Abstract
The dormant and durable spore form of Bacillus anthracis is an ideal biological weapon of mass destruction1,2. Once inhaled, spores are transported by alveolar macrophages to lymph nodes surrounding the lungs, where they germinate; subsequent vegetative expansion causes an overwhelming flood of bacteria and toxins into the blood, killing up to 99% of untreated victims. Natural and genetically engineered antibiotic-resistant bacilli amplify the threat of spores being used as weapons, and heighten the need for improved treatments and spore-detection methods after an intentional release. We exploited the inherent binding specificity and lytic action of bacteriophage enzymes called lysins for the rapid detection and killing of B. anthracis. Here we show that the PlyG lysin, isolated from the γ phage of B. anthracis, specifically kills B. anthracis isolates and other members of the B. anthracis ‘cluster’ of bacilli in vitro and in vivo. Both vegetative cells and germinating spores are susceptible. The lytic specificity of PlyG was also exploited as part of a rapid method for the identification of B. anthracis. We conclude that PlyG is a tool for the treatment and detection of B. anthracis.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Mock, M. & Fouet, A. Anthrax. Annu. Rev. Microbiol. 55, 647–671 (2001)
Inglesby, T. V. et al. Anthrax as a biological weapon, 2002: Updated recommendations for management. J. Am. Med. Assoc. 287, 2236–2252 (2002)
Wang, I. N., Smith, D. L. & Young, R. Holins: The protein clocks of bacteriophage infections. Annu. Rev. Microbiol. 54, 799–825 (2000)
Lopez, R., Garcia, E., Garcia, P. & Garcia, J. L. The pneumococcal cell wall degrading enzymes: A modular design to create new lysins? Microb. Drug Resist. 3, 199–211 (1997)
Loessner, M. J., Kramer, K., Ebel, F. & Scherer, S. C-terminal domains of Listeria monocytogenes bacteriophage murein hydrolases determine specific recognition and high-affinity binding to bacterial cell wall carbohydrates. Mol. Microbiol. 44, 335–349 (2002)
Loeffler, J. M., Nelson, D. & Fischetti, V. A. Rapid killing of Streptococcus pneumoniae with a bacteriophage cell wall hydrolase. Science 294, 2170–2172 (2001)
Nelson, D., Loomis, L. & Fischetti, V. A. Prevention and elimination of upper respiratory colonization of mice by group A streptococci by using a bacteriophage lytic enzyme. Proc. Natl Acad. Sci. USA 98, 4107–4112 (2001)
Redmond, C., Henderson, I., Turnbull, P. C. B. & Bowen, J. Phage from different strains of Bacillus anthracis. Salisbury Med. Bull. Spec. Suppl. 87, 60–63 (1996)
Brown, E. R. & Cherry, W. B. Specific identification of Bacillus anthracis by means of a variant bacteriophage. J. Infect. Dis. 96, 34–39 (1955)
Turnbull, P. C. B. Definitive identification of Bacillus anthracis—a review. J. Appl. Microbiol. 87, 237–240 (1999)
Helgason, E. et al. Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis—one species on the basis of genetic evidence. Appl. Environ. Microbiol. 66, 2627–2630 (2000)
Burdon, K. L., Davis, J. S. & Wende, R. D. Experimental infection of mice with Bacillus cereus: Studies of pathogenesis and pathologic changes. J. Infect. Dis. 117, 307–316 (1967)
Lamanna, C. & Jones, L. Lethality for mice of vegetative and spore forms of Bacillus cereus and Bacillus cereus-like insect pathogens injected intraperitoneally and subcutaneously. J. Bacteriol. 85, 532–535 (1963)
Fischetti, V. A. Phage antibacterials make a comeback. Nature Biotechnol. 19, 734–735 (2001)
Makino, S., Ito, N., Inoue, T., Miyata, S. & Moriyama, R. A spore-lytic enzyme released from Bacillus cereus spores during germination. Microbiology 140, 1403–1410 (1994)
Jenkinson, H. F., Kay, D. & Mandelstam, J. Temporal dissociation of late events in Bacillus subtilis sporulation from expression of genes that determine them. J. Bacteriol. 141, 793–805 (1980)
Henriques, A. O. & Moran, C. P. Jr. Structure and assembly of the bacterial endospore coat. Methods 20, 95–110 (2000)
Santo, L. Y. & Doi, R. H. Ultrastructural analysis during germination and outgrowth of Bacillus subtilis spores. J. Bacteriol. 120, 475–481 (1974)
Brussow, H. & Hendrix, R. W. Phage genomics: Small is beautiful. Cell 108, 13–16 (2002)
