Tolerance evaluation in Salmonella enterica serovar Typhimurium challenged with sublethal amounts of Rosmarinus officinalis L. essential oil or 1,8‐cineole in meat model

The induction of direct bacterial tolerance and cross‐tolerance (NaCl, acid pH, high temperature) in Salmonella Typhimurium ATTC 14028 following the exposure to sublethal amounts of the essential oil from Rosmarinus officinalis L. (ROEO), and its major component 1,8‐cineole (CIN) was evaluated in this study. Direct protection was not induced when cells were exposed to 1/2 MIC and 1/4 MIC of ROEO or CIN in meat broth and in previously irradiated meat ground‐beef. Cells exposed to ROEO or CIN at sublethal amounts did not present cross‐protection to high temperature, lactic acid and NaCl. Likewise, cells progressively subcultured in meat broth containing increasing amounts of ROEO or CIN were able to survive only up to 1/4 MIC for both tested substances. From these results, S. Typhimurium ATCC 14028 was not capable to develop direct or cross‐tolerance when exposed to ROEO or CIN in a meat‐based growth media and was not able to develop direct tolerance in a meat‐based model.     

Evidence of Viable But Non-Culturable state in Listeria monocytogenes by direct viable count and CTC-DAPI double staining

Viable but Non-culturable (VBNC) state in bacteria was detected originally in environmental microbiology studies. In particular, this state has been demonstrated for a number of human pathogens (Escherichia coli, Salmonella enteritidis, Vibrio cholerae, Legionella pneumophila and Campylobacter jejuni). The presence of VBNC cells poses a major public health problem since they cannot be detected by traditional culturing methods and moreover the cells remain potentially pathogenic under favourable conditions. But the VBNC state has not been yet described in Listeria monocytogenes. Production of VBNC L. monocytogenes cells was studied using four strains resuspended in microcosm water. Various strains of L. monocytogenes were resuspended in filtered sterilized water adjusted to pH 6·0, incubated at 20°C and 4°C with or without NaCl supplementation (0%, 7%), with gentle shaking at 100 rpm. The culturability of starved cells suspensions was determined by spread plate count (PCA). The cells activity was measured by a Direct Viable Count technique and by CTC-DAPI double staining. Two strains of four (CNL 895807, Scott A) exhibited a VBNC state. They lost their culturability and maintained a cellular activity, even after 10 weeks of starvation in microcosm water.     

Application of Central Composite Design to evaluate the antilisterial activity of hydro-alcohol berry extract of Myrtus communis L.

The antimicrobial activity of the hydro-alcohol extract of Myrtus communis L. (ME) berries was investigated against six Listeria monocytogenes strains (2 type strains and 4 isolates). Sub-lethal ME concentrations reduced L. monocytogenes counts by at least 2 log cycles. A Central Composite Design was used to investigate the combined effects of sub-lethal concentrations of ME (0.039–0.195 mL/100 mL), NaCl (0–2.0 g/100 mL) and pH (5.0–7.0) on strains growth. ME affected growth parameters, generally extending lag phase length and reducing maximum growth, sometimes with interactive effects with pH. The highest ME concentrations (0.117–0.195 mL/100 mL) combined with the lowest pH values (5.0–6.0) strongly reduced or even inhibited strains growth.     

Enhanced chemical stability,chromatic properties and regeneration of betalains in Rivina humilis L. berry juice

Betalains are hydrophilic colorants containing chromophore betalamic acid. Owing to poor stability, its usage is limited to low acidic short shelf-life, and frozen foods. In this report, effect of metals (inorganic Se⁴⁺, Zn²⁺, and Cu²⁺) on stability of betalains in Rivina humilis L. berry juice (RBJ) was studied in presence of 10 and 40 μg metal/mL with/without ascorbic acid (AA; 0.25 and 0.5 g/100 mL). Se bleached RBJ betacyanins mildly, whereas Zn, and Cu bleached the pigments significantly. AA protected the pigments from metal-induced bleaching, and stabilizing effect of 0.25 g AA/100 mL was higher (P < 0.05) than 0.5 g AA/100 mL. AA (0.25 g/100 mL) + Se (40 μg/mL) enhanced (five-fold) the half-life time of betacyanins. RBJ betacyanins degraded up to 95% and 96% on treatment for 36 min at 90 °C and storage for 48 days at 25 °C, respectively, whereas only 15% pigment was lost on storage (5 °C, 90 days). AA (0.25 g/100 mL) + Se (40 μg/mL) regenerated the pigments on storage at 5 °C after thermal degradation. Color values of AA (0.25 g/100 mL) + Se (40 μg/mL) containing samples indicated an orange tinge, whereas other samples turned brown yellow after thermal treatment.     

