F-specific coliphage detection by the Bluephage method

Abstract

F-specific coliphages have been proposed as viral indicators of fecal pollution. These intestinal phages infect cells through the F-pili of the host strains used for their detection, Escherichia. coli HS/FAmp in the US-EPA standard method and Salmonella enterica WG49 in the ISO method. The recently designed Bluephage protocol allows the rapid detection of as low as one somatic coliphage in a working day. The current study describes a new Bluephage method designed to exclusively detect F-specific phages. It employs two new host strains, CB14 and CB16, which detect the same number of F-specific phages as their respective parental strains HS and WG49. In the Bluephage method, when the strain is lysed by bacteriophage infection, the yellow medium turns blue. As low as one F-specific phage was detected in 3 to 5 h by this approach and when the sample contained high phage concentrations, results were obtained in less than 3 h. The F-specific Bluephage method can be used with different sample volumes and allows phage quantification by the most probable number technique. Strain CB14 performed more consistently than CB16, with comparable detection efficiency after increasing the incubation time to 50 min without shaking.

Keywords: Coliphages; F-specific coliphages; Fecal pollution; Glucuronidase; New tools; Viral indicator; Virus.