Genomic characterization of four novel bacteriophages infecting the clinical pathogen <i>Klebsiella pneumoniae</i>

Bonilla, Estrada; Costa, Ana Rita; Berg, Daan F van den; Rossum, Teunke van; Hagedoorn, Stefan; Walinga, Hielke; Xiao, Minfeng; Song, Wenchen; Haas, Pieter-Jan; Nóbrega, Franklin L.; Brouns, Stan J. J.

doi:10.1093/dnares/dsab013

Cited by 15 publications

(5 citation statements)

References 69 publications

(61 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“… 44 Second, for RBD8_KL102, we found that 55% of the predictions pointed to bacteria from the KL38. In this case, the RBD_KL102 representative sequence showed 35% amino acid identity and 84% query coverage with the Dpo of phage φKp34, which infects KL38, 45 suggesting cross-reactivity or an evolutionary association between the abilities to infect both CLTs. Next, clusters RB64 and RBD65 were both present in phages infecting KL23, but only prophages with identity to RBD65 were found in bacteria from KL23, suggesting that the latter was the active Dpo.…”

Section: Resultsmentioning

confidence: 93%

Genetic determinants of host tropism in Klebsiella phages

Beamud¹,

García-González²,

Gómez-Ortega³

et al. 2023

Cell Reports

View full text Add to dashboard Cite

Section: Resultsmentioning

confidence: 93%

Genetic determinants of host tropism in Klebsiella phages

Beamud¹,

García-González²,

Gómez-Ortega³

et al. 2023

Cell Reports

View full text Add to dashboard Cite

“…Rho-factor independent terminators and promoters were looked for by ARNOLD 65 and PHIRE servers 66 . The phage genome map was drawn using CGView ( http://wishart.biology.ualberta.ca/cgview/ ) and Linear Genome Plot 67 , 68 . To recognize virulence and antibiotic resistance genes, the ResFinder 4.0 and ARDB database 19 , 20 were employed.…”

Section: Methodsmentioning

confidence: 99%

Genomic analysis of vB_PaS-HSN4 bacteriophage and its antibacterial activity (in vivo and in vitro) against Pseudomonas aeruginosa isolated from burn

Rafiei,

Bouzari

2024

Sci Rep

View full text Add to dashboard Cite

The most frequent infections caused by Pseudomonas aeruginosa are local infections in soft tissues, including burns. Today, phage use is considered a suitable alternative to cure infections caused by multi-drug-resistant (MDR) and extensively drug-resistant (XDR) bacteria. We investigated the potential of a novel phage (vB_PaS-HSN4) belonging to Caudoviricetes class, against XDR and MDR P. aeruginosa strains in vivo and in vitro. Its biological and genetic characteristics were investigated. The phage burst size and latent were 119 and 20 min, respectively. It could tolerate a broad range of salt concentrations, pH values, and temperatures. The combination with ciprofloxacin significantly enhanced biofilm removal after 24 h. The genome was dsDNA with a size of 44,534 bp and encoded 61 ORFs with 3 tRNA and 5 promoters. No virulence factor was observed in the phage genome. In the in vivo infection model, treatment with vB_PaS-HSN4 increased Galleria mellonella larvae survival (80%, 66%, and 60%) (MOI 100) and (60%, 40%, and 26%) (MOI 1) in the pre-treatment, co-treatment, and post-treatment experiments, respectively. Based on these characteristics, it can be considered for the cure of infections of burns caused by P. aeruginosa.

show abstract

“…Pseudomonas phage PAXYB1 was obtained from the Fagenbank (Delft, Netherlands). Klebsiella phage φKp18 was isolated from sewage water using K. pneumoniae L0549, a clinical isolate from UMCU, using an enrichment procedure as previously described 50 . Phage amplification was carried out by infecting the host strain (PAO1 for the Pseudomonas phages, L0549 for φKp18) overnight at 37 °C in Lysogeny Broth (LB).…”

Section: Methodsmentioning

confidence: 99%

Monitoring phage-induced lysis of gram-negatives in real time using a fluorescent DNA dye

Egido

Toner-Bartelds

Costa

et al. 2023

Sci Rep

View full text Add to dashboard Cite

Bacteriophages (phages) are viruses that specifically attack bacteria. Their use as therapeutics, which constitutes a promising alternative to antibiotics, heavily relies on selecting effective lytic phages against the pathogen of interest. Current selection techniques are laborious and do not allow for direct visualization of phage infection dynamics. Here, we present a method that circumvents these limitations. It can be scaled for high-throughput and permits monitoring of the phage infection in real time via a fluorescence signal readout. This is achieved through the use of a membrane-impermeant nucleic acid dye that stains the DNA of damaged or lysed bacteria and new phage progeny. We have tested the method on Pseudomonas aeruginosa and Klebsiella pneumoniae and show that an increase in fluorescence reflects phage-mediated killing. This is confirmed by other techniques including spot tests, colony plating, flow cytometry and metabolic activity measurements. Furthermore, we illustrate how our method may be used to compare the activity of different phages and to screen the susceptibility of clinical isolates to phage. Altogether, we present a fast, reliable way of selecting phages against Gram-negative bacteria, which may be valuable in optimizing the process of selecting phages for therapeutic use.

show abstract

Genomic characterization of four novel bacteriophages infecting the clinical pathogen Klebsiella pneumoniae

Cited by 15 publications

References 69 publications

Genetic determinants of host tropism in Klebsiella phages

Genetic determinants of host tropism in Klebsiella phages

Genomic analysis of vB_PaS-HSN4 bacteriophage and its antibacterial activity (in vivo and in vitro) against Pseudomonas aeruginosa isolated from burn

Monitoring phage-induced lysis of gram-negatives in real time using a fluorescent DNA dye

Contact Info

Product

Resources

About