Acid Adaptation Enhances Tolerance of Escherichia coli O157:H7 to High Voltage Atmospheric Cold Plasma in Raw Pineapple Juice.

Pathogens that adapt to environmental stress can develop an increased tolerance to some physical or chemical antimicrobial treatments. The main objective of this study was to determine if acid adaptation increased the tolerance of Escherichia coli O157:H7 to high voltage atmospheric cold plasma (HVACP) in raw pineapple juice. Samples (10 mL) of juice were inoculated with non-acid-adapted (NAA) or acid-adapted (AA) E. coli to obtain a viable count of ~7.00 log10 CFU/mL. The samples were exposed to HVACP (70 kV) for 1-7 min, with inoculated non-HVACP-treated juice serving as a control. Juice samples were analyzed for survivors at 0.1 h and after 24 h of refrigeration (4 °C). Samples analyzed after 24 h exhibited significant decreases in viable NAA cells with sub-lethal injury detected in both NAA and AA survivors (p < 0.05). No NAA survivor in juice exposed to HVACP for 5 or 7 min was detected after 24 h. However, the number of AA survivors was 3.33 and 3.09 log10 CFU/mL in juice treated for 5 and 7 min, respectively (p < 0.05). These results indicate that acid adaptation increases the tolerance of E. coli to HVACP in pineapple juice. The potentially higher tolerance of AA E. coli O157:H7 to HVACP should be considered in developing safe juice processing parameters for this novel non-thermal technology.

A Review of Regulatory Standards and Advances in Essential Oils as Antimicrobials in Foods.

As essential oils (EOs) possess GRAS status, there is a strong interest in their application to food preservation. Trends in the food industry suggest consumers are drawn to environmentally friendly alternatives and less synthetic chemical preservatives. Although the use of EOs has increased over the years, adverse effects have limited their use. This review aims to address the regulatory standards for EO usage in food, techniques for delivery of EOs, essential oils commonly used to control pathogens and molds, and advances with new active compounds that overcome sensory effects for meat products, fresh fruits and vegetables, fruit and vegetable juices, seafood, dairy products, and other products. This review will show adverse sensory effects can be overcome in various products by the use of edible coatings containing encapsulated EOs to facilitate the controlled release of EOs. Depending on the method of cooking, the food product has been shown to mask flavors associated with EOs. In addition, using active packaging materials can decrease the diffusion rate of the EOs, thus controlling undesirable flavor characteristics while still preserving or prolonging the shelf life of food. The use of encapsulation in packaging film can control the release of volatile or active ingredients. Further, use of EOs in the vapor phase allows for contact indirectly, and use of nanoemulsion, coating, and film wrap allows for the controlled release of the EOs. Research has also shown that combining EOs can prevent adverse sensory effects. Essential oils continue to serve as a very beneficial way of controlling undesirable microorganisms in food systems.

Shiga Toxin-Producing Escherichia coli in the Long-Term Survival Phase Exhibit Higher Chlorine Tolerance and Less Sublethal Injury Following Chlorine Treatment of Romaine Lettuce.

The extent of chlorine inactivation and sublethal injury of stationary-phase (STAT) and long-term survival-phase (LTS) cells of Shiga toxin-producing Escherichia coli (STEC) in vitro and in a lettuce postharvest wash model was investigated. Four STEC strains were cultured in tryptic soy broth supplemented with 0.6% (w/v) yeast extract (TSBYE; 35°C) for 24 h and 21 d to obtain STAT and LTS cells, respectively. Minimum bactericidal concentration (MBC) and dose-response assays were performed to determine chlorine's antibacterial efficacy against STAT and LTS cells. Chlorine solutions (pH 6.5) and romaine lettuce were each inoculated with STAT and LTS cells to obtain initial populations of ∼7.8 log colony-forming units (CFU)/mL. Survivors in chlorine solutions were determined after 30 s. Inoculated lettuce samples were held at 22°C ± 1°C for 2 h or 20 h and then exposed to chlorine (10-40 ppm) for 60 s. Survivors were enumerated on nonselective and selective agar media following incubation (35°C, 48 h). The MBC for STAT and LTS cells was 0.04 and 0.08 ppm, respectively. Following exposure (30 s) to chlorine at 2.5, 5.0, and 10 ppm, STAT cells were reduced to <1.0 log CFU/mL, whereas LTS survivors were at 5.10 (2.5 ppm), 3.71 (5.0 ppm), and 2.55 (10 ppm) log CFU/mL. At 20 and 40 ppm chlorine, greater log CFU reductions of STAT cells (1.64 and 1.85) were observed compared with LTS cells (0.94 and 0.83) after 2 h of cell contact with lettuce (p < 0.05), but not after 20 h. Sublethal injury in STEC after chlorine (40 ppm) treatment was lower in LTS compared with STAT survivors (p < 0.05). Compared with STAT cells, LTS cells of STEC seem to have higher chlorine tolerance as planktonic cells and as attached cells depending on cell contact time on lettuce. In addition, a higher percentage of LTS cells, compared with STAT cells, survive in a noninjured state after chlorine (40 ppm) treatment of lettuce.

