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The mechanism of the antimicrobial action of Spanish oregano (Corydothymus capitatus), Chinese cinnamon (Cinnamomum cassia), and savory (Satureja montana) essential oils against cell membranes and walls of bacteria was studied by the measurement of the intracellular pH and ATP concentration, the release of cell constituents, and the electronic microscopy observations of the cells when these essential oils at their MICs were in contact with Escherichia coli O157:H7 and Listeria monocytogenes. E. coli O157:H7 and L. monocytogenes, two pathogenic foodborne bacteria, were used as gram-negative and gram-positive bacterial models, respectively. Treatment with these essential oils at their MICs affected the membrane integrity of bacteria and induced depletion of the intracellular ATP concentration. Spanish oregano and savory essential oils, however, induced more depletion than Chinese cinnamon oil. An increase of the extracellular ATP concentration was observed only when Spanish oregano and savory oils were in contact with E. coli O157:H7 and L. monocytogenes. Also, a significantly higher (P < or = 0.05) cell constituent release was observed in the supernatant when E. coli O157:H7 and L. monocytogenes cells were treated with Chinese cinnamon and Spanish oregano oils. Chinese cinnamon oil was more effective to reduce significantly the intracellular pH of E. coli O157:H7, whereas Chinese cinnamon and Spanish oregano decreased more significantly the intracellular pH of L. monocytogenes. Electronic microscopy observations revealed that the cell membrane of both treated bacteria was significantly damaged. These results suggest that the cytoplasmic membrane is involved in the toxic action of essential oils.

Milk protein-based edible films containing 1.0% (w/v) oregano, 1.0% (w/v) pimento, or 1.0% oregano-pimento (1:1) essential oils mix were applied on beef muscle slices to control the growth of pathogenic bacteria and increase the shelf life during storage at 4 degrees C. Meat and film were periodically tested during 7 days for microbial and biochemical analysis. The lipid oxidation potential of meat was evaluated by the determination of thiobarbituric reactive substances (TBARS). The availability of phenolic compounds from essential oils was evaluated by the determination of total phenolic compounds present in the films during storage. Antioxidant properties of films during storage were also evaluated following a modified procedure of the N,N-diethyl-p-phenylenediamine colorimetric method. Oregano-based films stabilized lipid oxidation in beef muscle samples, whereas pimento-based films presented the highest antioxidant activity. The application of bioactive films on meat surfaces containing 10(3) colony-forming units/cm2 of Escherichia coli O157:H7 or Pseudomonas spp. showed that film containing oregano was the most effective against both bacteria, whereas film containing pimento oils seems to be the least effective against these two bacteria. A 0.95 log reduction of Pseudomonas spp. level, as compared to samples without film, was observed at the end of storage in the presence of films containing oregano extracts. A 1.12 log reduction of E. coli O157:H7 level was noted in samples coated with oregano-based films.

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