Effect of preinoculation growth media and fat levels on thermal inactivation of a serotype 4b strain of Listeria monocytogenes in frankfurter slurries

Preinoculation growth conditions and fat levels were evaluated for effects on the heat resistance of Listeria monocytogenes strain MFS 102 in formulated frankfurter slurries and on frankfurter surfaces. Comparison of linear inactivation rates (D-values) for cells heated in frankfurter slurry showed that growth conditions were significant (P<0.05) factors affecting subsequent thermal resistance. The average D(60 degrees C)-values for the five preinoculation growth media tested from most resistant to least heat resistant were: tryptic soy broth with 0.6% yeast extract (TSBYE) (2.2 min) and 8.5% fat slurry (2.2 min), followed by 23% fat slurry (1.7 min) and 11% fat slurry (1.7 min), and then TSYBE with quaternary ammonium compounds added (TSBYE+Q) (1 min). The fat level in the frankfurter heating media also had a significant (P<0.05) effect on the thermal death rate of L. monocytogenes. Cells heated in 8.5% fat slurry had a significantly higher (P<0.05) D(60 degrees C)-value (2.2 min) than those heated in 11% fat (1.0 min) and 23% fat slurry (0.9 min). Growth media (TSBYE, 8.5% fat slurry, and TSBYE+Q), and fat level (15% and 20%), however, were not significant factors (P>0.05) affecting thermal inactivation rates on frankfurter surfaces. Heat inactivation rates were consistently higher on frankfurter surfaces compared to similar treatments done in frankfurter slurry. On frankfurter surfaces, a 2.3- to 5.1-log(10) reduction was achieved after 15 min depending on frankfurter surface type. The time necessary to achieve a 3-log(10) reduction using post-processing pasteurization of frankfurters in a hot water-bath at 60 degrees C almost doubled for cells grown in TSBYE and heated in 23% fat frankfurter slurry (19.6 min) versus cells grown and heated in 8.5% fat frankfurter slurry (10.8 min).     

Occurrence of foodborne pathogens and characterization of Staphylococcus aureus in cheese produced on farm-dairies

The objective of this study was to address knowledge gaps identified in an earlier risk assessment of Staphylococcus aureus and raw milk cheese. A survey of fresh and short-time ripened cheeses produced on farm-dairies in Sweden was conducted to investigate the occurrence and levels of S. aureus, Listeria monocytogenes and Escherichia coli, to characterize S. aureus isolates with special emphasis on enterotoxin genes, antibiotic resistance, bio-typing and genetic variation, and to collect information related to production practices. In general, the hygienic quality of farm-dairy cheeses appeared to be of an acceptable microbiological quality, e.g. L. monocytogenes and staphylococcal enterotoxin were not detected in cheese samples. However, E. coli and enterotoxigenic S. aureus were frequently found in raw milk cheeses and sometimes at levels that are of concern, especially in fresh cheese. Interestingly, levels in raw milk fresh cheese were significantly lower when starter cultures were used. Up to five S. aureus colonies per cheese, if possible, were characterized and about 70% of isolates carried one or more enterotoxin genes, most common were sec and sea. The Ovine biotype (73%) was most common among isolates from goat milk cheese and the Human biotype (60%) from cow milk cheese. Of all isolates, 39% showed decreased susceptibility to penicillin, but the proportion of isolates from cows' cheese (66%) compared to isolates from goats' cheese (27%) was significantly higher. S. aureus isolates with different properties were detected in cheese from the same farm and, sometimes even the same cheese. Isolates with the same pulsed-field gel electrophoresis (PFGE)-pattern were detected on geographically distant dairies. This indicates that multiple sources and routes of contamination are important. To improve the safety of these products efforts to raise awareness of the importance of hygiene barriers and raw milk quality as well as improved process control can be suggested, e.g. use of starter cultures and monitoring of fermentation with a pH-meter. For future safety assessments, a better understanding of factors determining toxin production in these cheeses is needed.     

