Isolation of Four Lytic Phages Infecting Klebsiella pneumoniae K22 Clinical Isolates from Spain

Domingo‐Calap, Pilar; Beamud, Beatriz; Vienne, Justine; Gónzález-Candelas, Fernando; Sanjuán, Rafael

doi:10.3390/ijms21020425

Cited by 24 publications

(31 citation statements)

References 39 publications

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“…This has been previously shown in other Klebsiella spp. [18] and in phages infecting other encapsulated bacteria, such as Escherichia coli or Acinetobacter baumannii [19,20]. The haloes of phage πVLC6 were wider than those of πVLC5, whereas lysed areas were larger for πVLC5, suggesting that these were two distinct phages.…”

Section: Isolation and Phenotypic Characterization Of Two New Klebsiementioning

confidence: 99%

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Isolation and Characterization of Two Klebsiella pneumoniae Phages Encoding Divergent Depolymerases

Domingo‐Calap

Beamud

Mora-Quilis

et al. 2020

IJMS

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The emergence of multidrug-resistant bacteria is a major global health concern. The search for new therapies has brought bacteriophages into the spotlight, and new phages are being described as possible therapeutic agents. Among the bacteria that are most extensively resistant to current antibiotics is Klebsiella pneumoniae, whose hypervariable extracellular capsule makes treatment particularly difficult. Here, we describe two new K. pneumoniae phages, πVLC5 and πVLC6, isolated from environmental samples. These phages belong to the genus Drulisvirus within the family Podoviridae. Both phages encode a similar tail spike protein with putative depolymerase activity, which is shared among other related phages and probably determines their ability to specifically infect K. pneumoniae capsular types K22 and K37. In addition, we found that phage πVLC6 also infects capsular type K13 and is capable of striping the capsules of K. pneumoniae KL2 and KL3, although the phage was not infectious in these two strains. Genome sequence analysis suggested that the extended tropism of phage πVLC6 is conferred by a second, divergent depolymerase. Phage πVLC5 encodes yet another putative depolymerase, but we found no activity of this phage against capsular types other than K22 and K37, after testing a panel of 77 reference strains. Overall, our results confirm that most phages productively infected one or few Klebsiella capsular types. This constitutes an important challenge for clinical applications.

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Section: Isolation and Phenotypic Characterization Of Two New Klebsiementioning

confidence: 99%

“…Recently, our group has isolated and characterized four new phages of K. pneumoniae (πVLC1 to πVLC4) from environmental samples in Valencia, Spain [18]. All of these were closely related phylogenetically, belonged to the genus Drulisvirus, encoded only one depolymerase, and showed a very narrow capsular type range.…”

Section: Introductionmentioning

confidence: 99%

Isolation and Characterization of Two Klebsiella pneumoniae Phages Encoding Divergent Depolymerases

Domingo‐Calap

Beamud

Mora-Quilis

et al. 2020

IJMS

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“…Morphology of plaques from Ganges water showed a halo around the clear zone, which might indicate depolymerase activity of KpG that could target the capsule of the strain (Fig. 1) (Domingo-Calap et al 2020). The phage displayed stability at the following pH 5.0, 7.0, 9.0, 11.0 with a survival rate of 86%, 100%, 91% and 77% respectively (Fig.…”

Section: Resultsmentioning

confidence: 95%

“…Another report showed that four lytic phages, belonging to the family Podoviridae, infecting K. pneumoniae capsular type K22, were isolated from environmental samples. They possessed narrow infectivity against a K. pneumoniae clinical isolates with K22 capsular type (Domingo-Calap et al 2020). In addition, the plaque morphology of the phages showed a halo around the lysis area, implying presence of depolymerase that lyses the capsule (Domingo-Calap et al…”

Section: Introductionmentioning

confidence: 99%

Phages from Ganges River curtail in vitro biofilms and planktonic growth of drug resistant Klebsiella pneumoniae in a zebrafish infection model

Sundaramoor

Thothathri

Bhaskaran

et al. 2020

Preprint

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Bacteriophages are a promising alternative for curtailing infections caused by multi drug resistant (MDR) bacteria. The objective of the present study is to evaluate phage populations from water bodies to inhibit planktonic and biofilm mode of growth of drug resistant Klebsiella pneumoniae in vitro and curtail planktonic growth in vivo in a zebrafish model. Phage specific to K. pneumoniae (MTCC 432) was isolated from Ganges river (designated as KpG). One-step growth curve, in vitro time kill curve study and in vivo infection model were performed to evaluate the efficacy of phage against planktonic growth. Crystal violet assay and colony biofilm assay was done to determine the action of phages on biofilms. KpG phages had a greater burst size, better bactericidal potential and enhanced inhibitory effect against biofilms formed at liquid air and solid air interfaces. In vivo injection of KpG phages revealed that it did not pose any toxicity to zebrafish as evidenced by liver/brain enzyme profiles and by histopathological analysis. In vitro time kill assay showed a 3 log decline and a 6 log decline in K. pneumoniae colony counts, when phages were administered individually and in combination with streptomycin, respectively. The muscle tissue of zebrafish, infected with K. pneumoniae and treated with KpG phages showed a significant 2 log decline in bacterial counts relative to untreated control. Our study reveals that KpG phages has the potential to curtail plantonic and biofilm mode of growth in vivo in higher animal models.

show abstract

Section: Resultsmentioning

confidence: 97%

Phages from Ganges River curtail in vitro biofilms and planktonic growth of drug resistant Klebsiella pneumoniae in a zebrafish infection model

Sundaramoor

Thothathri

Bhaskaran

et al. 2020

Preprint

View full text Add to dashboard Cite

Bacteriophages are a promising alternative for curtailing infections caused by multi drug resistant (MDR) bacteria. The objective of the present study is to evaluate phage populations from water bodies to inhibit planktonic and biofilm mode of growth of drug resistant Klebsiella pneumoniae in vitro and curtail planktonic growth in vivo in a zebrafish model. Phage specific to K. pneumoniae (MTCC 432) was isolated from Ganges river (designated as KpG). One-step growth curve, in vitro time kill curve study and in vivo infection model were performed to evaluate the efficacy of phage to curtail planktonic growth. Crystal violet assay and colony biofilm assay was done to determine the action of phages on biofilms. KpG phages had a greater burst size, better bactericidal potential and enhanced inhibitory effect against biofilms formed at liquid air and solid air interfaces. In vivo injection of KpG phages revealed that it did not pose any toxicity to zebrafish as evidenced by liver/brain enzyme profiles and by histopathological analysis. In vitro time kill assay showed a 3 log decline and a 6 log decline in K. pneumoniae colony counts, when phages were administered individually and in combination with streptomycin, respectively. The muscle tissue of zebrafish, infected with K. pneumoniae and treated with KpG phages showed a significant 2 log decline in bacterial counts relative to untreated control. Our study reveals that KpG phages has the potential to curtail plantonic and biofilm mode of growth in vivo in higher animal models.

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Isolation of Four Lytic Phages Infecting Klebsiella pneumoniae K22 Clinical Isolates from Spain

Cited by 24 publications

References 39 publications

Isolation and Characterization of Two Klebsiella pneumoniae Phages Encoding Divergent Depolymerases

Isolation and Characterization of Two Klebsiella pneumoniae Phages Encoding Divergent Depolymerases

Phages from Ganges River curtail in vitro biofilms and planktonic growth of drug resistant Klebsiella pneumoniae in a zebrafish infection model

Phages from Ganges River curtail in vitro biofilms and planktonic growth of drug resistant Klebsiella pneumoniae in a zebrafish infection model

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