Reduction of Listeria monocytogenes on frankfurters treated with lactic acid solutions of various temperatures

United States regulations require ready-to-eat meat and poultry processors to control Listeria monocytogenes using interventions which may include antimicrobials that reduce post-processing contamination by at least 1 log-cycle; if the treatment achieves ≥2 log reductions, the plant is subject to less frequent microbial testing. Lactic acid (LA) may be useful as a post-lethality intervention and its antimicrobial properties may increase with temperature of application. The aim of this study was to evaluate the effect of LA solution concentration and temperature on L. monocytogenes counts of inoculated frankfurters and to identify parameters (concentration, temperature, and time) that achieve 1 and 2 log-unit immediate reductions. Frankfurters were surface-inoculated with a 10-strain mixture of L. monocytogenes (4.4 ± 0.1 log CFU/cm²) and then immersed in distilled water or LA solutions (0–3%) of 4, 25, 40, or 55 °C for 0–120 s. A regression equation for L. monocytogenes reduction included significant (P < 0.05) effects by the terms of concentration, time, temperature, and the interaction of concentration and temperature; other tested parameters (other interactions, quadratic and cubic terms), within the experimental range examined, did not affect (P ≥ 0.05) the extent of reduction. Results indicated that the effectiveness of LA against L. monocytogenes, in addition to concentration, increased with solution temperature (in the range of 0.6–2.8 log CFU/cm²). The developed equation may allow processors to vary conditions of treatment with LA to achieve a 1 or 2 log-unit reduction of the pathogen and comply with United States regulations.     

Spray application of liquid smoke to reduce or eliminate Listeria monocytogenes surface inoculated on frankfurters

In a simulated post process contamination scenario liquid smoke was sprayed on the frankfurters after peeling, and then inoculated with Listeria monocytogenes (Lm). Samples that did not receive a liquid smoke spray remained at approximately 2 log cfu/cm² during the 48 h of storage while the levels on the liquid smoke treated frankfurters continued to decline until they were below detection level (1 cfu/100 cm²). A shelf-life study lasting 140 days indicated that liquid smoke suppressed the growth of Lm for up to 130 days. An application of 2 or 3 ml liquid smoke at packaging resulted in at least a 1 log reduction of Lm within 12 h post packaging.     

Effect of fat content on survival of Listeria monocytogenes during simulated digestion of inoculated beef frankfurters stored at 7 °C

Potential effects of the fat content of frankfurters on the gastrointestinal survival of Listeria monocytogenes were investigated. At various stages of storage (7 °C, up to 55 days), inoculated frankfurters of low (4.5%) and high (32.5%) fat content were exposed to a dynamic gastrointestinal model (37 °C) and L. monocytogenes counts were determined at intervals during exposure in each gastrointestinal compartment (gastric, GC; intestinal, IC). Bacterial survival curves in each compartment were fitted with the Baranyi and Roberts mathematical model. L. monocytogenes populations on low- and high-fat frankfurters exceeded 8.0 log CFU/g at 39 and 55 days of storage, respectively. Major declines in populations occurred after 60 min on low-fat frankfurters in the GC, with reductions of 2.6 to >7.2 log CFU/g at 120 min on days 1 and 39 of storage, respectively. L. monocytogenes reductions in high-fat frankfurters ranged from 1.6 (day-1) to 5.2 (day-55) log CFU/g. Gastric inactivation rates were 0.080–0.194 and 0.030–0.097 log CFU/g/min for low- and high-fat samples, respectively. Since gastric emptying began while the gastric pH was >5, initial counts (enumerated 30 min after ingestion) reaching the IC depended on initial contamination levels on each product, which increased during storage. Subsequent reductions during the intestinal challenge were 0.1–1.4 log CFU/g. Findings indicated protective effects of fat against gastric destruction of L. monocytogenes. However, since the effects of fat were observed mainly at later stages of gastric exposure, they did not influence numbers of viable cells reaching the IC.     