Effects of packaging type and storage temperature on the growth of foodborne pathogens on shredded ‘Romaine’ lettuce

Fresh produce can be a vehicle for the transmission of pathogens capable of causing human illnesses and some of them can grow on fresh-cut vegetables. The survival and growth of Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes inoculated onto shredded lettuce was determined under modified atmosphere packaging conditions, at various storage temperatures. We also monitored changes in pH and gas atmospheres within the packages and the growth of psychrotrophic and mesophilic microorganisms. After pathogen inoculation, shredded lettuce was packaged in films of different permeability and stored at 5 and 25 °C. After 10 days at 5 °C populations of E. coli O157:H7 and Salmonella decreased approximately 1.00 log unit while L. monocytogenes increased about 1.00 log unit, in all package films. Moreover, the pathogens level increased between 2.44 and 4.19 log units after 3 days at 25 °C. Psychrotrophic and mesophilic bacteria had similar growth at both temperatures with higher populations in air than in the other atmospheres. The composition of the storage atmosphere within the packaging of lettuce had no significant effect on the survival and growth of the pathogens used in this study at refrigeration temperatures. The results obtained can be considered as a warning indicator, which reinforces the necessity for corrective measures to avoid contamination of vegetables.     

Microbiological quality of retail cheeses made from raw, thermized or pasteurized milk in the UK

Two studies of retail fresh, ripened and semi-hard cheeses made from raw, thermized or pasteurized milk were undertaken in the UK during 2004 and 2005 to determine the microbiological quality of these products. Using microbiological criteria in European Commission Recommendations 2004/24/EC and 2005/175/EC, 2% of both raw, thermized (37/1819 samples) and pasteurized (51/2618 samples) milk cheeses were of unsatisfactory quality. Raw or thermized milk cheeses were of unsatisfactory quality due to levels of Staphylococcus aureus at 10(4)cfu g(-1), Escherichia coli at 10(5)cfu g(-1), and/or Listeria monocytogenes at 10(2)cfu g(-1), whereas pasteurized milk cheeses were of unsatisfactory quality due to S. aureus at 10(3)cfu g(-1) and/or E. coli at 10(3)cfu g(-1). Salmonella was not detected in any samples. Cheeses were of unsatisfactory quality more frequently when sampled from premises rated as having little or no confidence in management and control systems, and stored/displayed at above 8 degrees C. Raw or thermized milk cheeses were also more likely to be of unsatisfactory quality when they were unripened types, and pasteurized milk cheeses when they were: semi-hard types; from specialist cheese shops or delicatessens; cut to order. These results emphasize the need for applying and maintaining good hygiene practices throughout the food chain to prevent contamination and/or bacterial growth. Labelling of cheeses with clear information on whether the cheese was prepared from raw milk also requires improvement.     

Reduction of bacteria on spinach, lettuce, and surfaces in food service areas using neutral electrolyzed oxidizing water

Food safety issues and increases in food borne illnesses have promulgated the development of new sanitation methods to eliminate pathogenic organisms on foods and surfaces in food service areas. Electrolyzed oxidizing water (EO water) shows promise as an environmentally friendly broad spectrum microbial decontamination agent. EO water is generated by the passage of a dilute salt solution ( approximately 1% NaCl) through an electrochemical cell. This electrolytic process converts chloride ions and water molecules into chlorine oxidants (Cl(2), HOCl/ClO(-)). At a near-neutral pH (pH 6.3-6.5), the predominant chemical species is the highly biocidal hypochlorous acid species (HOCl) with the oxidation reduction potential (ORP) of the solution ranging from 800 to 900mV. The biocidal activity of near-neutral EO water was evaluated at 25 degrees C using pure cultures of Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis. Treatment of these organisms, in pure culture, with EO water at concentrations of 20, 50, 100, and 120ppm total residual chlorine (TRC) and 10min of contact time resulted in 100% inactivation of all five organisms (reduction of 6.1-6.7log(10)CFU/mL). Spray treatment of surfaces in food service areas with EO water containing 278-310ppm TRC (pH 6.38) resulted in a 79-100% reduction of microbial growth. Dip (10min) treatment of spinach at 100 and 120ppm TRC resulted in a 4.0-5.0log(10)CFU/mL reduction of bacterial counts for all organisms tested. Dipping (10min) of lettuce at 100 and 120ppm TRC reduced bacterial counts of E. coli by 0.24-0.25log(10)CFU/mL and reduced all other organisms by 2.43-3.81log(10)CFU/mL.     

