Isolation and characterization of bacteriophage to control multidrug-resistant Pseudomonas aeruginosa planktonic cells and biofilm

AbstractIn recent years, the use of bacteriophages (or 'phages') against multidrug-resistant (MDR) bacteria including Pseudomonas aeruginosa has drawn considerable attention, globally. In this work, we report the isolation and detailed characterization of a highly lytic Pseudomonasphage DRL-P1 isolated from wastewater. Under TEM, DRL-P1 appeared as a member of the phage family Myoviridae. DRL-P1 featured rapid adsorption (~ 5 min), short-latency (~ 30 min), and large burst size (~ 100 PFU per infected cell). DRL-P1 can withstand a wide temperature range (4 °C to 40 °C) and pH (5.0 to 10.0) conditions. The 66,243 bp DRL-P1 genome (MN564818) encodes at least 93 ORFs, of which 36 were functionally annotated based on homology with similar phage proteins available in the databases. Comparative analyses of related genomes suggest an independent evolutionary history and discrete taxonomic position of DRL-P1 within genus Pbunavirus. No toxin or antibiotic resistance genes was identified. DRL-P1 is tolerant to lyophilization and encapsulation techniques and retained lytic activity even after 18 months of storage. We also demonstrated decontaminating potentials of DRL-P1 in vitro, on an artificially contaminated cover-slip model. To the best of our knowledge, this is the first Pbunavirus to be reported from India. Our study suggests DRL-P1 as a potential candidate for various applications.

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Isolation and Characterization of T7-Like Lytic Bacteriophages Infecting Multidrug Resistant Pseudomonas aeruginosa Isolated from Egypt

Current Microbiology ◽

10.1007/s00284-015-0788-8 ◽

2015 ◽

Vol 70 (6) ◽

pp. 786-791 ◽

Cited By ~ 4

Author(s):

Gamal El Didamony ◽

Ahmed Askora ◽

Aya A. Shehata

Keyword(s):

Pseudomonas Aeruginosa ◽

Multidrug Resistant ◽

Isolation And Characterization

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Isolation and characterization of Pseudomonas aeruginosa bacteriophages — potential agents for phage therapy

10.47183/mes.2021.027 ◽

2021 ◽

Author(s):

MA Kornienko ◽

NS Kuptsov ◽

DI Danilov ◽

RB Gorodnichev ◽

MV Malakhova ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Phage Therapy ◽

Illumina Miseq ◽

Multidrug Resistant ◽

Open Reading Frames ◽

Lytic Activity ◽

Isolation And Characterization ◽

Double Stranded Dna ◽

Activity Spectrum

Pseudomonas aeruginosa — is one of the pathogens characterized by the critical number of multidrug-resistant (MDR) strains. Phage therapy is considered an alternative to antibiotics, especially in treatment of infections caused by MDR strains. The aim of this study was to isolate and characterize P. aeruginosa phages that could potentially be suitable for treating infectious diseases. To isolate the P. aeruginosa phages, enrichment cultures were used. The lytic activity spectrum was confirmed by spot testing on 40 P. aeruginosa strains. Whole-genome sequencing was performed using Illumina MiSeq instrument. Phylogenetic analysis was done using VICTOR tool. Isolated phages vB_PaeA-55-1w and vB_PaeM-198 from Autographiviridae and Myoviridae families, respectively, had a broad spectrum of lytic activity (about 50% each), including lysis of MDR strains. The genomes vB_PaeA-55-1w and vB_PaeM-198 comprise double-stranded DNA of 42.5 and 66.3 kbp in length, respectively. Open reading frames were annotated for both phages (52 for vB_PaeA-55-1w, and 95 for vB_PaeM-198), no integrases and toxins were detected. On a phylogenetic tree, vB_PaeA-55-1w phage was clustered with phages from the Phikmvvirus genus (Autographiviridae family), which are also used in phage therapy. vB_PaeM-198 phage was clustered with phages from the Pbunavirus genus (Myoviridae family). vB_PaeA-55-1w and vB_PaeM-198 phages could be considered as candidates for phage therapy and may be used to treat infections caused by MDR P. aeruginosa.