Growth characteristics of Listeria monocytogenes as affected by a native microflora in cooked ham under refrigerated and temperature abuse conditions

This study examined the growth characteristics of Listeria monocytogenes as affected by a native microflora in cooked ham at refrigerated and abuse temperatures. A five-strain mixture of L. monocytogenes and a native microflora, consisting of Brochothrix spp., isolated from cooked meat were inoculated alone (monocultured) or co-inoculated (co-cultured) onto cooked ham slices. The growth characteristics, lag phase duration (LPD, h), growth rate (GR, log₁₀ cfu/h), and maximum population density (MPD, log₁₀ cfu/g), of L. monocytogenes and the native microflora in vacuum-packed ham slices stored at 4, 6, 8, 10, and 12 °C for up to 5 weeks were determined. At 4-12 °C, the LPDs of co-cultured L. monocytogenes were not significantly different from those of monocultured L. monocytogenes in ham, indicating the LPDs of L. monocytogenes at 4-12 °C were not influenced by the presence of the native microflora. At 4-8 °C, the GRs of co-cultured L. monocytogenes (0.0114-0.0130 log₁₀ cfu/h) were statistically but marginally lower than those of monocultured L. monocytogenes (0.0132-0.0145 log₁₀ cfu/h), indicating the GRs of L. monocytogenes at 4-8 °C were reduced by the presence of the native microflora. The GRs of L. monocytogenes were reduced by 8-7% with the presence of the native microflora at 4-8 °C, whereas there was less influence of the native microflora on the GRs of L. monocytogenes at 10 and 12 °C. The MPDs of L. monocytogenes at 4-8 °C were also reduced by the presence of the native microflora. Data from this study provide additional information regarding the growth suppression of L. monocytogenes by the native microflora for assessing the survival and growth of L. monocytogenes in ready-to-eat meat products.     

Assessment of chemically characterised Rosmarinus officinalis L. essential oil and its major compounds as plant‐based preservative in food system based on their efficacy against food‐borne moulds and aflatoxin secretion and as antioxidant

The study explores antifungal, anti‐aflatoxigenic and antioxidant efficacy of Rosmarinus officinalis essential oil (ROEO) and its major compounds. In addition, the mode of action of ROEO and its practical efficacy as preservative have been assessed. GC‐MS analysis of ROEO identified 16 compounds; α‐pinene, 1,8‐cineole and camphor being the major compounds. The minimum concentration for inhibition of growth and aflatoxin B₁ secretion against A. flavus (LHP‐6) was found to be 1.5, >5.0, 4.0 and 3.0 μL mL^?1 and 1.25, >5.0, 3.5 and 3.0 μL mL^?1 for ROEO, α‐pinene, 1,8‐cineole and camphor, respectively. The IC₅₀ value through DPPH analysis and percentage inhibition of linoleic acid peroxidation of ROEO were 0.042 μL mL^?1 and 71.05%, respectively. The targeted site of antifungal action of ROEO was confirmed as plasma membrane through ergosterol measurement and TEM analysis. Moreover, ROEO significantly protected Piper nigrum fruits against mould infestation upto 6 months in in vivo trial.     

