Effect of Radio Frequency Heating on Nalidixic Acid-Adapted Shiga Toxin-Producing and Non-pathogenic <Emphasis Type="Italic">Escherichia coli</Emphasis> Strains in Buffer

This study aimed to evaluate the use of nalidixic acid-adapted strains of three major Shiga toxin-producing Escherichia coli (STEC) and non-pathogenic E. coli for the use of radio frequency (RF) heating using phosphate buffer saline (PBS). The effectiveness of RF was evaluated on cocktails of various STEC serotypes (O157:H7, O26:H11, O111) and non-pathogenic E. coli at different endpoint temperatures (55, 60, and 65 °C). All strains were successfully adapted to nalidixic acid (Nal). In general, the results indicated that Nal-adapted strains were not significantly different from Nal-sensitive strains evaluated either before treatment or at the endpoint temperatures. Nal-adapted strains were, therefore, confirmed to be effective as marker organisms in studies involving the use of RF in buffer. Results also showed that the thermal inactivation of strains was more effective as the treatment temperature increased, particularly at 65 °C, which showed a 6.0 log CFU/ml reduction. The results of the present study serve as a baseline to study RF as a potential intervention technology for non-intact beef products.     

Antibacterial activities of a cinnamon essential oil with cetylpyridinium chloride emulsion against <Emphasis Type="Italic">Escherichia coli</Emphasis> O157:H7 and <Emphasis Type="Italic">Salmonella</Emphasis> Typhimurium in basil leaves

This study examined the antibacterial activities of two different cinnamon essential oil emulsions against Escherichia coli O157:H7 and Salmonella Typhimurium on basil leaves. Cinnamon oil (0.25%) treatments containing CPC (0.05%) exhibited greater effects on the pathogenic bacteria than cinnamon oil treatment without this emulsifier (p < 0.05). Treatment with cinnamon bark and leaf oil emulsions (CBE and CLE, respectively) reduced the populations of E. coli O157:H7 by 4.10 and 5.10 log CFU/g, and S. Typhimurium by 2.71 and 2.82 log CFU/g, respectively. Scanning electron micrographs showed morphological changes in the two pathogenic bacteria following emulsion treatment. In addition, there was no difference in the color or ascorbic acid content of the basil leaves by the emulsion treatment. These results suggest that CBE or CLE treatment can be an effective way to ensure the microbial safety of minimally processed vegetables and a good alternative to chlorination treatment in the fresh produce industry.     

Antimicrobial Activity of Vanillin and Mixtures with Cinnamon and Clove Essential Oils in Controlling <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> and <Emphasis Type="Italic">Escherichia coli</Emphasis> O157:H7 in Milk

The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of vanillin against Listeria monocytogenes Scott A and Escherichia coli O157:H7 was determined in tripticase soy broth (TSB), pH 7 and 6, incubated at 35 °C/24 h and in semi-skim milk incubated at 35 °C/24 h and 7 °C/14 days. The influence of the fat content of milk on the antimicrobial activity of vanillin was tested in whole and skim milk incubated at 7 °C/14 days. Mixtures of clove and cinnamon with vanillin were also evaluated in semi skim milk incubated at 7 °C. The MICs for L. monocytogenes were 3,000 ppm in TSB (pH 7) and 2,800 ppm in TSB (pH 6). The MICs for E. coli O157:H7 were 2,800 ppm in TSB (pH 7) and 2,400 ppm in TSB (pH 6). The MBCs in TSB were 8,000 ppm for L. monocytogenes and 6,000 ppm for E. coli O157:H7. The pH values assayed did not influence significantly the MIC or MBC in TSB. The MICs in semi-skim milk for L. monocytogenes and E. coli O157:H7 were 4,000 and 3,000 ppm at 35 °C/24 h, and 2,500 and 1,000 ppm at 7 °C/7 days, respectively. The MBCs were 20,000 ppm for L. monocytogenes and 11,000 ppm for E. coli O157:H7. High incubation temperatures did not affect the MBC but increased the MIC of the vanillin in milk. This effect could be attributed to the increased membrane fluidity and to the membrane perturbing activity of vanillin at low temperatures. The fat in milk reduced significantly the antimicrobial activity of vanillin, probably due to effect protective of the fat molecules. Mixtures of clove and cinnamon leaves inhibited the growth of L. monocytogenes in a similar way that vanillin alone but had a synergistic effect on the E. coli O157:H7. Mixtures of cinnamon bark and vanillin had always a synergistic effect and some of the combination assayed showed bactericidal activity on the population of L. monocytogenes and E. coli O 157:H7.     

Quantification and Discrimination of Viable and Dead <Emphasis Type="Italic">Escherichia coli</Emphasis> O157:H7 Cells from Chicken Without Enrichment by Ethidium Bromide Monoazide Real-time Loop-Mediated Isothermal Amplification

In this study, a rapid and sensitive method of real-time loop-mediated isothermal amplification (Rti-LAMP) assays was developed for quantification and discrimination of viable and heat-killed E. coli O157:H7 cells treated with low concentration of ethidium bromide monoazide (EMA). Four micrograms per milliliter of EMA was chosen as the optimal concentration which did not inhibit DNA amplification derived from viable cells, but significantly increased the Tt values of dead cells in Rti-LAMP assays. When the DNA from 2.0?×?10³ viable CFU of E. coli O157:H7 was subjected to EMA-Rti-LAMP, the resulting Tt value was 17.73 min. In contrast, the DNA from 2.0?×?10³?CFU completely heat destroyed CFU of E. coli O157:H7 did not yield a positive amplification which Tt value was regarded as 60 min. When the DNA from viable plus heat-killed CFU at a ratio of 5:2995 was subjected to EMA-Rti-LAMP, the resulting Tt value was 23.06 min, which was statistically identical (P?<?0.05) to the Tt value of 24.07 min obtained with the DNA from only 5 viable CFU. The results indicate that even though 3.0?×?10³ dead cells yielded a negative amplification setting the Tt value as 60 min, low numbers of viable cells in the presence of much higher numbers of dead cells still yielded a linear plot for enumerating viable CFU from Tt values. Detection of E. coli O157:H7 derived from contaminated chicken samples, the EMA-Rti-LAMP could notably distinguish viable and heat-killed cells from 5.0?×?10¹ to 1.0?×?10⁴?CFU/g without enrichment.     

Modeling high pressure inactivation of <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Listeria innocua</Emphasis> in whole milk

The survival curves of Escherichia coli and Listeria innocua inactivated by high hydrostatic pressure (HHP) were obtained at room temperature (∼22 °C) and at five pressure levels (400, 450, 500, 550 and 600 MPa) in whole milk. These curves were described by the Weibull model and parameters of this model were reduced from two to one with slight loss of goodness-of-fit. The logarithm of the time constant parameter (δ) of the reduced Weibull model was described with respect to high pressure (P). This approach can be used to define a z _p value analogous to the modeling of the classical D value (increase in pressure that results in one log unit decrease of δ values). The development of accurate survival models under high pressure, as presented here, can be very beneficial to food industry for designing, evaluating and optimizing HHP processes as a new preservation technology.     

Ultraviolet Treatment of Orange Juice to Inactivate <Emphasis Type="Italic">E. coli</Emphasis> O157:H7 as Affected by Native Microflora

