Monocytogenes Growth Research Articles

Determination of antilisterial effect of some microbial isolates from traditional zlatar cheese during the fermentation of soft white cheese.

The aim of this study was to select autochthonous lactic acid bacteria (LAB) isolates with antilisterial activity from Zlatar cheese and to evaluate the ability of selected LAB to control Listeria monocytogenes growth during soft white cheese production. The genotype characterization of isolated LAB (n = 93) was done using PCR method by 16S rRNA sequencing. In this way, the following isolates were detected: Lactococcus lactis ssp. lactis (40 isolates), Enterococcus faecalis (30), Lactobacillus plantarum (12), Leuconostoc mesenteroides ssp. mesenteroides (3) Lactobacillus garvieae (3), Lactobacillus curvatus (2), Lactobacillus casei (1), Enterococcus faecium (1) and Staphylococcus hominis (1). Each isolated LAB was tested for bacteriocin-producing ability. It was determined that two LAB isolates had bactericidal properties: Lactococcus lactis ssp. lactis SRB/ZS/094 and Enterococcus faecalis SRB/ZS/090. Semi-purified of enterococcal bacteriocin (enterocin) was isolated using precipitation procedures with ammonium sulphate. Its properties were determined (strength and range of activities). Isolated enterocin and bacteriocin-producing Lactococcus strain showed significant antimicribial activity against Listeria monocytogenes, but still the inhibition activity of Staphylococcus aurues and Escherichia coli was not detected. Based on the obtained laboratory results, in the second phase of the research, the antilisterial effect of bacteriocin isolated from Enterococcus faecalis SRB/ZS/090 and cells Lactococcus lactis ssp. lactis SRB/ZS/094 were determined, that are added as additives in the production of soft white cheese through five variants. Cheese supplemented with enterocin (E2) had the lowest aerobic mesophilic bacteria count, indicating that enterocin (E2) play an important role for bio-preservation.

Effect of moderate intensity pulsed electric field on shelf-life of chicken breast meat

ABSTRACT 1. Moderate intensity pulsed electric field (MIPEF) is a fairly new research topic for microbial inactivation on muscle foods. In this study, indirect application of MIPEF (2.5 kV/cm, 4.67 kV/cm, 7 kV/cm) was evaluated in terms of the growth of some pathogenic bacteria and the quality characteristics of chicken breast meat during cold storage (4°C). 2. This study was conducted in two stages. In the first stage, pure cultures of S. enteritidis, P. aeruginosa, S. aureus, L. monocytogenes, E. coli and C. jejuni were inoculated in broth and stored in a refrigerator at 4°C under MIPEF. Both E. coli and C. jejuni showed a remarkable resistance to MIPEF under 4.67 kV/cm and 7 kV/cm, respectively. 3. In the second stage, MIPEF was applied to packaged chicken breast samples. The limit for total mesophilic aerobic bacteria (TMAB) count was exceeded in 4.67 and 7 kV/cm applied groups 2 d later than seen in the control group (C). Almost 2 log differences were determined for total coliform bacteria in 4.67 and 7 kV/cm applied groups as compared to C (P < 0.05). Inoculated P. aeruginosa count remained the same, whereas L. monocytogenes growth was promoted by 4.67 kV/cm (P < 0.05). 4. pH value, CIE L*, b*, C* colour values of chicken breast fillets were not affected by MIPEF application while ΔE values showed the maximum change in the C group. 5. This study demonstrated that improved shelf-life of chicken breast fillets was provided by moderate intensity DC-pulses in combination with cold storage.

