Potentially hazardous foods require time/temperature control for safety. According to the U.S. Food and Drug Administration Food Code, most cheeses are potentially hazardous foods based on pH and water activity, and a product assessment is required to evaluate safety of storage >6 h at 21°C. We tested the ability of 67 market cheeses to support growth of Listeria monocytogenes (LM), Salmonella spp. (SALM), Escherichia coli O157:H7 (EC), and Staphylococcus aureus (SA) over 15 days at 25°C. Hard (Asiago and Cheddar), semi-hard (Colby and Havarti), and soft cheeses (mozzarella and Mexican-style), and reduced-sodium or reduced-fat types were tested. Single-pathogen cocktails were prepared and individually inoculated onto cheese slices (∼10(5) CFU/g). Cocktails were 10 strains of L. monocytogenes, 6 of Salmonella spp., or 5 of E. coli O157:H7 or S. aureus. Inoculated slices were vacuum packaged and stored at 25°C for ≤ 15 days, with surviving inocula enumerated every 3 days. Percent salt-in-the-moisture phase, percent titratable acidity, pH, water activity, and levels of indigenous/starter bacteria were measured. Pathogens did not grow on 53 cheeses, while 14 cheeses supported growth of SA, 6 of SALM, 4 of LM, and 3 of EC. Of the cheeses supporting pathogen growth, all supported growth of SA, ranging from 0.57 to 3.08 log CFU/g (average 1.70 log CFU/g). Growth of SALM, LM, and EC ranged from 1.01 to 3.02 log CFU/g (average 2.05 log CFU/g), 0.60 to 2.68 log CFU/g (average 1.60 log CFU/g), and 0.41 to 2.90 log CFU/g (average 1.69 log CFU/g), respectively. Pathogen growth varied within cheese types or lots. Pathogen growth was influenced by pH and percent salt-in-the-moisture phase, and these two factors were used to establish growth/no-growth boundary conditions for safe, extended storage (≤25°C) of pasteurized milk cheeses. Pathogen growth/no-growth could not be predicted for Swiss-style cheeses, mold-ripened or bacterial surface-ripened cheeses, and cheeses made with nonbovine milk, as insufficient data were gathered. This challenge study data can support science-based decision making in a regulatory framework.
Biopreservatives are clean-label ingredients used to control pathogenic and spoilage microorganisms in ready-to-eat foods including cheese. The efficacies of six commercial biopreservatives in controlling L. monocytogenes growth at 4°C were tested in a high-moisture model cheese (pH 6.00, 56% moisture, 1.25% salt) made of cream, micellar casein, water, salt, lactose, lactic acid, and a single protective culture (PC-1, PC-2, or PC-3, 10 6 CFU/g target) or bacterial fermentate (CM-1 or CM-2 [cultured milk] or CSV-1 [cultured sugar-vinegar blend], 0.5% or 1.0% level). Cheeses were inoculated with 3-log CFU/g L. monocytogenes (5-strain cocktail), after which 25-g samples were vacuum-sealed and stored at 4°C for 8 weeks. L. monocytogenes populations from triplicate samples were enumerated weekly on Modified Oxford agar in duplicate trials. L. monocytogenes growth (≥1-log increase) was observed in approximately 1 week in control cheese and those formulated with 10 6 CFU/g PC-1 or PC-2. Growth was delayed to 2.5 weeks in model cheeses formulated with 10 6 CFU/g PC-3 or 0.5% CM-2 and to 3 weeks with 0.5% CM-1 or CSV-1. Growth was further delayed to 6.5-7.5 weeks in model cheeses formulated with 1.0% CM-1 or CM-2, while formulation with 1.0% CSV-1 inhibited L. monocytogenes growth for 8 weeks. In a second set of experiments, the combined effect of pH and 0.5% CSV-1 on L. monocytogenes inhibition was investigated. Incorporation of 0.5% CSV-1 delayed L. monocytogenes growth to 3, 6, and >10 weeks in cheeses of pH 6.00, 5.75, and 5.50, respectively, versus growth observed in 1, 1, and 3.5 weeks in control cheeses. These data suggest that certain fermentates have greater antilisterial activity than protective cultures in directly acidified cheeses with direct biopreservative incorporation and refrigerated storage. Further research is needed to optimize conditions to prevent listerial growth utilizing protective cultures in fresh, soft cheeses.
High-moisture, low-acid cheeses have been shown to support Listeria monocytogenes growth during refrigerated storage. Prior studies suggest that organic acids vary in their antilisterial activity and that cheeses of lower pH delay growth longer than those of higher pH; however, no standard pH value for Listeria control in cheese exists. The objective of this research was to create a predictive model to include the effects of acid type, pH, and moisture on the growth of L. monocytogenes in a model cheese system. Cream, micellar casein, water, lactose, salt, and acid (citric, lactic, acetic, or propionic) were combined in 32 formulations targeting 4 pH values (5.25, 5.50, 5.75, and 6.00) and two moisture levels (50 and 56%). Each was inoculated with 3 log CFU/g L. monocytogenes (five-strain mixture) after which 25-g samples were vacuum sealed and stored 8 weeks at 4°C. Triplicate samples were enumerated on modified Oxford agar weekly in duplicate trials. Model cheeses formulated with acetic and propionic acids inhibited growth (i.e., no observed increase in L. monocytogenes populations over 8 weeks) at pH ≤5.75, while those formulated with lactic acid inhibited growth at pH 5.25 only. In contrast, all model cheeses formulated with citric acid supported growth. Resulting growth curves were fitted for lag phase and growth rate before constructing models for each. The pH and acid type were found to significantly affect both growth parameters (P < 0.05), while moisture (50 to 56%) was not statistically significant in either model (P ≥ 0.05). The effects of acetic and propionic acid were not significantly different. In contrast, model cheeses made with citric acid had significantly shorter lag phases than the other acids tested, but growth rates after lag were statistically similar to model cheeses made with lactic acid. These data suggest propionic ∼ acetic > lactic > citric acids in antilisterial activity within the model cheese system developed and can be used in formulating safe high-moisture cheeses. HIGHLIGHTS
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