Behaviour of Listeria monocytogenes in packaged fresh mushrooms (Agaricus bisporus)

AIMS: The aim of this study was to evaluate the potential of Listeria monocytogenes to grow in mushrooms packaged in two different types of PVC films when stored at 4 degrees C and 10 degrees C. METHODS AND RESULTS: Mushrooms were packed in two polymeric films (perforated and nonperforated PVC) and stored at 4 degrees C and 10 degrees C. The carbon dioxide and oxygen content inside the packages, aerobic mesophiles, psychrotrophs, Pseudomonas spp., Listeria monocytogenes, faecal coliforms, Escherichia coli, anaerobic spores and major sensory factors were determined. The mushrooms packaged in nonperforated film and stored at 4 degrees C had the most desirable quality parameters (texture, development stage and absence of moulds). Listeria monocytogenes was able to grow at 4 degrees C and 10 degrees C in inoculated mushrooms packaged in perforated and nonperforated films between 1 and 2 log units during the first 48 h. After 10 d of storage, the populations of L. monocytogenes were higher in mushrooms packaged in nonperforated film and stored at 10 degrees C. CONCLUSIONS: MAP followed by storage at 4 degrees C or 10 degrees C extends the shelf life by maintaining an acceptable appearance, but allows the growth and survival of L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: According to this study additional hurdles must be studied in order to prevent the growth of L. monocytogenes.     

Identification of lactic acid bacteria from spoilage associations of cooked and brined shrimps stored under modified atmosphere between 0 degrees C and 25 degrees C

AIMS: To evaluate spoilage and identify lactic acid bacteria (LAB) from spoilage associations of cooked and brined shrimps stored under modified atmosphere packaging (MAP) at 0, 5, 8, 15 and 25 degrees C. METHODS AND RESULTS: Bacterial isolates (102) from spoilage associations of cooked and brined MAP shrimps were characterized by phenotypic tests and identified as lactic acid bacteria (78 isolates), other Gram-positive bacteria (13 isolates) and Gram-negative bacteria (11 isolates). A selection of 48 LAB isolates were further characterized and identified by phenotypic tests and SDS-PAGE electrophoresis of whole cell proteins. Selected clusters of LAB isolates were analysed by plasmid profiling, pulsed field gel electrophoresis and 16S rRNA gene sequencing. Enterococcus faecalis was identified in spoilage associations at 15 degrees C and 25 degrees C, and its metabolic activity corresponded to chemical changes in spoiled products. Carnobacterium divergens, a non-motile Carnobacterium sp. nov. and Lactobacillus curvatus were the LAB species observed in spoilage associations of products stored at 0 degrees C, 5 degrees C and 8 degrees C. CONCLUSIONS: Enterococcus spp. and Carnobacterium spp. were the dominant parts of spoilage associations of cooked and brined MAP shrimps stored at high and low temperatures, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: The SDS-PAGE technique and simple biochemical keys allowed the majority of LAB isolates from spoilage associations of cooked and brined MAP shrimps to be identified at the species level.     

Effects of high-pressure processing on Listeria monocytogenes, spoilage microflora and multiple compound quality indices in chilled cold-smoked salmon

AIMS: To evaluate the effect of high-pressure processing (HPP) on Listeria monocytogenes, microbial and chemical changes and shelf-life in chilled cold-smoked salmon (CSS). METHODS AND RESULTS: First, challenge tests with L. monocytogenes were carried out using HPP of the product at 0.1 (control), 150, 200 and 250 MPa. Secondly, storage trials with the naturally contaminated product and HPP at 0.1 (control) and 200 MPa were realized. Shelf-life, microbial changes and chemical changes were determined and existing predictive models and multiple compound quality indices evaluated. HPP with 250 MPa did not inactivate L. monocytogenes but significant lag phases of 17 and 10 days were observed at ca 5 and 10 degrees C, respectively. HPP with 200 MPa had a marked effect on both colour and texture of CSS. CONCLUSIONS: High-pressure processing was unable to prevent growth of L. monocytogenes or spoilage of chilled CSS. Existing mathematical models allowed growth rates of L. monocytogenes and shelf-life of samples without high-pressure treatments to be predicted. SIGNIFICANCE AND IMPACT OF THE STUDY: High-pressure processing seems more appropriate for new types of salmon products than for a classical product like CSS where consumers expect specific quality attributes.     

