The documented survival of pathogenic bacteria including Listeria monocytogenes (LM), shiga toxin-producing Escherichia coli (STEC), and Salmonella during the manufacture and aging of some cheeses highlights the need for additional interventions to enhance food safety. Unfortunately, few interventions are compliant with the Standards of Identity for cheese. Protective bacterial cultures (PC) represent actionable, natural interventions. However, supportive data for commercially produced PCs regarding their efficacy against pathogens and potential antagonism with each other and cheesemaking cultures are scant, thereby impeding their potential use by the cheese industry. The overall objective of this study was to identify commercially produced PCs that exert antimicrobial activity towards pathogens with minimal impact on beneficial cheese microbes. Direct antagonism and agar well diffusion assays were used to determine the impact of 10 commercially produced PCs on the growth of starter cultures and cultures of ripening bacteria and fungi. Deferred antagonism was used to evaluate the potential for antimicrobial effects against LM, STEC, and Salmonella. PCs and starter cultures were co-cultured in UHT milk to determine the effects of co-culture on starter acidification profiles when incubated according to a simulated cheese-making temperature profile (4 h at 35°C followed by 20 h at 20°C). Compatibility assays suggest that PC antagonism is microbe and strain specific. Only one PC negatively impacted the acidification of the starters tested. PC antagonism of ripening bacteria and fungi growth varied but was consistent within species. All PCs displayed deferred inhibition of LM, STEC, and Salmonella growth but to varying degrees. These data identify commercial PCs with potential for the control of pathogens and characterize their compatibility with cheesemaking cultures for future use by cheesemakers and investigations of their efficacy in the production of cheese.
Listeria selective enrichment media are designed to enhance the isolation of the organism and increase the chances of detection. Drawbacks include the requirements for prolonged sample incubation (48 to 72 h) and manual addition of selective agents, which may be a source of contamination. Modified Listeria recovery broth (mLRB) is a proprietary enrichment medium formulated to facilitate the recovery of injured cells; its selective agents are incorporated into a format that allows delayed release until 6 h of incubation. We evaluated the change in cell populations over time for acid- and nitrite-injured Listeria monocytogenes in mLRB with the selective agents added manually at 0 h (mLRBS0) and 6 h (mLRBS6). Recovery of injured cells in mLRB plus time-delayed tablets (mLRBTD) was also compared with that in enrichment media recommended by the U.S. Department of Agriculture (University of Vermont broth), the U.S. Food and Drug Administration (buffered Listeria enrichment broth), and the International Organization for Standardization (demi-Fraser broth). Nitrite- or acid-injured Listeria at approximately 10 CFU/ml were inoculated into each broth medium, and Listeria populations were enumerated at various times from 12 to 48 h of incubation at 37°C. Analysis of variance revealed that acid-injured Listeria populations in mLRBS6 at 24 h were significantly higher ( P < 0.05) than those in mLRBS0; however, the differences in populations on these two media were not significant for nitrite-injured cells. Cell populations of four strains of Listeria inoculated into mLRBTD were significantly higher at 24 h than when those strains were enriched in buffered Listeria enrichment broth, demi-Fraser broth, and University of Vermont broth. Comparison between artificially contaminated milk and meat samples with a four-strain cocktail of Listeria resulted in cell populations that were significantly higher ( P < 0.05) at 24 h on mLRBTD for contaminated meat than on mLRB for contaminated milk. Delivery of selective agents via time-delayed release tablets into mLRB maximizes recovery of acid- and nitrite-injured Listeria and saves analyst time during food sample analysis.