Modeling the release of antimicrobial agents (thymol and carvacrol) from two different encapsulation materials

Food Science and Biotechnology - The release of microencapsulated natural antimicrobial (AM) agents (thymol and carvacrol) from two encapsulating matrixes [maltodextrin (MD) and soy protein (SP)]...     

Inactivation of Escherichia coli O157:H7 on stainless steel upon exposure to Paenibacillus polymyxa biofilms

We investigated the potential use of biofilm formed by a competitive-exclusion (CE) microorganism to inactivate Escherichia coli O157:H7 on a stainless steel surface. Five microorganisms showing inhibitory activities against E. coli O157:H7 were isolated from vegetable seeds and sprouts. The microorganism with the greatest antimicrobial activity was identified as Paenibacillus polymyxa (strain T5). In tryptic soy broth (TSB), strain T5 reached a higher population at 25 °C than at 12 or 37 °C without losing inhibitory activity against E. coli O157:H7. When P. polymyxa (6 log CFU/mL) was co-cultured with E. coli O157:H7 (2, 3, 4, or 5 log CFU/mL) in TSB at 25 °C, the number of E. coli O157:H7 decreased significantly within 24 h. P. polymyxa formed a biofilm on stainless steel coupons (SSCs) in TSB at 25 °C within 24 h, and cells in biofilms, compared to attached cells without biofilm formation, showed significantly increased resistance to a dry environment (43% relative humidity [RH]). With the exception of an inoculum of 4 log CFU/coupon at 100% RH, upon exposure to biofilm formed by P. polymyxa on SSCs, populations of E. coli O157:H7 (2, 4, or 6 log CFU/coupon) were significantly reduced within 48 h. Most notably, when E. coli O157:H7 at 2 log CFU/coupon was applied to SSCs on which P. polymyxa biofilm had formed, it was inactivated within 1 h, regardless of RH. These results will be useful when developing strategies using biofilms produced by competitive exclusion microorganisms to inactivate foodborne pathogens in food processing environments.     

Essential oil composition,antimicrobial and antioxidant properties of Mosla chinensis Maxim

The essential oil of Mosla chinensis Maxim was analysed by gas chromatography/mass spectrometry, and its main components are carvacrol (57.08%), p-cymene (13.61%), thymol acetate (12.68%), thymol (6.67%), and γ-terpinene (2.46%). The essential oil exhibited great potential antimicrobial activity against all eight bacterial and nine fungal strains. Antioxidant activity was also tested, the essential oil showing significantly higher antioxidant activity than that of the methanol extract. In addition, the amounts of total phenol components in the plant methanol extract (47.3 ± 0.4 μg/mg) and the oil (80.7 ± 0.5 μg/mg) were determined. The results presented here indicate that the essential oil of M. chinensis has antimicrobial and antioxidant properties, and is therefore a potential source of antimicrobial and antioxidant agents for the food and pharmaceutical industries.     

Sucrose monolaurate improves the efficacy of sodium hypochlorite against Escherichia coli O157:H7 on spinach

It is well-recognized that chlorine has limited efficacy when applied to inactivate pathogens on fresh produce. One of the many factors limiting efficacy is the high interfacial tension of chlorine-based sanitizers that limits the access of chlorine to the microorganisms. In this work, we investigated the efficacy of sodium hypochlorite (200 ppm, pH 6.0) at 4 and 20 °C against Escherichia coli O157:H7 inoculated on baby spinach leaves as affected by the surfactant sucrose monolaurate (SML) at below (100 ppm), above (250 ppm), and well above (10,000 ppm) the critical micelle concentration (CMC) of ~ 200 ppm at 20 °C. The surfactant-containing chlorine treatments were compared to those with buffer only, surfactant only, and chlorine only. Significantly improved inactivation, as evidenced by survival of E. coli O157:H7 was achieved when 250 or 10,000 ppm SML was added with chlorine. This is attributed to the reduction of interfacial tension between the sanitizing solutions and spinach surface. Treatments at 20 °C resulted in greater mean inactivation than those at 4 °C but the difference was not significant. Comparisons of SML concentrations in treatment solutions before and after sanitization showed that SML decreased more at a lower temperature and when chlorine was present, resulting from adsorption of SML onto spinach matrix. Our work illustrates the importance of using surfactants at concentrations above the CMC to enhance the efficacy of chlorine sanitization.     