Fate of post-processing inoculated Listeria monocytogenes on vacuum-packaged pepperoni stored at 4, 12 or 25 °C

If present, Listeria monocytogenes may not be eliminated during processing of pepperoni or may be introduced during peeling, slicing, or packaging. We evaluated the fate of the pathogen on sliced inoculated pepperoni during vacuum-packaged storage, and potential differences in survival among three types of inocula, including nonacid-adapted, acid-adapted and pepperoni extract-habituated cultures. Commercial pepperoni (two replicates, three samples per treatment) was sliced and inoculated (3 to 4 log CFU/cm²), before vacuum-packaging and storage for up to 180 days at 4, 12 or 25 °C. Samples were periodically analyzed for pathogen counts (PALCAM agar) and total bacterial counts (tryptic soy agar with 0.6% yeast extract). The pH of the product was relatively stable (4.50–4.81) throughout storage. Overall, levels of the pathogen (all inocula) and total counts decreased continuously during storage at all temperatures. The pathogen died slower at 4 °C than at 12 and 25 °C, while at 12 and 25 °C the death rates were similar. Death rates depended on type of inoculum and generally decreased in the order: acid-adapted, extract-habituated and nonacid-adapted inoculum. At day 60, pathogen levels were below the detection limit and remained undetectable throughout the rest of the 180-day storage period, regardless of inoculum type and storage temperature. Therefore, storage of sliced vacuum-packaged pepperoni, especially at ambient temperature, prior to consumption may reduce the potential risk of listeriosis.     

Growth kinetics of Listeria monocytogenes and spoilage microorganisms in fresh-cut cantaloupe

The main objective of this study was to investigate the growth kinetics of Listeria monocytogenes and background microorganisms in fresh-cut cantaloupe. Fresh-cut cantaloupe samples, inoculated with three main serotypes (1/2a, 1/2b, and 4b) of L. monocytogenes, were incubated at different temperatures, ranging from 4 to 43 °C, to develop kinetic growth models. During storage studies, the population of both background microorganisms and L. monocytogenes began to increase almost immediately, with little or no lag phase for most growth curves. All growth curves, except for two growth curves of L. monocytogenes 1/2a at 4 °C, developed to full curves (containing exponential and stationary phases), and can be described by a 3-parameter logistic model. There was no significant difference (P = 0.28) in the growth behaviors and the specific growth rates of three different serotypes of L. monocytogenes inoculated to fresh-cut cantaloupe. The effect of temperature on the growth of L. monocytogenes and spoilage microorganisms was evaluated using three secondary models. For L. monocytogenes, the minimum and maximum growth temperatures were estimated by both the Ratkowsky square-root and Cardinal parameter models, and the optimum temperature and the optimum specific growth rate by the Cardinal parameter model. An Arrhenius-type model provided more accurate estimation of the specific growth rate of L. monocytogenes at temperatures <4 °C. The kinetic models developed in this study can be used by regulatory agencies and food processors for conducting risk assessment of L. monocytogenes in fresh-cut cantaloupe, and for estimating the shelf-life of fresh-cut products.     

Rhamnolipid and surfactin inhibit Listeria monocytogenes adhesion

The aim of this study was to assess the potential use of biosurfactants in inhibiting the Listeria monocytogenes strains adhesion to polystyrene surfaces. Surfactin and rhamnolipids were used. The adhesion profiles of 15 strains showed that most of these bacteria can be classified as moderate to strongly adherent. Both biosurfactants were able to reduce bacterial adhesion, and the effect was more pronounced against the strongly adherent strains. The most promising result was obtained for ATCC7644 strain, which showed an adhesion reduction of 84% for surfactin (30-h growth period). Rhamnolipids decreased the ATCC15313 adhesion by 82%. ATCC19112 adhesion was reduced by 53% for surfactin and purified rhamnolipid. Sodium dodecyl-sulphate was less effective than the biosurfactants, showing maximal adhesion reduction of 23% for 19112 and 7644. The purified rhamnolipid inhibited 100% of the growth of strongly adherent, suggesting that this surfactant can be exploited as a potential agent to control L. monocytogenes.     