Quantification of Listeria spp. contamination on shell and flesh of cooked black tiger prawns (Penaeus monodon)

AIMS: To quantify Listeria levels on the shell and flesh of artificially contaminated cooked prawns after peeling, and determine the efficacy of Listeria innocua as a model for L. monocytogenes in this system. METHODS AND RESULTS: A L. monocytogenes and L. innocua strain were inoculated separately onto cooked black tiger prawns using two protocols (immersion or swabbing with incubation). Prawns were peeled by two methods (gloved hand or scalpel and forceps) and numbers of Listeria on shells, flesh and whole prawn controls were determined. Prawns were exposed to crystal violet dye to assess the penetration of liquids. Regardless of preparation method or bacterial strain there were ca 1log10 CFU more Listeria per shell than per peeled prawn. Dye was able to penetrate to the flesh in all cases. CONCLUSIONS: Shell-on prawns may be only slightly safer than shell-off prawns. Listeria innocua is an acceptable model for L. monocytogenes in this system. SIGNIFICANCE AND IMPACT OF THE STUDY: Reduced risk from L. monocytogenes on prawns can only be assured by adequate hygiene or heating.     

Mild heat treatment of lettuce enhances growth of Listeria monocytogenes during subsequent storage at 5 degrees C or 15 degrees C

AIMS: The objective of this study was to determine the influence of mild heat treatment, storage temperature and storage time on the survival and growth of Listeria monocytogenes inoculated onto cut iceberg lettuce leaves. METHODS AND RESULTS: Before or after inoculation with L. monocytogenes, cut iceberg lettuce leaves were dipped in water (20 or 50 degrees C) containing or not 20 mg l(-1) chlorine, for 90 s, then stored at 5 degrees C for up to 18 days or 15 degrees C for up to 7 days. The presence of 20 mg l(-1) chlorine in the treatment water did not significantly (alpha=0.05) affect populations of the pathogen, regardless of other test parameters. The population of L. monocytogenes on lettuce treated at 50 degrees C steadily increased throughout storage at 5 degrees C for up to 18 days. At day 10 and thereafter, populations were 1.7-2.3 log10 cfu g(-1) higher on lettuce treated at 50 degrees C after inoculation compared with untreated lettuce or lettuce treated at 20 degrees C, regardless of chlorine treatment. The population of L. monocytogenes increased rapidly on lettuce stored at 15 degrees C. At 2 and 4 days, significantly higher populations were detected on lettuce that had been treated at 50 degrees C, compared with respective samples that had been treated at 20 degrees C, regardless of inoculation before or after treatment, or the presence of 20 mg l(-1) chlorine in the treatment water. CONCLUSIONS: The results clearly demonstrated that mild heat treatment of cut lettuce leaves enhances the growth of L. monocytogenes during subsequent storage at 5 or 15 degrees C. SIGNIFICANCE AND IMPACT OF THE STUDY: Mild heat treatment of cut lettuce may result in a prolonged shelf life as a result of delaying the development of brown discoloration. However, heat treatment also facilitates the growth of L. monocytogenes during storage at refrigeration temperature, thereby increasing the potential risk of causing listeriosis.     

Microbial spoilage and formation of biogenic amines in fresh and thawed modified atmosphere-packed salmon (Salmo salar) at 2 degrees C

AIMS: To evaluate the microbial spoilage, formation of biogenic amines and shelf life of chilled fresh and frozen/thawed salmon packed in a modified atmosphere and stored at 2 degrees C. METHODS AND RESULTS: The dominating microflora, formation of biogenic amines and shelf life were studied in two series of storage trials with naturally contaminated fresh and thawed modified atmosphere-packed (MAP) salmon at 2 degrees C. Photobacterium phosphoreum dominated the spoilage microflora of fresh MAP salmon at more than 10(6) cfu g(-1) and the activity of this specific spoilage organism (SSO) limited the shelf life of the product to ca 14 and 21 d in the two experiments. Despite the high levels of P. phosphoreum, less than 20 mg kg(-1) histamine was observed in fresh MAP salmon prior to sensory spoilage. Freezing eliminated P. phosphoreum and extended the shelf life of MAP salmon at 2 degrees C by 1-2 weeks. Carnobacterium piscicola dominated the spoilage microflora of thawed MAP salmon and probably produced the ca 40 mg kg(-1) tyramine detected in this product at the end of its shelf life. CONCLUSIONS: Photobacterium phosphoreum dominated the spoilage microflora of fresh MAP salmon but produced only small amounts of biogenic amines in this product. The elimination of P. phosphoreum by freezing allowed this bacteria to be identified as the SSO in fresh MAP salmon. SIGNIFICANCE AND IMPACT OF THE STUDY: The identification of P. phosphoreum as the SSO in fresh MAP salmon facilitates the development of methods to determine and predict the shelf life of this product, as previously shown with fresh MAP cod.     

Application of Leuconostoc carnosum for biopreservation of cooked meat products

AIMS: To optimize the practical use of the bacteriocin producing Leuconostoc carnosum 4010 in order to inhibit the growth of Listeria monocytogenes in sliced meat products. METHODS AND RESULTS: Four different methods for biopreservation using the partially purified bacteriocin or the living culture of Leuc. carnosum 4010 were evaluated. The methods using the living protective culture added to the sliced gas packed meat product were more effective in preventing growth of L. monocytogenes than the use of the partially purified leucocins 4010 or bacteriocin produced during fermentation before heat treatment of the saveloy. The application method giving the highest reduction in L. monocytogenes used nozzles for sprinkling the protective culture on all surfaces of each slice of the meat product. In the control samples without the protective culture, L. monocytogenes grew to ca. 107 CFU g(-1), whereas for the application method using nozzles for distributing the protective culture, counts of L. monocytogenes never exceeded 10 CFU g(-1) during 4 weeks of storage at 10 degrees C. CONCLUSIONS: The live cells of the bacteriocin producing Leuc. carnosum 4010 was the most efficient method as it inhibited the growth of L. monocytogenes in cooked, sliced and gas packed saveloy stored at 5 and 10 degrees C for 4 weeks. SIGNIFICANCE AND IMPACT OF THE STUDY: The results indicate that biopreservation with lactic acid bacteria is a suitable alternative to chemical preservatives. An even distribution of the protective culture was found to be essential for the efficacy of the protective culture in pilot plant trials.     

Inhibitory effects of raw carrots on Listeria monocytogenes.

The survival and growth of two strains of Listeria monocytogenes on raw and cooked carrots stored at 5 and 15 degrees C and in carrot juice media at 30 degrees C were investigated. The influence of shredding, chlorine treatment, and packaging under an atmosphere containing 3% O2 and 97% N2 on the behavior of L. monocytogenes and naturally occurring microflora was determined. Populations of viable L. monocytogenes decreased upon contact with whole and shredded raw carrots but not cooked carrots. Viable populations also decreased in cell suspensions in which raw carrots were dipped. Small populations of L. monocytogenes detected on whole carrots immediately after dipping were essentially nondetectable after 7 days of storage at 5 or 15 degrees C. After a lag of 7 days at 5 degrees C, significant (P less than or equal to 0.05) increases in populations were detected on shredded carrots after 24 days of storage. Carrots stored at 5 or 15 degrees C spoiled before L. monocytogenes grew. Populations of mesophilic aerobes, psychrophiles, and yeasts and molds increased throughout storage at 5 and 15 degrees C. Cutting treatment (whole or shredded carrots), chlorine treatment, and modified-atmosphere packaging had no effect on the survival or growth of L. monocytogenes or naturally occurring microflora. The presence of raw carrot juice in tryptic phosphate broth at a concentration as low as 1% substantially reduced the maximum population of L. monocytogenes reached after 24 h at 30 degrees C. The anti-Listeria effect of carrots was essentially eliminated when the carrots were cooked.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inhibitory effects of raw carrots on Listeria monocytogenes

