Dataset Information

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

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.

SUBMITTER: Danis-Wlodarczyk K

PROVIDER: S-EPMC4440721 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Publications

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

Danis-Wlodarczyk Katarzyna K Olszak Tomasz T Arabski Michal M Wasik Slawomir S Majkowska-Skrobek Grazyna G Augustyniak Daria D Gula Grzegorz G Briers Yves Y Jang Ho Bin HB Vandenheuvel Dieter D Duda Katarzyna Anna KA Lavigne Rob R Drulis-Kawa Zuzanna Z

PloS one 20150521 5

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 ...[more]

PMID: 25996839

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Project description:Diabetic foot infection has become one of the most important public health concerns and is a growing problem. Pseudomonas aeruginosa is an important opportunistic multidrug-resistant bacterium in diabetic foot infections. In the absence of antibiotics active against MDR strains of P. aeruginosa, phage therapy becomes a key way to deal with P. aeruginosa infections. Out of 185 samples collected from diabetic foot ulcers, 50 (27.02%) isolates were identified as P. aeruginosa. The incidence increases with older ages, and males (n=34, 68%) predominated in all age groups. The tested isolates showed maximum susceptibility towards colistin (80%), imipenem (72%), amikacin (66%), and piperacillin/tazobactam (62%), while these isolates showed moderate susceptibility towards ceftazidime (58%), cefepime (52%) and gentamicin (46%). However, it showed complete resistance (100%) to ampicillin, cefaclor, and sulphamethoxazole/trimethoprim and highly resistance to clindamycin (90%) and amoxicillin/clavulanic acid (84%). Two bacteriophages (ϕPAE1 and ϕPAE2) isolated from sewage samples showed a broad host range against P. aeruginosaa clinical strains. ϕPAE2 infected 74% (37/50) and ϕPAE2 58% (29/50). Furthermore, both phages were host-specific, infecting only P. aeruginosa strains and could not infect other bacterial species in the cross-infectivity studies. Both phages were found to be relatively heat stable as over a period of 1 h, after exposure to a temperature range of 37-50°C, no significant loss in phage activity was observed. On the other hand, the lowest activity was observed at 70°C (39.15%) for ϕPAE1 whereas it was inactivated at 75°C while the lowest activity was observed at 75°C (38.01%) for ϕPAE2 whereas it was inactivated at 80°C. Isolated phages were able to survive and lyse host bacteria over a wide pH range. The optimum pH range for infection was from 6 to 8. Furthermore, ϕPAE1 lost its ability to lyses at pH 2, 3, 11 and 12, whereas; ϕPAE2 lost its infectivity at pH 2, 3 and 12. Chloroform was the most effective solvent that reduced the infectivity of ϕPAE1 and ϕPAE2 to 63.27% and 77.88%, respectively. On the other hand, petroleum ether showed the lowest effect on the infectivity of ϕPAE1 and ϕPAE2; it was reduced to 96.4% and 97.48%, respectively, followed by acetone and ethyl alcohol. The ability of P. aeruginosa phages to form plaques after different storage temperatures (4°C, 30°C, 37°C and 44°C) for a month was slightly affected. The storage of ϕPAE1 and ϕPAE2 at 4ºC showed the least effect on its infectivity, and the storage at 44ºC showed the highest reduction in its infectivity. Moreover, Phage counts were slightly decreased by increasing storage period and temperature.

Project description:Pseudomonas aeruginosa is present throughout nature and is a common opportunistic pathogen in the human body. Carbapenem antibiotics are typically utilized as a last resort in the clinical treatment of multidrug-resistant infections caused by P. aeruginosa. The increase in carbapenem-resistant P. aeruginosa poses an immense challenge for the treatment of these infections. Bacteriophages have the potential to be used as antimicrobial agents for treating antibiotic-resistant bacteria. In this study, a new virulent P. aeruginosa phage, Phage_Pae01, was isolated from hospital sewage and shown to have broad-spectrum antibacterial activity against clinical P. aeruginosa isolates (83.6%). These clinical strains included multidrug-resistant P. aeruginosa and carbapenem-resistant P. aeruginosa. Transmission electron microscopy revealed that the phage possessed an icosahedral head of approximately 80 nm and a long tail about 110 m, indicating that it belongs to the Myoviridae family of the order Caudovirales. Biological characteristic analysis revealed that Phage_Pae01 could maintain stable activity in the temperature range of 4~ 60°C and pH range of 4 ~ 10. According to the in vitro lysis kinetics of the phage, Phage_Pae01 demonstrated strong antibacterial activity. The optimal multiplicity of infection was 0.01. The genome of Phage_Pae01 has a total length of 93,182 bp and contains 176 open reading frames (ORFs). The phage genome does not contain genes related to virulence or antibiotic resistance. In addition, Phage_Pae01 effectively prevented the formation of biofilms and eliminated established biofilms. When Phage_Pae01 was combined with gentamicin, it significantly disrupted established P. aeruginosa biofilms. We identified a novel P. aeruginosa phage and demonstrated its effective antimicrobial properties against P. aeruginosa in both the floating and biofilm states. These findings offer a promising approach for the treatment of drug-resistant bacterial infections in clinical settings.

Dataset Information

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

Publications

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

Similar Datasets

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