Prevalence of Multidrug-Resistant Listeria monocytogenes in Dairy Products with Reduction Trials Using Rosmarinic Acid, Ascorbic Acid, Clove, and Thyme Essential Oils

Continuous monitoring of Listeria spp., particularly Listeria monocytogenes, in foods is a mandatory task for food safety and microbiology sectors. This study aimed to determine the prevalence and antimicrobial resistance patterns of L. monocytogenes in milk and dairy products retailed in Egypt. Furthermore, an experimental trial was conducted to investigate the antilisterial effects of some phytochemicals. A total of 200 samples (market raw milk, Kareish cheese, Damietta cheese, and plain yoghurt, 50 each) were collected and examined for detection of Listeria spp. The results revealed that 8, 12, 1, and 0 samples of market raw milk, Damietta cheese, Kareish cheese, and plain yoghurt were contaminated with Listeria spp., respectively. Antimicrobial sensitivity testing revealed that all L. monocytogenes isolates (15) were resistant to streptomycin and erythromycin. Molecular analysis revealed that 86.67% of L. monocytogenes harbored hylA virulent gene. Use of rosmarinic acid, ascorbic acid, thyme, and clove essential oils significantly ( P &lt; 0.05 ) reduced L. monocytogenes growth in soft cheese—artificially contaminated with L. monocytogenes throughout a 4-week incubation period. In conclusion, strict hygienic conditions should be adopted during the preparation and distribution of dairy products. In addition, rosmarinic acid, ascorbic acid, clove, and thyme essential oils are good candidates as food preservatives with antilisterial activities.

Effects of temperature abuse on the growth and survival of Listeria monocytogenes on a wide variety of whole and fresh-cut fruits and vegetables during storage

Twenty types of fruits and vegetables representing a broad range of fresh produce categories were assessed for the growth potential and growth kinetics of Listeria monocytogenes under conditions reflecting commercial practices for storage and distribution and conditions reflecting sustained temperature abuse. Whole and fresh-cut produce was obtained from various commercial sources and inoculated with L. monocytogenes strains to evaluate the survival and growth for up to 25 days, depending on the projected commercial storage or retail shelf life for each commodity. The L.monocytogenes populations showed a sustained decline on all tested whole fruits, including avocado, blueberry, grape, mango, peach, green pepper, and tomato. Under “normal” storage conditions (defined as those reflective of common commercial practices), significant L. monocytogenes growth was only observed on fresh-cut cantaloupe (∼0.8 log) and fresh-cut mango (∼0.6 log). Exposure to temperature abuse conditions did not change the overall trends of L. monocytogenes survival on the whole fruits but did result in significant growth on several fresh-cut products, including celery, cauliflower, mango, onion, romaine lettuce, and cantaloupe. After inoculation on certain products (e.g., fresh-cut carrot and whole peach), L. monocytogenes seemed to lose cultivability rapidly but did retain viability as determined by the maintenance of cell membrane integrity.

Effect of Anolyte on S. Typhimurium and L. monocytogenes Growth in Minced Pork and Beef Cuts.

In this paper, anolyte is considered as a possible disinfectant for inhibiting the growth of bacteria in meat (beef cuts and minced pork). Meat cuts were contaminated with two concentrations of L. monocytogenes and S. Typhimurium, as these are the most common meat pathogens that are closely regulated by the EU, and treated with two different concentrations of anolyte: 20% for beef cuts and 18% for minced pork. Then, the total viable count (TVC), L. monocytogenes count and S. Typhimurium count were determined. In meat cuts and minced pork, anolyte was able to reduce TVC, S. Typhimurium and L. monocytogenes counts effectively, significantly decreasing L. monocytogenes and S. Typhimurium counts after spraying and throughout 29 days of incubation at 0–4 °C. TVC was reduced after spraying and for 10 days of incubation but later increased to be the same as before spraying with anolyte. Anolyte was effective when spraying beef cuts with a 20% solution for 60 s against pathogenic bacteria L. monocytogenes and Salmonella spp. and also when using it at a concentration of 18% from the minced meat mass. Initially, anolyte significantly decreased TVC, however during the storage period (10–29 days) TVC increased but remained significantly lower compared to control. Anolyte was effective in reducing L. monocytogenes and S. Typhimurium counts throughout the study, and after 29 days of incubation, these bacteria could not be detected in the samples treated with anolyte.

Logistic modeling to predict the minimum inhibitory concentration (MIC) of olive leaf extract (OLE) against Listeria monocytogenes.

