1987. Characterization of Streptococcus zooepidemicus (Lancefield group C) from human and selected animal infections. Epidemiol. Infect.98:171-182.

5. Blanchard, P. C., and K. M. Fiser. 1994. Streptococcus dysgalactiae polyarthritis in dairy goats. J. Am. Vet. Med. Assoc.205:739-741.

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