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Isolation of F-specific Inovirus filamentous phages from environmental sewage samples
Natasha Theriault1,2, Bradley W M Cook2, Steven S Theriault1,2
1University of Manitoba, Department of Microbiology, Winnipeg, Manitoba, Canada.
Abstract:
Bacteriophage (phage) isolation from various environmental sources is an important step in identifying phages suitable for therapeutic and/or research purposes. Most published protocols outlining phage isolation techniques focus on the identification and/or characterization of strictly lytic phages. Here, a simple, adapted protocol specific for the isolation of non-lytic, filamentous phages is described. This method can be used in laboratories with access to basic microbiological equipment. Briefly, the adapted protocol steps involve:•Phage enrichment with F-pilus expressing bacterial host and sewage filtrate•Dilution and agar overlay plating of enriched culture•Selection of hazy, turbid plaques followed by screening via PCR.
Insights
This study presents a simple protocol for isolating non-lytic, filamentous bacteriophages (phages) from environmental samples. The method uses basic equipment and focuses on identifying phages for research and therapeutic applications.
Area of Science:
- Microbiology
- Virology
- Molecular Biology
Background:
- Bacteriophage (phage) isolation is crucial for identifying phages for therapeutic and research use.
- Existing protocols often focus on lytic phages, neglecting non-lytic filamentous types.
- There is a need for accessible methods to isolate diverse phage types.
Purpose of the Study:
- To describe a simple, adapted protocol for the isolation of non-lytic, filamentous phages.
- To provide a method suitable for laboratories with basic microbiological equipment.
- To facilitate the identification of novel phages for potential applications.
Main Methods:
- Phage enrichment using F-pilus expressing bacterial hosts and sewage filtrate.
- Dilution and agar overlay plating of the enriched culture.
- Selection of hazy, turbid plaques followed by Polymerase Chain Reaction (PCR) screening.
Main Results:
- The described protocol successfully isolates non-lytic, filamentous phages.
- The method is adaptable for laboratories with standard microbiological equipment.
- The protocol enables the screening of environmental samples for specific phage types.
Conclusions:
- This adapted protocol offers a straightforward approach for isolating non-lytic, filamentous phages.
- The method enhances the scope of phage discovery for therapeutic and research purposes.
- Accessible phage isolation techniques are vital for advancing phage-based research.
