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A filamentous phage of Vibrio parahaemolyticus O3:K6 isolated in Laos
1Department of Bacteriology, Faculty of Medicine, University of the Ryukyus, Nishihara, Okinawa, Japan.
Abstract:
A filamentous phage, 'lvpf5,' of Vibrio parahaemolyticus O3:K6 strain LVP5 was isolated and characterized. The host range was not restricted to serotype O3:K6, but 7 of 99 V. parahaemolyticus strains with a variety of serotypes were susceptible to the phage. The phage was inactivated by heating at 80 C for 10 min and by treating with chloroform. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the phage exhibited a 3.8 kDa protein. The amino-terminal amino acid sequence of the coat protein was determined as AEGGAADPFEAIDLLGVATL. The phage genome consisted of a single-stranded DNA molecule. The activity of the phages was inhibited by anti-Na2 pili antibody.
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.