Satureja horvatii essential oil: In vitro antimicrobial and antiradical properties and in situ control of Listeria monocytogenes in pork meat

The dominant compounds in Satureja horvatii oil were p-cymene (33.14%), thymol (26.11%) and thymol methyl ether (15.08%). The minimum inhibitory concentration (MIC) varied from 0.03 to 0.57 mg/mL for bacteria, and from 0.56 to 2.23 mg/mL for yeast strains, while minimum bactericidal/yeast-cidal concentration (MBC/MYC) varied from 0.07 to 1.15 mg/mL and 1.11 to 5.57 mg/mL for bacteria and yeasts, respectively. The antiradical potential of the essential oil was evaluated using hydroxyl radical (•OH) generated in Fenton reaction. The meat preserving potential of essential oil from Satureja horvatii was investigated against L. monocytogenes. Essential oil successfully inhibited development of L. monocytogenes in pork meat. Sensorial evaluation on flavor and color of meat was performed. The color and flavor of meat treated with essential oil improved after 4 days of storage. S. horvatii essential oil can act as a potent inhibitor of food spoiling microorganisms, in meat products and also can be a useful source of natural antioxidants.     

The effect of pH and nitrite concentration on the antimicrobial impact of celery juice concentrate compared with conventional sodium nitrite on Listeria monocytogenes

The objectives of this study were to evaluate the impact of pH and nitrite from celery juice concentrate (CJ) on the growth of Listeria monocytogenes in broth and on ham slices, and to evaluate the impact of pH and nitrite from CJ on quality attributes of the ham. The pH of both broth and ham were increased by the addition of CJ. The CJ was less effective than conventional nitrite at 100 mg/kg nitrite in broth, but in ham, the CJ treatments at both 100 and 200 mg/kg resulted in growth of L. monocytogenes (p > 0.05) similar to that of the conventional nitrite at the same concentrations. Reducing the pH of CJ before addition to the ham had greater impact on L. monocytogenes growth at 200 mg/kg nitrite than at 100 mg/kg. Celery juice concentrate may increase meat product pH which could have implications for the antimicrobial impact of nitrite in some products.     

Sensory evaluation and inhibition of Listeria monocytogenes in bovine pâté added of chitosan from Mucor rouxii

This study aimed to assess the efficacy of chitosan from Mucor rouxii UCP 064 in inhibiting Listeria monocytogenes in bovine meat pâté at 4 °C. The influence of the chitosan addition on sensory aspects of the product was also evaluated. The addition of chitosan from M. rouxii at 5 mg/g to bovine meat pâté decreased the counts of L. monocytogenes from approximately 7 to 3 log cfu/g after 6 days of storage at 4 °C. In pâté without chitosan the counts were over 7 log cfu/g already after two days of storage. Sensory evaluation suggested that addition of chitosan in pâté would be acceptable to consumers, although some negative influence on flavor and taste was found. From these results, chitosan from fungi could be considered a possible alternative compound to control L. monocytogenes in ready-to-eat meat products.     

Antimicrobial Polylactic Acid Packaging Films against Listeria and Salmonella in Culture Medium and on Ready-to-Eat Meat

The contamination of Listeria monocytogenes and Salmonella spp. in ready-to-eat (RTE) meat products has been a concern for the meat industry. In this study, edible chitosan-acid solutions incorporating lauric arginate ester (LAE), sodium lactate (NaL), and sorbic acid (SA) alone or in combinations were developed and coated on polylactic acid (PLA) packaging films. Antimicrobial effects of coated PLA films on the growth of Listeria innocua, L. monocytogenes, and Salmonella Typhimurium in a culture medium (tryptic soy broth, TSB) and on the surface of meat samples were investigated. Antimicrobial PLA films containing 1.94 mg/cm² of chitosan and 1.94 μg/cm² of LAE were the most effective against both Listeria and Salmonella in TSB and reduced them to undetectable level (<0.69 log CFU/ml). The same PLA films with LAE significantly (p?L. innocua, L. monocytogenes, and S. Typhimurium on RTE meat during 3 and 5 weeks’ storage at 10 °C, achieving 2–3 log reduction of Listeria and 1–1.5 log reduction of Salmonella as compared with controls. PLA films coated with 1.94 mg/cm² of chitosan, 0.78 mg/cm² of NaL, and 0.12 mg/cm² of SA significantly reduced the growth of L. innocua but were less effective against Salmonella. The combination of NaL (0.78 mg/cm²) and SA (0.12 mg/cm²) with LAE (1.94 μg/cm²) did not generate additional or synergetic antimicrobial effect against Listeria or Salmonella on the meat surface. L. innocua had a similar sensitivity to the film treatments as L. monocytogenes, suggesting that L. innocua may be used as a surrogate of L. monocytogenes for further scaleup and validation studies. The film treatments were more effective against the microorganisms in TSB culture medium than in RTE meat, which suggests that in vivo studies are a necessary step to develop antimicrobial packaging for applications in foods.     

