Efficacy of Gaseous Ozone Application during Vacuum Cooling against Escherichia coli O157:H7 on Spinach Leaves as Influenced by Bacterium Population Size

Abstract

Foodborne disease outbreaks associated with the consumption of fresh produce pose a threat to public health, decrease consumer confidence in minimally processed foods, and negatively impact the sales of these commodities. The aim of the study was to determine the influence of population size of inoculated pathogen on its inactivation by gaseous ozone treatment during vacuum cooling. Spinach leaves were spot inoculated with Escherichia coli O157:H7 at approximate initial populations of 108, 107, and 105 CFU/g. Inoculated leaves were vacuum cooled (28.5 inHg; 4°C) in a custom-made vessel and then were subjected to a gaseous ozone treatment under the following conditions: 1.5 g of ozone per kg of gas mixture, vessel pressure at 10 lb/in2 gauge, 94 to 98% relative humidity, and 30 min of holding time at 9°C. Treatment of the leaves, having the aforementioned inocula, decreased E. coli populations by 0.2, 2.1, and 2.8 log CFU/g, respectively, compared with the inoculated untreated controls. Additionally, spinach leaves were inoculated at 1.4 × 103 CFU/g, which approximates natural contamination level, and the small populations remaining after ozone treatment were quantified using the most-probable-number (MPN) method. Vacuum and ozone sequential treatment decreased this E. coli O157:H7 population to <3 MPN/g (i.e., greater than 3-log reduction). Resulting log reductions were greater (P < 0.05) at the lower rather than the higher inoculum levels. In conclusion, treatment of spinach leaves with gaseous ozone is effective against pathogen loads comparable to those found in naturally contaminated fresh produce, but efficacy decreases as inoculum level increases.