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Characterization of the Newly Isolated Lytic Bacteriophages KTN6 and KT28 and Their Efficacy against Pseudomonas aeruginosa Biofilm
Author(s) -
Katarzyna Danis-Wlodarczyk,
Tomasz Olszak,
Michał Arabski,
Sławomir Wąsik,
Grażyna Majkowska-Skrobek,
Daria Augustyniak,
Grzegorz Guła,
Yves Briers,
Ho G. Jang,
Dieter Vandenheuvel,
Katarzyna Duda,
Rob Lavigne,
Zuzanna Drulis-Kawa
Publication year - 2015
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0127603
Subject(s) - myoviridae , biofilm , pseudomonas aeruginosa , lytic cycle , pyocyanin , microbiology and biotechnology , biology , bacteriophage , phage therapy , podoviridae , pseudomonas , bacteria , virology , virus , gene , escherichia coli , genetics , quorum sensing
We here describe two novel lytic phages, KT28 and KTN6, infecting Pseudomonas aeruginosa , isolated from a sewage sample from an irrigated field near Wroclaw, in Poland. Both viruses show characteristic features of Pbunalikevirus genus within the Myoviridae family with respect to shape and size of head/tail, as well as LPS host receptor recognition. Genome analysis confirmed the similarity to other PB1-related phages, ranging between 48 and 96%. Pseudomonas phage KT28 has a genome size of 66,381 bp and KTN6 of 65,994 bp. The latent period, burst size, stability and host range was determined for both viruses under standard laboratory conditions. Biofilm eradication efficacy was tested on peg-lid plate assay and PET membrane surface. Significant reduction of colony forming units was observed (70-90%) in 24 h to 72 h old Pseudomonas aeruginosa PAO1 biofilm cultures for both phages. Furthermore, a pyocyanin and pyoverdin reduction tests reveal that tested phages lowers the amount of both secreted dyes in 48-72 h old biofilms. Diffusion and goniometry experiments revealed the increase of diffusion rate through the biofilm matrix after phage application. These characteristics indicate these phages could be used to prevent Pseudomonas aeruginosa infections and biofilm formation. It was also shown, that PB1-related phage treatment of biofilm caused the emergence of stable phage-resistant mutants growing as small colony variants.

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