Quantifying Listeria monocytogenes prevalence and concentration in minced pork meat and estimating performance of three culture media from presence/absence microbiological testing using a deterministic and stochastic approach

Listeria monocytogenes poses a serious threat to public health, and the majority of cases of human listeriosis are associated with contaminated food. Reliable microbiological testing is needed for effective pathogen control by food industry and competent authorities. The aims of this work were to estimate the prevalence and concentration of L. monocytogenes in minced pork meat by the application of a Bayesian modeling approach, and also to determine the performance of three culture media commonly used for detecting L. monocytogenes in foods from a deterministic and stochastic perspective. Samples (n = 100) collected from local markets were tested for L. monocytogenes using in parallel the PALCAM, ALOA and RAPID'L.mono selective media according to ISO 11290-1:1996 and 11290-2:1998 methods. Presence of the pathogen was confirmed by conducting biochemical and molecular tests. Independent experiments (n = 10) for model validation purposes were performed. Performance attributes were calculated from the presence–absence microbiological test results by combining the results obtained from the culture media and confirmative tests. Dirichlet distribution, the multivariate expression of a Beta distribution, was used to analyze the performance data from a stochastic perspective. No L. monocytogenes was enumerated by direct-plating (<10 CFU/g), though the pathogen was detected in 22% of the samples. L. monocytogenes concentration was estimated at 14–17 CFU/kg. Validation showed good agreement between observed and predicted prevalence (error = −2.17%). The results showed that all media were best at ruling in L. monocytogenes presence than ruling it out. Sensitivity and specificity varied depending on the culture-dependent method. None of the culture media was perfect in detecting L. monocytogenes in minced pork meat alone. The use of at least two culture media in parallel enhanced the efficiency of L. monocytogenes detection. Bayesian modeling may reduce the time needed to draw conclusions regarding L. monocytogenes presence and the uncertainty of the results obtained. Furthermore, the problem of observing zero counts may be overcome by applying Bayesian analysis, making the determination of a test performance feasible.     

The effect of American cranberry (Vaccinium macrocarpon) constituents on the growth inhibition,membrane integrity,and injury of Escherichia coli O157:H7 and Listeria monocytogenes in comparison to Lactobacillus rhamnosus

The antimicrobial properties of the American cranberry were studied against Escherichia coli O157:H7, Listeria monocytogenes, and Lactobacillus rhamnosus to determine the effects on growth inhibition, membrane permeability, and injury. Cranberry powder was separated using a C-18 Sep-Pak cartridge into sugars plus organic acids (F1), monomeric phenolics (F2), and anthocyanins plus proanthocyanidins (F3). Fraction 3 was further separated into anthocyanins (F4) and proanthocyanidins (F5) using an LH-20 Sephadex column. Each fraction was diluted in the brain heart infusion (BHI) broth to determine the minimum inhibitory/bactericidal concentrations (MIC/MBC). L. monocytogenes was the most susceptible to cranberry fraction treatment with the lowest MIC/MBC for each treatment, followed by E. coli O157:H7 and L. rhamnosus. Membrane permeability and potential was studied using LIVE/DEAD viability assay and using Bis (1, 3-dibutylbarbituric acid) trimethine oxonol (DiBAC₄), respectively. L. rhamnosus demonstrated the highest permeability followed by E. coli O157:H7, and L. monocytogenes. L. rhamnosus demonstrated the highest recovery followed by E. coli O157:H7, and L. monocytogenes. Each cranberry fraction demonstrated membrane hyperpolarization at their native pH, while F2, F3, and F5 demonstrated membrane depolarization at neutral pH. With this knowledge cranberry compounds may be used to prevent maladies and potentially substitute for synthetic preservatives and antibiotics.     

