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Characterization of the Newly Isolated Lytic Bacteriophages KTN6 and KT28 and Their Efficacy against Pseudomonas aeruginosa Biofilm.

Danis-Wlodarczyk K, Olszak T, Arabski M, Wasik S, Majkowska-Skrobek G, Augustyniak D, Gula G, Briers Y, Jang HB, Vandenheuvel D, Duda KA, Lavigne R, Drulis-Kawa Z - PLoS ONE (2015)

Bottom Line: 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.It was also shown, that PB1-related phage treatment of biofilm caused the emergence of stable phage-resistant mutants growing as small colony variants.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wroclaw, Wroclaw, Poland; Division of Gene Technology, Catholic University of Leuven, Leuven, Belgium.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus

Transmission electron microscopic images of phage KT28 and KTN6.(A) KT28 has a isometric capsid with a diameter of 74 nm between opposite apices, a contractile tail 136 nm with a base plate. (B) KTN6 has a isometric capsid with diameter of 72 nm between opposite apices and a contractile tail 123 nm with a base plate. The scale bar represents 100nm.
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pone.0127603.g001: Transmission electron microscopic images of phage KT28 and KTN6.(A) KT28 has a isometric capsid with a diameter of 74 nm between opposite apices, a contractile tail 136 nm with a base plate. (B) KTN6 has a isometric capsid with diameter of 72 nm between opposite apices and a contractile tail 123 nm with a base plate. The scale bar represents 100nm.

Mentions: Two Pseudomonas lytic phages were isolated from sewage samples collected in natural waste-water treatment plant (irrigated fields) located in Wrocław, Poland. After purification phage titres were 109–1010 pfu/ml and caused 1.2–1.5 mm wide clear plaques with halo zone on 0.6% soft agar. The bacteriophages were examined by transmission electron microscopy (TEM) and classified on the basis of their morphological features to Pbunalikevirus (order Caudovirales, family Myoviridae) (Fig 1). The isolates were named as vB_PaeM_KT28 (KT28) and vB_PaeM_KTN6 (KTN6). The isometric head size was estimated to be 74 nm and 72 nm between opposite apices, and contractile tails 136 nm and 123 nm in the extended state, for KT28 and KTN6, respectively (S2 Table).


Characterization of the Newly Isolated Lytic Bacteriophages KTN6 and KT28 and Their Efficacy against Pseudomonas aeruginosa Biofilm.

Danis-Wlodarczyk K, Olszak T, Arabski M, Wasik S, Majkowska-Skrobek G, Augustyniak D, Gula G, Briers Y, Jang HB, Vandenheuvel D, Duda KA, Lavigne R, Drulis-Kawa Z - PLoS ONE (2015)

Transmission electron microscopic images of phage KT28 and KTN6.(A) KT28 has a isometric capsid with a diameter of 74 nm between opposite apices, a contractile tail 136 nm with a base plate. (B) KTN6 has a isometric capsid with diameter of 72 nm between opposite apices and a contractile tail 123 nm with a base plate. The scale bar represents 100nm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440721&req=5

pone.0127603.g001: Transmission electron microscopic images of phage KT28 and KTN6.(A) KT28 has a isometric capsid with a diameter of 74 nm between opposite apices, a contractile tail 136 nm with a base plate. (B) KTN6 has a isometric capsid with diameter of 72 nm between opposite apices and a contractile tail 123 nm with a base plate. The scale bar represents 100nm.
Mentions: Two Pseudomonas lytic phages were isolated from sewage samples collected in natural waste-water treatment plant (irrigated fields) located in Wrocław, Poland. After purification phage titres were 109–1010 pfu/ml and caused 1.2–1.5 mm wide clear plaques with halo zone on 0.6% soft agar. The bacteriophages were examined by transmission electron microscopy (TEM) and classified on the basis of their morphological features to Pbunalikevirus (order Caudovirales, family Myoviridae) (Fig 1). The isolates were named as vB_PaeM_KT28 (KT28) and vB_PaeM_KTN6 (KTN6). The isometric head size was estimated to be 74 nm and 72 nm between opposite apices, and contractile tails 136 nm and 123 nm in the extended state, for KT28 and KTN6, respectively (S2 Table).

Bottom Line: 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.It was also shown, that PB1-related phage treatment of biofilm caused the emergence of stable phage-resistant mutants growing as small colony variants.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wroclaw, Wroclaw, Poland; Division of Gene Technology, Catholic University of Leuven, Leuven, Belgium.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus