Growth potential of Listeria monocytogenes strains in mixed ready-to-eat salads

In this study, a microbiological challenge test in three artificially contaminated retail mixed mayonnaise-based ready-to-eat salads stored at refrigerator temperatures (3 °C and 7 °C) for 48 h was carried out. Shrimp-tomato salad, smoked ham salad and garlic cheese salad were separately contaminated by a suspension of particular Listeria monocytogenes strains. The number of L. monocytogenes, Enterobacteriaceae, staphylococci and total plate count (CFU/g) was determined. Listeria monocytogenes growth potential in the salads was calculated and evaluated.A significant increase in total plate count and L. monocytogenes count throughout storage of all three investigated salads was found. Enterobacteriaceae levels were high at the beginning in all salads but significantly (p < 0.05) decreased throughout the experiment depending on the temperature.All investigated L. monocytogenes strains demonstrated growth at both temperatures but expressed different growth potential. Especially garlic cheese salad and smoked ham salad were able to support the growth of Listeria. Shrimp-tomato salad supported growth the least. The growth potential increased with the increasing temperature and exceeded 0.5 log₁₀ CFU/g in many cases. If the potential for growth is > 0.5 log₁₀ CFU/g, food products can potentially endanger human health. Reference strain (ATCC 7644) showed the least growth potential almost in all cases in comparison with strains isolated from frozen pollock loins and from thermally treated specialty sausage containing preservatives. To eliminate the occurrence of microbiological risks, the shelf-life of the studied salads was estimated.     

A comparison between E-beam irradiation and high pressure treatment for cold-smoked salmon sanitation: microbiological aspects

The effectiveness of electron beam irradiation and high pressure treatment for the sanitation of cold-smoked salmon from two points of view, microbial safety and shelf-life extension, was compared. From the response of L. monocytogenes INIA H66a to irradiation, a D value of 0.51 kGy was calculated. For samples stored at 5 °C, 1.5 kGy would be sufficient to attain a Food Safety Objective (FSO) of 2 log₁₀cfu/g L. monocytogenes for a 35-day shelf-life, whereas 3 kGy would be needed in the case of a temperature abuse (5 °C + 8 °C). Pressurization at 450 MPa for 5 min was considered to be an insufficient treatment, since the FSO of 2 log₁₀cfu/g L. monocytogenes was only attained for a shelf-life of 21 days at 5 °C. However, treatment at 450 MPa for 10 min achieved this FSO for samples held during 35 days at 5 °C, or during 21 days under temperature abuse (5 °C + 8 °C) conditions. Irradiation at 2 kGy kept the microbial population of smoked salmon below 6 log₁₀cfu/g after 35 days at 5 °C, with negligible or very light changes in its odor. Pressurization at 450 MPa for 5 min also kept the microbial population below 6 log₁₀cfu/g after 35 days at 5 °C and did not alter odor, but affected negatively the visual aspect of smoked salmon.     

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.     

A multiplex magnetic capture hybridisation and multiplex Real-Time PCR protocol for pathogen detection in seafood

Seafood could become a source of bacterial pathogens by exposure to contaminated water or through processing practices, thus representing a public health hazard. Conventional culture-based analytical methods take several days to be completed, while the molecular rapid identification of bacterial pathogens is crucial for effective disease control. The developed application consist of a multiplex magnetic capture hybridisation (mMCH) assay for the simultaneous isolation of Salmonella spp. and Listeria monocytogenes DNA from seafood, using paramagnetic amino-modified nanoparticles with capture oligonucleotides, and a triplex Real-Time PCR with an Internal Amplification Control (IAC), in accordance with ISO 22174. The detection probability was 100% with 10 genome equivalents of each target species co-amplified in the same reaction. The complete molecular procedure was tested on raw and smoked salmon fillets artificially contaminated with known amounts of one or both target bacteria (1–10³ cfu/g), directly or after culture enrichment, and compared for equivalence with the standard methods. Results revealed a complete agreement between the two approaches, with a sensitivity of 1 cfu/g, in enriched samples, and higher sensitivity (10²–10³ cfu/g) of the molecular method in samples examined before culture enrichment. The proposed procedure was also able to identify a natural contamination by L. monocytogenes in smoked salmon with a considerable shortening of time.     

Effects of γ-irradiation on Listeria monocytogenes population,colour, texture and sensory properties of Feta cheese during cold storage

Feta, a white brine cheese, was produced and contaminated with Listeria monocytogenes. Contamination occurred either at the beginning (pre-process contamination) or at the end of Feta manufacturing (post-process contamination). In the first case the milk was contaminated with 10³ cfu/ml, and 2 months later, in the final product, the L. monocytogenes population was approximately 10⁵ cfu/g. In the second case, the brine (NaCl, 7% w/v), in which the Feta was packaged, was contaminated with 10³ cfu/ml. Contaminated Feta samples were vacuum-packaged and exposed to irradiation doses of 1.0, 2.5 and 4.7 kGy and stored at 4 °C for a month. In the pre-process contaminated samples none of the irradiation doses eliminated L. monocytogenes; however the highest dose reduced the viable population to a level which is in compliance with EC regulations. In the post-process contamination, the 2.5 kGy and 4.7 kGy doses reduced L. monocytogenes counts below the detection limit. Irradiation had no effect on the texture of Feta. Irradiation at 4.7 kGy increased Feta's redness and decreased its yellowness and lightness. Sensorial analyses showed that at the 4.7 kGy dose, the aroma profile of Feta was temporarily affected, since it was restored after 30 days of cold storage.     

Desiccation of adhering and biofilm Listeria monocytogenes on stainless steel: Survival and transfer to salmon products

The foodborne bacterial pathogen, Listeria monocytogenes, commonly contaminates foods during processing, where the microorganisms are potentially subjected to low relative humidity (RH) conditions for extended periods of time. The objective of this study was to examine survival during desiccation (43% RH and 15 °C) of biofilm L. monocytogenes N53-1 cells on stainless steel coupons and to assess subsequent transfer to salmon products. Formation of static biofilm (2 days at 100% RH and 15 °C) prior to desiccation for 23 days significantly (P < 0.05) improved survival of cells desiccated in initial low salt concentrations (0.5%) compared to the survival for non-biofilm cells also desiccated in low salt, indicating the protective effect of the biofilm matrix. Osmoadaptation of cells in 5% NaCl before formation of the static biofilm significantly (P < 0.05) increased long-term desiccation survival (49 days) irrespectively of the initial salt levels (0.5% and 5% NaCl). The efficiency of transfer (EOT) of desiccated biofilm cells was significantly (P < 0.05) lower than EOTs for desiccated non-biofilm bacteria, however, as biofilm formation enhanced desiccation survival more bacteria were still transferred to smoked and fresh salmon. In conclusion, the current work shows the protective effect of biofilm formation, salt and osmoadaptation on the desiccation survival of L. monocytogenes, which in turn increases the potential for cross-contamination during food processing.     

Loss of viability of Listeria monocytogenes in contaminated processed cheese during storage at 4, 12 and 22 °C

The behaviour of Listeria monocytogenes in a processed cheese product was evaluated over time by inoculating the product with three different L. monocytogenes strains (Scott A, CA and a strain isolated from processed cheese) at three different inoculation levels (ca. 6 × 10⁵, ca. 6 × 10³ and 10² CFU/g of cheese or less) and after storage of the contaminated products at 4, 12 or 22 °C. Growth of L. monocytogenes was not observed in any of the experimental trials (experiments involving different combinations of strain, inoculum level and storage temperature) throughout the storage period. L. monocytogenes populations decreased over time with a rate that was strain- and storage temperature-dependent. Nonetheless, for cheeses that had been inoculated with the higher inoculum and stored at 4 °C viable populations of L. monocytogenes could be detected for up to nine months post-inoculation. The L. monocytogenes survival curves obtained from the different trials were characterised by a post-inoculation phase during which the populations remained essentially unchanged (lag phase) followed by a phase of logarithmic decline. The duration of the lag phase and the rate of inactivation of L. monocytogenes in the different trials were estimated based on data from the linear descending portions of the survival curves. In addition, a non-linear Weibull-type equation was fitted to the data from each survival curve with satisfactory results. The results of the present study emphasize that, according to the definition laid down in the European Union Regulation 1441/2007, the processed cheese product tested in this work should be considered and classified as one that does not support the growth of L. monocytogenes under reasonable foreseeable conditions of distribution and storage. However, post-processing contamination of the product should be austerely avoided as the pathogen can survive in the product for extended periods of time, particularly under refrigerated storage (4 °C).     

