Determination of Listeria monocytogenes growth potential on new fresh salmon preparations

The aim of this work was to evaluate the increase of Listeria monocytogenes on new salmon preparations (salt–sugar–pepper–dill salmon) in comparison with that obtained on cold-smoked salmon. Salmon preparations were inoculated with L. monocytogenes and were analyzed during storage at 4 °C then 8 °C. At 8 °C, the bacteria growth was of 4.53 log CFU g⁻¹ in cold-smoked salmon and of 2.06 log CFU g⁻¹ in salt–sugar–pepper–dill salmon without background microflora. The growth of L. monocytogenes was different in new salmon preparation because the mixture salt–sugar–pepper had an anti-Listeria activity and its presence could inhibitory to the growth. It is difficult to generalize findings observed with cold-smoked salmon to a new salmon preparation.     

Influence of phenolic compounds from wines on the growth of Listeria monocytogenes

The anti-microbial properties against Listeria monocytogenes of pure flavonoids rutin, catechin and quercetin; non-flavonoids gallic, vanillic, protocatechuic and caffeic acids and total polyphenols of three Argentinean wines, Cabernet Sauvignon, Malbec and Merlot varieties were investigated. The non-flavonoid caffeic acid and the flavonoids rutin and quercetin were the compounds with higher inhibitory activities on L. monocytogenes growth. The knowledge of the anti-listerial effect of different wines varieties could be the basis to demonstrate if the wine consumption with a meal may collaborate in the health protection against some foodborne organisms such as L. monocytogenes.     

The role of the consumer in the reduction of Listeria monocytogenes in lettuces by washing at home

Lettuce is highly appreciated for its nutritional properties; however microbial contamination through the food chain and its raw consumption may jeopardize these known benefits to the diet. The objective of this study was to determine the role of the consumer at the stage of washing at home, in relation to the probability of illness due to the presence of Listeria monocytogenes in lettuce. Survival curves of L. monocytogenes after washing (dipping with and without addition of bleach, and washing under a running tap) were studied. A mathematical model for each washing method was calculated by fitting experimental data. The obtained models were used to estimate the probability of illness after washing at home. Results show that although consumers can only deal with low loads of L. monocytogenes, their role is essential to reduce the normal contamination level of lettuces and ensure their safety.     

Occurrence of Listeria spp. in Brazilian fresh sausage and control of Listeria monocytogenes using bacteriophage P100

Since the 1980s, an increase in outbreaks of human listeriosis linked to contaminated food has been a concern of health authorities. Intensively manipulated foods, such as Brazilian fresh sausage, are frequently responsible for food-borne diseases. In this work the occurrence of Listeria monocytogenes and the efficacy of bacteriophage P100 (LISTEX?) to control the microorganism was evaluated in Brazilian fresh sausage. Eighty samples were analyzed, 40 each of swine and chicken Brazilian fresh sausage. Listeria spp. were isolated from 12 samples (15%), of which three (3.75%) were positive for L. monocytogenes. L. monocytogenes strains isolated belonged to serotype 1/2a. L. monocytogenes 1/2a was inoculated in Brazilian fresh sausage (2.1 × 10⁴ cfu/g) with the bacteriophage added thereafter (3.0 × 10⁷ pfu/g). Samples were analysed immediately (day zero) and then stored at 4 °C for 10 days. The bacteriophage P100 reduced L. monocytogenes counts by 2.5 log units at both 0 and 10 days compared to controls without bacteriophage. In spite of this, the populations of L. moncytogenes increased over the 10 day storage. Our data demonstrate that in one of the samples the use of the bacteriophage dropped the bacteria count below the level of direct detection. This study demonstrates a new alternative for pathogen control in the food industry, especially in the processes used to produce Brazilian fresh sausage.     