Application of an active alginate coating to control the growth of Listeria monocytogenes on poached and deli turkey products

The relatively high prevalence of Listeria monocytogenes in ready-to-eat (RTE) turkey products is of great concern. The overall objective of this study was to develop antimicrobial edible coating formulations to effectively control the growth of this pathogen. The antimicrobials studied were nisin (500 IU/g), Novagard CB 1 (0.25%), Guardian NR100 (500 ppm), sodium lactate (SL, 2.4%), sodium diacetate (SD, 0.25%), and potassium sorbate (PS, 0.3%). These were incorporated alone or in binary combinations into five edible coatings: alginate, κ-carrageenan, pectin, xanthan gum, and starch. The coatings were applied onto the surface of home-style poached and processed deli turkey discs inoculated with ~ 3 log CFU/g of L. monocytogenes. The turkey samples were then stored at 22 °C for 7 days. For poached and processed deli turkey, the coatings were found to be equally effective, with pectin being slightly less effective than the others. The most effective poached turkey treatments seemed to be SL (2.4%)/SD (0.25%) and Nisin (500 IU/g)/SL (2.4%), which yielded final populations of 3.0 and 4.9 log CFU/g respectively compared to the control which was 7.9 log CFU/g. For processed deli turkey, the most effective antimicrobial treatments seemed to be Nisin (500 IU/g)/SD (0.25%) and Nisin (500 IU/g)/SL (2.4%) with final populations of 1.5 and 1.7 log CFU/g respectively compared to the control which was 6.5 log CFU/g. In the second phase of the study, home-style poached and store-purchased roasted (deli) turkey inoculated with the pathogen at a level of ~ 3 log CFU/g were coated with alginate incorporating selected antimicrobial combinations and stored for 8 weeks at 4 °C. Alginate coatings supplemented with SL (2.4%)/PS (0.3%) delayed the growth of L. monocytogenes with final counts reaching 4.3 log CFU/g (home-style poached turkey) and 6.5 log CFU/g (roasted deli turkey) respectively while the counts in their untreated counterparts were significantly higher (P < 0.05) reaching 9.9 and 7.9 log CFU/g, respectively. This study therefore demonstrates the effectiveness of using alginate-based antimicrobial coatings to enhance the microbiological safety and quality of RTE poultry products during chilled storage.     

Fate of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella on fresh-cut celery

Illnesses from Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella have been associated with the consumption of numerous produce items. Little is known about the effect of consumer handling practices on the fate of these pathogens on celery. The objective of this study was to determine pathogen behavior at different temperatures under different storage conditions. Commercial fresh-cut celery was inoculated at ca. 3 log CFU/g onto either freshly cut or outer uncut surfaces and stored in either sealed polyethylene bags or closed containers. Samples were enumerated following storage for 0, 1, 3, 5, and 7 days when held at 4 °C or 12 °C, and after 0, 8, and 17 h, and 1, and 2 days when held at 22 °C. At 4 °C, all populations declined by 0.5–1.0 log CFU/g over 7 days. At 12 °C, E. coli O157:H7 and Salmonella populations did not change, while L. monocytogenes populations increased by ca. 0.5 log CFU/g over 7 days. At 22 °C, E. coli O157:H7, Salmonella, and L. monocytogenes populations increased by ca. 1, 2, or 0.3 log CFU/g, respectively, with the majority of growth occurring during the first 17 h. On occasion, populations on cut surfaces were significantly higher than those on uncut surfaces. Results indicate that populations are reduced under refrigeration, but survive and may grow at elevated temperatures.     