The survival and growth of two strains of Listeria monocytogenes on raw and cooked carrots stored at 5 and 15 degrees C and in carrot juice media at 30 degrees C were investigated. The influence of shredding, chlorine treatment, and packaging under an atmosphere containing 3% O2 and 97% N2 on the behavior of L. monocytogenes and naturally occurring microflora was determined. Populations of viable L. monocytogenes decreased upon contact with whole and shredded raw carrots but not cooked carrots. Viable populations also decreased in cell suspensions in which raw carrots were dipped. Small populations of L. monocytogenes detected on whole carrots immediately after dipping were essentially nondetectable after 7 days of storage at 5 or 15 degrees C. After a lag of 7 days at 5 degrees C, significant (P less than or equal to 0.05) increases in populations were detected on shredded carrots after 24 days of storage. Carrots stored at 5 or 15 degrees C spoiled before L. monocytogenes grew. Populations of mesophilic aerobes, psychrophiles, and yeasts and molds increased throughout storage at 5 and 15 degrees C. Cutting treatment (whole or shredded carrots), chlorine treatment, and modified-atmosphere packaging had no effect on the survival or growth of L. monocytogenes or naturally occurring microflora. The presence of raw carrot juice in tryptic phosphate broth at a concentration as low as 1% substantially reduced the maximum population of L. monocytogenes reached after 24 h at 30 degrees C. The anti-Listeria effect of carrots was essentially eliminated when the carrots were cooked.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of the pH-dependent,stationary-phase acid tolerance in Listeria monocytogenes and Salmonella Typhimurium DT104 induced by culturing in media with 1% glucose: a comparative study with Escherichia coli O157:H7

AIMS: To comparatively evaluate the adaptive stationary-phase acid tolerance response (ATR) in food-borne pathogens induced by culturing in glucose-containing media, as affected by strain variability and antibiotic resistance, growth temperature, challenge pH and type of acidulant. METHODS AND RESULTS: Antibiotic resistant or sensitive strains of Listeria monocytogenes, Salmonella including S. Typhimurium DT104, and Escherichia coli O157:H7 were cultured (30 degrees C for 24 h; 10 degrees C for up to 14 days) in trypticase soya broth with yeast extract (TSBYE) with 1% or without glucose to induce or prevent acid adaptation, respectively. Cultures were subsequently exposed to pH 3.5 or 3.7 with lactic or acetic acid at 25 degrees C for 120 min. Acid-adapted cultures were more acid tolerant than nonadapted cultures, particularly those of L. monocytogenes and Salmonella. No consistent, positive or negative, influence of antibiotic resistance on the pH-inducible ATR or acid resistance (AR) was observed. Compared with 30 degrees C cultures, growth and acid adaptation of L. monocytogenes and S. Typhimurium DT104 at 10 degrees C markedly reduced their ATR and AR in stationary phase. E. coli O157:H7 had the greatest AR, relying less on acid adaptation. A 0.2 unit difference in challenge pH (3.5-3.7) caused great variations in survival of acid-adapted and nonadapted cells. CONCLUSIONS: Culturing L. monocytogenes and Salmonella to stationary phase in media with 1% glucose induces a pH-dependent ATR and enhances their survival to organic acids; thus, this method is suitable for producing acid-adapted cultures for use in food challenge studies. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacterial pathogens may become acid-adapted in foods containing glucose or other fermentable carbohydrates. Low storage temperatures may substantially decrease the stationary-phase ATR of L. monocytogenes and S. Typhimurium DT104, but their effect on ATR of E. coli O157:H7 appears to be far less dramatic.     

Development of polythene films for food packaging activated with an antilisterial bacteriocin from Lactobacillus curvatus 32Y