Thyme Oil Enhances the Inactivation of Salmonella enterica on Raw Chicken Breast Meat During Marination in Lemon Juice With Added Yucca schidigera Extract.

Enteric pathogens such as Salmonella enterica can survive in low pH conditions and pose a food safety threat during marinating of raw poultry meat. A study was conducted to investigate the effectiveness of thyme oil for killing S. enterica on raw chicken during marination in lemon juice containing yucca extract. Samples of raw chicken breast were inoculated with a five-serovar mixture of S. enterica (~108 CFU/mL) and immersed for 2, 4, 6, and 8 h in four lemon-based marinades at 22°C: lemon juice alone (L), L with added 0.5% yucca extract (L + Y), L + Y and 0.5% thyme oil (L + Y + 0.5% TO) and L + Y + 1.0% TO. The L and L + Y served as controls. Survivors were determined by surface plating chicken homogenates on xylose-lysine tergitol-4 (XLT4) agar and XLT4 agar overlaid with non-selective agar (TAL) and counting bacterial colonies after 48 h of incubation (35°C). Marinades containing Y and TO significantly reduced initial viable populations of S. enterica compared to control (L and L + Y) solutions (P < 0.05). Based on S. enterica survivors on TAL medium, the L and L + Y reduced initial populations by 1.12 and 1.42 Log CFU/sample, respectively, after 8 h whereas, Log reductions caused by L + Y + 0.5% TO and L + Y + 1.0% TO, respectively, were 2.62 and 3.91 (P < 0.05). Numbers of survivors were higher on TAL compared to XLT4 agar (P < 0.05); however, the extent of sub-lethal injury caused by the marinades was not statistically significant (P > 0.05). The death rate of S. enterica increased significantly (P < 0.05) in the marinades containing TO (0.5 or 1.0%) compared to control (L + Y). Based on these results, thyme oil has good potential to increase the antimicrobial efficacy of lemon juice marinade against Salmonella on raw chicken breast and enhance the microbial safety of this popular poultry product.

Long-Term Survival Phase Cells of Salmonella Typhimurium ATCC 14028 Have Significantly Greater Resistance to Ultraviolet Radiation in 0.85% Saline and Apple Juice.

Nonendospore-forming pathogenic bacteria in the long-term survival (LTS) phase can remain viable for months or years and may show reduced susceptibility to various antimicrobial interventions. In the present study, we investigated the response of LTS phase Salmonella enterica serovar Typhimurium (ATCC 14028) to ultraviolet (UV) radiation in 0.85% (w/v) saline and apple juice and the extent of sublethal injury in LTS phase survivors. The LTS-phase Salmonella Typhimurium cells were cultured at 35°C for 14 days in tryptic soy broth with 0.6% (w/v) yeast extract (TSBYE). Exponential- and stationary-phase cells, cultured in TSBYE (35°C) for 2.5 and 18 h, respectively, served as control samples. Cells (107 CFU [colony-forming unit]/mL) from each physiological state were exposed to UV light in saline (80 µW/cm2) and apple juice (1500 µW/cm2). The Salmonella Typhimurium survivors were plated for enumeration on either tryptic soy agar with 0.6% yeast extract or xylose-lysine-tergitol 4 (XLT4) agar and colonies counted after incubation (35°C, 24 h). Of all the growth phases tested, LTS phase cells were consistently impacted the least by UV treatment (p < 0.05). In saline, D-values of exponential, stationary, and LTS Salmonella Typhimurium were 0.35, 0.38, and 0.49 min, respectively. D-values in apple juice at pH 3.63 and pH 5.65 were 2.52, 3.19, and 3.57 min and 3.24, 3.50, and 4.18 min, respectively. UV radiation (80 µW/cm2) of Salmonella Typhimurium in saline for 2.5 min reduced the number of exponential- and stationary-phase cells by ∼7.19 and 6.30 log10 CFU/mL, respectively. In contrast, LTS cells were only reduced by 5.08 log10 CFU/mL. Among the three physiological states, LTS phase cells had the least sublethal injury in the surviving population (p < 0.05). These results indicate that the LTS state cross-protects Salmonella Typhimurium against UV radiation and should be considered in determination of the UV radiation D-value for this pathogen.