Isolation of F-specific Inovirus filamentous phages from environmental sewage samples

TL;DR

This paper describes a simple method to isolate non-lytic filamentous phages from sewage using basic lab equipment.

Contribution

A new protocol for isolating non-lytic F-specific inovirus phages from environmental samples is introduced.

Findings

The protocol uses F-pilus expressing bacteria and sewage filtrate for phage enrichment.

Hazy plaques were identified as potential filamentous phages and confirmed via PCR.

The method is accessible for labs with basic microbiological tools.

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

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

Image, graphical abstract

Figures