High-throughput method for a kinetics analysis of the high-pressure inactivation of microorganisms using microplates

Using microplates as pressure and cultivation vessels, a high-throughput method was developed for analyzing the high-pressure inactivation kinetics of microorganisms. The loss of viability from a high-pressure treatment, measured based on the growth delay during microplate cultivation, showed reproducibility with the conventional agar plate method and was applicable for the kinetics analysis.     

Effects and interactions of sodium lactate, sodium diacetate, and pediocin on the thermal inactivation of starved Listeria monocytogenes on bologna

The effects and interactions of temperature (56.3-60 °C), sodium lactate (SL; 0-4.8%), sodium diacetate (SDA; 0-2.5%), and pediocin (0-10,000 AU) on starved Listeria monocytogenes (10⁷ CFU/g) on bologna were investigated. Bologna slices containing SL and SDA in the formulation were dipped in pediocin, surface inoculated, and treated at various temperatures using combinations of parameters determined by central composite design. D-values were calculated. The observed D-values ranged from 2.8 min at 60 °C to 24.61 min at 56.3 °C. Injury ranged from 9.1 to 76% under various conditions. The observed D-values were analyzed using second order response surface regression for temperature, SL, SDA, and pediocin, and a predictive model was developed. Predicted D-values were calculated and ranged from 3.7 to 19 min for various combinations of parameters. Temperature alone reduced the predicted D-values from 33.96 min at 56.3 °C to 11.51 min at 60 °C. Addition of SL showed a protective effect. Other combination treatments either reduced or increased D-values depending on temperature. The combination of SL and SDA was effective at lower temperatures, however, higher levels of SDA at higher temperatures made the organism more heat resistant. Pediocin (up to 5000 AU) with increasing temperature and SDA reduced D-values. Depending on temperature and concentration, the interactions between various additives can affect thermal inactivation of L. monocytogenes on bologna. Starvation rendered L. monocytogenes more susceptible to heat and additives.     

A mathematical model of inactivation kinetics for a four-strain composite of Salmonella Enteritidis and Oranienburg in commercial liquid egg yolk

The goal of this study was to develop a general model of inactivation of salmonellae in commercial liquid egg yolk for temperatures ranging from 58 °C to 66 °C by studying the inactivation kinetics of Salmonella in liquid egg yolk. Heat-resistant salmonellae (three serovars of Enteritidis [two of phage type 8 and one PT 13] and one Oranienburg) were grown to stationary phase in Tryptic Soy Broth and concentrated 10-fold by centrifugation. Each inoculum was added to liquid egg yolk and mixed thoroughly, resulting in a final population of ca. 7 log CFU/ml egg yolk. Inoculated yolk was injected into sterile glass capillary tubes, flame-sealed and heated in a water bath at 58, 60, 62, 64, and 66 °C. Capillary tubes were ethanol sanitized, rinsed, and contents were extracted. Yolk was diluted, surface plated onto Tryptic Soy Agar + 0.1% sodium pyruvate and 50 μg/ml nalidixic acid and incubated at 37 °C for 24 h before colonies were enumerated. Decimal reduction values were calculated from survivor curves with a minimum inactivation of 6 log CFU/ml at each temperature. Survival curves (except for 66 °C) featured initial lag periods before first order linear inactivation. Estimated asymptotic D-values were 1.83 min at 58 °C, 0.69 min at 60 °C, 0.26 min at 62 °C, 0.096 min at 64 °C and 0.036 min at 66 °C. The estimate of the asymptotic z-value was ca. 4.7 °C with standard error of 0.07 °C. A linear relationship between the log₁₀ of the lag times and temperature was observed. A general kinetic model of inactivation was developed. The results of the study provide information that can be used by processors to aid in producing safe pasteurized egg yolk products and for satisfying pasteurization performance standards and developing industry guidance.     

