The T7-related Pseudomonas putida phage φ15 displays virion-associated biofilm degradation properties.

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 Ps...

Full description

Bibliographic Details
Main Authors: Anneleen Cornelissen, Pieter-Jan Ceyssens, Jeroen T'Syen, Helena Van Praet, Jean-Paul Noben, Olga V Shaburova, Victor N Krylov, Guido Volckaert, Rob Lavigne
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2011-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3079711?pdf=render
_version_ 1828389671667761152
author Anneleen Cornelissen
Pieter-Jan Ceyssens
Jeroen T'Syen
Helena Van Praet
Jean-Paul Noben
Olga V Shaburova
Victor N Krylov
Guido Volckaert
Rob Lavigne
author_facet Anneleen Cornelissen
Pieter-Jan Ceyssens
Jeroen T'Syen
Helena Van Praet
Jean-Paul Noben
Olga V Shaburova
Victor N Krylov
Guido Volckaert
Rob Lavigne
author_sort Anneleen Cornelissen
collection DOAJ
description 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 ϕ15, a 'T7-like virus' with a virion-associated exopolysaccharide (EPS) depolymerase. Phage ϕ15 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 ϕ15-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 ϕ15 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.
first_indexed 2024-12-10T06:32:42Z
format Article
id doaj.art-69d51ecd52394b21bf214efa7a9a05d4
institution Directory Open Access Journal
issn 1932-6203
language English
last_indexed 2024-12-10T06:32:42Z
publishDate 2011-01-01
publisher Public Library of Science (PLoS)
record_format Article
series PLoS ONE
spelling doaj.art-69d51ecd52394b21bf214efa7a9a05d42022-12-22T01:59:00ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0164e1859710.1371/journal.pone.0018597The T7-related Pseudomonas putida phage φ15 displays virion-associated biofilm degradation properties.Anneleen CornelissenPieter-Jan CeyssensJeroen T'SyenHelena Van PraetJean-Paul NobenOlga V ShaburovaVictor N KrylovGuido VolckaertRob LavigneFormation 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 ϕ15, a 'T7-like virus' with a virion-associated exopolysaccharide (EPS) depolymerase. Phage ϕ15 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 ϕ15-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 ϕ15 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.http://europepmc.org/articles/PMC3079711?pdf=render
spellingShingle Anneleen Cornelissen
Pieter-Jan Ceyssens
Jeroen T'Syen
Helena Van Praet
Jean-Paul Noben
Olga V Shaburova
Victor N Krylov
Guido Volckaert
Rob Lavigne
The T7-related Pseudomonas putida phage φ15 displays virion-associated biofilm degradation properties.
PLoS ONE
title The T7-related Pseudomonas putida phage φ15 displays virion-associated biofilm degradation properties.
title_full The T7-related Pseudomonas putida phage φ15 displays virion-associated biofilm degradation properties.
title_fullStr The T7-related Pseudomonas putida phage φ15 displays virion-associated biofilm degradation properties.
title_full_unstemmed The T7-related Pseudomonas putida phage φ15 displays virion-associated biofilm degradation properties.
title_short The T7-related Pseudomonas putida phage φ15 displays virion-associated biofilm degradation properties.
title_sort t7 related pseudomonas putida phage φ15 displays virion associated biofilm degradation properties
url http://europepmc.org/articles/PMC3079711?pdf=render
work_keys_str_mv AT anneleencornelissen thet7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT pieterjanceyssens thet7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT jeroentsyen thet7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT helenavanpraet thet7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT jeanpaulnoben thet7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT olgavshaburova thet7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT victornkrylov thet7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT guidovolckaert thet7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT roblavigne thet7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT anneleencornelissen t7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT pieterjanceyssens t7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT jeroentsyen t7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT helenavanpraet t7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT jeanpaulnoben t7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT olgavshaburova t7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT victornkrylov t7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT guidovolckaert t7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties
AT roblavigne t7relatedpseudomonasputidaphageph15displaysvirionassociatedbiofilmdegradationproperties