This research investigated the efficacy of gaseous ozone for the inactivation of Escherichia coli ATCC 25922 and NCTC 12900 strains in orange juice. Orange juice inoculated with E. coli (10 6 CFU mL -1 ) as a challenge microorganism was treated with ozone at 75-78µg mL -1 for different time periods (0-18 min). The efficacy of ozone for inactivation of both strains of E. coli was evaluated as a function of different juice types: model orange juice, fresh unfiltered juice, juice without pulp, and juice filtered through 500m or 1mm sieves. Fast inactivation rates for total reduction of E. coli were achieved in model orange juice (60 seconds) and in juice with low pulp content (6 min). However, in unfiltered juice inactivation was achieved after 15-18 min. This indicated that juice organic matter interferes with antibacterial activity of gaseous ozone. The effect of prior acid (pH 5.0) exposure of E. coli strains on the inactivation efficacy of ozone treatment was also investigated. There was a strain effect observed, where prior acid exposure resulted in higher inactivation times in some cases by comparison with the control cells.However, the overarching influence on inactivation efficacy of ozone was related to the pulp content. Generally, the applied gaseous ozone treatment of orange juice resulted in a population reduction of 5 log cycles.Key words: Escherichia coli, ozone, non-thermal inactivation, acid exposure, orange juice, microbial kinetics Industrial relevance: To facilitate the preservation of unstable nutrients many juice processors have investigated alternatives to thermal pasteurisation, including unpasteurised short shelf life juices with high retail value. This trend has continued within the European Union. However within the US recent regulations by the FDA have 3 required processors to achieve a 5-log reduction in the numbers of the most resistant pathogens in their finished products. Pathogenic E. coli may survive in acid environments such as fruit juices for long periods. This study demonstrates that the use of ozone as a non-thermal technology is effective for inactivation of E. coli and acid exposed E. coli in orange juice. Information on the design of the ozone treatment for inactivation of E. coli which results into safe juice products is also among the main outputs of this work. Ozone auto-decomposition makes this technology safe for fruit juice processing.4