Antagonistic effect of Pseudomonas graminis CPA-7 against foodborne pathogens in fresh-cut apples under simulated commercial conditions

Recently, we reported that the application of the strain CPA-7 of Pseudomonas graminis, previously isolated from apple, could reduce the population of foodborne pathogens on minimally processed (MP) apples and peaches under laboratory conditions. Therefore, the objective of the present work was to find an antioxidant treatment and a packaging atmosphere condition to improve CPA-7 efficacy in reducing a cocktail of four Salmonella and five Listeria monocytogenes strains on MP apples under simulated commercial processing. The effect of CPA-7 application on apple quality and its survival to simulated gastric stress were also evaluated. Ascorbic acid (2%, w/v) and N-acetyl-l-cysteine (1%, w/v) as antioxidant treatments reduced Salmonella, L. monocytogenes and CPA-7 recovery, meanwhile no reduction was observed with NatureSeal^® AS1 (NS, 6%, w/v). The antagonistic strain was effective on NS-treated apple wedges stored at 10 °C with or without modified atmosphere packaging (MAP). Then, in a semi-commercial assay, efficacy of CPA-7 inoculated at 10⁵ and 10⁷ cfu mL⁻¹ against Salmonella and L. monocytogenes strains on MP apples with NS and MAP and stored at 5 and 10 °C was evaluated. Although high CPA-7 concentrations/populations avoided Salmonella growth at 10 °C and lowered L. monocytogenes population increases were observed at both temperatures, the effect was not instantaneous. No effect on apple quality was detected and CPA-7 did not survived to simulated gastric stress throughout storage. Therefore, CPA-7 could avoid pathogens growth on MP apples during storage when use as part of a hurdle technology in combination with disinfection techniques, low storage temperature and MAP.     

Thermal inactivation kinetics of Shiga toxin-producing Escherichia coli in buffalo Mozzarella curd

The use of raw milk in the processing of buffalo Mozzarella cheese is permitted, but the heat treatment used for stretching the curd must ensure that the final product does not contain pathogens such as Shiga toxin-producing Escherichia coli (STEC) that may be present on buffalo dairy farms. This study carried out challenge tests at temperatures between 68°C and 80°C for 2 to 10 min to simulate curd temperatures during the stretching phase. Curd samples were inoculated with 2 STEC strains (serotypes O157 and O26), and their inactivation rates were assessed in the different challenge tests. The curd samples were digested with papain to ensure a homogeneous dispersion of bacteria. The STEC cells were counted after inoculation (range 7.1–8.7 log cfu/g) and after heat treatments using the most probable number (MPN) technique. A plot of log MPN/g versus time was created for each separate experiment. The log linear model with tail was used to provide a reasonable fit to observed data. Maximum inactivation rate (k_max, min⁻¹), residual population (log MPN/g), decimal reduction time (min), and time for a 4D (4-log₁₀) reduction (min) were estimated at each temperature tested. A 4D reduction of the O26 STEC strain was achieved when curd was heated at 68°C for 2.6 to 6.3 min or at 80°C for 2.1 to 2.3 min. Greater resistance was observed for the O157 strain at 68°C because k_max was 1.48 min⁻¹. The model estimates can support cheesemakers in defining appropriate process criteria needed to control possible STEC contamination in raw milk intended for the production of Mozzarella.     

