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A filamentous phage of Vibrio parahaemolyticus O3:K6 isolated in Laos

N Nakasone1, M Ikema, N Higa

  • 1Department of Bacteriology, Faculty of Medicine, University of the Ryukyus, Nishihara, Okinawa, Japan.

Microbiology and Immunology
|June 29, 1999
PubMed

Insights

A novel filamentous phage, lvpf5, was isolated from Vibrio parahaemolyticus. This phage infects various serotypes and its activity is inhibited by anti-Na2 pili antibody.

Area of Science:

  • Microbiology
  • Virology
  • Bacteriology

Background:

  • Vibrio parahaemolyticus is a significant foodborne pathogen.
  • Bacteriophages are viruses that infect bacteria and can be used for phage therapy.
  • Characterization of phages provides insights into bacterial-phage interactions.

Purpose of the Study:

  • To isolate and characterize a novel filamentous phage, designated lvpf5, from Vibrio parahaemolyticus O3:K6 strain LVP5.
  • To determine the host range, stability, protein composition, and genome type of the phage.
  • To investigate potential mechanisms of phage-host interaction.

Main Methods:

  • Isolation and purification of filamentous phage lvpf5.
  • Host range determination across various Vibrio parahaemolyticus serotypes.
  • Stability assays (heat, chloroform).
  • Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for protein analysis.
  • Amino-terminal amino acid sequencing of the coat protein.
  • Genome analysis (single-stranded DNA).
  • Inhibition assays using anti-Na2 pili antibody.

Main Results:

  • Filamentous phage lvpf5 was successfully isolated from Vibrio parahaemolyticus O3:K6.
  • The phage exhibited a broader host range than initially expected, infecting 7 out of 99 tested strains across different serotypes.
  • Phage lvpf5 was inactivated by heat (80°C for 10 min) and chloroform.
  • SDS-PAGE revealed a 3.8 kDa coat protein with a determined amino-terminal sequence (AEGGAADPFEAIDLLGVATL).
  • The phage genome comprises single-stranded DNA.
  • Phage activity was inhibited by anti-Na2 pili antibody, suggesting a role for pili in infection.

Conclusions:

  • Filamentous phage lvpf5 represents a new phage isolate of Vibrio parahaemolyticus with potential applications in controlling this pathogen.
  • The phage's broad host range and specific protein components warrant further investigation for therapeutic or diagnostic purposes.
  • The inhibition by anti-Na2 pili antibody indicates that these structures are likely involved in the phage's attachment or entry mechanism.

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