Antimicrobial activity of Satureja montana L. essential oil against Clostridium perfringens type A inoculated in mortadella-type sausages formulated with different levels of sodium nitrite

This research evaluated the antimicrobial effect of the winter savory (Satureja montana L.) essential oil (EO) against Clostridium perfringens type A (ATCC 3624) inoculated in mortadella-type sausages formulated with different levels of sodium nitrite (NaNO₂: 0 ppm, 100 ppm and 200 ppm) in addition to EO at concentrations of 0.0%, 0.78%, 1.56% and 3.125% stored at 25 °C for 30 days. The EO extracted by hydrodistillation and analyzed by gas chromatography-mass spectrometry (CG-MS) was tested in vitro using an agar well diffusion method for determination of minimum inhibitory concentration (MIC) on C. perfringens. According to compositional analysis of the winter savory EO, 26 chemical compounds were identified, and the major constituents were thymol (28.99%), p-cymene (12.00%), linalool (11.00%) and carvacrol (10.71%). The results obtained showed that EO applied at a concentration of 1.56%, which was defined as the MIC, exhibited antimicrobial activity against C. perfringens in the in vitro assays, and the transmission electron microscopy (TEM) revealed structural damage and cell lysis of C. perfringens caused by EO treatment. A synergistic effect between NaNO₂ and EO was observed. In mortadella-type sausages formulated with 100 ppm of NaNO₂ and EO at all concentrations tested, the population of target microorganisms was reduced (p ≤ 0.05) compared to control samples during all storage period. This data suggests the potential combined use of savory EO and minimal amounts of the synthetic additive, NaNO₂ to control C. perfringens in mortadella, which goes according to current market trends, where consumers are requesting natural products.     

Efficacy of adding detergents to sanitizer solutions for inactivation of Escherichia coli O157:H7 on Romaine lettuce

Numerous Escherichia coli O157:H7 outbreaks have been linked to consumption of fresh lettuce. The development of effective and easily implemented wash treatment could reduce such incidents. The purpose of this study was to evaluate the addition of food-grade detergents to sanitizer solutions for inactivation of E. coli O157:H7 on Romaine lettuce. Freshly-cut leaves of Romaine lettuce were dip-inoculated to achieve a final cell concentration of 7.8 ± 0.2 log CFU/g, air-dried for 2 h, and stored overnight at 4 °C. Leaves were then washed for 2 min in an experimental short chain fatty acid formulation (SCFA) or in one of the following solutions with or without 0.2% dodecylbenzenesulfonic acid or 0.2% sodium 2-ethyl hexyl sulfate: 1) deionized water; 2) 100 ppm chlorine dioxide; 3) 100 ppm chlorine; and 4) 200 ppm chlorine. Following wash treatment, samples were blended in neutralizing buffer (1:3) and surface plated on the selective media CT-SMAC. The efficacy of wash treatments, with or without the detergents, in inactivating E. coli O157:H7 cells on lettuce leaves were not significantly different. The most effective wash solution was SCFA, which was capable of reducing E. coli O157:H7 populations by more than 5 log CFU/g. The rest of the wash treatments resulted in a population reduction of less than 1 log CFU/g. The effectiveness of SCFA surpasses that of other sanitizer treatments tested in this study and requires further research to optimize treatments to preserve lettuce quality. Conventional detergents did not enhance the efficacy of any of the wash treatments tested during this study.     