Surface enhanced Raman scattering (SERS) with biopolymer encapsulated silver nanosubstrates for rapid detection of foodborne pathogens

A biopolymer encapsulated with silver nanoparticles was prepared using silver nitrate, polyvinyl alcohol (PVA) solution, and trisodium citrate. It was deposited on a mica sheet to use as SERS substrate. Fresh cultures of Salmonella Typhimurium, Escherichia coli, Staphylococcus aureus and Listeria innocua were washed from chicken rinse and suspended in 10 ml of sterile deionized water. Approximately 5 μl of the bacterial suspensions was placed on the substrate individually and exposed to 785 nm HeNe laser excitation. SERS spectral data were recorded over the Raman shift between 400 and 1800 cm^− 1 from 15 different spots on the substrate for each sample; and three replicates were done on each bacteria type. Principal component analysis (PCA) model was developed to classify foodborne bacteria types. PC1 identified 96% of the variation among the given bacteria specimen, and PC2 identified 3%, resulted in a total of 99% classification accuracy. Soft Independent Modeling of Class Analogies (SIMCA) of validation set gave an overall correct classification of 97%. Comparison of the SERS spectra of different types of gram-negative and gram-positive bacteria indicated that all of them have similar cell walls and cell membrane structures. Conversely, major differences were noted around the nucleic acid and amino acid structure information between 1200 cm^− 1 and 1700 cm^− 1 and at the finger print region between 400 cm^− 1 and 700 cm^− 1. Silver biopolymer nanoparticle substrate could be a promising SERS tool for pathogen detection. Also this study indicates that SERS technology could be used for reliable and rapid detection and classification of food borne pathogens.     

Functionality of liquid smoke as an all-natural antimicrobial in food preservation

The smoking of foods, especially meats, has been used as a preservation technique for centuries. Today, smoking methods often involve the use of wood smoke condensates, commonly known as liquid smoke. Liquid smoke is produced by condensing wood smoke created by the pyrolysis of sawdust or wood chips followed by removal of the carcinogenic polyaromatic hydrocarbons. The main products of wood pyrolysis are phenols, carbonyls and organic acids which are responsible for the flavor, color and antimicrobial properties of liquid smoke. Several common food-borne pathogens such as Listeria monocytogenes, Salmonella, pathogenic Escherichia coli and Staphylococcus have shown sensitivity to liquid smoke in vitro and in food systems. Therefore liquid smoke has potential for use as an all-natural antimicrobial in commercial applications where smoke flavor is desired. This review will cover the application and effectiveness of liquid smoke and fractions of liquid smoke as an all-natural food preservative. This review will be valuable for the industrial and research communities in the food science and technology areas.     

Staphylococcus aureus and Listeria monocytogenes in Norwegian raw milk cheese production

The aim of this study was to survey the presence of Staphylococcus aureus and Listeria monocytogenes during the cheese making process in small-scale raw milk cheese production in Norway.The prevalence of S. aureus in bovine and caprine raw milk samples was 47.3% and 98.8%, respectively. An increase in contamination during the first 2-3 h resulted in a 73.6% prevalence of contamination in the bovine curd, and 23 out of 38 S. aureus-negative bovine milk samples gave rise to S. aureus-positive curds. The highest contamination levels of S. aureus were reached in both caprine and bovine cheese after 5-6 h (after the first pressing). There was no contamination of L. monocytogenes in caprine cheeses and only one (1.4%) contaminated bovine cheese.This work has increased our knowledge about S. aureus and L. monocytogenes contamination during the process of raw milk cheese production and gives an account of the hygiene status during the manufacture of Norwegian raw milk cheeses.     

A numerical approach for predicting volumetric inactivation of food borne microorganisms in liquid substrates by pulsed light treatment

Pulsed light (PL) technology is able to effectively destroy a wide variety of food spoilage and pathogenic microorganisms. However, the effectiveness of PL treatment depends on direct exposure of the target microorganisms to the short, high energy pulses of light. The complex physical and chemical properties of foods affect the way light propagates through a given food substrate, and thus there is a real potential for insufficiently or non-uniformly treated products. The objective of this work was to develop a method for predicting levels and spatial distribution of microbial inactivation in PL treatment of liquid substrates, and to validate the predictions with experimental data. Three liquids with different composition and optical properties (BPB, TSB, apple juice) were inoculated with either Escherichia coli ATCC 25922 or Listeria innocua FSL C2-008 and treated with PL, in two different geometries. The Weibull model was used to describe the microbial inactivation kinetics for each organism. The kinetic equations were coupled with previously determined equations describing either the total fluence (F_total) or UV fluence (F_UV) distribution in each of the liquids, for either cylindrical or rectangular prismatic geometries. COMSOL simulation software was used to generate maps of spatial distribution of microbial inactivation and to predict the average volumetric inactivation for each substrate. The model that used F_total provided gross over-estimations for microbial inactivation, while using F_UV as the treatment dose yielded reasonably good predictions of microbial inactivation, especially for the more opaque and turbid substrates. This approach can help processors determine which substrates would be suitable for PL treatment, and to design highly effective and uniform PL treatments.     

