Characterizing the Biology of Lytic Bacteriophage vB_EaeM_φEap-3 Infecting Multidrug-Resistant Enterobacter aerogenes

Multidrug-resistantKlebsiella pneumoniae(MRKP) has steadily grown beyond antibiotic control. However, a bacteriophage is considered to be a potential antibiotic alternative for treating bacterial infections. In this study, a lytic bacteriophage, phage 1513, was isolated using a clinical MRKP isolate KP 1513 as the host and was characterized. It produced a clear plaque with a halo and was classified as Siphoviridae. It had a short latent period of 30 min, a burst size of 264 and could inhibit KP 1513 growthin vitrowith a dose-dependent pattern. Intranasal administration of a single dose of 2 × 109 PFU/mouse 2 h after KP 1513 inoculation was able to protect mice against lethal pneumonia. In a sublethal pneumonia model, phage-treated mice exhibited a lower level ofK. pneumoniaeburden in the lungs as compared to the untreated control. These mice lost less body weight and exhibited lower levels of inflammatory cytokines in their lungs. Lung lesion conditions were obviously improved by phage therapy. Therefore, phage 1513 has a great effectin vitroandin vivo, which has potential to be used as an alternative to an antibiotic treatment of pneumonia that is caused by the multidrug-resistantK. pneumoniae.

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5-Arylideneimidazolones with Amine at Position 3 as Potential Antibiotic Adjuvants against Multidrug Resistant Bacteria

Molecules ◽

10.3390/molecules24030438 ◽

2019 ◽

Vol 24 (3) ◽

pp. 438 ◽

Cited By ~ 3

Author(s):

Aneta Kaczor ◽

Karolina Witek ◽

Sabina Podlewska ◽

Joanna Czekajewska ◽

Annamaria Lubelska ◽

...

Keyword(s):

Efflux Pump ◽

Enterobacter Aerogenes ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Aromatic Rings ◽

Efflux Pump Inhibitor ◽

Fold Reduction ◽

Dual Action ◽

New Compounds

Searching for new chemosensitizers of bacterial multidrug resistance (MDR), chemical modifications of (Z)-5-(4-chlorobenzylidene)-2-(4-methylpiperazin-1-yl)-3H-imidazol-4(5H)-one (6) were performed. New compounds (7–17), with fused aromatic rings at position 5, were designed and synthesized. Crystallographic X-ray analysis proved that the final compounds (7–17) were substituted with tertiary amine-propyl moiety at position 3 and primary amine group at 2 due to intramolecular Dimroth rearrangement. New compounds were evaluated on their antibiotic adjuvant properties in either Gram-positive or Gram-negative bacteria. Efflux pump inhibitor (EPI) properties towards the AcrAB-TolC pump in Enterobacter aerogenes (EA289) were investigated in the real-time efflux (RTE) assay. Docking and molecular dynamics were applied to estimate an interaction of compounds 6–17 with penicillin binding protein (PBP2a). In vitro ADME-Tox properties were evaluated for compound 9. Most of the tested compounds reduced significantly (4-32-fold) oxacillin MIC in highly resistant MRSA HEMSA 5 strain. The anthracene-morpholine derivative (16) was the most potent (32-fold reduction). The tested compounds displayed significant EPI properties during RTE assay (37–97%). The naphthyl-methylpiperazine derivative 9 showed the most potent “dual action” of both oxacillin adjuvant (MRSA) and EPI (E. aerogenes). Molecular modeling results suggested the allosteric mechanism of action of the imidazolones, which improved binding of oxacillin in the PBP2a active site in MRSA.

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Inhibitors of Antibiotic Efflux in Resistant Enterobacter aerogenes and Klebsiella pneumoniae Strains

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.48.3.1043-1046.2004 ◽

2004 ◽

Vol 48 (3) ◽

pp. 1043-1046 ◽

Cited By ~ 54

Author(s):

Jacqueline Chevalier ◽

Jérôme Bredin ◽

Abdallah Mahamoud ◽

Monique Malléa ◽

Jacques Barbe ◽

...

Keyword(s):

Multidrug Resistance ◽

Klebsiella Pneumoniae ◽

Efflux Pump ◽

Enterobacter Aerogenes ◽

Drug Susceptibility ◽

Multidrug Resistant ◽

Resistance Phenotype ◽

Pump Mechanism ◽

Energy Dependent ◽

Antibiotic Efflux

ABSTRACT In Enterobacter aerogenes and Klebsiella pneumoniae, efflux provides efficient extrusion of antibiotics and contributes to the multidrug resistance phenotype. One of the alkoxyquinoline derivatives studied here, 2,8-dimethyl-4-(2′-pyrrolidinoethyl)-oxyquinoline, restores noticeable drug susceptibility to resistant clinical strains. Analyses of energy-dependent chloramphenicol efflux indicate that this compound inhibits the efflux pump mechanism and improves the activity of structurally unrelated antibiotics on multidrug-resistant E. aerogenes and K. pneumoniae isolates.