Olive leaf extract (OLE) has been increasingly recognized as a natural and effective antimicrobial against a host of foodborne pathogens. This study attempts to predict the minimum inhibitory concentration (MIC) of OLE against Listeria monocytogenes F2365 by utilizing the asymptotic deceleration point (PDA) in a logistic model (LM), namely MIC-PDA. The experimental data obtained from the inhibitory rate (IR) versus OLE concentration against L. monocytogenes were sufficiently fitted (R2 = 0.88957). Five significant critical points were derived by taking the multi-order derivatives of the LM function: the inflection point (PI), the maximum acceleration point (PAM), the maximum deceleration point (PDM), the absolute acceleration point (PAA), and the asymptotic deceleration point (PDA). The PDA ([OLE] = 37.055 mg/mL) was employed to approximate the MIC-PDA. This MIC value was decreased by over 42% compared to the experimental MIC of 64.0 mg/mL, obtained using the conventional 2-fold dilution method (i.e., MIC-2fold). The accuracy of MIC-PDA was evaluated by an in vitro L. monocytogenes growth inhibition assay. Finally, the logistic modeling method was independently validated using our previously published inhibition data of OLE against the growths of Escherichia coli O157:H7 and Salmonella enteritidis. The MIC-PDA (for [OLE]) values were estimated to be 41.083 and 35.313 mg/mL, respectively, compared to the experimental value of 62.5 mg/mL. Taken together, MIC-PDA, as estimated from the logistic modeling, holds the potential to shorten the time and reduce cost when OLE is used as an antimicrobial in the food industry.

Growth and Biocontrol of Listeria monocytogenes in Greek Anthotyros Whey Cheese without or with a Crude Enterocin A-B-P Extract: Interactive Effects of the Native Spoilage Microbiota during Vacuum-Packed Storage at 4 °C.

Effective biopreservation measures are needed to control the growth of postprocess Listeria monocytogenes contamination in fresh whey cheeses stored under refrigeration. This study assessed growth and biocontrol of inoculated (3 log10 CFU/g) L. monocytogenes in vacuum-packed, fresh (1-day-old) or ‘aged’ (15-day-old) Anthotyros whey cheeses, without or with 5% of a crude enterocin A-B-P extract (CEntE), during storage at 4 °C. Regardless of CEntE addition, the pathogen increased by an average of 2.0 log10 CFU/g in fresh cheeses on day 15. Gram-negative spoilage bacteria also increased by an average of 2.5 log10 CFU/g. However, from day 15 to the sell-by date (days 35–40), L. monocytogenes growth ceased, and progressively, the populations of the pathogen declined in most cheeses. This was due to an unmonitored, batch-dependent natural acidification by spoilage lactic acid bacteria, predominantly Leuconostoc mesenteroides, which reduced the cheese pH to 5.5, and finally to ≤5.0. The pH reductions and associated declines in pathogen viability were greater in the CEntE-treated samples within each batch. L. monocytogenes failed to grow in cheeses previously ‘aged’ in retail for 15 days. Overall, high population levels (>7.5 log10 CFU/g) of psychrotrophic Enterobacteriaceae, particularly Hafnia alvei, were associated with an extended growth and increased survival of L. monocytogenes during storage.

Analysis of the Bioprotective Potential of Different Lactic Acid Bacteria Against Listeria monocytogenes in Cold-Smoked Sea Bass, a New Product Packaged Under Vacuum and Stored at 6 ± 2°C.

