Isolation and characterization of a novel O157:H7-specific phage as a biocontrol agent.

PURPOSE

O157:H7 is one of the major foodborne pathogens of global public concern. Bacteriophages (phages) have emerged as a promising alternative to antibiotics for controlling pathogenic bacteria. Here, a lytic O157:H7-specific phage (KFS-EC) was isolated, identified, and characterized to evaluate its potential as a biocontrol agent for O157:H7.

METHODS

KFS-EC was isolated from slaughterhouse in Korea. Morphological analysis, genomic analysis and several physiological tests were performed to identify and characterize the KFS-EC.

RESULTS

A specificity test indicated KFS-EC was strictly specific to O157:H7 strains among 60 bacterial strains tested. Morphological and phylogenetic analyses confirmed that KFS-EC belongs to the genus, subfamily, and the family of phages. KFS-EC genome consists of 164,725 bp and a total of 270 coding sequence features, of which 114 open reading frames (ORFs) were identified as phage functional genes. KFS-EC does not contain genes encoding lysogenic property and pathogenicity, which ensure its safe application. KFS-EC was relatively stable (~1 log decrease) under stressed conditions such as temperatures (20 °C-50 °C), pHs (3-11), organic solvents (ethanol and chloroform), and biocides (0.1% citric acid, 1% citric acid, and 0.1% peracetic acid). KFS-EC was able to inhibit O157:H7 efficiently at a multiplicity of infection (MOI) of 0.01 for 8 h with greater inhibitory effect and durability and was stable at 4 °C and 22 °C over a 12-week storage period.

CONCLUSIONS

Our results suggest that KFS-EC could be used as a biocontrol agent to O157:H7.