Defining treatment conditions for pulsed electric field pasteurization of apple juice

The influence of temperature and the presence of N^α-lauroyl ethylester (ethyl lauroyl arginate, LAE) on the inactivation caused by continuous pulsed electric field treatments (PEF) in Escherichia coli O157:H7 suspended in apple juice have been investigated to define treatment conditions applicable at industrial scale that promote an equivalent safety level when compared with thermal processing. In the range of experimental conditions investigated (outlet temperature: 20-40 °C, electric field strength: 20-30 kV, treatment time: 5-125 μs) at outlet temperatures equal or lower than 55 ± 1 °C, the inactivation of E. coli O157:H7 treated in apple juice ranged from 0.4 to 3.6 Log₁₀ cycles reduction and treated in apple juice supplemented with LAE (50 ppm) ranged from 0.9 to 6.7 Log₁₀ cycles reduction.An empirical mathematical model was developed to estimate the treatment time and total specific energy input to obtain 5 Log₁₀ cycles reduction in the population of E. coli O157:H7 suspended in apple juice supplemented with 50 ppm of LAE at different electric field strengths and inlet temperatures. Treatment conditions established for E. coli O157:H7 were validated with other PEF resistant Gram-positive (Listeria monocytogenes, and Staphylococcus aureus) and Gram-negative (Salmonella enterica serovar Typhimurium) strains. When the treatment was applied to the apple juice, a treatment of 25 kV/cm for 63 μs corresponding with an outlet temperature of 65 °C and input energy of 125 kJ/kg was required to achieve more than 5 Log₁₀ cycles in the four strains investigated. The addition of LAE reduced the treatment time required to obtain an equivalent inactivation (> 5 Log₁₀ cycles) in the four microorganisms to 38.4 μs, the outlet temperature to 55 °C, and the input energy to 83.2 kJ/kg.     

Efficacy of polyethylene-based antimicrobial films containing principal constituents of basil

The feasibility of low-density polyethylene (LDPE)-based films containing linalool or methylchavicol as antimicrobial (AM) packages to retard microbial growth on food surfaces was investigated. The AM LDPE-based films were tested for inhibition against selected microorganisms. Both compounds retained their AM activity, after an extrusion film-blowing process, against Escherichia coli in solid medium. Cheddar cheese was wrapped with the AM films and the packaged cheese samples were stored at 4 °C. The changes in the mesophilic aerobic bacteria and coliform, as well as yeast and mould counts were monitored. In addition, cheese samples inoculated with E. coli or Listeria innocua were wrapped with the AM films, stored at refrigerated (4 °C) or at abuse (12 °C) temperatures and the count of these microorganisms was monitored as a function of time. The results showed an inhibitory effect of these AM films against microbial growth in naturally contaminated cheese and in inoculated samples. The effect on suppression of E. coli and L. innocua growth was more pronounced at the abuse temperature. Methylchavicol-LDPE-based film exhibited a higher efficacy of inhibition than that of linalool-LDPE-based film. In addition, a sensory evaluation was performed with regards to possible taint in the flavour of the cheese. Taint in flavour as affected by linalool or methylchavicol was not significantly detectable by the panelists at the end of the storage period of 6 weeks. This study shows the potential use of polymeric films containing the principal constituents of basil as the AM components for enhancing quality and safety of cheeses.     

Antimicrobial activity of binary combinations of natural and synthetic phenolic antioxidants against Enterococcus faecalis

This study investigated the antimicrobial activity of 3 natural (thymol, carvacrol, and gallic acid) and 2 synthetic [butylated hydroxyanisole (BHA) and octyl gallate] phenolic compounds, individually and in binary combinations, on 4 dairy isolates of Enterococcus faecalis with different virulence factors (β-hemolytic, gelatinase, or trypsin activities; acquired resistance to erythromycin or tetracycline; and natural resistance to gentamicin). A checkerboard technique and a microdilution standardized method were used. All compounds individually tested exhibited antimicrobial activity against E. faecalis, with minimal inhibitory concentrations (MIC) ranging from 30 μg/mL (octyl gallate) to 3,150 μg/mL (gallic acid), although no significant differences were detected among strains to each phenolic compound. Carvacrol in combination with thymol or gallic acid, and gallic acid combined with octyl gallate showed partial synergistic inhibition of all E. faecalis strains. The most effective combinations were thymol + carvacrol and gallic acid + octyl gallate, as the MIC for each of these compounds was reduced by 67 to 75% compared with their respective individual MIC. These results highlight the possibility of using combinations of these phenolic compounds to inhibit the growth of potential virulent or spoilage E. faecalis strains by reducing the total amount of additives used in dairy foods.     