The aim of the work was to monitor the presence of Listeria monocytogenes in cold-smoked fish products (trout, salmon, and sea bass) marketed in Italy. Cold-smoked sea bass is a new product that has not yet been commercialized and was collected from the production facility. Monitoring data have shown that cold-smoked products can be contaminated by L. monocytogenes, the presence of which has been highlighted mainly by enrichment culture (presence in 25 g). The isolated Listeria were serotyped and belonged mainly to low-virulence serotypes (1/2c), followed by serotypes 1/2a, 1/2b, and 4b. Furthermore, considering the ability of L. monocytogenes to grow in these products due to their chemical–physical characteristics (pH > 6.0, Aw > 0.97) and long shelf life at 4°C, an additional aim was to verify the activity of different bioprotective starters, including Lactilactobacillus sakei (LAK-23, Sacco srl, Via Alessandro Manzoni 29/A, 22071 Cadorago, CO, Italy), Carnobacterium spp., Lacticaseibacillus casei (SAL 106), and Lacticaseibacillus paracasei (SAL 211), in cold-smoked sea bass. All starters were bacteriocin producers. For this experiment, smoked sea bass samples were intentionally inoculated with a mixture of three different strains of L. monocytogenes and of each starter culture. After inoculation, the smoked sea bass were vacuum-packed and stored at 6 ± 2°C for 60 days, simulating the typical abuse storage temperature of markets and home refrigerators. At 0, 15, 30, 45, and 60 days, the sea bass samples were analyzed to evaluate the effectiveness of the starters against L. monocytogenes. Listeria monocytogenes growth was prevented only by the addition of the LAK-23 starter. Indeed, at the end of the shelf life, the amount of L. monocytogenes observed was similar to that in the inoculum. Consequently, the use of this starter can allow the inclusion of cold-smoked sea bass or smoked fish products in category 1.3 of Regolamento CE 2073/2005, which are products that do not support the growth of this microorganism. Finally, the activity of the LAK-23 starter did not produce an off flavor or off odor in the smoked sea bass.

Control of Listeria monocytogenes growth and virulence in a traditional soft cheese model system based on lactic acid bacteria and a whey protein hydrolysate with antimicrobial activity

“Torta del Casar” is a Spanish soft-ripened cheese made with sheep's raw milk and subjected to a short ripening process, which favors the growth of pathogenic microorganisms including Listeria monocytogenes. The development of strategies to control pathogens and minimize health risks associated with the presence of L. monocytogenes in these products is of great interest. In this regard, the anti-Listeria activity of a whey protein hydrolysate (ProH) alone or combined with six lactic acid bacteria strains isolated from cheese was evaluated in this study as a biocontrol strategy using a “Torta del Casar” cheese-based medium. The most active combinations of lactic acid bacteria assayed induced a reduction higher than two logarithmic units in the growth of L. monocytogenes (serotype 4b) compared to their respective control when they were co-inoculated in “Torta del Casar” cheese-based medium at 7 °C for 7 days. In addition, the observed downregulation of some key virulence genes of L. monocytogenes suggests that the strain Lactiplantibacillus plantarum B2 alone and combined with the strain Lactiplantibacillus spp. B4 are good candidates to be used as biocontrol agents against L. monocytogenes growth in traditional soft cheeses based on raw milk during their storage at refrigeration temperatures.

Wet versus Dry Inoculation Methods Have a Significant Effect of Listeria monocytogenes Growth on Many Types of Whole Intact Fresh Produce

Listeria monocytogenes causes relatively few outbreaks linked to whole fresh produce but triggers recalls each year in the United States. There are limited data on the influence of wet versus dry inoculation methods on pathogen growth on whole produce. A cocktail of five L. monocytogenes strains that included clinical, food, and environmental isolates associated with foodborne outbreaks and recalls was used. Cultures were combined to target a final wet inoculum concentration of 4 to 5 log CFU/mL. The dry inoculum was prepared by mixing wet inoculum with 100 g of sterile sand and drying for 24 h. Produce investigated belonged to major commodity families: Ericaceae (blackberry, raspberry, and blueberry), Rutaceae (lemon and mandarin orange), Rosaceae (sweet cherry), Solanaceae (tomato), Brassaceae (cauliflower and broccoli), and Apiaceae (carrot). Whole intact, inoculated fruit and vegetable commodities were incubated at 2, 12, 22, and 35 ± 2°C. Commodities were sampled for up to 28 days, and the experiment was replicated six times. The average maximum growth increase was obtained by measuring the maximum absolute increase for each replicate within a specific commodity, temperature, and inoculation method. Data for each commodity, replicate, and temperature were used to create primary growth or survival models describing the lag phase and growth or shoulder and decline as a function of time. Use of a liquid inoculum (versus dry inoculum) resulted in a markedly increased L. monocytogenes growth rate and growth magnitude on whole produce surfaces. Temperature highly influenced this difference: a greater effect seen with more commodities at higher temperatures (22 and 35°C) versus lower temperatures (2 and 12°C). These findings need to be explored for other commodities and pathogens. The degree to which wet or dry inoculation techniques more realistically mimic contamination conditions throughout the supply chain (e.g., production, harvest, postharvest, transportation, or retail) should be investigated.