Survival and growth of Listeria monocytogenes in lubricants used in the food industry

The survival and growth of three Listeria monocytogenes strains in 10 lubricants (synthetic and mineral-oil based) used in the food industry, and rapeseed oil, was investigated at room temperature (20 °C) and refrigerated (5 °C). Additionally, the transfer of L. monocytogenes from lubricants to stainless steel surfaces and vice versa was investigated. Though the amount of L. monocytogenes in most lubricants, both pure and soiled, decreased significantly (p < 0.05) during the 14 d test period, lubricants may act as sources of contamination on the basis of the results obtained on the survival of L. monocytogenes. In general, temperature had significant effect (p < 0.05) on listericidal effect of lubricants contrary to soiling (p > 0.05), however the effect of both factors was dependent on lubricant (p < 0.05). The results clearly showed that L. monocytogenes survived in synthetic conveyer belt lubricant diluted in water. In addition, L. monocytogenes was transferred significantly (p < 0.05) from stainless steel surfaces into conveyer-belt lubricants and into mineral-oil based hydraulic oil.     

A review of minimal and defined media for growth of Listeria monocytogenes

Listeria monocytogenes is known to be an important foodborne pathogen and is the causative agent of one of the deadliest foodborne illnesses. The organism has a wide range of environmental conditions under which it will survive and grow, and often contaminates processing plants and retail environments in which ready-to-eat foods are manufactured, prepared and served. Although L. monocytogenes is not a fastidious organism and can grow in a variety of rich media, defined and minimal media are necessary to elucidate the minimal environmental nutrients that are required for the survival and growth of this organism. In addition, many of the virulence factors required for L. monocytogenes to invade and multiply in mammalian host cells are not produced in rich media and thus induction of these factors is best studied in a minimal medium. This review covers the historical development of minimal media for Listeria spp. and explores the various factors required for survival and growth of this organism. To the best of our knowledge, this is the first time that these media have been compared side by side. In order to better compare studies using different chemical substrates such as a different carbon source, all concentrations of components in each medium have been converted to molar concentrations.     

Lactic acid bacteria in an alginate film inhibit Listeria monocytogenes growth on smoked salmon

Antimicrobial packaging with lactic acid bacteria incorporated into the film matrix is a novel approach for controlling the growth of food-borne pathogens in ready-to-eat food. The overall objective of this study was to assess the effect of two strains of lactic acid bacteria (LAB) and nisin trapped in an alginate matrix, on Listeria monocytogenes growth on vacuum packed cold-smoked salmon. A film was formulated containing two LAB strains and nisin (100 IU/mL). LAB viability and bacteriocin like substance production (BLS) were assessed using the plate antagonism technique. To check the film antagonistic activity, pieces of salmon (4.0 × 4.0 cm²), inoculated with L. monocytogenes at a final concentration of 10⁴ CFU/cm², were covered with film containing both LAB strains plus nisin and stored at 4 °C. L. monocytogenes colonies on OXA agar were counted after 0, 7, 14, 21 and 28 days to evaluate pathogen inhibition. All treatments led to effective diffusion of the BLS that inhibited L. monocytogenes for 20 days after film preparation, with inhibition zones of 5.7 cm² for film coupons of 8 mm in diameter. After 28 days, salmon pieces covered with the film without inhibitors showed an increase of 2.4 log cycles in L. monocytogenes growth. In contrast, films with either LAB strain or a combination of both strains and nisin had a bacteriostatic effect on the pathogen over a period of 28 days, which exceeds the industrial standard shelf life for smoked salmon. The results demonstrate that these films inhibit L. monocytogenes growth on salmon during refrigerated storage.     

