Background
Current Clostridium difficile infection (CDI) antibiotic regimens have become increasingly ineffective at achieving
cure and preventing recurrence. A recently developed alternative to conventional antibiotics
are phage tail-like particles (PTLPs), which are proteins that are morphologically
similar to bacteriophages and are produced by C difficile. This study examines the in vitro killing spectrum of a previously unreported PTLP
isolated from a clinical isolate of C difficile.
Methods
Using patient-derived samples from an institutional review board-approved C difficile tissue bank, a ribotype 078 C difficile isolate was anaerobically incubated on blood agar plates that were preswabbed with
norfloxacin to induce the production of PTLPs. Concentrated PTLP populations were
confirmed using transmission electron microscopy. Using a standard lawn spot approach,
bactericidal activity was assessed as indicated by a clearing within the bacterial
lawn. The PTLP genomic cluster was also fully sequenced and open reading frames were
annotated according to predicted function.
Results
PTLPs were assessed using 64 patient-derived C difficile isolates of varying ribotypes. PTLPs demonstrated complete bactericidal activity
in 21 of 25 ribotype 027 isolates with partial activity in 2 of the 25. Complete bactericidal
activity was not demonstrated against any other ribotype or non-difficile bacteria, suggesting a species and ribotype specificity. Functional genes, which
may be necessary for killing, were identified within the PTLP genetic locus.
Conclusion
PTLPs demonstrate capability in eradicating C difficile in vitro, and with further development, may represent an organism-specific, microbiome-sparing
therapy for CDI.
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to SurgeryAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Guidelines for diagnosis, treatment, and prevention of Clostridium difficile infections.Am J Gastroenterol. 2013; 108: 478-498
- Treatment of Clostridium difficile-associated disease: old therapies and new strategies.Lancet Infect Dis. 2005; 5: 549-557
- Breaking the cycle: treatment strategies for 163 cases of recurrent Clostridium difficile disease.Am J Gastroenterol. 2002; 97: 1769-1775
- Reproducible community dynamics of the gastrointestinal microbiota following antibiotic perturbation.Infect Immun. 2009; 77: 2367-2375
- Antibiotic administration alters the community structure of the gastrointestinal micobiota.Gut Microbes. 2010; 1: 279-284
- Fecal microbiota transplantation for Clostridium difficile infection: systematic review and meta-analysis.Am J Gastroenterol. 2013; 108: 500-508
- Antibacterial efficacy of R-type pyocins towards Pseudomonas aeruginosa in a murine peritonitis model.Antimicrob Agents Chemother. 2008; 52: 1647-1652
- An engineered R-type pyocin is a highly specific and sensitive bactericidal agent for the food-borne pathogen Escherichia coli O157:H7.Antimicrob Agents Chemother. 2009; 53: 3074-3080
- The R-type pyocin of Pseudomonas aeruginosa is related to P2 phage, and the F-type is related to lambda phage.Mol Microbiol. 2000; 38: 213-231
- The defective prophage pool of Escherichia coli O157: prophage-prophage interactions potentiate horizontal transfer of virulence determinants.PLoS Pathog. 2009; 5: e1000408
- Lipopolysaccharide as Shield and Receptor for R-Pyocin-Mediated Killing in Pseudomonas aeruginosa.J Bacteriol. 2010; 192: 1921-1928
- Horizontal gene transfer converts non-toxigenic Clostridium difficile strains into toxin producers.Nat Commun. 2013; 4: 2601
- Morphological and genetic diversity of temperate phages in Clostridium difficile.Appl Environ Microbiol. 2007; 73: 7358-7366
- Diverse Temperate Bacteriophage Carriage in Clostridium difficile 027 Strains.PLoS One. 2012; 7: e37263
- Novel high-molecular-weight, R-type bacteriocins of Clostridium difficile.J Bacteriol. 2012; 194: 6240-6247
- Prophage carriage and diversity within clinically relevant strains of Clostridium difficile.Appl Environ Microbiol. 2012; 78: 6027-6034
- Production of a bacteriophage, a phage tail-like bacteriocin and an antibiotic by Bacillus azotofixans.An Acad Bras Cienc. 1990; 62: 85-94
- Characterization of cryptic prophages (monocins) in Listeria and sequence analysis of a holin/endolysin gene.Microbiology. 1995; 141: 2577-2584
- Phage tail-like (high-molecular-weight) bacteriocins of Budvicia aquatica and Pragia fontium (Enterobacteriaceae).Appl Environ Microbiol. 2005; 71: 8970-8973
- Characterization of enterocoliticin, a phage tail-like bacteriocin, and its effect on pathogenic Yersinia enterocolitica strains.Appl Environ Microbiol. 2001; 67: 5634-5642
- Pyocin S2 (Sa) kills Pseudomonas aeruginosa strains via the FpvA type I ferripyoverdine receptor.J Bacteriol. 2007; 189: 7663-7668
- The pyocins of Pseudomonas aeruginosa.Biochimie. 2002; 84: 499-510
- Developmental pathways for the temperate phage: lysis vs lysogeny.Annu Rev Genet. 1972; 6: 157-190
- Toxin production by an emerging strain of Clostridium difficile associated with outbreaks of severe disease in North America and Europe.Lancet. 2005; 366: 1079-1084
- Gut flora in health and disease.Lancet. 2003; 361: 512-519
- Retargeting R-type pyocins to generate novel bactericidal protein complexes.Appl Environ Microbiol. 2008; 74: 3868-3876
- Protein cell surface display in Gram-positive bacteria: from single protein to macromolecular protein structure.FEMS Microbiol Lett. 2006; 256: 1-15
- Systematic analysis of an amidase domain CHAP in 12 Staphylococcus aureus genomes and 44 staphylococcal phage genomes.Comput Biol Chem. 2010; 34: 251-257
- C-terminal domains of Listeria monocytogenes bacteriophage murein hydrolases determine specific recognition and high-affinity binding to bacterial cell wall carbohydrates.Mol Microbiol. 2002; 44: 335-349
Article info
Publication history
Published online: July 22, 2014
Accepted:
June 19,
2014
Footnotes
Funding Source: No internal or external financial support was used for this report.
Disclosures: The authors' have no conflicts of interest or financial ties to disclose.
Outside support: AvidBiotics Corp. (South San Francisco, CA) provided a control reagent that was used during this experiment. AvidBiotics is supported by a SBIR II grant (#1R43 AI098186) from National Institute of Allergy and Infectious Diseases.
Identification
Copyright
© 2015 Elsevier Inc. Published by Elsevier Inc. All rights reserved.