The effect of yeast concentration on ultraviolet (UV) inactivation of five strains of Escherichia coli O157:H7 from different sources, inoculated both individually and simultaneously in orange juice, was analyzed and mathematically modeled. The presence of yeast cells in orange juice decreases the performance of UV radiation on E. coli inactivation. UV absorption coefficients in the juice increased with increasing yeast concentration, and higher UV doses were necessary to inactivate bacterial strains. UV intensities of I = 3.00 ± 0.3 mW/cm² and exposure times (t) between 0 and 10 min were applied; radiation doses (energy, E = I × t) ranging between 0 and 2 J/cm² were measured using a UV digital radiometer. All the tested individual strains showed higher resistance to the treatment when UV radiation was applied at 4 °C in comparison to 20 °C. UV inactivation of E. coli O157:H7 individual strain was satisfactory fitted with a first order kinetic model. A linear relationship was found between UV absorptivities and D values (radiation doses required to decrease microbial population by 90%) for each strain. The dose required to reach 5-log reduction for the most unfavorable conditions that is the most UV resistant strain, and maximum background yeast concentration was 2.19 J/cm² at 4 °C (corresponding to 11 min of UV treatment) and 2.09 J/cm² at 20 °C (corresponding to 10.55 min of UV treatment). When a cocktail of strains was inoculated in orange juice, the logistic equation was the best model that fits the experimental results due to the deviation from the log-linear kinetics. The UV resistance between strain cocktail and single strain were mathematically compared. Slopes of the decline curves for strain cocktail at high UV doses were lower than the slopes of the log-linear equation calculated for the individual strains, even for the most resistant one. Therefore, microbial inactivation tests using a cocktail of strains are particularly important to determine the performance of the UV inactivation treatment.     

Biofilm Formation by Shiga Toxin-Producing Escherichia coli O157:H7 and Non-O157 Strains and Their Tolerance to Sanitizers Commonly Used in the Food Processing Environment

Shiga toxin-producing Escherichia coli (STEC) strains are important foodborne pathogens. Among these, E. coli O157:H7 is the most frequently isolated STEC serotype responsible for foodborne diseases. However, the non-O157 serotypes have been associated with serious outbreaks and sporadic diseases as well. It has been shown that various STEC serotypes are capable of forming biofilms on different food or food contact surfaces that, when detached, may lead to cross-contamination. Bacterial cells at biofilm stage also are more tolerant to sanitizers compared with their planktonic counterparts, which makes STEC biofilms a serious food safety concern. In the present study, we evaluated the potency of biofilm formation by a variety of STEC strains from serotypes O157:H7, O26:H11, and O111:H8; we also compared biofilm tolerance with two types of common sanitizers, a quaternary ammonium chloride-based sanitizer and chlorine. Our results demonstrated that biofilm formation by various STEC serotypes on a polystyrene surface was highly strain-dependent, whereas the two non-O157 serotypes showed a higher potency of pellicle formation at air-liquid interfaces on a glass surface compared with serotype O157:H7. Significant reductions of viable biofilm cells were achieved with sanitizer treatments. STEC biofilm tolerance to sanitization was strain-dependent regardless of the serotypes. Curli expression appeared to play a critical role in STEC biofilm formation and tolerance to sanitizers. Our data indicated that multiple factors, including bacterial serotype and strain, surface materials, and other environmental conditions, could significantly affect STEC biofilm formation. The high potential for biofilm formation by various STEC serotypes, especially the strong potency of pellicle formation by the curli-positive non-O157 strains with high sanitization tolerance, might contribute to bacterial colonization on food contact surfaces, which may result in downstream product contamination.     

Differences of Bactericidal Efficacy on <Emphasis Type="Italic">Escherichia coli</Emphasis>, <Emphasis Type="Italic">Staphylococcus aureus</Emphasis>, and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> of Slightly and Strongly Acidic Electrolyzed Water

In the present study, the disinfection efficacy of slightly acidic electrolyzed water (SAEW) and strongly acidic electrolyzed water (AEW) was tested on three bacteria, Escherichia coli, Staphylococcus aureus, and Bacillus subtilis and the disinfection mechanism was discussed. The results showed that SAEW had a stronger antibacterial efficacy against these tested bacteria in comparison with AEW. The results also showed that both SAEW and AEW treatments could damage the cell membrane, which was demonstrated microcosmically by scanning electron microscopy (SEM), thus causing leakages of protein, DNA, RNA, and ATP, resulting in the death of microbes. Moreover, AEW treatment could not cause the degradations of DNA and RNA, and nucleic acids including DNA and RNA are not the target point of its bactericidal efficacy. However, SAEW could maybe cause the degradation of RNA, and RNA may be the target in its antibacterial activity. We suggested that the differences in antibacterial efficacy between SAEW and AEW could be explained by the different impacts on RNA of tested strains.     

