The combined effects of acetic acid and mustard flour were investigated to ascertain their impact on Escherichia coli O157:H7 stored at 5 and 22°C. Samples were prepared with various concentrations of acetic acid (0, 0.25, 0.5, 0.75, and 1% [vol/vol]) combined with 10% (wt/vol) Baltimore or Coleman mustard flour and 2% (fixed; wt/vol) sodium chloride. An acid-adapted mixture of three E. coli O157:H7 strains (106 to 107 CFU/ml) was inoculated into prepared mustard samples that were stored at 5 and 22°C, and samples were assayed periodically for the survival of E. coli O157:H7. The numbers of E. coli O157:H7 were reduced much more rapidly at 22°C than at 5°C. E. coli O157:H7 was rapidly reduced to below the detection limit (<0.3 log10 CFU/ml) after 1 day at 22°C, whereas it survived for up to 5 days at 5°C. There was no synergistic or additive effect with regard to the killing of E. coli O157:H7 with the addition of small amounts of acetic acid to the mustard flour. When stored at 5°C, mustard in combination with 0.25 (M-0.25), 0.5 (M-0.5), and 0.75% (M-0.75) acetic acid exerted less antimicrobial activity than the control (M-0). The order of lethality at 5°C was generally M-0.25 = M-0.5 < M-0.75 = M-0 < M-1. The addition of small amounts of acetic acid (<0.75%) to mustard retards the reduction of E. coli O157:H7. Statistical reduction in populations of E. coli O157:H7 (P < 0.05) was enhanced relative to that of the control (mustard alone) only with the addition of 1% acetic acid. This information may help mustard manufacturers to understand the antimicrobial activity associated with use of mustard flour in combination with acetic acid.

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