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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/11927

Title: The T7-Related Pseudomonas putida Phage phi 15 Displays Virion-Associated Biofilm Degradation Properties
Authors: Cornelissen, Anneleen
Ceyssens, Pieter-Jan
T'Syen, Jeroen
Van Praet, Helena
NOBEN, Jean-Paul
Shaburova, Olga V.
Krylov, Victor N.
Volckaert, Guido
Lavigne, Rob
Issue Date: 2011
Citation: PLOS ONE, 6 (4)
Abstract: Formation of a protected biofilm environment is recognized as one of the major causes of the increasing antibiotic resistance development and emphasizes the need to develop alternative antibacterial strategies, like phage therapy. This study investigates the in vitro degradation of single-species Pseudomonas putida biofilms, PpG1 and RD5PR2, by the novel phage phi 15, a 'T7-like virus' with a virion-associated exopolysaccharide (EPS) depolymerase. Phage phi 15 forms plaques surrounded by growing opaque halo zones, indicative for EPS degradation, on seven out of 53 P. putida strains. The absence of haloes on infection resistant strains suggests that the EPS probably act as a primary bacterial receptor for phage infection. Independent of bacterial strain or biofilm age, a time and dose dependent response of phi 15-mediated biofilm degradation was observed with generally a maximum biofilm degradation 8 h after addition of the higher phage doses (10(4) and 10(6) pfu) and resistance development after 24 h. Biofilm age, an in vivo very variable parameter, reduced markedly phage-mediated degradation of PpG1 biofilms, while degradation of RD5PR2 biofilms and phi 15 amplification were unaffected. Killing of the planktonic culture occurred in parallel with but was always more pronounced than biofilm degradation, accentuating the need for evaluating phages for therapeutic purposes in biofilm conditions. EPS degrading activity of recombinantly expressed viral tail spike was confirmed by capsule staining. These data suggests that the addition of high initial titers of specifically selected phages with a proper EPS depolymerase are crucial criteria in the development of phage therapy.
Notes: [Cornelissen, Anneleen; Ceyssens, Pieter-Jan; T'Syen, Jeroen; Van Praet, Helena; Volckaert, Guido; Lavigne, Rob] Katholieke Univ Leuven, Lab Gene Technol, Louvain, Belgium. [Noben, Jean-Paul] Hasselt Univ, Sch Life Sci, Biomed Res Inst, Diepenbeek, Belgium. [Noben, Jean-Paul] Hasselt Univ, Transnat Univ Limburg, Diepenbeek, Belgium. [Shaburova, Olga V.; Krylov, Victor N.] State Inst Genet Ind Microorganisms, Moscow, Russia. rob.lavigne@biw.kuleuven.be
URI: http://hdl.handle.net/1942/11927
DOI: 10.1371/journal.pone.0018597
ISI #: 000289671100010
ISSN: 1932-6203
Category: A1
Type: Journal Contribution
Validation: ecoom, 2012
Appears in Collections: Research publications

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