Morphological and physiological responses of Campylobacter jejuni to stress

Under conditions of stress, cells of Campylobacter assume a coccoid shape that may be an evolutionary strategy evolved by the organism to enable survival between hosts. However, the physiology of Campylobacter as it devolves from spiral to coccoid-shaped morphology is poorly understood. In this study, conditions influencing the survival of Campylobacter jejuni ATCC 35921 in broth were determined. Cells in late log phase were resuspended in broth at 4 or 60 degrees C. The culturability of these cold- or heat-stressed cell suspensions was determined by spread plate counts and the activity of cells by the direct viable count technique and 5-cyano-2,3-ditolyltetrazolium chloride staining. C. jejuni changed form completely from culturable to viable but nonculturable cells (VBNC) within 25 days at 4 degrees C, and 15 min at 60 degrees C. Light microscopy of C. jejuni VBNC cells showed that the spiral-shaped cells became coccoid, and transmission electron microscopy of C. jejuni VBNC cells showed that the outer membrane was lost in aging cell suspensions. Furthermore, a limited proteomic study was carried out to compare C. jejuni proteins that exhibited increased or decreased synthesis on exposure to 60 degrees C.     

Lactose oxidase: An enzymatic approach to inhibit Listeria monocytogenes in milk

Listeria monocytogenes is a ubiquitous pathogen that can cause morbidity and mortality in immunocompromised individuals. Growth of L. monocytogenes is possible at refrigeration temperatures due to its psychrotrophic nature. The use of antimicrobials in dairy products is a potential way to control L. monocytogenes growth in processes with no thermal kill step, thereby enhancing the safety of such products. Microbial-based enzymes offer a clean-label approach for control of L. monocytogenes outgrowth. Lactose oxidase (LO) is a microbial-derived enzyme with antimicrobial properties. It oxidizes lactose into lactobionic acid and reduces oxygen, generating H₂O₂. This study investigated the effects of LO in UHT skim milk using different L. monocytogenes contamination scenarios. These LO treatments were then applied to raw milk with various modifications; higher levels of LO as well as supplementation with thiocyanate were added to activate the lactoperoxidase system, a natural antimicrobial system present in milk. In UHT skim milk, concentrations of 0.0060, 0.012, and 0.12 g/L LO each reduced L. monocytogenes counts to below the limit of detection between 14 and 21 d of refrigerated storage, dependent on the concentration of LO. In the 48-h trials in UHT skim milk, LO treatments were effective in a concentration-dependent fashion. The highest concentration of LO in the 21-d trials, 0.12 g/L, did not show great inhibition over 48 h, so concentrations were increased for these experiments. In the lower inoculum, after 48 h, a 12 g/L LO treatment reached levels of 1.7 log cfu/mL, a reduction of 1.3 log cfu/mL from the initial inoculum, whereas the control grew out to approximately 4 log cfu/mL, an increase of 1 log cfu/mL from the inoculum on d 0. When a higher challenge inoculum of 5 log cfu/mL was used, the 0.12 g/L and 1.2 g/L treatments reduced the levels by 0.2 to 0.3 log cfu/mL below the initial inoculum and the 12 g/L treatment by >1 log cfu/mL below the initial inoculum by hour 48 of storage at refrigeration temperatures. After the efficacy of LO was determined in UHT skim milk, LO treatments were applied to raw milk. Concentrations of LO were increased, and the addition of thiocyanate was investigated to supplement the effect of the lactoperoxidase system against L. monocytogenes. When raw milk was inoculated with 2 log cfu/mL, 1.2 g/L LO alone and combined with sodium thiocyanate reduced ~0.8 log cfu/mL from the initial inoculum on d 7 of storage, whereas the control grew out to >1 log cfu/mL from the initial inoculum. Furthermore, in the higher inoculum, 1.2 g/L LO combined with sodium thiocyanate reduced L. monocytogenes counts from the initial inoculum by >1 log cfu/mL, whereas the control grew out 2 log cfu/mL from the initial inoculum. Results from this study suggest that LO is inhibitory against L. monocytogenes in UHT skim milk and in raw milk. Therefore, LO may be an effective treatment to prevent L. monocytogenes outgrowth, increase the safety of raw milk, and be used as an effective agent to prevent L. monocytogenes proliferation in fresh cheese and other dairy products. This enzymatic approach is a novel application to control the foodborne pathogen L. monocytogenes in dairy products.     

