The effect of CO2 and diacetyl, alone or in combination, on spoilage microflora in ground beef was determined. Ground beef was treated with 20, 30, or 40% CO2 for 22 days (study I); 20, 50, or 100 μg/g diacetyl for 26 days (study II); or a combination of 20% CO2 and 100 μg/g diacetyl for 40 days (study III). Antimicrobial effectiveness was determined by aerobic plate counts (log10 CFU/g) using plate count agar (total aerobic bacteria), deMan Rogosa Sharpe (MRS) Lactobacillus agar (gram-positive bacteria), MacConkey agar (gram-negative bacteria), pH, and informal organoleptic assessments (by appearance and by odor). In study I, total bacteria and pH increased by day 4 in control meat samples. For all CO2 levels, gram-negative bacteria decreased and gram-positive bacteria increased compared with untreated controls. The pH remained constant for CO2-treated meat. Control samples had an off-odor and a brown appearance, while CO2-treated samples had no off-odor but did have a brown appearance. For samples treated with diacetyl (study II), spoilage was evident by day 7 for samples treated with 0, 20, and 50 μg/g diacetyl for all parameters examined. Ground beef treated with 100 μg/g diacetyl was spoiled on day 12. Diacetyl was detected (by odor) in samples that were treated with 100 μg/g diacetyl and had a brown appearance. Meat samples treated with the combination of CO2 and diacetyl (study III) showed that the addition of diacetyl did not have an additive effect on microbial growth. Combination-treated meat maintained a red appearance and no off-odor. Diacetyl and CO2 could be used in combination to maintain a red color and inhibit spoilage microorganisms.

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