Abstract

A novel phage, Φ241, specific for Escherichia coli O157:H7 was isolated from an industrial cucumber fermentation where both acidity (pH ≤ 3.7) and salinity (≥5% NaCl) were high. The phage belongs to the Myoviridae family. Its latent period was 15 min and average burst size was 53 phage particles per infected cell. The phage was able to lyse 48 E. coli O157:H7 strains, but none of the 18 non-O157 strains (including E. coli O104:H7) or the 2 O antigen-negative mutants of O157:H7 strain, 43895Δper (also lacking H7 antigen) and F12 (still expressing H7 antigen). However, the phage was able to lyse a per-complemented strain (43895ΔperComp) which expresses O157 antigen. These results indicated that phage Φ241 is specific for O157 antigen, and E. coli strains lacking O157 antigen were resistant to the phage infection, regardless of the presence or absence of H7 antigen. SDS-PAGE profile revealed at least 13 structural proteins of the phage. The phage DNA was resistant to many commonly used restriction endonucleases, suggesting the presence of modified nucleotides in the phage genome. At the multiplicity of infection of 10, 3, or 0.3, the phage caused a rapid cell lysis within 1 or 2 h, resulting in 3.5- or 4.5-log-unit reduction in cell concentration. The high lytic activity, specificity and tolerance to low pH and high salinity make phage Φ241 a potentially ideal biocontrol agent of E. coli O157:H7 in various foods. To our knowledge, this is the first report on E. coli O157:H7 phage isolated from high acidity and salinity environment.

Highlights

  • Escherichia coli O157:H7 has emerged as one of the major foodborne pathogens

  • A total of 46 E. coli O157:H7 strains, and 18 E. coli nonO157:H7 strains from various sources were obtained from the culture collection of USDA Agricultural Research Service located at North Carolina State University

  • Soft tryptic soy agar (TSA) agar used in plaque assay was prepared with tryptic soy broth (TSB) broth supplemented with 0.6% agar

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Summary

Introduction

Escherichia coli O157:H7 has emerged as one of the major foodborne pathogens Each year, it causes more than 73,000 illnesses, 2,100 hospitalizations, and 60 deaths in the U.S (Mead and Griffin, 1998; Mead et al, 1999; Rangel et al, 2005). Human infection by E. coli O157:H7 can frequently be traced to the food or water contaminated with cattle manure (Gyles, 2007). The infection by this pathogen can result in severe hemorrhagic colitis and life-threatening hemolytic uremic syndrome (Remis et al, 1984; Cleary, 1988; Tarr, 1995; Nataro and Kaper, 1998). Acid resistance is especially crucial for food-borne pathogens that must survive the hostile acidic condition in the stomach before entering and colonizing the small intestines or colon (Berk et al, 2005; Chen and Jiang, 2014)

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