Helgason, E., Caugant, D. A., Olsen, I. & Kolsto, A. B. Genetic structure of population of Bacillus cereus and B. thuringiensis isolates associated with periodontitis and other human infections. J. Clin. Microbiol. 38, 1615–1622 (2000)
Pannucci, J., Okinaka, R. T., Sabin, R. & Kuske, C. R. Bacillus anthracis pXO1 plasmid sequence conservation among closely related bacterial species. J. Bacteriol. 184, 134–141 (2002)
Ticknor, L. O. et al. Fluorescent amplified fragment length polymorphism analysis of norwegian Bacillus cereus and Bacillus thuringiensis soil isolates. Appl. Environ. Microbiol. 67, 4863–4873 (2001)
Schmitt, C. K. et al. Absence of all components of the flagellar export and synthesis machinery differentially alters virulence of Salmonella enterica serovar Typhimurium in models of typhoid fever, survival in macrophages, tissue culture invasiveness, and calf enterocolitis. Infect. Immun. 69, 5619–5625 (2001)
Jackson, P. J. et al. Characterization of the variable-number tandem repeats in vrrA from different Bacillus anthracis isolates. Appl. Environ. Microbiol. 63, 1400–1405 (1997)
Guzman, L. M., Belin, D., Carson, M. J. & Beckwith, J. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J. Bacteriol. 177, 4121–4130 (1995)
Loessner, M. J., Wendlinger, G. & Scherer, S. Heterogeneous endolysins in Listeria monocytogenes bacteriophages: A new class of enzymes and evidence for conserved holin genes within the siphoviral lysis cassettes. Mol. Microbiol. 16, 1231–1241 (1995)
Stopa, P. J., Tieman, D., Coon, P. A., Milton, M. M. & Paterno, D. Detection of biological aerosols by luminescence techniques. Field Anal. Chem. Technol. 3, 283–290 (1999)
Corran, J. The induction of supersuppressor mutants of Bacillus subtilis by ethyl methanesulphonate and the posttreatment modification of mutation yield. Mol. Gen. Genet. 103, 42–57 (1968)
Mazas, M., Martinez, S., Lopez, M., Alvarez, A. B. & Martin, R. Thermal inactivation of Bacillus cereus spores affected by the solutes used to control water activity of the heating medium. Int. J. Food Microbiol. 53, 61–67 (1999)
Keim, P. et al. Multiple-locus variable-number tandem repeat analysis reveals genetic relationships within Bacillus anthracis. J. Bacteriol. 182, 2928–2936 (2000)
Acknowledgements
We thank P. J. Piggot, L. W. Mayer, A. L. Turetsky, A. Aronson, A. Keynan, H.-W. Ackerman, R. J. McNall and T. A. Kokjohn for their gifts of strains; E. Sphicas at the Bio-imaging Resource Center at The Rockefeller University for help with electron microscopy; R. L. Russell and S. Zhu for technical help; New Horizons Diagnostics for their luminometer and reagents and P. Model and members of the V.A.F. laboratory for reviewing this manuscript. We also thank A. Keynan for reviewing this manuscript and for advice regarding spore preparation and germination. This work was supported by a grant from the Defense Advanced Research Projects Agency (DARPA).
Author information
Authors and Affiliations
Ethics declarations
Competing interests
The authors declare that they have no competing financial interests.
Rights and permissions
About this article
Cite this article
Schuch, R., Nelson, D. & Fischetti, V. A bacteriolytic agent that detects and kills Bacillus anthracis. Nature 418, 884–889 (2002). https://doi.org/10.1038/nature01026
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nature01026
This article is cited by
-
Endolysin EN572-5 as an alternative to treat urinary tract infection caused by Streptococcus agalactiae
Applied Microbiology and Biotechnology (2024)
-
New bacteriophage-derived lysins, LysJ and LysF, with the potential to control Bacillus anthracis
Applied Microbiology and Biotechnology (2024)
-
Therapeutic potential of bacteriophage endolysins for infections caused by Gram-positive bacteria
Journal of Biomedical Science (2023)
-
Composition and functions of bacterial membrane vesicles
Nature Reviews Microbiology (2023)
-
Substantiation of propitious “Enzybiotic” from two novel bacteriophages isolated from a wastewater treatment plant in Qatar
Scientific Reports (2022)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.