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Open Access Highly Accessed Research

Characterising the biology of novel lytic bacteriophages infecting multidrug resistant Klebsiella pneumoniae

Agata Kęsik-Szeloch1, Zuzanna Drulis-Kawa1*, Beata Weber-Dąbrowska2, Jerzy Kassner1, Grażyna Majkowska-Skrobek1, Daria Augustyniak1, Marzanna Łusiak-Szelachowska2, Maciej Żaczek2, Andrzej Górski23 and Andrew M Kropinski45

Author Affiliations

1 Institute of Genetics and Microbiology, University of Wroclaw, Przybyszewskiego 63/77, Wroclaw, 51-148, Poland

2 L. Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Centre of Excellence, Weigla 12, Wroclaw, 53-114, Poland

3 Department of Clinical Immunology, The Medical University of Warsaw, Nowogrodzka 59, Warszawa, 02-006, Poland

4 Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, 110 Stone Road West, Guelph, ON, N1G 3 W4, Canada

5 Department of Molecular & Cellular Biology, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2 W1, Canada

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Virology Journal 2013, 10:100  doi:10.1186/1743-422X-10-100

Published: 28 March 2013

Abstract

Background

Members of the genus Klebsiella are among the leading microbial pathogens associated with nosocomial infection. The increased incidence of antimicrobial resistance in these species has propelled the need for alternate/combination therapeutic regimens to aid clinical treatment. Bacteriophage therapy forms one of these alternate strategies.

Methods

Electron microscopy, burst size, host range, sensitivity of phage particles to temperature, chloroform, pH, and restriction digestion of phage DNA were used to characterize Klebsiella phages.

Results and conclusions

Of the 32 isolated phages eight belonged to the family Myoviridae, eight to the Siphoviridae whilst the remaining 16 belonged to the Podoviridae. The host range of these phages was characterised against 254 clinical Enterobacteriaceae strains including multidrug resistant Klebsiella isolates producing extended-spectrum beta-lactamases (ESBLs). Based on their lytic potential, six of the phages were further characterised for burst size, physicochemical properties and sensitivity to restriction endonuclease digestion. In addition, five were fully sequenced. Multiple phage-encoded host resistance mechanisms were identified. The Siphoviridae phage genomes (KP16 and KP36) contained low numbers of host restriction sites similar to the strategy found in T7-like phages (KP32). In addition, phage KP36 encoded its own DNA adenine methyltransferase. The φKMV-like KP34 phage was sensitive to all endonucleases used in this study. Dam methylation of KP34 DNA was detected although this was in the absence of an identifiable phage encoded methyltransferase. The Myoviridae phages KP15 and KP27 both carried Dam and Dcm methyltransferase genes and other anti-restriction mechanisms elucidated in previous studies. No other anti-restriction mechanisms were found, e.g. atypical nucleotides (hmC or glucosyl hmC), although Myoviridae phage KP27 encodes an unknown anti-restriction mechanism that needs further investigation.

Keywords:
Bacteriophage; Klebsiella spp.; Multidrug resistance; Restriction endonuclease patterns; Myoviridae; Siphoviridae; Podoviridae