Efficacy of Acidified Oils against Salmonella in Low-Moisture Environments.

When processing low-moisture, high-fat foods such as peanut butter and nuts, water-based sanitization is unsuitable due to the immiscible nature of water and fats. Dry sanitization mainly uses flammable compounds such as isopropanol, requiring equipment cooling before application. The use of oils to deliver antimicrobials against foodborne pathogens enables the use of elevated temperatures, thus eliminating processing downtimes associated with dry sanitization. This study delivered organic acids and medium-chain fatty acids (100, 250, and 500 mM) in peanut oil against Salmonella enterica serovar Enteritidis desiccated at 75% relative humidity (RH). Acetic acid in peanut oil (AO) at 45°C was the most effective food-grade acid, causing a 4.4-log reduction in S. Enteritidis at 500 mM. AO caused cellular injury and was effective against a variety of S. Enteritidis strains. Confocal microscopy demonstrated that cells treated with 50 mM and 250 mM AO had significant membrane damage and reduced cellular respiration compared to untreated controls. Treatment efficacy increased with the increase in acid concentration, treatment duration, and treatment temperature from 20 to 45°C. Transmission electron microscopy after treatment with 100 and 250 mM AO revealed membrane ruffling and leakage in cell membranes, especially at 45°C. Reduction of the RH to 33% during desiccation of S. Enteritidis caused a decrease in AO efficacy compared to that at 75% RH, while at a higher RH of 90%, there was an increase in the efficacy of AO. Acidified oils can serve as robust, cost-effective replacements for dry-sanitation methods and improve safety of low moisture foods. IMPORTANCE Currently, dry sanitization products used during food processing often contain flammable compounds which require processing to stop and equipment to cool before application. This leads to processing downtimes and consequently, economic losses. This challenge is compounded by exposure to dryness which frequently renders Salmonella resistant to heat and different antimicrobials. Thus, the development of heat-tolerant oil-based antimicrobial compounds is a novel approach for sanitizing in low-moisture (dry) environments such as those found in peanut butter, tree nuts, and chocolate manufacturing. This study shows that acidified oils, especially acetic acid in peanut oil at elevated temperatures (45°C), was highly effective against desiccated Salmonella. Acidified oils have the potential to replace dry sanitizers, increasing the frequency of sanitization, leading to an improvement in food safety.

Modeling the effect of simultaneous use of allyl isothiocyanate and cinnamaldehyde on high hydrostatic pressure inactivation of Uropathogenic and Shiga toxin-producing Escherichia coli in ground chicken.

BACKGROUND: A combination of high-pressure processing (HPP) and antimicrobials is a well-known approach for enhancing the microbiological safety of foods. However, few studies have applied multiple antimicrobials simultaneously with HPP, which could be an additional hurdle for microbial inactivation. The present study applied a full factorial design to investigate the impact of HPP (225-325 MPa; 10-20 min), allyl isothiocyanate (AITC) (0.3-0.9 g kg-1 ) and trans-cinnamaldehyde (tCinn) (1.0-2.0 g kg-1 ) on the inactivation of Shiga toxin-producing Escherichia coli (STEC) O157:H7 and uropathogenic E. coli (UPEC) in ground chicken meat. RESULTS: The regulatory requirement of 5-log reduction was achieved at 305 MPa, 18 min, 0.8 g kg-1 AITC and 1.7 g kg-1 tCinn for STEC O157:H7 and at 293 MPa, 16 min, 0.6 g kg-1 AITC and 1.6 g kg-1 tCinn for UPEC, as specified by response surface analysis and verified via experiments. The surviving population was eliminated by post-treatment storage of 9 days at 10 °C. The developed linear regression models showed r2 > 0.9 for the E. coli inactivation. The developed dimensionless non-linear regression models covered a factorial range slightly wider than the original experimental limit, with probability Pr > F (< 0.0001). CONCLUSION: Simultaneous use of AITC and tCinn reduced not only the necessary concentration of each compound, but also the intensity of high-pressure treatments, at the same time achieving a similar level of microbial inactivation. STEC O157:H7 was found to be more resistant than UPEC to the HPP-AITC-tCinn stress. The developed models may be applied in commercial application to enhance the microbiological safety of ground chicken meat. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.

Efficacy of acidified water-in-oil emulsions against desiccated Salmonella as a function of acid carbon chain-length and membrane viscosity.

