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Bacteriophage PEV20 and Ciprofloxacin Combination Treatment Enhances Removal of Pseudomonas aeruginosa Biofilm Isolated from Cystic Fibrosis and Wound Patients

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Abstract

Antibiotic resistance in Pseudomonas aeruginosa biofilms necessitates the need for novel antimicrobial therapy with anti-biofilm properties. Bacteriophages (phages) are recognized as an ideal biopharmaceutical for combating antibiotic-resistant bacteria especially when used in combination with antibiotics. However, previous studies primarily focused on using phages against of P. aeruginosa biofilms of laboratory strains. In the present study, biofilms of six P. aeruginosa isolated from cystic fibrosis and wound patients, and one laboratory strain was treated singly and with combinations of anti-Pseudomonas phage PEV20 and ciprofloxacin. Of these strains, three were highly susceptible to the phage, while one was partially resistant and one was completely resistant. Combination treatment with PEV20 and ciprofloxacin enhanced biofilm eradication compared with single treatment. Phage and ciprofloxacin synergy was found to depend on phage-resistance profile of the target bacteria. Furthermore, phage and ciprofloxacin combination formulation protected the lung epithelial and fibroblast cells from P. aeruginosa and promoted cell growth. The results demonstrated that thorough screening of phage-resistance is crucial for designing phage-antibiotic formulation. The addition of highly effective phage could reduce the ciprofloxacin concentration required to combat P. aeruginosa infections associated with biofilm in cystic fibrosis and wound patients.

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Abbreviations

CF:

Cystic fibrosis

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Acknowledgments

We thank Prof Craig Winstanley of the University of Liverpool, UK, for strains LESB58 and MANC3733. Research reported in this publication was supported by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under Award Number R33AI121627 (H-K C.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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Author notes

  1. Rachel Yoon Kyung Chang and Theerthankar Das contributed equally to this work.

Authors and Affiliations

  1. Advanced Drug Delivery Group, School of Pharmacy, The University of Sydney, Sydney, NSW, 2006, Australia

    Rachel Yoon Kyung Chang & Hak-Kim Chan

  2. Department of Infectious Diseases and Immunology, Sydney Medical School, The University of Sydney, Sydney, NSW, Australia

    Theerthankar Das & Jim Manos

  3. The Evergreen State College, Olympia, Washington, 98502, USA

    Elizabeth Kutter

  4. AmpliPhi Biosciences AU, 7/27 Dale Street, Brookvale, Sydney, NSW, 2100, Australia

    Sandra Morales

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  1. Rachel Yoon Kyung Chang
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Correspondence to Hak-Kim Chan.

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Supplemental 1

Percentage biofilm viability in 48-h-old biofilm after 24-h treatment with tobramycin alone (MIC, 2MIC, and 3MIC), PEV20 alone (108 PFU/mL), or antibiotics (1/2MIC, MIC, and 2MIC) combined with PEV20 (108 PFU/mL). Error bars represent standard deviations from multiple cultures (n  =  4). * indicates statistically significant differences (P < 0.05) in percentage biofilm viability of the treated groups in comparison with non-treated control. (PNG 510 kb)

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Chang, R.Y.K., Das, T., Manos, J. et al. Bacteriophage PEV20 and Ciprofloxacin Combination Treatment Enhances Removal of Pseudomonas aeruginosa Biofilm Isolated from Cystic Fibrosis and Wound Patients. AAPS J 21, 49 (2019). https://doi.org/10.1208/s12248-019-0315-0

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