The probability of growth of Listeria monocytogenes in cooked salmon and tryptic soy broth as affected by salt,smoke compound,and storage temperature

The objectives of this study were to examine and model the probability of growth of Listeria monocytogenes in cooked salmon containing salt and smoke (phenol) compound and stored at various temperatures. A growth probability model was developed, and the model was compared to a model developed from tryptic soy broth (TSB) to assess the possibility of using TSB as a substitute for salmon. A 6-strain mixture of L. monocytogenes was inoculated into minced cooked salmon and TSB containing 0–10% NaCl and 0–34 ppm phenol to levels of 10^2–3 cfu/g, and the samples were vacuum-packed and stored at 0-–25 °C for up to 42 days. A total 32 treatments, each with 16 samples, selected by central composite designs were tested. A logistic regression was used to model the probability of growth of L. monocytogenes as a function of concentrations of salt and phenol, and storage temperature. Resulted models showed that the probabilities of growth of L. monocytogenes in both salmon and TSB decreased when the salt and/or phenol concentrations increased, and at lower storage temperatures. In general, the growth probabilities of L. monocytogenes were affected more profoundly by salt and storage temperature than by phenol. The growth probabilities of L. monocytogenes estimated by the TSB model were higher than those by the salmon model at the same salt/phenol concentrations and storage temperatures. The growth probabilities predicted by the salmon and TSB models were comparable at higher storage temperatures, indicating the potential use of TSB as a model system to substitute salmon in studying the growth behavior of L. monocytogenes may only be suitable when the temperatures of interest are in higher storage temperatures (e.g., >12 °C). The model for salmon demonstrated the effects of salt, phenol, and storage temperature and their interactions on the growth probabilities of L. monocytogenes, and may be used to determine the growth probability of L. monocytogenes in smoked seafood.     

Survival and death of Listeria monocytogenes on cooked bacon at three storage temperatures

Survival of Listeria monocytogenes on cooked bacon cubes (a_w 0.910 ± 0.080), strips (a_w 0.726 ± 0.054), and bits (a_w 0.620 ± 0.038) was determined during a 25 week storage period at −20, 4.4, and 22 °C. Selective enrichment and subsequent enzyme-linked fluorescent antibody (ELFA) detection were used to asses survival on samples inoculated at ca. 1-log₁₀ CFU/g (LI). Samples inoculated at ca. 5.5-log₁₀ CFU/g (HI) were analyzed over time by direct plating on modified Oxford medium (MOX). The Baranyi model was fitted to the inactivation curves of HI samples using the DMFit program. At −20 °C, a decline of about 1-log₁₀ CFU/g occurred on all HI cooked bacon types by 14 weeks, although most LI samples remained positive by the ELFA detection method for 25 weeks. At 4.4 and 22 °C, some strips and bits LI samples were negative for the pathogen within 3 weeks, and >1.5 log₁₀ CFU/g reductions occurred on HI strips and bits by 8 weeks. Reductions on cubes at refrigeration and ambient temperature were ca. 0.5 log₁₀ CFU/g, and cubes remained positive on LI samples for 25 weeks. Rate parameter estimates indicated that the population declined fastest on strips and bits at 22 °C compared to all other product and temperature combinations. This study demonstrates that cooked bacon does not support the growth of L. monocytogenes and that the pathogen gradually dies off during storage.     

Smearing of soft cheese with Enterococcus faecium WHE 81, a multi-bacteriocin producer,against Listeria monocytogenes

Enterococcus faecium WHE 81, a multi-bacteriocin producer, was tested for its antimicrobial activity on Listeria monocytogenes in Munster cheese, a red smear soft cheese. The naturally delayed and superficial contamination of this type of cheese allowed the use of E. faecium WHE 81 at the beginning of the ripening as a surface culture. A brine solution inoculated at 10⁵ CFU of E. faecium WHE 81 per mL was sprayed on the cheese surface during the first smearing operation. On day 7, smearing of cheese samples with a brine solution at 10² CFU of L. monocytogenes per mL yielded initial cell counts of approximately 50 CFU g⁻¹ of the pathogen on the cheese surface. Although, in some instances, L. monocytogenes could survive (<50 CFU g⁻¹) in the presence of E. faecium WHE 81, it was unable to initiate growth. In control samples however, L. monocytogenes counts often exceeded 10⁴ CFU g⁻¹. In other respects, E. faecium WHE 81, which naturally existed in Munster cheese, did not adversely impact on the ripening process.     