Application of caffeine, 1,3,7-trimethylxanthine,to control Escherichia coli O157:H7

The objective of this research was to determine the effectiveness of caffeine on inactivation of Escherichia coli O157:H7 in brain heart infusion (BHI) broth. Overnight samples of five E. coli O157:H7 strains of (E0019, F4546, H1730, 944 and Cider) were used in this study. These strains were individually inoculated at an initial inoculum level of 2 log CFU/ml into BHI broth containing caffeine at different concentrations (0.00%, 0.25%, 0.50%, 0.75%, 1.00%, 1.25%, 1.50%, 1.75%, and 2.00%). Samples were then incubated at 37 °C for 24 h. Bacterial growth was monitored at different time intervals by measuring turbidity at 610 nm using a spectrophotometer. Results revealed that the addition of caffeine inhibited the growth of E. coli O157:H7. Significant growth inhibition was observed with concentration levels of 0.50% and higher. These results indicate that caffeine has potential as an antimicrobial agent for the treatment of E. coli O157:H7 infection and should be investigated further as a food additive to increase biosafety of consumable food products.     

Antimicrobial activity of some plant extracts and essential oils against foodborne pathogens in vitro and on the fate of inoculated pathogens in chocolate

The efficacy of commercially available plant extracts and essential oils used extensively as flavour ingredients in confectionery products were used as antimicrobials in laboratory media against the following microorganisms: Escherichia coli O157:H7, Salmonella Enteritidis, Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus. Using the disc diffusion method, inhibition zones in diameter >20 mm were observed by adding 10 μl of each antimicrobial substance on the following microorganisms: lemon flavour applied on E. coli O157:H7, lemongrass essences against S. aureus, plum using a B. cereus strain and strawberry flavour using a L. monocytogenes strain. E. coli O157:H7 strains were the most susceptible microorganisms inhibited by 18 extracts, followed by S. Typhimurium and S. aureus which were inhibited by 17 extracts. Lemon flavour, lemongrass essences, pineapple and strawberry flavour inhibited the foodborne pathogens at the lowest concentration (5 ml/100 ml). Plant extracts and essential oils with potent antimicrobial activities were tested in chocolate held at different temperatures (7 and 20 °C) in dry or humidified environment, which resulted in different a_w values of the product (i.e. 0.340, 0.450, and 0.822), in order to determine their efficacy on the fate of the inoculated pathogens. The most inhibitory action was observed by lemon flavour applied on chocolate inoculated with E. coli cocktail culture after storage at 20 °C for 9 days. Plant extracts tested on chocolate show an enhanced inhibitory effect during storage at 20 °C indicating that their application may provide protection in case of storage at the above temperature or even higher.     

Escherichia coli O157:H7 survival,biofilm formation and acid tolerance under simulated slaughter plant moist and dry conditions

Microorganisms persisting in slaughter plant environments may develop acid resistance and be translocated to other environmental surfaces or products. The objective of this study was to evaluate the potential of Escherichia coli O157:H7 to form biofilms and maintain acid resistance, under different culture habituation scenarios, on stainless steel coupons (2 × 5 × 0.08 cm), in the presence of beef carcass decontamination runoff fluids (washings). Coupons were stored in test tubes with unsterilized water washings (WW; pH 6.94) or lactic acid washings (LAW; pH 4.98), which were inoculated with E. coli O157:H7 (10³–10⁴ CFU/ml) and incubated at 15 (24 or 48 h) or 35 °C (7 or 24 h), simulating different habituation scenarios on sites of a slaughter plant, including sanitation and overnight drying, during consecutive operational shifts. Acid resistance (AR) of planktonic and detached E. coli O157:H7 cells was assessed in tryptic soy broth adjusted to pH 3.5 with lactic acid. The highest pre-drying attachment and AR of E. coli O157:H7 were observed after 24 h at 35 °C and 48 h at 15 °C. Drying reduced (P < 0.05) recovery of attached E. coli O157:H7 cells; however, exposure of dried coupons to uninoculated washings allowed recovery of attached E. coli O157:H7, which restored AR, especially under conditions that favored post-drying growth. Exposure of attached cells to 50 ppm PAA for 45 s before drying, as well as habituation in LAW, reduced the recovery and AR of E. coli O157:H7. Therefore, incomplete removal of biofilms may result in cells of increased AR, especially in sites within a slaughter plant, in which liquid meat wastes may remain for long periods of time.     