Effect of freezing on the physicochemical, textural and sensorial characteristics of salmon (Salmo salar) smoked with a liquid smoke flavouring

This study reports the effect of different refrigeration/freezing treatments on the physicochemical, textural and sensorial properties of farmed Atlantic salmon (Salmo salar) treated with a commercial liquid smoke flavouring. Observations were made on three groups of fillets - group RFS: salted, smoked and stored at 4 °C; group BFS: frozen at −25 °C for 24 h, thawed, salted, smoked and stored at 4 °C; and group AFS: salted, smoked and frozen at −25 °C for 24 h and stored at −18 °C - over a period of 45 days. Scores (on a scale of 1-9) were provided for different sensorial attributes by a panel of 10 trained tasters. Sixty percent of the panellists consistently preferred the AFS fillets. The maximum shelf life associated with each treatment was defined as the last sampling day on which a mean score of ≤5 was awarded for the fillet sensorial attributes by ≥50% of the panellists. Freezing the salmon for 24 h before smoking (BFS) did not increase its shelf life (30 days) over that of refrigerated smoked salmon (RFS). In addition, the former treatment had a negative effect on the adhesiveness, cohesiveness, smoke odour intensity and colour intensity of the flesh. However, maintaining the fish frozen at −18 °C (AFS) increased its shelf life (>45 days) and invested the flesh with greater firmness, cohesiveness and colour intensity.     

Bactericidal activity of lauric arginate in milk and Queso Fresco cheese against Listeria monocytogenes cold growth

Lauric arginate (LAE) at concentrations of 200 ppm and 800 ppm was evaluated for its effectiveness in reducing cold growth of Listeria monocytogenes in whole milk, skim milk, and Queso Fresco cheese (QFC) at 4°C for 15 to 28 d. Use of 200 ppm of LAE reduced 4 log cfu/mL of L. monocytogenes to a nondetectable level within 30 min at 4°C in tryptic soy broth. In contrast, when 4 log cfu/mL of L. monocytogenes was inoculated in whole milk or skim milk, the reduction of L. monocytogenes was approximately 1 log cfu/mL after 24 h with 200 ppm of LAE. When 800 ppm of LAE was added to whole or skim milk, the initial 4 log cfu/mL of L. monocytogenes was nondetectable following 24 h, and no growth of L. monocytogenes was observed for 15 d at 4°C. With surface treatment of 200 or 800 ppm of LAE on vacuum-packaged QFC, the reductions of L. monocytogenes within 24 h at 4°C were 1.2 and 3.0 log cfu/g, respectively. In addition, the overall growth of L. monocytogenes in QFC was decreased by 0.3 to 2.6 and by 2.3 to 5.0 log cfu/g with 200 and 800 ppm of LAE, respectively, compared with untreated controls over 28 d at 4°C. Sensory tests revealed that consumers could not determine a difference between QFC samples that were treated with 0 and 200 ppm of LAE, the FDA-approved level of LAE use in foods. In addition, no differences existed between treatments with respect to flavor, texture, and overall acceptability of the QFC. Lauric arginate shows promise for potential use in QFC because it exerts initial bactericidal activity against L. monocytogenes at 4°C without affecting sensory quality.     

Control of Growth of L. monocytogenes in Fresh Salmon using Microgard and Nisin

Pathogens like Listeria monocytogenes in fresh, or even in cold smoked salmon, have become a major public health concern for the salmon processing industry and government agencies. The effect of bacteriocin solutions (Microgard^™ and Nisin) on reducing total microbial counts, inhibiting Listeria monocytogenes, and prolonging the shelf-life was evaluated. Listeria monocytogenes was inoculated onto chilled and on frozen and thawed salmon samples. The combination of Nisin and Microgard reduced the total aerobic bacteria populations of fresh chilled salmon by 2 log (P<0.05) and increased its shelf-life, at 6 °C, by 3-4 d, as compared with the control. The above bacteriocin combination also reduced the growth of inoculated Listeria monocytogenes in frozen-thawed salmon and increased its shelf-life from 5 to 10 d at 6 °C. The bacteriocin treatment did not affect surface pH values or color of the fish.     