Rapid detection of Listeria monocytogenes in food using culture enrichment combined with real-time PCR

A rapid method for the detection of Listeria monocytogenes in foods combining culture enrichment and real-time PCR was compared to the ISO 11290-1 standard method. The culture enrichment component of the rapid method is based on the ISO standard and includes 24 h incubation in half-Fraser broth, 4 h incubation in Fraser broth followed by DNA extraction and real-time PCR detection of the ssrA gene of L. monocytogenes. An internal amplification control, which is co-amplified with the same primers as the L. monocytogenes DNA, was also included in the assay. The method has a limit of detection of 1–5 CFU/25 g food sample and can be performed in 2 working days compared to up to 7 days for the ISO standard. A variety of food samples from retail outlets and food processing plants (n = 175) and controls (n = 31) were tested using rapid and conventional methods. The rapid method was 99.44% specific, 96.15% sensitive and 99.03% accurate when compared to the standard method. This method has the potential to be used as an alternative to the standard method for food quality assurance providing rapid detection of L. monocytogenes in food.     

Efficacy of bacteriocin-containing cell-free culture supernatants from lactic acid bacteria to control Listeria monocytogenes in food

Consumer demands have led to an increased interest in the use of natural antimicrobials for food protection. With the objective of developing novel products for enhancing the microbial safety of food, we have tested cell-free culture supernatants (CFS's) of eight antagonistic bacterial strains for their efficacy to inhibit Listeria monocytogenes in different food matrices. The antagonistic strains represented different members of the order Lactobacillales as well as one isolate of Staphylococcus sciuri and all showed strong inhibition of L. monocytogenes on agar plates. Cell-free supernatants were obtained after growing the bacteria in a yeast extract-glucose broth. In six of the CFS's, different class IIa bacteriocins, namely leucocin A, leucocin B, mundticin L, pediocin PA-1, sakacin A, and sakacin X, were identified as the major anti-listerial compounds. For the other two strains, the active substances could not be ascertained conclusively. The minimal effective concentration (MEC) of the individual CFS's to achieve a 2.3 log₁₀ reduction of L. monocytogenes was determined in culture broth, whole milk, and ground beef at 4 °C. While all bacteriocin-containing CFS's were effective in broth at concentrations from 52 to 205 AU/ml, significant higher concentrations were needed when applied in food. Best results were obtained using CFS's containing pediocin PA-1, that displayed only three- and ten-times higher MEC's in milk (307 AU/ml) and ground meat (1024 AU/g) compared to broth, respectively. A twenty-fold increase in the MEC (2048 AU/ml) was observed for a mundticin L-containing fermentate, and a CFS containing leucocin A and B was inactivated more than fifty-fold (> 1280 AU/ml) in both food matrices. Remarkably, the sakacin A and sakacin X containing CFS's displayed very selective inactivation rates, in which sakacin A was only effective in meat (512 AU/g), while sakacin X was only effective in milk (2048 AU/ml). In all cases, inhibition of L. monocytogenes was only transient and surviving or resistant bacteria started growing after prolonged storage. These results highlight the importance of careful testing the effectiveness of bacteriocins in the food systems for which they are intended to be applied against the selected target and non-target bacteria. Furthermore, the outgrowth of surviving or resistant bacterial populations points out that the tested bacteriocins are not suited to assure full inhibition of L. monocytogenes in a food product, if not applied in combination with additional preservative measures.     

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.     

Prato cheese containing Lactobacillus casei 01 fails to prevent dextran sodium sulphate-induced colitis

The role of experimental probiotic Prato cheese containing Lactobacillus casei 01 in the prevention of dextran sodium sulphate (DSS)-induced ulcerative colitis in mice was evaluated. For the DSS in vivo model, mice were divided into six groups. Groups 1–3 represented noninflamed groups and groups 4–6 received a DSS (2%) solution. Mice from groups 1 and 4 intragastrically received 500 μL of phosphate-buffered saline (control groups), while mice from groups 2 and 5 intragastrically received 500 μL of conventional Prato cheese (control groups). Finally, mice from group 3 and 6 intragastrically received 500 μL of Prato probiotic cheese containing L. casei 01 (9.47 log cfu mL⁻¹). Groups treated with probiotic Prato cheese exhibited reduced weight loss caused by the consumption of DSS solution. However, the probiotic cheese failed to reduce the inflammation scores in DSS-induced colitis group. Overall, probiotic Prato cheese was not effective to ameliorate the symptoms of DSS-induced colitis.     