The effect of sage herbal dust products on Listeria monocytogenes growth in minced pork

Valorization of waste materials became an important economical and ecological aspect of sustainable development. Sage herb has been well known for its antimicrobial properties, so its use as an antimicrobial agent has a lot of potential in the food industry. In this work, sage herbal dust essential oil obtained by hydrodistillation and supercritical fluid extraction, and supercritical fluid extract were screened for antimicrobial effect against Listeria monocytogenes ATCC 15313 in minced pork. Application of supercritical fluid extract had a lower antimicrobial effect of 1 log CFU/ml in a meat simulation medium, compared to the essential oils (2.5–3.5 log CFU/ml). On the other hand, all analyzed samples showed a statistically significant reduction of the L. monocytogenes growth in minced pork. Sage herbal dust essential oil and supercritical fluid extract have the potential for control of L. monocytogenes growth in minced pork during storage up to 6 days at refrigerating temperature. Practical applications The use of natural antimicrobials as food additives can have an economic impact due to the reduction of losses caused by spoilage. The novelty of conducted research relies on the use of waste material from tea factory, for example, sage herbal dust. The application of sage herbal dust essential oil and extract can prolong the shelf life of meat products.

Dynamic model to describe kinetic behavior of Listeria monocytogenes in smoked salmon

AbstractIn this study, the dynamic model was developed to describe cell counts of Listeria monocytogenes in smoked salmon under distribution conditions. A 9‐strain mixture of L. monocytogenes was inoculated in smoked salmon at 3–4 Log CFU/g. The inoculated samples were stored at 4°C, 10°C, and 20°C for up to 16 days, and the bacterial cell counts were enumerated on PALCAM agar. The Baranyi model was fitted to the L. monocytogenes cell counts to calculate the lag phase duration (LPD; hr) and maximum growth rate (μmax; Log CFU/g/hr). LPD was 74.22 hr at 4°C, and the value was rapidly decreased to 4.84 hr at 20°C as temperature increased. μmax ranged from 0.01 to 0.13 Log CFU/g/hr, depending on the storage temperature. The kinetic parameters were fitted with secondary models as a function of temperature. Cell counts predicted by the developed model were compared with the predicted cell counts obtained at 8°C and 15°C, and the differences between predicted and observed cell counts were indicated by root mean square error (RMSE). The dynamic model was then developed with the kinetic parameters derived from the secondary model and simulated the L. monocytogenes growth. RMSE (0.368) indicates that the model performance was appropriate. These results suggest that the developed kinetic models can be useful in predicting the fates of L. monocytogenes in smoked salmon under the dynamic conditions of storage temperature and time.

Antimicrobial mechanism of 4-hydroxyphenylacetic acid on Listeria monocytogenes membrane and virulence

4-Hydroxyphenylacetic acid (4-HPCA) is the major intestinal metabolite of kaempferol and polymeric proanthocyanidins whereas the effect of 4-HPCA on Listeria monocytogenes remains unknown. In this study, we investigated the effect and mechanism of action of 4-HPCA on the highly lethal foodborne pathogen Listeria monocytogenes. Our results indicated that 4-HPCA inhibited L. monocytogenes growth and proliferation in a dose-dependent manner. In particular, L. monocytogenes displayed negligible growth or proliferation after 4-HPCA treatment (15.61 mM) for 24 h. The impact of 4-HPCA on cell membrane structure and function was investigated in terms of fluorometric cell membrane integrity, zeta potential and relative electrical conductivity. We observed an approximately 15 % fluorescence reduction in the cell membrane after MIC treatment. The zeta potential of the bacteria shifted significantly from −49.74 to −43.70 mV, −36.65 mV and −37.97 mV after treatment with 4-HPCA at the MIC for 0 h, 3 h and 12 h, respectively. The absolute value of the relative electrical conductivities increased significantly following 3 h, 6 h, 9 h and 15 h of 4-HPCA treatment at the MIC level. The results of scanning electron microscopy (SEM) showed that cells treated with 4-HPCA displayed a wrinkled morphology and irregular shapes. Moreover, 4-HPCA obviously decreased the expression of three virulence genes (hlyA, prfA, and inlA) in L. monocytogenes after 12 h of treatment. All these results verified that 4-HPCA, as an effective antibacterial compound against L. monocytogenes, could cause cell death through cell membrane damage and decrease the expression of three virulence factors.