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Erratum: Corrigendum: Isolation and characterization of a bacteriophage phiEap-2 infecting multidrug resistant Enterobacter aerogenes

Scientific Reports ◽

10.1038/srep46805 ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 1

Author(s):

Erna Li ◽

Xiao Wei ◽

Yanyan Ma ◽

Zhe Yin ◽

Huan Li ◽

...

Keyword(s):

Enterobacter Aerogenes ◽

Multidrug Resistant ◽

Isolation And Characterization

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Isolation and characterization of a lytic bacteriophage against Pseudomonas aeruginosa

Scientific Reports ◽

10.1038/s41598-021-98457-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Sonika Sharma ◽

Sibnarayan Datta ◽

Soumya Chatterjee ◽

Moumita Dutta ◽

Jhuma Samanta ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Burst Size ◽

Antibiotic Resistance Genes ◽

Multidrug Resistant ◽

Potential Candidate ◽

Lytic Bacteriophage ◽

Isolation And Characterization ◽

Phage Proteins

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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Lytic Bacteriophage Screening Strategies for Multidrug-Resistant Bloodstream Infections in a Burn Intensive Care Unit

Medical Science Monitor ◽

10.12659/msm.917706 ◽

2019 ◽

Vol 25 ◽

pp. 8352-8362

Author(s):

Zichen Yang ◽

Yunlong Shi ◽

Cheng Zhang ◽

Xiaoqiang Luo ◽

Yu Chen ◽

...

Keyword(s):

Intensive Care Unit ◽

Intensive Care ◽

Bloodstream Infections ◽

Multidrug Resistant ◽

Lytic Bacteriophage ◽

Screening Strategies

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Characterization and protective effects of lytic bacteriophage pAh6.2TG against a pathogenic multidrug-resistant Aeromonas hydrophila in Nile tilapia ( Oreochromis niloticus )

10.22541/au.162366557.74509707/v1 ◽

2021 ◽

Author(s):

Le Thanh Dien ◽

Le Buu Ky ◽

Bui The Huy ◽

Muhammad Fadhlullah Mursalim ◽

Pattanapon Kayansamruaj ◽

...

Keyword(s):

Aeromonas Hydrophila ◽

Nile Tilapia ◽

Protective Efficacy ◽

Lethal Dose ◽

Multidrug Resistant ◽

Lytic Bacteriophage ◽

Protective Effects ◽

Bacterial Counts ◽

Treatment Groups ◽

Phage Treatment

Bacteriophage is considered an alternative to antibiotics and environmentally friendly approach to tackle antimicrobial resistance (AMR) in aquaculture. Here, we reported isolation, morphology and genomic characterizations of a newly isolated lytic bacteriophage, designated pAh6.2TG. Host range and stability of pAh6.2TG in different environmental conditions, and protective efficacy against a pathogenic multidrug-resistant (MDR) Aeromonas hydrophila in Nile tilapia were subsequently evaluated. The results showed that pAh6.2TG is a member of the family Myoviridae which has genome size of 51,780 bp, encoding 65 putative open reading frames (ORFs), and is most closely related to Aeromonas phage PVN02 (99.33% nucleotide identity). The pAh6.2TG was highly specific to A. hydrophila and infected 83.3% tested strains of MDR A. hydrophila (10 out of 12) with relative stability at pH 7 9, temperature 0 40 C and salinity 0 40 ppt. In experimental challenge, pAh6.2TG treatments significantly improved survivability of Nile tilapia exposed to a lethal dose of the pathogenic MDR A. hydrophila, with relative percent survival (RPS) of 73.3% and 50% for phage multiplicity of infection (MOI) 1.0 and 0.1, respectively. Significant reduction of bacterial counts in rearing water at 3 h (6.7 ± 0.5 to 18.1 ± 6.98 folds) and in fish liver at 48 h post-treatment (2.7 ± 0.24 to 34.08 ± 26.4 folds) was observed in phage treatment groups while opposite pattern for bacterial counts was observed in untreated control. Interestingly, the surviving fish provoked specific antibody (IgM) against the challenged A. hydrophila. These results might explain the higher survival in phage treatment groups. In summary, the findings suggested that the lytic bacteriophage pAh6.2TG is an effective alternative to antibiotics to control MDR A. hydrophila in tilapia and possibly other freshwater fish.

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Virulent Drexlervirial Bacteriophage MSK, Morphological and Genome Resemblance With Rtp Bacteriophage Inhibits the Multidrug-Resistant Bacteria

Frontiers in Microbiology ◽

10.3389/fmicb.2021.706700 ◽

2021 ◽

Vol 12 ◽

Author(s):

Muhammad Saleem Iqbal Khan ◽

Xiangzheng Gao ◽

Keying Liang ◽

Shengsheng Mei ◽

Jinbiao Zhan

Keyword(s):