Sanitizing low-moisture food (LMF) processing equipment is challenging due to the increased heat resistance of Salmonella spp. in low-water activity (aw) environments. Food-grade oils mixed with acetic acid have been shown effective against desiccated Salmonella. In this study, different hydrocarbon chain-length (Cn) organic acids were tested against desiccated Salmonella by using 1% v/v water-in-oil (W/O) emulsion as the delivery system for 200 mM acid. Fluorescence lifetime imaging microscopy (FLIM) was utilized with a BODIPY-based molecular rotor to evaluate membrane viscosity under environmental conditions such as desiccation and temperature elevation. Drying hydrated Salmonella cells to 75% equilibrium relative humidity (ERH) increased the membrane viscosity from 1,199 to 1,309 mPa·s (cP) at 22°C. Heating to 45°C decreased the membrane viscosity of hydrated cells from 1,199 to 1,082 mPa·s, and decreased that of the desiccated cells from 1,309 to 1,245 mPa·s. At both 22°C and 45°C, desiccated Salmonella was highly susceptible (>6.5 microbial log reduction (MLR) per stainless-steel coupon) to a 30-min treatment with the W/O emulsions formulated with short carbon chain acids (C1-3). By comparison, the emulsion formulations with longer carbon chain acids (C4-12) showed little to no MLR at 22°C, but achieved >6.5 MLR at 45°C. Based upon the decreased Salmonella membrane viscosity and the increased antimicrobial efficacy of C4-12 W/O emulsions with increasing temperature, we propose that heating can make the membrane more fluid which may allow the longer carbon chain acids (C4-12) to permeate or disrupt membrane structures.

Oil-Based Sanitization in Low-Moisture Environments: Delivery of Acetic Acid with Water-in-Oil Emulsions.

Contamination with Salmonella spp. and Listeria monocytogenes is concerning across low-moisture food (LMF)-processing environments due to the pronounced survival of these organisms under dry conditions. This study treated desiccated bacteria with acetic acid delivered by oil with and without water-in-oil (W/O) emulsion. The influences of cellular desiccation, emulsion water concentration, water activity (aw), and treatment temperature were investigated. Acetic acid dissolved in oil (i.e., acidified oil) showed low levels of antimicrobial efficacy. After treatment with acidified oil (200 mM acetic acid at 22°C for 30 min), Salmonella enterica serovar Enteritidis phage type 30 cells desiccated to 75% equilibrium relative humidity (ERH) and 33% ERH were reduced by 0.69 and 0.05 log CFU/coupon, respectively. The dispersion of a low level of water (≥0.3%, vol/vol) within the acidified oil with the surfactant (i.e., acidified W/O emulsion) significantly enhanced the antimicrobial efficacy. After treatment with the acidified W/O emulsion (200 mM acetic acid at 22°C for 20 min), desiccated Salmonella (4-strain cocktail) and L. monocytogenes (3-strain cocktail) cells were reduced by >6.52 log most probable number (MPN)/coupon, regardless of the desiccation levels. Increased efficacy was observed with temperature elevation. Reduced efficacy was observed when glycerol was added to the aqueous phase of the emulsion to decrease the solution aw, indicating that the enhanced efficacy of the acidified W/O emulsion was associated with differential osmotic pressure. The antimicrobial mechanism may be due to the membrane disruption induced by acetic acid, in combination with the hypoosmotic stress provided by W/O emulsion, creating cellular lysis, as illustrated by electron micrographs. IMPORTANCE Aqueous-based cleaning and sanitation are undesirable in processing facilities that manufacture low-moisture foods such as peanut butter and chocolate. Alcohol-based sanitization is advantageous because it leaves no residue on the contact surface but requires the processing facility to close temporarily due to flammability. At >6.52 log kill of desiccated Salmonella and Listeria monocytogenes cells, the developed oil-based formulation has the potential to be an effective dry sanitation method.

Synergistic effect of high hydrostatic pressure, allyl isothiocyanate, and acetic acid on the inactivation and survival of pathogenic Escherichia coli in ground chicken.