Effect of nanovesicle-encapsulated nisin on growth of Listeria monocytogenes in milk

Commercial nisin was encapsulated in nanovesicles (mean diameter 140 nm) prepared from partially purified soy lecithin. Nisin-loaded liposomes and unencapsulated (free) nisin were initially tested in BHI medium and skim milk inoculated with Listeria monocytogenes and incubated for 48 h at 30 °C. At such abuse temperature conditions, free nisin showed better inhibitory than the liposomal counterparts. Subsequently, the effect of encapsulated or free nisin was evaluated in combination with refrigeration (7 ± 1 °C) in both whole (3.25% fat) and skim (0% fat) milk for up to 14 day. A decrease of 3–4 log cycles in L. monocytogenes counts was observed for free and encapsulated nisin at 0.5 mg/ml concentration. Liposome encapsulation of antimicrobial peptides may be important to overcome stability issues and interaction with food components. The utilization of nanovesicle-encapsulated nisin in combination with low temperatures appeared to be effective to control L. monocytogenes in milk, emphasizing the importance of hurdle technology to assure food safety.     

Fat content increases the lethality of ultra-high-pressure homogenization on Listeria monocytogenes in milk

Listeria monocytogenes CCUG 15526 was inoculated at a concentration of approximately 7.0 log₁₀ cfu/mL in milk samples with 0.3, 3.6, 10, and 15% fat contents. Milk samples with 0.3 and 3.6% fat content were also inoculated with a lower load of approximately 3.0 log₁₀ cfu/mL. Inoculated milk samples were subjected to a single cycle of ultra-high-pressure homogenization (UHPH) treatment at 200, 300, and 400 MPa. Microbiological analyses were performed 2 h after the UHPH treatments and after 5, 8, and 15 d of storage at 4°C. Maximum lethality values were observed in samples treated at 400 MPa with 15 and 10% fat (7.95 and 7.46 log₁₀ cfu/mL), respectively. However, in skimmed and 3.6% fat milk samples, complete inactivation was not achieved and, during the subsequent 15 d of storage at 4°C, L. monocytogenes was able to recover and replicate until achieving initial counts. In milk samples with 10 and 15% fat, L. monocytogenes recovered to the level of initial counts only in the milk samples treated at 200 MPa but not in the milk samples treated at 300 and 400 MPa. When the load of L. monocytogenes was approximately 3.0 log₁₀ cfu/mL in milk samples with 0.3 and 3.6% fat, complete inactivation was not achieved and L. monocytogenes was able to recover and grow during the subsequent cold storage. Fat content increased the maximum temperature reached during UHPH treatment; this could have contributed to the lethal effect achieved, but the amount of fat of the milk had a stronger effect than the temperature on obtaining a higher death rate of L. monocytogenes.     

Molecular subtyping and virulence gene analysis of Listeria monocytogenes isolates from food

A total of 67 Listeria monocytogenes isolates from 698 raw meat samples were characterized for molecular serogroup identification and antimicrobial susceptibility. Approximately one third (32.8%) of the isolates belonged to molecular serogroup 1/2a, 3a, followed by 1/2c, 3c (26.9%), 1/2b, 3b, 7 (22.4%), 4b, 4d, 4e (16.4%) and 4a, 4c (1.5%). Most of the L. monocytogenes isolates were susceptible to 14 antimicrobials tested but several were resistant to tetracycline, ciprofloxacin and nitrofurantoin. An additional 30 L. monocytogenes isolates from chicken and produce in our collection were also included to determine the presence of significant virulence markers. All 97 isolates carried inlC and inlJ except for a lineage III isolate 110-1. Most Listeriolysin S (LLS)-carrying isolates (11/12) belonged to lineage I, whereas the remaining one isolate belonged to lineage III. Five 4b, 4d, 4e isolates including two from turkey and three from produce belonged to Epidemic Clone I (ECI). Four molecular serogroup associated mutation types that lead to premature stop codons (PMSCs) in inlA were identified. PFGE and inlA sequence analysis results were concordant, and different virulence potential within 1/2a, 3a and 4b, 4d, 4e isolates were observed. The study revealed that a subset of isolates from meat and produce belonged to ECI, harbored inlC, inlJ and LLS, and produced full length InlA, suggesting that they be capable of causing human illness.     

