Characterization of Siphoviridae phage Z and studying its efficacy against multidrug-resistant Klebsiella pneumoniae planktonic cells and biofilm

J Med Microbiol. 2015 Apr;64(Pt 4):454-462. doi: 10.1099/jmm.0.000040. Epub 2015 Feb 13.

Abstract

Biofilm has many serious consequences for public health and is a major virulence factor contributing to the chronicity of infections. The aim of the current study was to isolate and characterize a bacteriophage that inhibits multidrug-resistant Klebsiella pneumonia (M) in planktonic form as well as biofilm. This phage, designated bacteriophage Z, was isolated from wastewater. Its adsorption rate to its host bacterium was significantly enhanced by MgCl2 and CaCl2. It has a wide range of pH and heat stability. From its one-step growth, latent time and burst size were determined to be 24 min and about 320 virions per cell, respectively. As analysed by transmission electron microscopy, phage Z had an icosahedral head of width 76±10 nm, length 92±14 nm and icosahedron side 38 nm, and a non-contractile tail 200±15 nm long and 14-29 nm wide. It belongs to the family Siphoviridae in the order Caudovirales. Six structural proteins ranging from 18 to 65 kDa in size were revealed by SDS-PAGE. The genome was found to comprise double-stranded DNA with an approximate size of 36 kb. Bacteria were grown in suspension and as biofilms to compare the susceptibility of both phenotypes to the phage lytic action. Phage Z was effective in reducing biofilm biomass after 24 and 48 h, showing more than twofold and threefold reduction, respectively. Biofilm cells and stationary-phase planktonic bacteria were killed at a lower rate than exponential-phase planktonic bacteria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophages / growth & development*
  • Bacteriophages / isolation & purification
  • Bacteriophages / physiology
  • Bacteriophages / ultrastructure
  • Biofilms / growth & development*
  • Calcium Chloride / metabolism
  • DNA, Viral / genetics
  • Drug Resistance, Multiple, Bacterial*
  • Electrophoresis, Polyacrylamide Gel
  • Hydrogen-Ion Concentration
  • Klebsiella pneumoniae / physiology*
  • Klebsiella pneumoniae / virology*
  • Magnesium Chloride / metabolism
  • Microbial Viability / drug effects
  • Microbial Viability / radiation effects
  • Microscopy, Electron, Transmission
  • Molecular Weight
  • Siphoviridae / growth & development*
  • Siphoviridae / isolation & purification
  • Siphoviridae / physiology
  • Siphoviridae / ultrastructure
  • Temperature
  • Viral Proteins / chemistry
  • Viral Proteins / isolation & purification
  • Virus Attachment / drug effects
  • Wastewater / virology

Substances

  • DNA, Viral
  • Viral Proteins
  • Waste Water
  • Magnesium Chloride
  • Calcium Chloride