The effect of diets supplemented with thyme essential oils and rosemary extract on the deterioration of farmed gilthead seabream (Sparus aurata) during storage on ice

The effect on quality were assessed for gilthead seabream fed five different diets: control (basal diet); BHT (basal diet with 200 mg kg⁻¹ of butylated hydroxytoluene); rosemary (basal diet with 600 mg kg⁻¹ of rosemary extract - Rosmarinus officinalis); carvacrol (basal diet with 500 mg kg⁻¹ of essential oil of Thymbra capitata, carvacrol chemotype); and thymol (basal diet with 500 mg g⁻¹ of essential oil of Thymus zygis, subspecies gracilis, thymol chemotype). After 18 weeks of experimentation, the animals were stored on ice at 4 °C for 0, 7, 14, and 21 days. Physical-chemical, microbiological and sensory analyses were carried out at each sampling point to determine the degree of deterioration in the gilthead seabream. Lower indices of oxidation were observed in animals who were administered feeds supplemented with BHT, carvacrol and (to a lesser degree) rosemary. Lower bacteria counts were observed for the BHT and thymol groups, in addition to a slower deterioration in terms of sensory perception. Accordingly, the addition of natural antioxidants to the diet may have an added effect on fish quality, delaying post mortem deterioration.     

Assessment of Escherichia coli O157:H7-specific bacteriophages e11/2 and e4/1c in model broth and hide environments

The efficacy of bacteriophages e11/2 and e4/1c as potential biocontrol agents for Escherichia coli O157:H7 in food applications was assessed under conditions relevant to the food chain environment. The stability of each phage was determined following exposure to varying environmental conditions (pH, temperature, water activity, and sodium chloride) and the ability of each phage to infect and reduce E. coli O157:H7 numbers under selected conditions was also examined. Both e11/2 and e4/1c significantly (p < 0.05) reduced numbers of E. coli O157:H7 when exposed to pH values ranging from pH > 4 to pH 9, temperatures from 4 °C to 37 °C, water activity values of 0.87 or 0.91 to 1.00 and NaCl concentrations of 1% to 2.5%. Subsequently, a cocktail of both phages was used (e11/2 and e4/1c) to assess reduction of E. coli O157:H7 on cattle hide pieces. This involved inoculating pieces of hide (20 × 20 cm) with E. coli O157:H7 (approximately 10⁶ cfu/cm²) which were subsequently treated with either a suspension of a phage cocktail, consisting of e11/2 and e4/1c (multiplicity of infection of 1000 and 10,000, respectively) or water or not treated. Two different investigations were carried out; immediately or 1 h after treatment application was performed in different experiments. Swab samples taken immediately after phage treatment showed no significant (p > 0.05) reduction of E. coli O157:H7 numbers compared to the water treated or untreated samples. However, an extended exposure time of 1 h following phage application revealed a significant reduction (p < 0.05) (1.5 log₁₀ cfu/cm² reduction) in E. coli O157:H7 numbers compared to the numbers recovered on samples treated with water only. These findings demonstrate the potential use of e11/2 and e4/1c phages as a biocontrol agent for E. coli O157:H7 within various stages of the food chain, including on cattle hide.     

