Development and validation of a surrogate strain cocktail to evaluate bactericidal effects of pressure on verotoxigenic Escherichia coli

Abstract

Many strains of verotoxigenic Escherichia coli (VTEC) are highly resistant to pressure. To facilitate future studies to improve the elimination of VTEC by pressure processing of food, this study developed and validated a cocktail of non-pathogenic strains of E. coli with equal or higher resistance to pressure when compared to pressure resistant strains of VTEC. Strains of E. coli obtained from a beef processing plant were screened for their resistance to heat and pressure. Treatments were carried out in LB broth. Cell counts of 3 out of 16 strains were reduced by 5–6 log (cfu/mL) after 30min at 60°C, and cell counts of 10 out of 16 strains were reduced by 5–6 log (cfu/mL) after 30min at 40°C and 400MPa. All highly heat resistant strains were also pressure resistant but not all pressure resistant strains were also heat resistant. Pressure resistant and -sensitive strains of E. coli were treated in presence of 0 or 2% NaCl and at 3, 20, or 40°C. The effect of these parameters on the lethality of pressure treatments was comparable for all strains. The addition of 2% NaCl did not increase pressure resistance. The bactericidal effect of treatments at 3 and 20°C and 600MPa was comparable but inactivation of E. coli was faster at 40°C and 600MPa. The resistance to treatment with 600MPa at 20°C of a cocktail of 5 non-pathogenic strains of E. coli was compared to a 5 strain cocktail of pressure resistant VTEC. Treatments were performed in ground beef containing 15% fat. Survival and sublethal injury of the two cocktails was comparable; cell counts of beef inoculated with either cocktail were reduced by about 4 log (cfu/mL) after 30min of treatment. In conclusion, this study validated a cocktail of non-pathogenic strains of E. coli for use as surrogate organisms in studies on the elimination of E. coli by pressure.