Anti-listerial properties of garlic shoot juice at growth and morphology of Listeria monocytogenes

The anti-listerial effect of garlic shoot juice (GSJ) was investigated against the four strains of Listeria monocytogenes ATCC 19116, 19118, 19166 and 15313. Various concentrations of (1%, 2.5% and 5%) were used and applied for 0, 4, 7, 10 and 14 days at 4 °C and 0, 6, 12, 18 and 24 h at 37 °C. 5% GSJ showed the strongest anti-listerial effect of 3–4 log cfu/ml against all the bacterial strains tested as compared to control at 37 °C. At 4 °C, 2.5% GSJ showed a strong growth inhibitory effect of about 2–3 log cfu/ml against all the bacterial strains when compared to control after 14 days, and 5% GSJ rather decreased the number of bacterial cell when compared to initial polulation and showed general lysis of cytoplasm, lysis of cell wall and cellular swelling, and on scanning electron microscopy (SEM) observation, strain L. monocytogenes ATCC 19118 showed swelling, partially distorted shape, pore formation and empty cell formation inside the cell.     

The determination of ready-to-eat foods into Listeria monocytogenes growth and no growth categories by challenge tests

To assess the potential for a food to allow the growth of Listeria monocytogenes throughout its shelf life and be compliant with European Commission regulations, a challenge test was designed by the EU Community Reference Laboratory. The procedure for the determination of the growth potential includes the following: product characteristics, shelf-life of the product, number of batches, choice of the strain(s), preparation of the inoculum, preparation and inoculation of the test units, storage conditions, measurement of physico-chemical characteristics of the food and microbiological analysis to measure any increase in the pathogen load. The results are then used to assign a ready-to-eat food into a growth or no growth category, and if the food is able to support the growth of L. monocytogenes, to quantify the behavior of this bacteria in a food between production and consumption.     

Characterisation of the resistance and the growth variability of Listeria monocytogenes after high hydrostatic pressure treatments

The use of non-thermal methods for food preservation is due to consumer demands for microbiological safe products, without changes in the sensory and nutritional qualities of the product. High hydrostatic pressure (HHP) has emerged as an alternative to traditional thermal processing methods for foods.Listeria monocytogenes CECT 5672 was treated under HHPs (350, 400 and 450 MPa) for 3, 16 and 23 min. The effects of pH (5, 6 and 7) and sodium chloride concentration (0, 0.5 and 1.0) of the recovery medium were studied on L. monocytogenes. The kinetic parameters were estimated by the method described by Métris, George, and Baranyi (2006). From results obtained, histograms of the lag phase were generated and distributions were fitted. The duration of the lag phase of HHP damaged cells increased with the application of additional stresses. Histograms showed a shift to longer lag phases and an increase in variability with high stress levels in the recovery medium.Using a primary model together with Monte Carlo simulation, predictions of time to growth (100 cfu/g) of L. monocytogenes were established and they were compared with deterministic predictions. It was evidenced that deterministic predictions do not give a good indication of the probability of a certain level of growth.     

Lactic acid bacteria biodiversity in Italian marinated seafood salad and their interactions on the growth of Listeria monocytogenes

The aim of this research was to study the biodiversity of lactic acid bacteria (LAB) in marinated seafood salad (pH 5.0) and their interaction on the growth of Listeria monocytogenes. LAB were highly present in the samples considered in this study, reaching values of 8.0 log cfu/g at the end of product’s shelf-life. A high biodiversity in terms of LAB species and strains was detected by means of RAPD–PCR within the 171 bacterial isolates collected. Among them Lactobacillus curvatus, Lactobacillus sanfranciscensis and Enterococcus were present in all the salad batches considered. Three challenge tests against L. monocytogenes were carried out in order to assess the growth of this pathogen in the presence of dominant populations of LAB. L. monocytogenes tended to decrease in time, suggesting that a stable concentration of LAB inhibits the development of this pathogenic micro-organism.     

Listeria innocua growth in fresh cut mixed leafy salads packaged in modified atmosphere

To investigate Listeria innocua fate in fresh cut mixed leafy salads, packaged both in ordinary (OA) and modified atmosphere (MA), the type strain DSM 20649 was inoculated and its behaviour was monitored by conventional and molecular approaches, during storage at 4 °C.Results indicated that: fresh cut salads packaged under MA (at initial conditions of 6% CO₂ and 3% O₂) supported the growth of Listeria spp.; conventional plating technique could not detect L. innocua even into inoculated samples, whereas; PCR–DGGE analysis showed that the L. innocua became one of the dominant species in samples packaged in MA, starting from the 3rd day of storage.The study confirms that modified atmosphere has to be applied together with other preservative techniques in order to assure the inhibition of pathogenic micro-organisms in fresh cut vegetables.     