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A Novel N4-Like Bacteriophage Isolated from a Wastewater Source in South India with Activity against Several Multidrug-Resistant Clinical Pseudomonas aeruginosa Isolates

mSphere ◽

10.1128/msphere.01215-20 ◽

2021 ◽

Vol 6 (1) ◽

Author(s):

Nitasha D. Menon ◽

Megha S. Kumar ◽

T. G. Satheesh Babu ◽

Sucharita Bose ◽

Gayathri Vijayakumar ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

South India ◽

Bacterial Load ◽

Multidrug Resistant ◽

Lytic Bacteriophage ◽

Bacteriophage Therapy ◽

Content Type ◽

Isolation And Characterization ◽

Environmental Treatment

ABSTRACT Multidrug-resistant community-acquired infections caused by the opportunistic human pathogen Pseudomonas aeruginosa are increasingly reported in India and other locations globally. Since this organism is ubiquitous in the environment, samples such as sewage and wastewater are rich reservoirs of P. aeruginosa bacteriophages. In this study, we report the isolation and characterization of a novel P. aeruginosa N4-like lytic bacteriophage, vB_Pae_AM.P2 (AM.P2), from wastewater in Kerala, India. AM.P2 is a double-stranded DNA podovirus that efficiently lyses the model strain, PAO1, at a multiplicity of infection as low as 0.1 phage per bacterium and resistance frequency of 6.59 × 10−4. Synergy in bactericidal activity was observed between AM.P2 and subinhibitory concentrations of the antibiotic ciprofloxacin. Genome sequencing of AM.P2 revealed features similar to those of the N4-like P. aeruginosa phages LUZ7 and KPP21. As judged by two independent assay methods, spot tests and growth inhibition, AM.P2 successfully inhibited the growth of almost 30% of strains from a contemporary collection of multidrug-resistant P. aeruginosa clinical isolates from South India. Thus, AM.P2 may represent an intriguing candidate for inclusion in bacteriophage cocktails developed for various applications, including water decontamination and clinical bacteriophage therapy. IMPORTANCE In India, multidrug resistance determinants are much more abundant in community-associated bacterial pathogens due to the improper treatment of domestic and industrial effluents. In particular, a high bacterial load of the opportunistic pathogen P. aeruginosa in sewage and water bodies in India is well documented. The isolation and characterization of bacteriophages that could target emerging P. aeruginosa strains, representing possible epicenters for community-acquired infections, could serve as a useful alternative tool for various applications, such as phage therapy and environmental treatment. Continuing to supplement the repertoire of broad-spectrum bacteriophages is an essential tool in confronting this problem.

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Isolation and characterization of Pseudomonas aeruginosa exotoxin A binding glycoprotein from mouse LM cells

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)52256-0 ◽

1991 ◽

Vol 266 (4) ◽

pp. 2390-2396 ◽

Cited By ~ 3

Author(s):

M R Thompson ◽

J Forristal ◽

P Kauffmann ◽

T Madden ◽

K Kozak ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Exotoxin A ◽

Isolation And Characterization ◽

Pseudomonas Aeruginosa Exotoxin ◽

Binding Glycoprotein

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Isolation and Characterization of Two Lytic Bacteriophages Infecting a Multi-Drug Resistant Salmonella Typhimurium and Their Efficacy to Combat Salmonellosis in Ready-to-Use Foods

Microorganisms ◽

10.3390/microorganisms9020423 ◽

2021 ◽

Vol 9 (2) ◽

pp. 423

Author(s):

Ahmed Esmael ◽

Ehab Azab ◽

Adil A. Gobouri ◽

Mohamed A. Nasr-Eldin ◽

Mahmoud M. A. Moustafa ◽

...

Keyword(s):

Salmonella Typhimurium ◽

Multidrug Resistant ◽

Viable Count ◽

Food Storage ◽

Chicken Breast ◽

Whole Genome Analysis ◽

Isolation And Characterization ◽

Phage Cocktail ◽

Global Threat

Foodborne salmonellosis is a global threat to public health. In the current study, we describe the isolation and characterization of two broad-spectrum, lytic Salmonella phages: SPHG1 and SPHG3 infecting a multidrug-resistant Salmonella Typhimurium EG.SmT3. Electron microscopy and whole genome analysis identified SPHG1 as a Myovirus, while SPHG3 as a new member of the genus “Kuttervirus” within the family Ackermannviridae. SPHG1 and SPHG3 had a lysis time of 60 min. with burst sizes of 104 and 138 PFU/cell, respectively. The two phages were robust at variable temperatures and pH ranges that match the corresponding values of most of the food storage and processing conditions. A phage cocktail containing the two phages was stable in the tested food articles for up to 48 h. The application of the phage cocktail at MOIs of 1000 or 100 resulted in a significant reduction in the viable count of S. Typhimurium by 4.2 log10/sample in milk, water, and on chicken breast. Additionally, the phage cocktail showed a prospective ability to eradicate and reduce the biofilm that formed by S. Typhimurium EG.SmT3. A phage cocktail of SPHG1 and SPHG3 is considered as a promising candidate as a biocontrol agent against foodborne salmonellosis due to its broad host ranges, highly lytic activities, and the absence of any virulence or lysogeny-related genes in their genomes.

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