AIMS: The aims of this work were to (i) use a bacteriocin produced by Lactobacillus curvatus 32Y active against Listeria monocytogenes to activate polythene films by different methods, (ii) implement a large-scale process for antilisterial polythene films production and (iii) verify the efficacy of the developed films in inhibiting the growth of L. monocytogenes during the storage of meat products. METHODS AND RESULTS: The film was made active by using the antilisterial bacteriocin 32Y by Lact. curvatus with three different procedures: soaking, spraying and coating. The antimicrobial activity of the activated films was tested in plate assays against the indicator strain L. monocytogenes V7. All the used procedures yielded active polythene films although the quality of the inhibition was different. The coating was therefore employed to develop active polythene films in an industrial plant. The antimicrobial activity of the industrially produced films was tested in experiments of food packaging involving pork steak and ground beef contaminated by L. monocytogenes V7 at roughly 10(3) CFU cm(-2) and gram respectively. The results of the challenge tests showed the highest antimicrobial activity after 24 h at 4 degrees C, with a decrease of about 1 log of the L. monocytogenes population. CONCLUSIONS: Antimicrobial packaging can play an important role in reducing the risk of pathogen development, as well as extending the shelf life of foods. SIGNIFICANCE AND IMPACT OF THE STUDY: Studies of new food-grade bacteriocins as preservatives and development of suitable systems of bacteriocin treatment of plastic films for food packaging are important issues in applied microbiology and biotechnology, both for implementing and improving effective hurdle technologies for a better preservation of food products.     

Biogenic amine formation and microbial spoilage in chilled garfish (Belone belone belone)--effect of modified atmosphere packaging and previous frozen storage

AIMS: To evaluate biogenic amine formation and microbial spoilage in fresh and thawed chilled garfish. METHODS AND RESULTS: Storage trials were carried out with fresh and thawed garfish fillets at 0 or 5 degrees C in air or in modified atmosphere packaging (MAP: 40% CO2 and 60% N2). During storage, sensory, chemical and microbial changes were recorded and histamine formation by isolates from the spoilage microflora was evaluated at 5 degrees C. Photobacterium phosphoreum was responsible for histamine formation (>1000 ppm) in chilled fresh garfish. The use of MAP did not reduce the histamine formation. Strongly histamine-producing P. phosphoreum isolates formed 2080-4490 ppm at 5 degrees C, whereas below 60 ppm was formed by other P. phosphoreum isolates. Frozen storage inactivated P. phosphoreum and consequently reduced histamine formation in thawed garfish at 5 degrees C markedly. CONCLUSIONS: Photobacterium phosphoreum can produce above 1000 ppm of histamine in chilled fresh garfish stored both in air and in MAP. Freezing inactivates P. phosphoreum, extends shelf life and markedly reduces histamine formation in thawed MAP garfish during chilled storage. SIGNIFICANCE AND IMPACT OF THE STUDY: At 5 degrees C, more than 1000 ppm of histamine was formed in garfish; thus even when it is chilled this product represents a histamine fish-poisoning risk.     

The glutamate decarboxylase acid resistance mechanism affects survival of Listeria monocytogenes LO28 in modified atmosphere-packaged foods

AIMS: The contribution of the glutamate decarboxylase (GAD) acid resistance system to survival and growth of Listeria monocytogenes LO28 in modified atmosphere-packaged foods was examined. METHODS AND RESULTS: The survival and growth of the wild-type LO28 and four GAD deletion mutants (DeltagadA, DeltagadB, DeltagadC, DeltagadAB) in packaged foods (minced beef, lettuce, dry coleslaw mix) during storage at 4, 8 and 15 degrees C were studied. Survival and growth patterns varied with strain, product type, gas atmosphere and storage temperature. In minced beef, the wild-type LO28 survived better (P < 0.05) than the GAD mutant strains at 8 and 15 degrees C. In both packaged vegetables at all storage temperatures, the wild-type strain survived better (P < 0.05) than the double mutant DeltagadAB. The requirement for the individual gad genes varied depending on the packaged food. In the case of lettuce, gadA played the most important role, while the gadB and gadC genes played the greatest role in packaged coleslaw (at 15 degrees C). CONCLUSIONS: This work demonstrates that elements of the GAD system play significant roles in survival of L. monocytogenes LO28 during storage in modified atmosphere-packaged foods. SIGNIFICANCE AND IMPACT OF THE STUDY: A better understanding of how L. monocytogenes behaves in modified atmosphere-packaged foods, and how it responds to elevated carbon dioxide atmospheres.     