Temperature distribution in Spanish domestic refrigerators and its effect on Listeria monocytogenes growth in sliced ready-to-eat ham

In order to evaluate the increase of L. monocytogenes concentration during home storage, a low level of the pathogen (<10 CFU/g) was inoculated in sliced ham. Two storage temperatures (5 °C and 9 °C) were selected from a previous study in 33 home refrigerators. Three days of storage or less were necessary to reach the regulated limited concentration (100 CFU/g), at both temperatures. When storage time was extended to 5 days, the pathogen achieved risky values (>10³ CFU/g).     

Non starter lactic acid bacteria biofilms: A means to control the growth of Listeria monocytogenes in soft cheese

The possibility to consider non starter lactic acid bacteria (NSLAB) biofilms as a means to control the growth of Listeria monocytogenes in soft cheeses was evaluated. In particular, the aptitude to form biofilm of four NSLAB strains (Lactobacillus plantarum DSM1055, Lactobacillus casei DSM20011, Lactobacillus curvatus DSM20019 and Lactobacillus paracasei DSM20207) was investigated. All tested strains were able to form biofilm on stainless steel and the highest quantities were produced when NSLAB were present simultaneously (about 6 Log CFU cm^?2). Then, experimental cheeses were made in presence of chips containing 7-days NSLAB biofilms and inoculated with L. monocytogenes (about 2 Log CFU g^?1). Results demonstrated that NSLAB biofilms can be considered a useful means to delay the growth of L. monocytogenes in soft cheeses: the maximum cell load attained at the stationary phase was about 6 Log CFU g^?1 in experimental cheeses against about 7 Log CFU g^?1 observed in control samples.     

Growth capacity of Listeria monocytogenes in ingredients of ready-to-eat salads

Raw minimally processed vegetables and fruits as well as boiled pasta and potato are the main ingredients of ready-to-eat (RTE) salads. These ingredients are diverse in their ability to support growth of Listeria monocytogenes. The objective of this study was to determine to what extent different ingredients can contribute to growth of L. monocytogenes in RTE salads. This was done by assessing the growth capacity of L. monocytogenes in ingredients of RTE salads by means of challenge tests and determination of the effect of product characteristics and presence of competitive flora. The majority of the tested products did not support or hardly supported growth of L. monocytogenes. The calculated maximum increase of 3.4 log CFU/g was exceeded in Galia melon, potato and pasta products. In RTE dinner salads, growth of L. monocytogenes was supported by the carbohydrate component. Galia melon, which is often used as ingredient of fruit salads, can contribute to growth of L. monocytogenes in these products. Growth inhibition, or differences in relative increase, could not always be explained by a_w, pH or presence of competitive flora. The effect of competitive flora, especially lactic acid bacteria, on growth inhibition was observed in non-pasteurized potato, white cabbage and mango.     

Surveillance of listeriosis and its causative pathogen,Listeria monocytogenes

To manage the problem of foodborne listeriosis requires an understanding of the burden of the disease on a worldwide scale as foods that are prone to contamination are eaten widely domestically and many are traded globally. Surveillance of the disease, caused by Listeria monocytogenes, is typically restricted to developed countries, but many of these do not consider listeriosis as a notifiable disease and estimate the numbers by other means. Incidence rates range from 0.3 to 1.3 per 100,000, but most are in the 0.3–0.5 range, irrespective of the regulatory system and industry control programmes that have been in place. Ready-to-eat foods are the vehicle for transmission of the Listeria through contamination somewhere in the food chain. Meat, poultry and dairy products have been most frequently implicated, but other foods including produce may also have been vehicles of transmission. Large outbreaks are usually linked to errors in food processing plants, such as contaminated slicing machines, followed by opportunities for growth of the pathogen. Less is known about home-generated illnesses but incorrect use of refrigerators can allow cross-contamination and growth of the pathogen to levels that can cause infections. In the U.S., door-to-door salesmen have sold contaminated Hispanic soft cheeses that have led to outbreaks and stillbirths. In addition to outbreak investigation, case-control studies, and the use of experts, risk assessments, and food attribution studies can help focus on areas of greatest risk for prevention and control measures throughout the food chain.     