Multidrug Resistant ◽

The Other ◽

Structural Proteins ◽

Resistant Bacteria ◽

Tail Fiber ◽

Lytic Bacteriophage ◽

Tail Fiber Protein ◽

Multidrug Resistant Bacteria ◽

Genome Database ◽

Fiber Protein

Phage-host interactions are likely to have the most critical aspect of phage biology. Phages are the most abundant and ubiquitous infectious acellular entities in the biosphere, where their presence remains elusive. Here, the novel Escherichia coli lytic bacteriophage, named MSK, was isolated from the lysed culture of E. coli C (phix174 host). The genome of phage MSK was sequenced, comprising 45,053 bp with 44.8% G + C composition. In total, 73 open reading frames (ORFs) were predicted, out of which 24 showed a close homology with known functional proteins, including one tRNA-arg; however, the other 49 proteins with no proven function in the genome database were called hypothetical. Electron Microscopy and genome characterization have revealed that MSK phage has a rosette-like tail tip. There were, in total, 46 ORFs which were homologous to the Rtp genome. Among these ORFs, the tail fiber protein with a locus tag of MSK_000019 was homologous to Rtp 43 protein, which determines the host specificity. The other protein, MSK_000046, encodes lipoprotein (cor gene); that protein resembles Rtp 45, responsible for preventing adsorption during cell lysis. Thirteen MSK structural proteins were identified by SDS-PAGE analysis. Out of these, 12 were vital structural proteins, and one was a hypothetical protein. Among these, the protein terminase large (MSK_000072) subunit, which may be involved in DNA packaging and proposed packaging strategy of MSK bacteriophage genome, takes place through headful packaging using the pac-sites. Biosafety assessment of highly stable phage MSK genome analysis has revealed that the phage did not possess virulence genes, which indicates proper phage therapy. MSK phage potentially could be used to inhibit the multidrug-resistant bacteria, including AMP, TCN, and Colistin. Further, a comparative genome and lifestyle study of MSK phage confirmed the highest similarity level (87.18% ANI). These findings suggest it to be a new lytic isolated phage species. Finally, Blast and phylogenetic analysis of the large terminase subunit and tail fiber protein put it in Rtp viruses’ genus of family Drexlerviridae.

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Imipenem Resistance of Enterobacter aerogenes Mediated by Outer Membrane Permeability

Journal of Clinical Microbiology ◽

10.1128/jcm.38.3.1048-1052.2000 ◽

2000 ◽

Vol 38 (3) ◽

pp. 1048-1052 ◽

Cited By ~ 83

Author(s):

Charléric Bornet ◽

Anne Davin-Regli ◽

Claude Bosi ◽

Jean-Marie Pages ◽

Claude Bollet

Keyword(s):

Membrane Permeability ◽

Clinical Course ◽

Enterobacter Aerogenes ◽

Multidrug Resistant ◽

Fatality Rate ◽

Evolution Of Resistance ◽

Resistant Strains ◽

Imipenem Resistance ◽

Outer Membrane Permeability ◽

Cessation Of Treatment

Multidrug-resistant Enterobacter aerogenes strains are increasingly isolated in Europe and especially in France. Treatment leads to imipenem resistance, because of a lack of porin. We studied the evolution of resistance in 29 strains isolated from four patients during their clinical course. These strains belonged to the prevalent epidemiological type observed in France in previous studies (C. Bosi, et al., J. Clin. Microbiol. 37:2165–2169, 1999; A. Davin-Regli et al., J. Clin. Microbiol. 34:1474–1480, 1996). They also harbored a TEM-24 extended-spectrum β-lactamase-coding gene. Thirteen strains were susceptible to gentamicin and resistant to imipenem and cefepime. All of the patients showed E. aerogenes strains with this resistance after an imipenem treatment. One patient showed resistance to imipenem after a treatment with cefpirome. Twelve of these 13 strains showed a lack of porin. Cessation of treatment with imipenem for three patients was followed by reversion of susceptibility to this antibiotic and the reappearance of porins, except in one case. For one patient, we observed three times in the same day the coexistence of resistant strains lacking porin and susceptible strains possessing porin. The emergence of multidrug-resistant E. aerogenes strains is very disquieting. In our study, infection by E. aerogenesincreased the severity of the patients' illnesses, causing a 100% fatality rate.

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Complete Genome Sequence of the Myoviral Bacteriophage YS35, Which Causes the Lysis of a Multidrug-Resistant Pseudomonas aeruginosa Strain

Genome Announcements ◽

10.1128/genomea.01395-17 ◽

2018 ◽

Vol 6 (6) ◽

pp. e01395-17 ◽

Cited By ~ 1

Author(s):

Sheng Yu ◽

Honglan Huang ◽

Yuchong Hao ◽

Hongyan Shi ◽

Chunyan Zhao ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Genome Sequence ◽

Complete Genome Sequence ◽

Complete Genome ◽

Multidrug Resistant ◽

Sewage Sample ◽

Lytic Bacteriophage ◽

Double Stranded Dna

ABSTRACT The lytic bacteriophage YS35, which is capable of lysing multidrug-resistant Pseudomonas aeruginosa strains, was isolated from a sewage sample. Here, we describe the complete genome sequence of this myoviral bacteriophage, which contains 93,296 bp of double-stranded DNA and has a 49.4% G+C content.

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