Control of Listeriamonocytogenes on commercially-produced frankfurters prepared with and without potassium lactate and sodium diacetate and surface treated with lauric arginate using the Sprayed Lethality in Container (SLIC®) delivery method

Viability of Listeriamonocytogenes was monitored on frankfurters formulated with or without potassium lactate and sodium diacetate at a ratio of ca. 7:1 and treated with lauric arginate (LAE; 22 or 44 ppm) using the Sprayed Lethality in Container (SLIC®) delivery method. Without antimicrobials, pathogen numbers remained relatively constant at ca. 3.3 log CFU/package for ca. 30 d, but then increased to ca. 8.4 log CFU/package over 120 d. Regardless of whether or not lactate and diacetate were included, when treated with LAE, pathogen numbers decreased from ca. 3.3 log CFU/package to ca. 1.5 log CFU/package within 2 h, but then increased to 7.3 and 6.7 log CFU/package, respectively, after 120 d. When frankfurters were formulated with lactate and diacetate and treated with LAE, pathogen numbers decreased by ca. 2.0 log CFU/package within 2 h and remained relatively unchanged over the 120 d. These data confirm that LAE provides an initial lethality towards L. monocytogenes and when used in combination with reduced levels/ratio of lactate and diacetate as an ingredient for frankfurters provides inhibition throughout shelf life.     

A predictive model for heat inactivation of Listeria monocytogenes biofilm on buna-N rubber

The purpose of this study was to develop a predictive model for the heat inactivation of Listeria monocytogenes in monoculture (strains Scott A and 3990) and with competing bacteria (Pseudomonas sp. and Pantoea agglomerans) formed on buna-N rubber with and without the presence of food-derived soil. Biofilms were produced on rubber disks in dilute Tryptic Soy broth (dTSB) with incubation for 48 h at 25 °C. Duplicate biofilm samples were heat treated for 1, 3, 5, and 15 min at 70, 72, 75, 77 and 80 °C and tested for survivors using enrichment media. The experiment was repeated six times. A predictive model was developed and plots were generated showing the percent probability of L. monocytogenes inactivation in biofilms after heat treatment. For example, to achieve a 95% probability level of complete inactivation required heat treatment of 76 °C for 6 min. The predicted model was validated using a five-strain cocktail of L. monocytogenes. The validated prediction model indicates that with proper maintenance of the time/temperature controls L. monocytogenes in biofilms on rubber surfaces will be inactivated. This model can be used as a tool in the selection of hot water sanitation processes for rubber surfaces.     

Listeria monocytogenes and ready-to-eat seafood in Spain: Study of prevalence and temperatures at retail

The aim of this study was to obtain data from refrigerated ready-to-eat seafood products at retail in Spain (young eels, crabstick and smoked salmon), regarding prevalence and levels of Listeria monocytogenes, storage temperatures and the impact of transport conditions (type of bag) on the temperature of the product. The one-year surveillance period was carried out according to the EC Regulation No. 2073/2005, taking 5 units/batch and analyzing 250 samples following ISO 11290-1/A1 and ISO 11290-2/A methodologies. Low prevalence of L. monocytogenes was observed in surimi products, while 4.8% of smoked salmon samples were positive for Listeria with low levels (<10 cfu/g) and uneven pathogen distribution. A single company was responsible for 80% of the positive lots. All purchased products showed values higher than 4 °C at retail and an average increase of 2.5 °C or up to 6.2 °C was recorded when isothermal or plastic shopping bags were used for transport, respectively. To avoid noncompliance of the Food Safety Objective for L. monocytogenes in seafood RTE products more efforts from all stakeholders are needed, with special attention so as to improve control and maintenance of refrigerators at retail and to enhance consumer education regarding food safety practices.     