The challenge of setting risk-based microbiological criteria for Listeria monocytogenes

After more than 20 years of work with discussing the setting of microbiological criteria for Listeria monocytogenes in foods, Codex Alimentarius on Food Hygiene has finalised a proposal that was recently adopted by the Codex Alimentarius Commission. The effort of developing procedures for making the microbiological criteria risk-based to the greatest extent possible has challenged scientists and managers during this long time period. Yet, the establishment of microbiological criteria for L. monocytogenes is still being discussed and several approaches are possible. Setting of microbiological criteria continues to be a risk management decision – even when using mathematical modelling to enhance the scientific level of a risk-based approach.     

Effects of high pressure homogenization on the activity of lysozyme and lactoferrin against Listeria monocytogenes

The effects of high pressure homogenization treatment at 100 MPa (HPH), in comparison to different heat treatments, 70 °C for 30 s, 70 °C for 5 min or 100 °C for 5 min, on the activity of lysozyme and lactoferrin, were studied. The antimicrobial activities of lysozyme and lactoferrin were tested on Listeria monocytogenes inoculated in milk or cultural medium.The results indicated that antimicrobial activities of lactoferrin and lysozyme were enhanced and/or accelerated by HPH treatment. Particularly, the highest immediate inactivation values were recorded when L. monocytogenes cells were added to HPH-treated lactoferrin, processed simultaneously or separately with the target microorganism. Although to a lesser extent than HPH treatment the heat treatments applied also were able to increase the antimicrobial activity of lysozyme.     

Susceptibility of Listeria monocytogenes from traditional cheese-dairies to in-use sanitizers

The minimal inhibitory concentrations (MICs) and the minimal bactericidal concentrations (MBCs) of five sanitizers (alkyl amine acetate, phosphoric acid, anionic tension active and two chlorine/alkaline) against 15 isolates of Listeria monocytogenes (including isolates found persistently in two dairies) were determined. Alkyl amine acetate was the only agent whose efficacy was poorly affected by the presence of organic matter. The values of MICs and MBCs of cells pre-exposed to sub-inhibitory and sub-lethal concentrations of the disinfectants, did not suggest adaptation. It is possible that factors other than resistance or adaptation to sanitizers could account for the persistence of particular clones of L. monocytogenes in these dairies.     

Adaptive response of Listeria monocytogenes to heat and its impact on food safety

Listeria monocytogenes is an important food associated pathogen because of its relatively high heat resistance and ability to multiply in refrigeration temperatures. Its thermotolerance can be increased when its cells are subjected to heat shock. One- to eight-fold increase of D values of L. monocytogenes have been reported, depending on the heat shock duration, the temperature and the heating menstrum. This acquisition of heat tolerance is related to the induction of the synthesis of heat shock proteins (HSPs).The adaptive response of food pathogens has important consequences on the safety of thermally processed foods. It is believed that this is responsible for the frequent occurrence of deviations (tails and shoulders) during heat treatments that are observed in the exponential model of microbial inactivation. These deviations from log-linear kinetic especially encountered under mild heat treatments, mean that prediction of food safety can no longer rely upon D and z values. Adaptive response to heat must be considered when quantifying and modeling microbial inactivation during thermal processing in order to achieve microbiologically safe products without overly conservative heat processes. Therefore a more mechanistic approach is needed for more accurate predictions of thermal inactivation. Prerequisite to this model are thorough studies to understand how L. monocytogenes and other pathogens adapt their cellular physiology to overcome heat and other stresses.