Influence of interactions between temperature,ferric ammonium citrate and glycine betaine on the growth of Listeria monocytogenes in a defined medium

AIMS: To investigate interactions, if any, between temperature, ferric ammonium citrate and glycine betaine on the growth of Listeria monocytogenes in modified Pine's medium (Pine et al. 1989). METHODS AND RESULTS: Modified Pine's medium containing 0, 0.044, 0.088 or 0.176 g l(-1) ferric ammonium citrate, and 0 or 1 mM glycine betaine, was inoculated with each of two L. monocytogenes strains and incubated at 4, 25 or 37 degrees C. The optical density at 600 nm, and cell numbers, were determined at appropriate time intervals. At 4 degrees C, but not other temperatures, increasing ferric ammonium citrate resulted in improved growth in the absence, but not the presence, of glycine betaine. The presence of glycine betaine was inhibitory at 25 and 37 degrees C, but not at 4 degrees C. CONCLUSIONS: Interactions affecting the growth kinetics of L. monocytogenes were apparent between the parameters investigated. SIGNIFICANCE AND IMPACT OF THE STUDY: Limitations on the use of modified Pine's medium, and the significance of iron metabolism at lower temperatures, were revealed.     

The differential adherence capabilities of two Listeria monocytogenes strains in monoculture and multispecies biofilms as a function of temperature

AIMS: To determine the differential adherence capabilities at three different temperatures of Listeria monocytogenes Scott A, a clinical food pathogen, and L. monocytogenes FM876, a persistent strain from a milk-processing environment, to stainless steel. METHODS AND RESULTS: Differential adherence was investigated by submerging stainless steel coupons in both 48-h Listeria monocultures and mixed cultures additionally containing Staphylococcus xylosus DP5H and Pseudomonas fragi ATCC 4973. Immunofluorescent microscopy and image analysis techniques were utilized to identify and quantify the L. monocytogenes cells adhering to the steel at 4 degrees C, 18 degrees C and 30 degrees C. The monoculture biofilms consistently contained greater L. monocytogenes numbers than the multispecies biofilms, with the persistent strain FM876 showing significantly greater adherence than strain Scott A. Optimum adherence occurred at 18 degrees C in monoculture biofilms. CONCLUSION: L. monocytogenes strains exhibit differential, temperature-dependent, adherence to stainless steel. SIGNIFICANCE AND IMPACT OF THE STUDY: These results demonstrate temperature dependent biofilm adherence and support previous findings that persistent strains exhibit increased adherence capability.     

Environmental factors influencing the inactivation of Listeria monocytogenes by pulsed electric fields

AIMS: To investigate the influence of the growth phase, growth temperature, storage time, pH and aw of the treatment medium on the resistance of Listeria monocytogenes to pulsed electric fields (PEF). METHODS AND RESULTS: Square wave pulses of 2 micros at a frequency of 1 Hz and 25 and 28 kV cm(-1) were used. Cells were more PEF resistant in the stationary than in the exponential phase at both incubation temperatures investigated (4 and 35 degrees C). Cells grown at 4 degrees C were more PEF sensitive than cells grown at 35 degrees C independent of the growth phase. After a treatment of 25 kV cm(-1) and 800 micros, 1.48, 3.86 and 5.09 log10 cycles of inactivation were obtained at pH 7.0, 5.4 and 3.8, respectively. A reduction in the aw of the treatment medium protected cells against PEF treatments. CONCLUSIONS: The PEF resistance of L. monocytogenes depended on different environmental factors. The influence of growth conditions and treatment medium characteristics should be known and controlled to obtain reproducible and reliable PEF inactivation data. SIGNIFICANCE AND IMPACT OF THE STUDY: Erroneous conclusions and misinterpretation of results are possible if factors affecting the PEF resistance of L. monocytogenes are not considered during PEF inactivation studies.     

The growth of Listeria monocytogenes in cheese packed under a modified atmosphere

The effect of modified atmosphere Packaging (MAP) on the growth of Listeria monocytogenes in mould ripened cheeses was studied at refrigeration temperatures (2-8.3 degrees C) over a storage period of 6 weeks. Control experiments in cling film with no atmospheric modification produced a lag time before growth of up to 1 week and rapid subsequent growth. MAP with a CO2 concentration of less than 20% allowed growth to occur but when O2 was incorporated; the lag time was reduced from 3 to 2 weeks and subsequent growth was also faster, producing an increase in cell numbers of 1.4 log cycles over the incubation period. N2-MAP in the absence of O2 increased the lag time to 3 weeks and slowed growth, while the inclusion of CO2 extended the lag to 3 weeks and slowed subsequent growth even more. In MAP with 80:10:10 (v/v/v) N2:CO2:O2, there was a lag period of 2-3 weeks before growth of L. monocytogenes occurred, while the total viable aerobic count (TVAC) decreased by 2-3 log cycles and the total Lactobacillus count showed little change. It was concluded that MAP was not suitable for preventing the growth of L. monocytogenes in such cheeses.     