Modelling the Effect of Salt Concentration on the Fate of Listeria monocytogenes Isolated from Costa Rican Fresh Cheeses.

“Turrialba cheese” is a Costa Rican fresh cheese highly appreciated due to its sensory characteristics and artisanal production. As a ready-to-eat dairy product, its formulation could support Listeria monocytogenes growth. L. monocytogenes was isolated from 14.06% of the samples and the pathogen was able to grow under all tested conditions. Due to the increasing demand for low-salt products, the objective of this study was to determine the effect of salt concentration on the growth of pathogen isolates obtained from local cheese. Products from retail outlets in Costa Rica were analyzed for L. monocytogenes. These isolates were used to determine growth at 4 °C for different salt concentration (0.5–5.2%). Kinetic curves were built and primary and secondary models developed. Finally, a validation study was performed using literature data. The R2 and Standard Error of fit of primary models were ranked from 0.964–0.993, and 0.197–0.443, respectively. An inverse relationship was observed between growth rate and salt concentration. A secondary model was obtained, with R2 = 0.962. The model was validated, and all values were Bf > 1, thus providing fail-safe estimations. These data were added to the free and easy-to-use predictive microbiology software “microHibro” which is used by food producers and regulators to assist in decision-making.

Investigating the Roles of Listeria monocytogenes Peroxidases in Growth and Virulence.

ABSTRACTBacteria have necessarily evolved a protective arsenal of proteins to contend with peroxides and other reactive oxygen species generated in aerobic environments. Listeria monocytogenes encounters an onslaught of peroxide both in the environment and during infection of the mammalian host, where it is the causative agent of the foodborne illness listeriosis. Despite the importance of peroxide for the immune response to bacterial infection, the strategy by which L. monocytogenes protects against peroxide toxicity has yet to be illuminated. Here, we investigated the expression and essentiality of all the peroxidase-encoding genes during L. monocytogenes growth in vitro and during infection of murine cells in tissue culture. We found that chdC and kat were required for aerobic growth in vitro, and fri and ahpA were each required for L. monocytogenes to survive acute peroxide stress. Despite increased expression of fri, ahpA, and kat during infection of macrophages, only fri proved necessary for cytosolic growth. In contrast, the proteins encoded by lmo0367, lmo0983, tpx, lmo1609, and ohrA were dispensable for aerobic growth, acute peroxide detoxification, and infection. Together, our results provide insight into the multifaceted L. monocytogenes peroxide detoxification strategy and demonstrate that L. monocytogenes encodes a functionally diverse set of peroxidase enzymes.IMPORTANCE Listeria monocytogenes is a facultative intracellular pathogen and the causative agent of the foodborne illness listeriosis. L. monocytogenes must contend with reactive oxygen species generated extracellularly during aerobic growth and intracellularly by the host immune system. However, the mechanisms by which L. monocytogenes defends against peroxide toxicity have not yet been defined. Here, we investigated the roles of each of the peroxidase-encoding genes in L. monocytogenes growth, peroxide stress response, and virulence in mammalian cells.