Efficiency of pulse pressure treatment for inactivation of <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Listeria innocua</Emphasis> in whole milk

Escherichia coli ATCC 11775 and Listeria innocua ATCC 33090 in whole milk were inactivated by single- and multi-pulsed (up to 10 pulses) high hydrostatic pressure (HHP) treatments. Both bacteria showed similar resistance to single- and multi-pulsed HHP. The efficiency of pulsed pressure treatment depends on the combination of holding time of each pulse and number of pulses. It was observed that multi-pulsed pressure treatment instead of traditional single-pulsed HHP could be used to pasteurize milk at a lower pressure level. Nevertheless, an optimization is necessary between the pulse holding time, number of pulses, and pressure levels to reach the desirable log-reduction of microorganisms compatible with industrial application. This study was partly presented in Joint 21st AIRAPT and 45th EHPRG International Conference on High Pressure Science and Technology held September 17–21, 2007 in Catania (Italy).     

A New Multiplexed Real-Time PCR Assay to Detect <Emphasis Type="Italic">Campylobacter jejuni</Emphasis>, <Emphasis Type="Italic">C. coli</Emphasis>, <Emphasis Type="Italic">C. lari</Emphasis>, and <Emphasis Type="Italic">C. upsaliensis</Emphasis>

A new rapid method based on real-time PCR was developed to detect four thermophilic Campylobacter species (Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis) in food samples. The assay targeted the bipA gene for C. upsaliensis and C. lari, whereas the gene encoding the ATP-binding protein CJE0832 was used to detect C. coli and C. jejuni. These genes were chosen for this assay due to their low variability and mutation rate at a species level. The multiplex PCR showed 100% inclusivity for all 25 thermophilic Campylobacter strains tested and 100% exclusivity for 38 non-targeted strains belonging to closely related species. The newly developed real-time PCR could detect down to 10² genomes/reaction and displayed efficiency above 97% for all species except for C. upsaliensis (90.1%). The method proved to be a reliable tool for food analysis, showing 100% sensitivity, 96% efficiency, and 92.45% specificity when validated against the gold standard method UNE-EN ISO 10272:2006 using 200 diverse food samples (meat, fish, fruits and vegetables, and raw milk). In artificially spiked samples, the detection limit of the method was 10 cfu/g in salad, 5 cfu/g in turkey meat, and 1 cfu/g in the rest of meat samples tested. Consequently, the newly designed molecular tool represents a quick and safe alternative to obtain reliable results concerning the presence/absence of the main thermophilic Campylobacter in any food sample.     

Influence of Exposure Time,Shear Stress,and Surfactants on Detachment of <Emphasis Type="Italic">Escherichia coli</Emphasis> O157:H7 from Fresh Lettuce Leaf Surfaces During Washing Process

Washing and sanitation of fresh produce are critical for detachment and inactivation of pathogenic and spoilage microorganisms. The overall goal of this study was to evaluate the influence of processing conditions such as washing time, shear stress, and a food grade surfactant on the detachment of Escherichia coli O157:H7 from lettuce leaf surfaces during a simulated washing process. A benchtop scale rotating disk washing device was designed to simulate the processing conditions used in commercial washing systems. Numerical simulations were carried out to calculate the flow profile and shear stress near the leaf surface. Removal of E. coli O157:H7 as a function of processing conditions in the rotating disk washing device was evaluated by enumeration of bacterial population using plate counting and changes in bioluminescence intensity and further confirmed by bioluminescence imaging. The results showed that shear stress was necessary to remove attached bacteria from inoculated leaf samples and simple incubation of inoculated leaves with wash water was not effective. Furthermore, detachment of bacteria from inoculated leaf samples increased with increasing shear stress and incubation time. Shear stress values in the range of 0 – 300 mPa were adequate to induce removal of approximately 1.5 log CFU/cm² of bacteria from inoculated samples. The addition of a surfactant increased the removal of bacteria from inoculated leaf samples by 0.5 log CFU/cm². Overall, these results may guide the design of new produce wash systems as well as current industrial practice related to flow conditions and use of surfactants during washing of fresh produce.     