Detection of E. coli O157:H7 in raw ground beef by Pathatrix™ immunomagnetic-separation, real-time PCR and cultural methods

Detection of Escherichia coli O157:H7 by conventional cultural methods can be difficult in food matrices with a high background flora such as raw ground beef. Raw ground beef samples, artificially contaminated separately with five strains of E. coli O157:H7 at low (~ 0.2 cfu/g) and high (~ 2 cfu/g) levels, were enriched by two enrichment protocols; buffered peptone water (BPW) at 37 °C for 5 h and 24 h and modified buffered peptone water with pyruvate (mBPWp) for 5 h at 37 °C followed by adding selective agents and incubating at 42 °C to 24 h. Detection of added E. coli O157:H7 by real-time PCR (RTiPCR) and recovery on isolation agars was performed before and after PATHATRIX™ immunomagnetic separation (IMS). RTiPCR detection and cultural recovery of inoculated E. coli O157:H7 after 5 h enrichment were poor at 0.21-0.24 cfu/g. The addition of IMS after 5 h enrichment did not improve RTiPCR detection but markedly improved recovery by culturing. By extending enrichment to 24 h, RTiPCR detection improved to 76% for either enrichment protocol without IMS. When 24 h enrichment was followed by IMS, RTiPCR detection was also further improved. Cultural recovery after 24 h enrichment was 56% and 84% without IMS and 100% and 92% after IMS for BPW and mBPWp respectively. Extended enrichment to 24 h followed by IMS was found to be sensitive and reliable for detection and cultural recovery from raw ground beef using either enrichment method.     

Effects of cooking methods and chemical tenderizers on survival of Escherichia coli O157:H7 in ground beef patties

This study evaluated chemical tenderizers and cooking methods to inactivate Escherichia coli O157:H7 in ground beef patties (model system for non-intact beef). Ground beef was inoculated with E. coli O157:H7 and mixed with (i) nothing (control), (ii) calcium chloride (CC) and flavoring agents (FA), (iii) CC, FA, and acetic acid (AA), (iv) sodium chloride (SC), sodium tripolyphosphate (ST), and potassium lactate (PL), and (v) the combination of SC, ST, PL, and AA. Patties were stored in aerobic or vacuum bags at − 20, 4, and 12 °C. Samples were grilled, broiled, or pan-fried to 60 or 65 °C. Total bacterial and E. coli O157:H7 populations remained unchanged during storage. Broiling was more effective in reducing E. coli O157:H7 than grilling and pan-frying, and acidified tenderizers reduced E. coli O157:H7 more than non-acidified tenderizers in broiling. Higher reductions were observed at 65 °C than 60 °C in broiled and grilled samples. These results indicate that acidified tenderizers and broiling may be useful in non-intact beef safety.     

GC/MS analysis and in vitro antioxidant activity of essential oil and methanol extracts of Thymus caramanicus Jalas and its main constituent carvacrol

Chemical composition of the essential oil, antioxidant activity (DPPH and β-carotene/linoleic acid assays), and total phenolic content (Folin–Ciocalteu assay) of aerial parts of Thymus caramanicus were determined. The highest radical-scavenging activity (DPPH test) was shown by the polar subfraction of the methanol extract (IC₅₀ = 43.0 μg/ml) which was also higher than that of butylated hydroxytoluene (BHT, IC₅₀ = 19.7 μg/ml). However, it was the nonpolar subfraction of the methanol extract that showed the highest inhibition (84.4%), as assessed by the β-carotene/linoleic acid assay, which was only slightly lower than that shown by BHT (93.3%). The antioxidant activities of the essential oil main component (carvacrol) were also evaluated for comparison. Total phenolic content of the polar subfraction, as gallic acid equivalents, was 124.3 μg/mg. Essential oil extracted from the aerial parts by hydrodistillation was analysed by GC and GC/MS. Fifteen constituents, representing 99.3% of the oil, were identified, of which the major ones, carvacrol (85.9%), thymol (3.3%), p-cymene (3.2%), γ-terpinene (1.8%) and borneol (1.3%), accounted for 95.6% of the oil.     