High throughput screening of technological and biopreservation traits of a large set of wild lactic acid bacteria from Brazilian artisanal cheeses

This study aimed to evaluate technological (acidification, proteolysis, lipolysis, resistance to low pH, NaCl, and bile salts) and biopreservation (antimicrobial activity against foodborne pathogens) features of 1002 LAB by high throughput screening (HTS) methods. The LAB was isolated from 11 types of Brazilian artisanal cheeses (BAC) marketed in the main 5 producing regions. Remarkable intra-species variability in acidification rates have been found, which was most pronounced between isolates from Mina's artisanal cheeses, Caipira and Coalho cheeses. Lacticaseibacillus paracasei and Levilactobacillus brevis showed the fastest acidification rate; however, all isolates showed slower acidification rates than a lactococcal control strain (4.3 × lower). When testing inhibitory effects, > 75% of LAB isolates could inhibit the growth of Staphylococcus aureus ATCC 19095 and Listeria monocytogenes ATCC 7644. Two of these isolates, identified as Lactiplantibacillus plantarum and Lentilactobacillus buchneri, the sterile and neutral supernatants alone, were sufficient to inhibit L. monocytogenes growth. Principal component analysis (PCA) allowed the identification of functional groups based on proteolytic and lipolytic activity, osmotic stress resistance, and inhibition of L. monocytogenes. The type of cheese the isolates were recovered from influenced properties such as anti-listerial compounds and lipolytic enzyme production. The use of HTS and multivariate statistics allowed insights into a diverse set of LAB technological and biopreservation properties. These findings allow a profound knowledge of the heterogeneity of a large set of isolates, which can be further used to design starter cultures with varied and combined properties, such as biopreservation and technological features. Besides that, HTS makes it possible to analyze a vast panel of LAB strains, reducing costs and time within laboratory analysis, while avoiding the loss of information once all LAB are tested at the same time (differently from the traditional labor-intensive approach, in which a few numbers of strains is tested per time).

Study of the microbial diversity of a panel of Belgian artisanal cheeses associated with challenge studies for Listeria monocytogenes

High throughput sequencing could become a powerful tool in food safety. This study was the first to investigate artisanal cheeses from Belgium (31 batches) using metagenetics, in relation to Listeria monocytogenes growth data acquired during a previous project. Five cheese types were considered, namely unripened acid-curd cheeses, smear- and mold-ripened soft cheeses, and Gouda-type and Saint-Paulin-type cheeses. Each batch was analyzed in triplicate the first and the last days of storage at 8 °C. Globally, 2697 OTUs belonging to 277 genera and to 15 phyla were identified. Lactococcus was dominant in all types, but Streptococcus was co-dominant in smear-ripened soft cheeses and Saint-Paulin-type cheeses. The dominant population was not always associated with added starter cultures. Bacterial richness and diversity were significantly higher in both types of soft cheeses than in other categories, including particular genera like Prevotella, Faecalibacterium and Hafnia-Obesumbacterium in mold-ripened cheeses and Brevibacterium, Brachybacterium, Microbacterium, Bacteroides, Corynebacterium, Marinilactibacillus, Fusobacterium, Halomonas and Psychrobacter in smear-ripened soft cheeses. A strong correlation was observed between no growth of L. monocytogenes in a smear-ripened cheese and the presence of an unknown Fusobacterium (relative abundance around 10%). This in silico correlation should be confirmed by further experiments in vitro and in situ.

Effect of Commercial Protective Cultures and Bacterial Fermentates on Listeria monocytogenes Growth in a Refrigerated High-Moisture Model Cheese