Estrogenic effects of various extracts from <Emphasis Type="Italic">Chamdanggui</Emphasis> (<Emphasis Type="Italic">Angelica gigas</Emphasis> Nakai) and <Emphasis Type="Italic">sogdan</Emphasis> (<Emphasis Type="Italic">Phlomis umbrosa</Emphasis> Turcz)

The various extracts from chamdanggui (Angelica gigas Nakai) and sogdan (Phlomis umbrosa Turcz) were evaluated for estrogenic activity and characterized according to HPLC profile. Chamdanggui and sogdan were individually extracted with 4 solvents (hot water, 70% ethanol, n-butanol, and dichloromethane) of differing polarities. Estrogenic activity was determined by E-screen using an estrogen-dependent MCF-7 BUS cell. Although almost all extracts showed estrogenic effects in a concentrationdependent manner, the hot water extract from chamdanggui (250 μg/mL) had the higher effect (138%). Among 90 fractions using HPLC separation of the hot water extract from chamdanggui, fraction 21 and 28 produced the highest estrogenic effects of 178 and 163% at 10 μg/mL, respectively. The results imply that the hot water extract from chamdanggui could be useful as an alternative hormone replacement therapy.     

Effect of Low-pressure Cold Plasma (LPCP) on the Wettability and the Inactivation of <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Listeria innocua</Emphasis> on Fresh-Cut Apple (<Emphasis Type="Italic">Granny Smith</Emphasis>) Skin

The aim of this study was to investigate the effect of low-pressure cold plasma (LPCP) on the inactivation of Escherichia coli and Listeria innocua on fresh-cut apple skin and its influence on wettability. Cold plasma treatments have shown to be effective to decontaminate foods, but their effect on the wettability has not been well studied. Surface-inoculated apple samples were treated with argon (Ar), nitrogen (N2), oxygen (O₂), and argon-oxygen (Ar-O₂) cold plasma using a commercial LPCP unit. Three different models were used to fit bacterial survival curves after the LPCP treatments. Changes in surface wettability were also determined by measuring the contact angle. The LPCP treatments using Ar, O₂, or Ar-O₂ mixture for 20 min were the most effective to inactivate E. coli with O₂, while the LPCP treatment with N2 for 20 min reduced L. innocua the most for (p?<?0.05). The highest increase in surface wettability was observed in samples treated for 20 min with O₂ and Ar-O₂. Different LPCP treatments have not great effectivity on the inactivation of E. coli and L. innocua on fresh-cut apple surface, but the all treatments changed the surface wettability of apples, making it more hydrophilic. This can be considered as a negative effect of the LPCP treatment because it can facilitate the adhesion and proliferation of re-contaminating microorganisms. More research should be undertaken to explore the use of other gases and complex surfaces.     

Effect of Tannin Extracts on Biofilms and Attachment of <Emphasis Type="Italic">Escherichia coli</Emphasis> on Lettuce Leaves

Lettuce is often involved in foodborne outbreaks caused by pathogenic Escherichia coli. Current control strategies have often proved ineffective to ensure safe food production. For that reason, the present study compared the efficacy of tannin extracts and chlorine treatments on the reduction of E. coli ATCC 25922 adhered to lettuce leaves. E. coli was inoculated artificially on leaf surfaces of fresh crisp lettuce. Effectiveness of water, chlorine (200 mg/L), and three commercial available tannin extracts from Acacia mearnsii De Wild. (tannin AQ (2 %, w/v), tannin SG (1 %, v/v) and tannin SM (1 %, v/v)) treatments was evaluated using the viable plate count method and scanning electron microscopy (SEM). SEM results revealed that bacterial cells are attached as individual cells and in clusters to the leaf surface after 2 h of incubation. Biofilm formation was observed after 24 h of incubation. The tannin SM treatment was able to reduce counts in approximately 2 log CFU/cm² on leaf segments. However, treatment was less effective in the reduction of E. coli counts after 24 h of incubation when compared to 2 h incubation of the same extract. The results suggest that the tannin SM extract diminishes E. coli counts adhered to and under biofilm formation on lettuce leaves and its effect is similar to the use of chlorine solutions.     