Sodium lactate,sodium diacetate and pediocin: Effects and interactions on the thermal inactivation of Listeria monocytogenes on bologna

The effects and interactions of temperature (56.3–60 °C), sodium lactate (SL; 0–4.8%), sodium diacetate (SD; 0–0.25%) and pediocin (0–10,000 AU) on Listeria monocytogenes on bologna were studied and a predictive inactivation model was developed. Bologna was manufactured with different SL/SD concentrations in the formulation, dipped in pediocin solution and treated at different temperatures using combinations of parameters determined by central composite design. D-values were calculated and analyzed using second order response regression. Predicted D-values were also calculated. The observed D-values for L. monocytogenes on bologna ranged from 2.10 to 35.59 min. Temperature alone decreased predicted D-values from 99.02 min at 56.3 °C to 44.71 min at 60.0 °C. Adding SL decreased D-values (85.43–22.71 min) further; however, heat and SD combined was the most effective for reducing L. monocytogenes on bologna. An SD level of 0.25% at 58.2 °C had the overall lowest predicted D-value (15.95 min). Combination treatments increased or decreased D-values, depending on the temperature. Pediocin (2500 and 5000 AU) and heat decreased D-values, but exhibited a protective effect at higher concentrations (≥7500 AU). The results showed that interactions between additives in formulations can vary at different temperatures/concentrations, thereby affecting thermal inactivation of foodborne pathogens in meat products. Hence, food processors should modify food formulations carefully, and verify with adequate testing so that product safety is not compromised.     

Factors affecting growth of foodborne pathogens on minimally processed apples

Escherichia coli O157:H7, Salmonella and Listeria innocua increased by more than 2 log₁₀ units over a 24 h period on fresh-cut ‘Golden Delicious’ apple plugs stored at 25 and 20 °C. L. innocua reached the same final population level at 10 °C meanwhile E. coli and Salmonella only increased 1.3 log₁₀ units after 6 days. Only L. innocua was able to grow at 5 °C. No significant differences were observed between the growth of foodborne pathogens on fresh-cut ‘Golden Delicious’, ‘Granny Smith’ and ‘Shampion’ apples stored at 25 and 5 °C. The treatment of ‘Golden Delicious’ and ‘Granny Smith’ apple plugs with the antioxidants, ascorbic acid (2%) and NatureSeal^® (6%), did not affect pathogen growth. The effect of passive modified atmosphere packaging (MAP) on the growth of E. coli, Salmonella and L. innocua on ‘Golden Delicious’ apple slices was also tested. There were no significant differences in growth of pathogens in MAP conditions compared with air packaging of ‘Golden Delicious’ apple plugs, but the growth of mesophilic and psychrotrophic microorganisms was inhibited. These results highlight the importance of avoiding contamination of fresh-cut fruit with foodborne pathogens and the maintenance of the cold chain during storage until consumption.     