Listeria monocytogenes in genussfertigen Lebensmitteln: eine Auswertung der amtlichen Untersuchungen in der Schweiz der Jahre 2006–2008

In the years 2006–2008, 13.978 ready-to-eat foodstuffs were tested by official laboratories of food control in Switzerland for compliance with legal limits for Listeria monocytogenes. Totally, the pathogen could be detected in 67 foods (0,5 % of all samples). Most frequently, raw meat cured sausages (proportion of positive samples 3,9 %) were contaminated followed by smoked fishes (1,4 %) and semi-hard cheeses (1,1 %). For soft cheese, a rather low contamination frequency of 0,3 % was shown. Quantification of L. monocytogenes was possible in 18 ready-to-eat foods from the market and six out of them showed high counts of >1.000 CFU per gram. Concerned were a sandwich with smoked salmon and other components (250.000 CFU/g), smoked salmon (180.000 CFU/g), smoked trout (13.000 CFU/g), semi-hard cheese (9.200 CFU/g) and salami (5.500 and 1.850 CFU/g). In connection with cases of listeriosis, the highest measured count (5 × 10⁷ CFU/ml) was found in liquid cream of a private household where it was probably contaminated and not adequately stored. Surprisingly, in 931 desserts and confectioneries, in 384 ice-creams, in 3.567 pre-cooked foods and in 806 samples of raw fruits or vegetables, L. monocytogenes was never isolated and in 720 delicatessen salads only once (Celery salad with <100 CFU/g). The evaluation of a high number of laboratory data allowed identifying the current focal points of risk, an information which is important for a risk-based design of future control activities.     

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.     

Heat resistance of Listeria species to liquid whole egg ultrapasteurization treatment

The lethality of ultrapasteurization treatments (70 °C/1.5 min.) applied at constant temperature (isothermal condition) and at a constantly raising temperature of 2 °C/min (non-isothermal condition) in liquid whole egg (LWE) against two strains of Listeria monocytogenes (STCC 5672 and 4032) and one of Listeria innocua has been investigated. Isothermal survival curves up to 71 °C were obtained, which followed first-order inactivation kinetics. The obtained D_t values indicated that L. innocua was significantly (p < 0.05) more heat resistant than L. monocytogenes strains. Non-significant (p > 0.05) differences were observed among z values (12.4 ± 0.4 °C, 13.1 ± 0.4 °C and 12.2 ± 0.7 °C for L. innocua and L. monocytogenes 5672 and 4032, respectively). Based on obtained D_t and z values, isothermal ultrapasteurization treatment (70 °C/1.5 min.) would provide 3.5-, 5.0-, and 6.5-Log₁₀ cycles of L. innocua and L. monocytogenes 5672 and 4032, respectively. Non-isothermal heating lag phase increased the thermotolerance of Listeria species in LWE. The simulated industrial pasteurization treatment for LWE (heating-up phase from 25 to 70 °C followed by 1.5 min. at 70 °C) would attain 5-Log₁₀ reductions of L. monocytogenes 5672 and 4032, and 3.7-Log₁₀ reductions of L. innocua. Therefore, the safety level of industrial ultrapasteurization concerning L. monocytogenes could be lower than that estimated with data obtained under isothermal conditions.     

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.     

Antibacterial effects of natural tenderizing enzymes on different strains of Escherichia coli O157:H7 and Listeria monocytogenes on beef

This study determined the efficacy of actinidin and papain on reducing Listeria monocytogenes and three mixed strains of Escherichia coli O157:H7 populations on beef. The average reduction of E. coli O157:H7 was greater than that of L. monocytogenes and higher concentrations of either protease yielded greater reduction in bacterial populations. For instance, actinidin at 700 mg/ml significantly (p ≤ 0.05) reduced the population of L. monocytogenes by 1.49 log cfu/ml meat rinse after 3 h at 25 & 35 °C, and by 1.45 log cfu/ml rinse after 24 h at 5 °C, while the same actinidin concentration significantly reduced the populations of three mixed strains of E. coli O157:H7 by 1.81 log cfu/ml rinse after 3 h at 25 & 35 °C, and 1.94 log cfu/ml rinse after 24 h at 5 °C. These findings suggest that, in addition to improving the sensory attributes of beef, proteolytic enzymes can enhance meat safety when stored at suitable temperatures.     