Inactivation and kinetic model for the <Emphasis Type="Italic">Escherichia coli</Emphasis> treated by a co-axial pulsed electric field

Inactivation of Escherichia coli CGMCC1.90 inoculated into carrot juice by a co-axial pulsed electric field was investigated and the fitting of its inactivation kinetics was performed by the Hülsheger and Peleg models. The average electric field strength ranged from 5 to 25 kV/cm and the number of pulses was from 207 to 1449 pulses. The level of E. coli inactivation increased with the increment of the electric field strength and the number of pulses. At the same specific energy input level, higher electric field strength caused higher microbial reduction. As the number of pulses increased, the kinetic constants b _E and calculated based on Hülsheger model varied from 0.3116 to 0.3790 and from 4.0565 to 1.6121 kV/cm, obtained by Peleg model from 8.0412 to 2.5676 kV/cm, but the k value changed little from 2.4213 to 2.6624, respectively. The model performance evaluation was assessed by using a series of indices including accuracy factor, bias factor, the sum of the squares of the differences of the natural logarithm of observed and predicted values, root mean square error and correlation coefficient R ² between observed and predicted values. According to these parameters, Hülsheger model fitted better to the inactivation by PEF than Peleg model in the study.     

Analysis of Intracellular pH in <Emphasis Type=Italic>Escherichia coli</Emphasis> O157:H7 to Determine the Effect of Chlorine Dioxide Decontamination

The aim of this study was to investigate the response of Escherichia coli O157:H7 when exposed to different concentrations of sanitation agent chlorine dioxide (ClO₂) by determining intracellular pH (pH_i). For this purpose, fluorescence ratio imaging microscopy was used together with pH-sensitive, ratiometric green fluorescent protein that was introduced in E. coli O157:H7 cells. Along with pH_i, colony counts were determined during the treatment with ClO₂. Results revealed several post-treatment subpopulations with different physiological states, as judged by their pH_i. The fraction of cells with no pH gradient increased, and the colony count decreased as the concentration of ClO₂ increased.     

Relation Between Near-Infrared Spectroscopy and Physicochemical Parameters for Discrimination of Honey Samples from <Emphasis Type="Italic">Jatai weyrauchi</Emphasis> and <Emphasis Type="Italic">Jatai angustula</Emphasis> Bees

This work aims to relate near-infrared (NIR) vibration bands to physicochemical (PC) parameters in order to discriminate honey samples. To the study, 14 Jatai weyrauchi honey samples from Roraima State (northern Brazil) and 15 Jatai angustula honey samples from Paraná State (southern Brazil) were evaluated by analyzing physicochemical parameters (moisture, pH, acidity, formaldehyde index, ashes, electric conductivity, color, hydroxymethylfurfural, protein, reducing sugars, total reducing sugars and sucrose, viscosity, and water activity), besides being submitted to spectral measurements in the near-infrared range from 900 to 1700 nm. By applying ComDim multiblock analysis, it was possible to simultaneously evaluate the results from NIR spectroscopy and the evaluated PC parameters. This tool allowed to highlight the relation between NIR spectra and PC parameters by producing informative graphs showing the relationship among the samples through the scores, the saliences of the blocks, and the loadings of the variables responsible for the similarities observed in the samples scores. Based on the principal component analysis, it was possible to conclude that the PC parameters can be replaced by NIR spectroscopy in the discrimination of honey samples, improving its evaluation regarding on the speed and decreasing the amount of reagents used for this purpose.     

Modeling the Inactivation of <Emphasis Type="Italic">Listeria innocua</Emphasis> and <Emphasis Type="Italic">Escherichia coli</Emphasis> in Fresh-Cut Tomato Treated with Pulsed Light

