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Applied and Environmental Microbiology
Vol. 80, No. 2, 20
14; Page: 515 - 524

Vibrio vulnificus Bacteriophage SSP002 as a Possible Biocontrol Agent

Hyun Sung Lee, Slae Choi, Hakdong Shin, Ju-Hoon Lee and Sang Ho Choi

Department of Agricultural Biotechnology, Center for Food Safety and Toxicology, and Center for Food and Bioconvergence, Seoul National University, Seoul, South Korea

Abstract

A novel Vibrio vulnificus-infecting bacteriophage, SSP002, belonging to theSiphoviridae family, was isolated from the coastal area of the Yellow Sea of South Korea. Host range analysis revealed that the growth inhibition of phage SSP002 is relatively specific to V. vulnificus strains from both clinical and environmental samples. In addition, a one-step growth curve analysis and a bacteriophage stability test revealed a latent period of 65 min, a burst size of 23 ± 2 PFU, as well as broad temperature (20°C to 60°C) and pH stability (pH 3 to 12) ranges. A Tn5random transposon mutation of V. vulnificus and partial DNA sequencing of the inserted Tn5 regions revealed that the flhAflhBfliF, and fleQ mutants are resistant to SSP002 phage infection, suggesting that the flagellum may be the host receptor for infection. The subsequent construction of specific gene-inactivated mutants (flhAflhBfliF, and fleQ) and complementation experiments substantiated this. Previously, the genome of phage SSP002 was completely sequenced and analyzed. Comparative genomic analysis of phage SSP002 andVibrio parahaemolyticus phage vB_VpaS_MAR10 showed differences among their tail-related genes, supporting different host ranges at the species level, even though their genome sequences are highly similar. An additional mouse survival test showed that the administration of phage SSP002 at a multiplicity of infection of 1,000 significantly protects mice from infection by V. vulnificus for up to 2 months, suggesting that this phage may be a good candidate for the development of biocontrol agents against V. vulnificus infection.

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