Control of foodborne pathogens on fresh-cut fruit by a novel strain of Pseudomonas graminis

The consumption of fresh-cut fruit has substantially risen over the last few years, leading to an increase in the number of outbreaks associated with fruit. Moreover, consumers are currently demanding wholesome, fresh-like, safe foods without added chemicals. As a response, the aim of this study was to determine if the naturally occurring microorganisms on fruit are “competitive with” or “antagonistic to” potentially encountered pathogens. Of the 97 and 107 isolates tested by co-inoculation with Escherichia coli O157:H7, Salmonella and Listeria innocua on fresh-cut apple and peach, respectively, and stored at 20 °C, seven showed a strong antagonistic capacity (more than 1-log unit reduction). One of the isolates, CPA-7, achieved the best reduction values (from 2.8 to 5.9-log units) and was the only isolate able to inhibit E. coli O157:H7 at refrigeration temperatures on both fruits. Therefore, CPA-7 was selected for further assays. Dose-response assays showed that CPA-7 should be present in at least the same amount as the pathogen to adequately reduce the numbers of the pathogen. From the results obtained in in vitro assays, competition seemed to be CPA-7's mode of action against E. coli O157:H7. The CPA-7 strain was identified as Pseudomonas graminis. Thus, the results support the potential use of CPA-7 as a bioprotective agent against foodborne pathogens in minimally processed fruit.     

Comparing predicting models for heat inactivation of Listeria monocytogenes and Pseudomonas aeruginosa at different pH

Under the same experimental conditions it has been demonstrated that whereas survival curves of Listeria monocytogenes in the range of temperatures from 54 to 62 °C followed a first-order kinetic, those of Pseudomonas aeruginosa in the range of temperatures from 50 to 56 °C were not linear showing a shoulder followed by a linear region. The first order kinetic model did not describe survival curves of P. aeruginosa. A model based on the Weibull distribution (Log₁₀(N_t/N₀)=(1/−2.303)*(t/b)ⁿ)) accurately described the inactivation kinetics of both microorganisms at the three pHs of 4, 5.5, 7.4 investigated. For both microorganisms, the b value depended on the treatment temperature and the pH of the treatment medium. Whereas for L. monocytogenes the n value was independent of the treatment conditions, for P. aeruginosa the n value depended on the pH of the treatment medium.The model based on the Weibull distribution was capable of accurately predicting the treatment time to inactivate five Log₁₀ cycles of both microorganisms at the three pHs investigated.     

Effects of different sanitizing treatments on biofilms and attachment of Escherichia coli and Listeria monocytogenes on green leaf lettuce

The effects of ozone (2 mg/L), chlorine (100 mg/L) and organic acid (0.25 g/100 g citric acid plus 0.50 g/100 g ascorbic acid) treatments at 10 °C for 2 min on the removal of Escherichia coli and Listeria monocytogenes cells embedded inside biofilms on the surface of lettuce leaves were studied. None of the sanitizing treatments were found effective in removing the bacterial biofilms. Initiation of biofilms was observed after 24 h of incubation. Bacterial cells appeared as individual cells, rather than clusters after 6 h incubation, thus 99.9% reductions in both E. coli and L. monocytogenes counts were achieved with all the three treatments. However, after 48 h incubation, none of the treatments resulted in higher than 90% reduction in microbial counts. Biofilm formation was demonstrated for the 48 h incubated samples with SEM images.     

Combined effect of enterocin AS-48 and high hydrostatic pressure to control food-borne pathogens inoculated in low acid fermented sausages

The single and combined effects of enterocin AS-48 and high hydrostatic pressure (HHP) on Listeria monocytogenes, Salmonellaenterica, and Staphylococcus aureus was investigated in fuet (a low acid fermented sausage) during ripening and storage at 7 °C or at room temperature. AS-48 (148 AU g⁻¹) caused a drastic 5.5 log cfu g⁻¹ decrease in L. monocytogenes (P < 0.001) and a significant (P < 0.01) inhibition (1.79 logs) for Salmonella at the end of ripening (10 d). After pressurization (400 MPa) and storage Listeria counts remained below 5 cfu g⁻¹ in all fuets containing AS-48 (pressurized or not). HHP alone had no anti-Listeria effect. HHP treatment significantly reduced Salmonella counts, with lowest levels in pressurized fuets with AS-48. S. aureus showed similar growth for all treatments and storage conditions. These results indicate that AS-48 can be applied alone to control L. monocytogenes and combined with HHP treatment to control Salmonella in fuets.