The effectiveness of pulsed light (PL) treatments to inhibit microorganisms on fresh-cut tomatoes (Lycopersicon esculentum Mill., cv. Daniela) was investigated. Tomato slices inoculated with Escherichia coli or Listeria innocua were exposed to PL treatments (4, 6, or 8 J cm^?2 fluence) and kept cold at 4 °C for 20 days. L. innocua and E. coli counts, gases in the headspace of the containers (O₂ and CO₂), pH, titratable acidity, and soluble solid content were monitored throughout the cold storage. The PL treatments reduced significantly (p < 0.05) initial loads of both microbes. The effect of the PL fluence on the survival number of microoganisms was described by a log-linear model (R ² = 0.849–0.999). At any fixed time within the cold storing, the microbial counts for untreated samples were always higher than those cut tomatoes that had been previously PL-treated. The behavior of L. innocua and E. coli during the storage were well adjusted (R ² > 0.930) by Gompertzian models; the studied microorganisms exhibited different patterns during the storage period. On the other hand, O₂ and CO₂ partial pressures in containers with fresh-cut tomatoes were also significantly affected by PL treatments (p < 0.05). The highest PL fluence caused the greatest changes of O₂ and CO₂ contents. In addition, the application of PL triggered an acceleration of the O₂ consumption during the cold stage. PL treatments might be used to effectively extend the safety of fresh-cut tomatoes over 12 days of storage against E. coli and L. innocua growth.     

Evaluation of Lactic Acid as an Initial and Secondary Subprimal Intervention for Escherichia coli O157:H7, Non-O157 Shiga Toxin-Producing E. coli, and a Nonpathogenic E. coli Surrogate for E. coli O157:H7

Lactic acid can reduce microbial contamination on beef carcass surfaces when used as a food safety intervention, but effectiveness when applied to the surface of chilled beef subprimal sections is not well documented. Studies characterizing bacterial reduction on subprimals after lactic acid treatment would be useful for validations of hazard analysis critical control point (HACCP) systems. The objective of this study was to validate initial use of lactic acid as a subprimal intervention during beef fabrication followed by a secondary application to vacuum-packaged product that was applied at industry operating parameters. Chilled beef subprimal sections (100 cm(2)) were either left uninoculated or were inoculated with 6 log CFU/cm(2) of a 5-strain mixture of Escherichia coli O157:H7, a 12-strain mixture of non-O157 Shiga toxin-producing E. coli (STEC), or a 5-strain mixture of nonpathogenic (biotype I) E. coli that are considered surrogates for E. coli O157:H7. Uninoculated and inoculated subprimal sections received only an initial or an initial and a second "rework" application of lactic acid in a custombuilt spray cabinet at 1 of 16 application parameters. After the initial spray, total inoculum counts were reduced from 6.0 log CFU/cm(2) to 3.6, 4.4, and 4.4 log CFU/cm(2) for the E. coli surrogates, E. coli O157:H7, and non-O157 STEC inoculation groups, respectively. After the second (rework) application, total inoculum counts were 2.6, 3.2, and 3.6 log CFU/cm(2) for the E. coli surrogates, E. coli O157:H7, and non-O157 STEC inoculation groups, respectively. Both the initial and secondary lactic acid treatments effectively reduced counts of pathogenic and nonpathogenic strains of E. coli and natural microflora on beef subprimals. These data will be useful to the meat industry as part of the HACCP validation process.     

A novel process for obtaining phenylpropanoic acid precursor using <Emphasis Type="Italic">Escherichia coli</Emphasis> with a constitutive expression system

Phenylpropanoids are widely used in food supplements, pharmaceuticals, and cosmetics with diverse benefits to human health. Trans-cinnamic acid or p-coumaric acid is usually used as the starting precursor to produce phenylpropanoids. Synthetic bioengineering of microbial cell factories offers a sustainable and flexible alternative method for obtaining these compounds. In this study, a constitutive expression system consisting of Rhodotorula glutinis phenylalanine/tyrosine ammonia lyase was developed to produce a phenylpropanoic acid precursor in Escherichia coli. To improve trans-cinnamic acid and p-coumaric acid production, BioBrick optimization was investigated, causing a 7.2- and 14.2-fold increase in the yield of these compounds, respectively. The optimum strain was capable of de novo producing 78.81 mg/L of trans-cinnamic acid and 34.67 mg/L of p-coumaric acid in a shake flask culture. The work presented here paves the way for the development of a sustainable and economical process for microbial production of a phenylpropanoic acid precursor.