Microbiological and organoleptic characteristics of beef trim and ground beef treated with acetic acid, lactic acid, acidified sodium chlorite, or sterile water in a simulated commercial processing environment to reduce Escherichia coli O157:H7 and Salmonella

The objective of this study was to validate the effectiveness of acetic and lactic acids (2% and 5%), acidified sodium chlorite (1000 ppm), and sterile water in reducing Escherichia coli O157:H7 and Salmonella Typhimurium in inoculated beef trim in a simulated processing environment. Samples were collected to assess microbial characteristics at three processing points. Results from this study indicate that all treatments, including sterile water, reduced pathogen concentrations (P < 0.05) of both E. coli O157:H7 and Salmonella Typhimurium in ground beef up to 0.5 and 0.6 log by 24 h, respectively. In some cases, there were no significant differences between the antimicrobial treatments and the sterile water using this application method. Triangle sensory test results of non-inoculated beef indicated there were no differences (P < 0.05) in the means of correct responses between controls or antimicrobial treatments at 6 or 24 h. While interventions are important for beef trim, use of the interventions must be validated under industry conditions to ensure proper effectiveness.     

Inactivation of different strains of Escherichia coli O157:H7 in various apple ciders treated with dimethyl dicarbonate (DMDC) and sulfur dioxide (SO2) as an alternative method

Escherichia coli has been identified as the causative agent in numerous foodborne illness outbreaks associated with the consumption of fresh apple cider. Apple cider has a pH which is normally below 4.0 and would not be considered a medium capable of supporting the growth of foodborne pathogens. The association of unpasteurized apple cider with foodborne illness due to E. coli O157:H7 has however, led to increased interest in potential alternative methods to produce pathogen free cider. Apple cider was prepared from eight different apple cultivars, inoculated with approximately 10⁶–10⁷ CFU of three strains of E. coli O157:H7 per ml (933, ATCC 43889, and ATCC 43895) and tested to determine the effectiveness of sulfur dioxide (SO₂) and dimethyl dicarbonate (DMDC). Bacterial populations for treated and untreated samples were then enumerated by using non-selective media. Eight different ciders were treated with DMDC (125 and 250 ppm) and SO₂ (25, 50, 75, 100 ppm). Greater than a 5-log reduction was achieved at room temperature with 250 ppm of DMDC and 50 ppm of SO₂ after the incubation time of 6 h and 24 h, respectively. Addition of DMDC and/or SO₂ may offer an inexpensive alternative to thermal pasteurization for the production of safe apple cider for small apple cider producers.     

Enhancing the thermal destruction of Escherichia coli O157:H7 in ground beef patties by trans-cinnamaldehyde

The effect of trans-cinnamaldehyde (TC) on the inactivation of Escherichia coli O157:H7 in undercooked ground beef patties was investigated. A five-strain mixture of E. coli O157:H7 was inoculated into ground beef (7.0 log CFU/g), followed by addition of TC (0, 0.15, and 0.3%). The meat was formed into patties and stored at 4 °C for 5 days or at −18 °C for 7 days. The patties were cooked to an internal temperature of 60 or 65 °C, and E. coli O157:H7 was enumerated. The numbers of E. coli O157:H7 did not decline during storage of patties. However, cooking of patties containing TC significantly reduced (P < 0.05) E. coli O157:H7 counts, by >5.0 log CFU/g, relative to the reduction in controls cooked to the same temperatures. The D-values at 60 and 65 °C of E. coli O157:H7 in TC-treated patties (1.85 and 0.08 min, respectively) were significantly lower (P < 0.05) than the corresponding D-values for the organism in control patties (2.70 and 0.29 min, respectively). TC-treated patties were more color stable and showed significantly lower lipid oxidation (P < 0.05) than control samples. TC enhanced the heat sensitivity of E. coli O157:H7 and could potentially be used as an antimicrobial for ensuring pathogen inactivation in undercooked patties. However detailed sensory studies will be necessary to determine the acceptability to consumers of TC in ground beef patties.