Use of Carnobacterium maltaromaticum cultures and hydroalcoholic extract of Lippia sidoides Cham. against Listeria monocytogenes in fish model systems

Minimally processed refrigerated ready-to-eat fishes may offer health risk of severe infection to susceptible individuals due to contamination by the psychrotolerant bacterium L. monocytogenes. In this work, inhibition of L. monocytogenes by a plant extract and lactic acid bacteria (LAB) was studied in model fish systems kept at 5 °C for 35 days. For that, fillets of tropical fish “surubim” (Pseudoplatystoma sp.) and hydroalcoholic extract of the plant Lippia sidoides Cham. (“alecrim pimenta”) were used. Fish peptone broth (FPB), “surubim” broth and “surubim” homogenate were inoculated with combinations of L. monocytogenes and bacteriocin-producing Carnobacterium maltaromaticum (C2 and A9b⁺) and non bacteriocin-producing C. maltaromaticum (A9b^-), in the presence or absence of extract of “alecrim pimenta” (EAP). In all model systems, monocultures of L. monocytogenes and carnobacteria reached final populations ≥ 10⁸ CFU/ml after 35 days, except for L. monocytogenes in “surubim” homogenate (10⁴ CFU/ml). In FPB, EAP alone and combined with cultures of LAB inhibited L. monocytogenes but carnobacteria without EAP were only weakly antilisterial. In “surubim” broth, EAP alone did not prevent L. monocytogenes growth but cultures of carnobacteria combined or not with EAP inhibited L. monocytogenes, with more pronounced effect being observed for C. maltaromaticum C2, which produced bacteriocin. In “surubim” homogenate, EAP alone and combined with cultures of C. maltaromaticum A9b⁻ and A9b⁺ were strongly inhibitory to L. monocytogenes, while C. maltaromaticum C2 with EAP caused transient inhibition of L. monocytogenes. No significant inhibition of L. monocytogenes was observed for carnobacteria in “surubim” homogenate without EAP. In conclusion, it was observed that the use of EAP and cultures of carnobacteria have potential to inhibit L. monocytogenes in fish systems and the applications should be carefully studied, considering the influence of food matrix.     

Antimicrobial activity of Thymus zygis essential oil against Listeria monocytogenes and its application as food preservative

Thymus zygis is an aromatic plant used in folk medicine. This work aimed to evaluate the anti-Listeria monocytogenes activity of T. zygis essential oil (EO), whose thymol is its major compound. Furthermore, the attenuation of this bacterium's virulence, namely by the inhibition of biofilm formation, motility and invasion of human cells, and the possible application of the EO in food were evaluated. The T. zygis EO showed antibacterial activity against L. monocytogenes with a minimum inhibitory concentration (MIC) of 0.05%, while showing a bactericidal effect. The EO significantly reduced the biofilm formation (inhibition from 16.85 to 89.86%) and motility (halos between 6.66 and 10.98 mm, compared to controls 13.12 to 17.22 mm), and not inducing cross-resistance to antibiotics, such as ampicillin, cefotaxime, erythromycin, gentamicin, tetracycline, and vancomycin. L. monocytogenes counts (initial inoculum of ~10⁶ CFU/mL) were lowered by the use of 2× MIC of T. zygis EO in the chicken juice (1.53 log CFU/mL) and lettuce model (to below the detection limit) after two days of storage. The use of EO (0.2% (v/v)) for sanitizing fresh vegetables, reduce L. monocytogenes and natural microbiota for values below the detection limit of the method for iceberg lettuce after an immersion of 5 min. For the spinach, L. monocytogenes was reduced in 4.35 log CFU/mL and the natural microbiota was diminished in a range of 4.47 to 5.94 log CFU/mL, when compared with the washing with water. Overall, the T. zygis EO has demonstrated a promising antimicrobial activity and these findings point to the potential of the EO as a natural food preservative or sanitizer for controlling L. monocytogenes in food products.