Single cell variability of L. monocytogenes grown on liver pâté and cooked ham at 7 degrees C: comparing challenge test data to predictive simulations

AIMS: The variability in growth between individual Listeria monocytogenes cells was investigated on liver paté and cooked ham. These results were compared to Monte Carlo simulations based on data collected previously in broths (Francois et al., submitted for publication). METHODS AND RESULTS: Single cells were isolated by a dilution protocol and inoculated on 15 g samples of liver paté and cooked ham, pasteurized in the packaging. Of each product, 250 samples were inoculated, of which 50 samples were analysed for L. monocytogenes on each analysis day. Results were compared to simulations, based on distributions that describe the variability of the individual cell lag phases and generation times of L. monocytogenes cultivated in broths. Based on the same simulation techniques, the variability effect was investigated for different inoculum levels (10, 100, 10,00 and 10,000 cells). It was demonstrated that the expected variability of the outgrowth of L. monocytogenes in a challenge test is very high for low inoculum levels. CONCLUSIONS: The variability in growth characteristics observed between different single L. monocytogenes cells on foods is very large. The simulations based on the previously collected optical density data in broths, could be confirmed by foods inoculated with single L. monocytogenes cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The large variability between different individual L. monocytogenes cells has serious consequences for the experimental design of a challenge test. One thousand cells per portion are necessary in order to reduce the variability to acceptable levels and quantify the behaviour of the pathogen consistently with a reasonable number of challenge tests.     

Minimum growth temperatures of Listeria monocytogenes and non-haemolytic Listeria

Minimum growth temperatures and those of decreased growth were determined for 100 strains of listerias. The ability of 78 strains of Listeria monocytogenes isolated from animals and 22 non-haemolytic strains to grow at low temperatures was studied, using a flooding technique, in a plate-type continuous temperature gradient incubator at temperatures between -1.6 and 14.5 degrees C. The mean minimum temperature for L. monocytogenes was +1.7 +/- 0.5 degrees C. The growth of non-haemolytic listerias was unobservable at +1.7 +/- 0.5 degrees C. The L. monocytogenes strains grew at about 0.6 degrees C lower than the non-pathogenic strains. No differences in growth temperatures were observed among L. monocytogenes strains isolated from different sources. The serovars with the OI antigen grew at lower temperatures (+1.0 +/- 0.3 degrees C) than the other common serovar 4b (+1.3 +/- 0.4 degrees C). The results indicate that L. monocytogenes grows better than non-haemolytic strains under cold conditions. The possible role of haemolysins as growth factors is also discussed.     

Evidence of an antilisterial factor induced by wounding of iceberg lettuce tissues

AIMS: To examine the influence of wound-associated reactions in cut iceberg lettuce (Lactuca sativa L.) tissues on the fate of Listeria monocytogenes. METHODS AND RESULTS: Aqueous extracts prepared from shredded iceberg lettuce before and after storage in high oxygen permeability film were inoculated with L. monocytogenes. Listeria monocytogenes grew in extracts prepared from fresh lettuce. In contrast, inhibition ranging from arrested growth to a decline in cell viability was observed in extracts prepared from samples stored for 1-3 days. Similar behaviour was evident in lettuce shreds inoculated with 10(5) CFU g(-1)L. monocytogenes immediately after processing or after 3 days in storage. Heat treatment of the cut tissues at 47 degrees C for 3 min before storage diminished the inhibitory effect. CONCLUSIONS: The results provided evidence that an antilisterial factor or factors are released by wounded iceberg lettuce tissues. Antilisterial activity was mitigated by heat treatment of the lettuce. SIGNIFICANCE AND IMPACT OF STUDY: This study indicates that intrinsic factors associated with plant metabolism could play a significant role in the ecology of human pathogens in packaged horticultural products.