Efficacy of bacteriophage LISTEXP100 combined with chemical antimicrobials in reducing Listeria monocytogenes in cooked turkey and roast beef

The aim of this study was to verify the effectiveness of the commercially available anti-Listeria phage preparation LISTEX^™P100 in reducing Listeria monocytogenes on ready-to-eat (RTE) roast beef and cooked turkey in the presence or absence of the chemical antimicrobials potassium lactate (PL) and sodium diacetate (SD). Sliced RTE meat cores at 4 and 10 °C were inoculated with cold-adapted L. monocytogenes to result in a surface contamination level of 10³ CFU/cm². LISTEX^TMP100 was applied at 10⁷ PFU/cm² and samples taken at regular time intervals during the RTE product's shelf life to enumerate viable L. monocytogenes. LISTEX^™P100 was effective during incubation at 4 °C with initial reductions of L. monocytogenes of 2.1 log₁₀ CFU/cm² and 1.7 log₁₀ CFU/cm², respectively, for cooked turkey and roast beef without chemical antimicrobials (there was no significant difference to the initial L. monocytogenes reductions in the presence of LISTEX^TMP100 for cooked turkey containing PL and roast beef containing SD-PL). In the samples containing no chemical antimicrobials, the presence of phage resulted in lower L. monocytogenes numbers, relative to the untreated control, of about 2 log CFU/cm² over a 28-day storage period at 4 °C. An initial L. monocytogenes cell reduction of 1.5 log₁₀ CFU/cm² and 1.7 log₁₀ CFU/cm², respectively, for cooked turkey and roast beef containing no chemical antimicrobials was achieved by the phage at 10 °C (abusive temperature). At this temperature, the L. monocytogenes cell numbers of samples treated with LISTEX™ P100 remained below those of the untreated control only during the first 14 days of the experiment for roast beef samples with and without antimicrobials. On day 28, the L. monocytogenes numbers on samples containing chemical antimicrobials and treated with LISTEX^TMP100 stored at 4 and 10 °C were 4.5 log₁₀ CFU/cm² and 7.5 log₁₀ CFU/cm², respectively, for cooked turkey, and 1.2 log₁₀ CFU/cm² and 7.2 log₁₀ CFU/cm², respectively, for roast beef. In both cooked turkey samples with and without chemical antimicrobials stored at 10 °C, the phage-treated samples had significantly lower numbers of L. monocytogenes when compared to the untreated controls throughout the 28-day storage period (P < 0.0001). For roast beef and cooked turkey containing chemical antimicrobials treated with LISTEX^TMP100 and stored at 4 °C, no more than a 2 log CFU/cm² increase of L. monocytogenes was observed throughout the stated shelf life of the product. This study shows that LISTEX^™P100 causes an initial reduction of L. monocytogenes numbers and can serve as an additional hurdle to enhance the safety of RTE meats when used in combination with chemical antimicrobials.     

Determination of the effect of plant essential oils obtained by supercritical fluid extraction on the growth and viability of Listeria monocytogenes in broth and food systems using flow cytometry

The antimicrobial properties of oregano, rosemary and laurel extracts obtained by supercritical fluid extraction were investigated by examining their influence on the growth and viability of Listeria monocytogenes in laboratory medium and broccoli juice at 30 and 8 °C. Important decreases in the L. monocytogenes population were shown in presence of all the extracts obtained from rosemary and one oregano extract. The counts were reduced below the level of detection after 4 h of exposure at 30 °C in laboratory medium. A bactericidal effect was observed also when L. monocytogenes was exposed to rosemary at 30 °C and 8 °C in broccoli juice. Significant reductions in growth rate and an increase in lag phase of L. monocytogenes were observed in presence of some of the laurel and oregano extracts at both temperatures.Flow cytometry was used as a rapid method to determine the antibacterial effect of supercritical extracts and the physiological state of L. monocytogenes. Bacterial viability performed by dual staining of L. monocytogenes with SYTO 9 and propidium iodide revealed three different cell populations, specifically, living, dead and compromised cells. Live cell percentage decreased with the time of exposure, whereas the percentage of compromised cells remained constant and the dead cells increased in the same period.     

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.     

Effects of brine concentration on shelf-life of hot-smoked tilapia (Oreochromis niloticus) stored at 4 °C

This work evaluated the effect of brine concentration on the shelf-life of hot-smoked tilapia (Oreochromis niloticus) stored at 4 °C. The fish were brined in solutions of 5%, 10%, and 15% NaCl and unsalted fish were used as controls. The fish were then smoked, cooled and stored at 4 °C. Oxidative rancidity measured by the peroxide value (PV), and thiobarbituric acid number (TBA) showed increases with the storage time and also as a result of the increasing salt content in fish muscle. Hot smoked tilapia can be stored safely under refrigerated conditions for over 35 days, and 5% brine was found to be optimal for storage.