Ozone (O3) is a strong antimicrobial agent with numerous potential applications in the food industry. High reactivity, penetrability, and spontaneous decomposition to a nontoxic product (i.e., O2) make ozone a viable disinfectant for ensuring the microbiological safety of food products. Ozone has been used for decades in many countries and recently, the generally recognized as safe (GRAS) status of this gas has been reaffirmed in the United States. Ozone, in the gaseous or aqueous phases, is effective against the majority of microorganisms tested by numerous research groups. Relatively low concentrations of ozone and short contact time are sufficient to inactivate bacteria, molds, yeasts, parasites, and viruses. However, rates of inactivation are greater in ozone demand-free systems than when the medium contains oxidizable organic substances. Susceptibility of microorganisms to ozone also varies with the physiological state of the culture, pH of the medium, temperature, humidity, and presence of additives (e.g., acids, surfactants, and sugars). Ozone applications in the food industry are mostly related to decontamination of product surface and water treatment. Ozone has been used with mixed success to inactivate contaminant microflora on meat, poultry, eggs, fish, fruits, vegetables, and dry foods. The gas also is useful in detoxification and elimination of mycotoxins and pesticide residues from some agricultural products. Excessive use of ozone, however, may cause oxidation of some ingredients on food surface. This usually results in discoloration and deterioration of food flavor. Additional research is needed to elucidate the kinetics and mechanisms of microbial inactivation by ozone and to optimize its use in food applications.

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When ozone (1.3 mM) was bubbled for 3 min in a mixture of shredded lettuce and water, counts of mesophilic andpsychrotrophic bacteria decreased 1.4 and 1.8 log,, cfu/g, respectively. Counts of these microorganisms on lettuce, from a different batch, decreased 3.9 and 4.6 log, respectively, during 5 min of ozone treatment. Shredded lettuce was treated with gaseous ozone, or mixed with aqueous solution of ozone (1:20 w/w) with or without bubbles. For effective delivery of ozone, stirring (low and high speed), sonication or stomaching was applied during the ozonation. Washing the lettuce with water only decreased total count on shredded lettuce by 0.74-1.0 log cfug. When lettuce in a treatment chamber was flushed with gaseous ozone, the total count decreased 0.85 log cfu/g, but when vacuum was applied before the ozoneflush, the total count decreased 0.96 log cfu/g. Bubbling ozone in water-lettuce mixture while sonicating, high-speed stirring, or before stomaching inactivated I .4, 1.9 and 1.9 log cfu/g, respectively.In conclusion, bubbling gaseous ozone in water is the most effective ozonation method. Eficient ozone delivery to microorganisms on lettuce requires a combination of ozone bubbling and high-speed stir.'Corresponding author.

Pulsed electric field (PEF) and ozone technologies are nonthermal processing methods with potential applications in the food industry. This research was performed to explore the potential synergy between ozone and PEF treatments against selected foodborne bacteria. Cells of Lactobacillus leichmannii ATCC 4797, Escherichia coli O157:H7 ATCC 35150, and Listeria monocytogenes Scott A were suspended in 0.1% NaCl and treated with ozone, PEF, and ozone plus PEE Cells were treated with 0.25 to 1.00 microg of ozone per ml of cell suspension, PEF at 10 to 30 kV/cm, and selected combinations of ozone and PEF. Synergy between ozone and PEF varied with the treatment level and the bacterium treated. L. leichmannii treated with PEF (20 kV/cm) after exposure to 0.75 and 1.00 microg/ml of ozone was inactivated by 7.1 and 7.2 log10 CFU/ml, respectively; however, ozone at 0.75 and 1.00 microg/ml and PEF at 20 kV/cm inactivated 2.2, 3.6, and 1.3 log10 CFU/ml, respectively. Similarly, ozone at 0.5 and 0.75 microg/ml inactivated 0.5 and 1.8 log10 CFU/ml of E. coli, PEF at 15 kV/cm inactivated 1.8 log10 CFU/ml, and ozone at 0.5 and 0.75 microg/ml followed by PEF (15 kV/cm) inactivated 2.9 and 3.6 log10 CFU/ml, respectively. Populations of L. monocytogenes decreased 0.1, 0.5, 3.0, 3.9, and 0.8 log10 CFU/ml when treated with 0.25, 0.5, 0.75, and 1.0 microg/ml of ozone and PEF (15 kV/cm), respectively; however, when the bacterium was treated with 15 kV/cm, after exposure to 0.25, 0.5, and 0.75 microg/ml of ozone, 1.7, 2.0, and 3.9 log10 CFU/ml were killed, respectively. In conclusion, exposure of L. leichmannii, E. coli, and L. monocytogenes to ozone followed by the PEF treatment showed a synergistic bactericidal effect. This synergy was most apparent with mild doses of ozone against L. leichmannii.

Sabouraud dextrose agar, containing up to 300 ppm of eugenol, isoeugenol or monolaurin, was prepared. Eight strains of mycotoxin and non‐mycotoxin‐producing Aspergillus spp. and Penicillium spp. were tested. Spore suspensions were spotted in the center of agar plates and the linear (radial) growth rate was measured during incubation. All agents inhibited mold growth, but isoeugenol and eugenol were more effective than monolaurin. The average decreases in linear growth rate in the presence of eugenol, isoeugenol and monolaurin were 56, 43 and 11% at 100 ppm and 81, 85 and 15% at 200 ppm, respectively. At 300 ppm eugenol and isoeugenol, all fungi were inhibited completely; however, monolaurin (up to 2000 ppm) did not prevent mold growth. Except for P. viridicatum NRRL 6062, the molds tested did not vary significantly (at p=0.05) in degree of inhibition by any particular antifungal agent. A. parasiticus was grown in a yeast extract dextrose broth containing 100 ppm of the antifungal agents. Decreases in mycelial dry weight in the presence of eugenol, isoeugenol and monolaurin were 5.3, and 7.9, 10.5%, respectively. The corresponding reductions in aflatoxin B1 synthesized were 58, 57 and 25%. In conclusion, eugenol and isoeugenol have strong antifungal properties which are best manifested when the surface growth of molds on solid media is monitored.