Meat and poultry are prone to contamination with foodborne pathogens sourced from the livestock or introduced from the processing environments. In this study, for retention of meat quality while assuring microbial food safety, mild levels of high hydrostatic pressure were hurdled with food-grade additives (i.e., allyl isothiocyanate [AITC] and acetic acid [AA], functioned as antimicrobials) to inactivate pathogenic Escherichia coli in ground chicken. The reductions of Shiga toxin-producing E. coli (STEC) O157:H7 and uropathogenic E. coli (UPEC) were described as a function of high hydrostatic pressure (200-350 MPa), process-holding time (10-25 min), AITC concentration (0.05-0.20% w/w), and AA concentration (0.10--0.30% w/w) using a full factorial design. The antimicrobials had little influence on bacterial inactivation without high pressure. Without the antimicrobials, a high-pressure treatment at 300 MPa and 4°C for 15 min reduced E. coli O157:H7 and UPEC by 1.52 and 2.52 log, respectively. A 5-log reduction was achieved when AITC and AA were combined with high pressure, indicating a synergistic effect. The survivors were further reduced to below the detection limit of 1 log CFU/g after subsequent storage tests at 4 and 10°C for 10 days. The STEC O157:H7 was found slightly more resistant than UPEC in our test matrix. The developed models showed good fits with experimental data (R2  > 0.95 for linear models; Pr > F (<0.0001) for dimensionless nonlinear models); which may help processors find/optimize the processing parameters to achieve target foodborne pathogens reduction for food safety requirement. PRACTICAL APPLICATION: Models were developed to predict the inactivation of pathogenic Escherichia coli in ground chicken by high-pressure processing (HPP) in combination with natural antimicrobial compounds. These models can be used to estimate/determine the HPP operation parameters and antimicrobial usage levels (i.e., allyl isothiocyanate and acetic acid) needed to achieve a specific microbial log reduction within the selected factor ranges. The operation parameters and clean-label ingredients are of interest in the food industry, which may benefit from the application of the models in achieving microbial safety, process optimization, and operation cost reduction.

Survival Evaluation of Salmonella and Listeria monocytogenes on Selective and Nonselective Media in Ground Chicken Meat Subjected to High Hydrostatic Pressure and Carvacrol.

High pressure processing (HPP) and treatment with the essential oil extract carvacrol had synergistic inactivation effects on Salmonella and Listeria monocytogenes in fresh ground chicken meat. Seven days after HPP treatment at 350 MPa for 10 min, Salmonella treated with 0.75% carvacrol was reduced to below the detection limit (1 log CFU/g) at 4°C and was reduced by ca. 6 log CFU at 10°C. L. monocytogenes was more sensitive to these imposed stressors, remaining below the detection limit during storage at both 4 and 10°C after HPP treatment at 350 MPa for 10 min following treatment with 0.45% carvacrol. However, pressure-injured bacterial cells may recover and lead to an overestimation of process lethality when a selective medium is used without proper justification. For HPP-stressed Salmonella, a 1- to 2-log difference was found between viable counts on xylose lysine Tergitol 4 agar and aerobic plate counts, but no significant difference was found for HPP-stressed L. monocytogenes between polymyxin-acriflavine-lithium chloride-ceftazidime-esculin-mannitol (PALCAM) agar and aerobic plate counts. HPP-induced bacterial injury and its recovery have been investigated by comparing selective and nonselective agar plate counts; however, few investigations have addressed this issue in the presence of essential oil extracts, taking into account the effect of high pressure and natural antimicrobial compounds (e.g., carvacrol) on bacterial survival in various growth media. Use of selective media may overestimate the efficacy of bacterial inactivation in food processing evaluation and validation studies, and the effects of various media should be systematically investigated.

Recycling of harbor sediment as lightweight aggregate.

Sediment sampled from Taichung Harbor was mixed with local reservoir sediment at different weight ratios to prepare lightweight aggregate at 1050, 1100, and 1150 degrees C. A pressure of 3000 or 5000 psi was used to shape the powder mixtures into pellets before the heating processes. The results indicate that the leaching levels of trace metals from the lightweight aggregate samples are considerably reduced to levels less than Taiwan Environmental Protection Administration regulatory limits. Increasing final process temperature tends to reduce the bulk density and crushing intensity of lightweight aggregate with a concomitant increase in water sorption capability. Lightweight aggregate with the lowest bulk density, 0.49 g cm(-3) for the 5000 psi sample, was obtained with the heating process to 1150 degrees C. Based on the X-ray absorption near edge structure results, FeSO(4) decomposition with a concomitant release of SO(x) (x = 2,3) is suggested to play an important role for the bloating process in present study.