Effect of high-pressure processing on reduction of Listeria monocytogenes in packaged Queso Fresco

The effect of high-hydrostatic-pressure processing (HPP) on the survival of a 5-strain rifampicin-resistant cocktail of Listeria monocytogenes in Queso Fresco (QF) was evaluated as a postpackaging intervention. Queso Fresco was made using pasteurized, homogenized milk, and was starter-free and not pressed. In phase 1, QF slices (12.7 × 7.6 × 1 cm), weighing from 52 to 66 g, were surface inoculated with L. monocytogenes (ca. 5.0 log₁₀ cfu/g) and individually double vacuum packaged. The slices were then warmed to either 20 or 40°C and HPP treated at 200, 400, and 600 MPa for hold times of 5, 10, 15, or 20 min. Treatment at 600 MPa was most effective in reducing L. monocytogenes to below the detection level of 0.91 log₁₀ cfu/g at all hold times and temperatures. High-hydrostatic-pressure processing at 40°C, 400 MPa, and hold time ≥15 min was effective but resulted in wheying-off and textural changes. In phase 2, L. monocytogenes was inoculated either on the slices (ca. 5.0 log₁₀ cfu/g; ON) or in the curds (ca. 7.0 log₁₀ cfu/g; IN) before the cheese block was formed and sliced. The slices were treated at 20°C and 600 MPa at hold times of 3, 10, and 20 min, and then stored at 4 and 10°C for 60 d. For both treatments, L. monocytogenes became less resistant to pressure as hold time increased, with greater percentages of injured cells at 3 and 10 min than at 20 min, at which the lethality of the process increased. For the IN treatment, with hold times of 3 and 10 min, growth of L. monocytogenes increased the first week of storage, but was delayed for 1 wk, with a hold time of 20 min. Longer lag times in growth of L. monocytogenes during storage at 4°C were observed for the ON treatment at hold times of 10 and 20 min, indicating that the IN treatment may have provided a more protective environment with less injury to the cells than the ON treatment. Similarly, HPP treatment for 10 min followed by storage at 4°C was the best method for suppressing the growth of the endogenous microflora with bacterial counts remaining below the level of detection for 2 out of the 3 QF samples for up to 84 d. Lag times in growth were not observed during storage of QF at 10°C. Although HPP reduced L. monocytogenes immediately after processing, a second preservation technique is necessary to control growth of L. monocytogenes during cold storage. However, the results also showed that HPP would be effective for slowing the growth of microorganisms that can shorten the shelf life of QF.     

Foci of contamination of Listeria monocytogenes in different cheese processing plants

Listeria monocytogenes is a ubiquitous bacterium widely distributed in the environment that can cause a severe disease in humans when contaminated foods are ingested. Cheese has been implicated in sporadic cases and in outbreaks of listeriosis worldwide. Environmental contamination, in several occasions by persistent strains, has been considered an important source of finished product contamination. The objectives of this research were to (i) evaluate the presence of L. monocytogenes within the factory environments and cheeses of three processing plants, artisanal producer of raw ewe's milk cheeses (APC), small-scale industrial cheese producer (SSI) and industrial cheese producer (ICP) each producing a distinct style of cheese, all with history of contamination by L. monocytogenes (ii) and identify possible sources of contamination using different typing methods (arsenic and cadmium susceptibility, geno-serotyping, PFGE). The presence of markers specific for 3 epidemic clones (ECI–ECIII) of L. monocytogenes was also investigated. Samples were collected from raw milk (n = 179), whey (n = 3), cheese brining solution (n = 7), cheese brine sludge (n = 505), finished product (n = 3016), and environment (n = 2560) during, at least, a four-year period. Listeria monocytogenes was detected in environmental, raw milk and cheese samples, respectively, at 15.4%, 1.1% and 13.6% in APC; at 8.9%, 2.9% and 3.4% in SSI; and at 0%, 21.1% and 0.2% in ICP. Typing of isolates revealed that raw ewe's milk and the dairy plant environment are important sources of contamination, and that some strains persisted for at least four years in the environment. Although cheeses produced in the three plants investigated were never associated with any case or outbreak of listeriosis, some L. monocytogenes belonging to specific PFGE types that caused disease (including putative epidemic clone strains isolated from final products) were found in this study.