Biopreservatives are clean-label ingredients used to control pathogenic and spoilage microorganisms in ready-to-eat foods, including cheese. In a first set of experiments, the efficacies of six commercial biopreservatives in controlling Listeria monocytogenes growth at 4°C were tested in a high-moisture model cheese (pH 6.00, 56% moisture, and 1.25% salt) made of cream, micellar casein, water, salt, lactose, lactic acid, and a single protective culture (PC-1, PC-2, or PC-3 at 106 CFU/g [target]) or bacterial fermentate (CM-1 or CM-2 [cultured milk] or CSV-1 [cultured sugar-vinegar blend], 0.5 or 1.0% target level). Cheeses were inoculated with 3 log CFU/g L. monocytogenes (5-strain cocktail), after which 25-g samples were vacuum sealed and stored at 4°C for 8 weeks. L. monocytogenes populations from triplicate samples were enumerated weekly on modified Oxford agar in duplicate trials. L. monocytogenes growth (≥1-log increase) was observed in approximately 1 week in control cheese and those formulated with 106 CFU of PC-1 or PC-2 per g. Growth was delayed to 2.5 weeks in model cheeses formulated with 106 CFU of PC-3 per g or 0.5% CM-2 and to 3 weeks with 0.5% CM-1 or CSV-1. Growth was further delayed to 6.5 to 7.5 weeks in model cheeses formulated with 1.0% CM-1 or CM-2, while formulation with 1.0% CSV-1 inhibited L. monocytogenes growth for 8 weeks. In a second set of experiments, the combined effects of pH and 0.5% CSV-1 on L. monocytogenes inhibition were investigated. Incorporation of 0.5% CSV-1 delayed L. monocytogenes growth to 3, 6, and >10 weeks in cheeses of pH 6.00, 5.75, and 5.50, respectively, versus growth observed in 1, 1, and 3.5 weeks in control cheeses. These data suggest that certain fermentates have greater antilisterial activity than protective cultures in directly acidified cheeses with direct biopreservative incorporation and refrigerated storage. Further research is needed to optimize the conditions to prevent listerial growth by utilizing protective cultures in fresh, soft cheeses.

Evaluation of growth potential and growth dynamics of Listeria monocytogenes on ready-to-eat fresh fruit.

The consumption of fresh or RTE fruits is increasing every year and Listeria monocytogenes has been identified on raw or minimally processed fruits. A food product can become contaminated with L. monocytogenes anywhere along the pathway of food production during planting, harvesting, packaging, distribution and serving. The aim of this work was to assess the microbiological risks associated with consumption of ready- to- eat fruit such as melon, pineapple, coconut and fruit salad. The presence of Escherichia coli, Salmonella spp. and L. monocytogenes was also evaluated. Microbiological challenge tests were carried out for the evaluation of the L. monocytogenes growth potential in RTE fruit stored at 4 and 8°C. E. coli counts resulted under the detection limit of 10 CFU g-1, Salmonella and L. monocytogenes were not detected (absence in 25g). The growth potential values in coconut and melon (δ>0.5) showed the growth capacity of Listeria at the temperatures considered. A low initial load, also derived from good hygiene practices, and correct storage temperatures are essential to reduce bacterial growth in RTE fruit. The challenge test showed how each type of RTE fruit has a different commercial life based on its specific growth potential and that food should be stored at temperatures not higher than 4°C for a short period.

Occurrence of Listeria monocytogenes in Counter-Sliced Turkey Meat Samples from Independent Delis in New York City

Research suggests that small, independent delicatessens are less likely to follow proper sanitation procedures, including slicer inspection, which could lead to a higher likelihood of these delis being a reservoir for Listeria monocytogenes growth and cross-contamination. This study was undertaken to determine the incidence of L. monocytogenes in counter-sliced turkey deli meat obtained from independent delis in an urban city. Deli meat, counter-sliced on site, was collected from 118 independent delis in New York City. The samples were analyzed for L. monocytogenes using the U.S. Department of Agriculture Microbiology Laboratory Guidebook methodology for isolation and confirmation. The selection criteria for delis included using the city's restaurant inspection and grading system. Two samples, from separate delis, were confirmed as positive for L. monocytogenes (1.69%). Analysis of the genomic sequences of one of the samples revealed a close match to a cluster of six clinical isolates, which were part of an ongoing multistate listeriosis outbreak spanning four states. The sequence of the second isolate matched a clinical isolate in a neighboring state. Both isolates were obtained from delis that did not have the top inspection grade. Although a snapshot of one urban area, this study is the first report on the current incidence of L. monocytogenes on counter-sliced deli meat from independent deli establishments. This study suggests that these delis can potentially serve as sources of L. monocytogenes contamination or contribute to downstream foodborne listeriosis. Information provided by city inspection and grading systems, in addition to the letter grade, may serve as a tool to identify delis with potential L. monocytogenes contamination issues and a basis for product and environmental sampling by public health authorities.