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.     

Growth of Listeria monocytogenes in refrigerated ready-to-eat foods in Japan

The ability of L. monocytogenes to grow in a series of Japanese ready-to-eat (RTE) foods, including boiled baby sardine and Japanese pickle, was tested at two different refrigeration temperatures. In RTE foods in which L. monocytogenes can grow, growth was significantly higher at 10°C than that at 4°C during their shelf lives and growth patterns varied extensively among the different types of foods. However, growth did not occur at 4°C within the shelf life of certain RTE foods, such as broiled squid. The patterns of growth were varied extensively with different sample types. These results suggest that some types of traditional Japanese RTE foods stored at 10°C may be potential sources of listeriosis. To reduce the risk of food-borne listeriosis, studies to determine the contamination levels in RTE foods and the effects of storage temperature on their shelf lives are needed.     

Prevalence of Listeria monocytogenes and Salmonella in ready-to-eat food in Catalonia, Spain

Listeria monocytogenes and Salmonella are pathogenic bacteria that can contaminate food products during or after processing. Ready-to-eat (RTE) food does not undergo any treatment to ensure its safety before consumption, and therefore risk of foodborne disease must be considered if these pathogens are present in the food. To evaluate the prevalence of these pathogens in RTE food, 140 RTE fish product samples, 501 RTE meat product samples, 462 RTE dairy samples, and 123 RTE dishes and desserts, providing a total of 1,226 samples, were collected from retail stores and food industry and analyzed for the presence of L. monocytogenes. A total of 1,379 samples consisting of 187 RTE fish products and 569 RTE meat products, 484 RTE dairy products, and 139 RTE dishes and desserts were collected and analyzed for the presence of Salmonella. L. monocytogenes was isolated from 20% of frozen Atlantic bonito small pies, 7.9% of smoked salmon samples, 11.1% of the pork luncheon meat samples, 6.2% of frozen chicken croquettes, 16.9% of cured dried sausage samples, 12.5% of cooked ham samples, and 20% of cooked turkey breast samples. L. monocytogenes was also found to be present in 1.3% of fresh salty cheese samples and 15.1% of frozen cannelloni samples. Salmonella was isolated from 1.2% of smoked salmon samples, 1.5% of frozen chicken croquettes, 2% of cooked ham samples, and 11.1% of cured dried sausage samples. Overall, occurrence of these pathogens in RTE foods was similar to that previously reported in the literature.     

Predictions of growth for Listeria monocytogenes and Salmonella during fluctuating temperature

We studied the predictive performance of a dynamic modelling approach, combined with predictions from the Food MicroModel software, applied to the growth of Listeria monocytogenes and Salmonella in pasteurised milk, chicken liver paté and minced chicken, under constant as well as fluctuating temperatures. We found that, in general, the accuracy of a prediction under fluctuation temperature was similar to that under constant temperature. Generally, there was a good agreement between predictions and observations. However, the growth of Listeria monocytogenes in pasteurised milk was inhibited largely by the natural flora present.     

Contamination profile of Listeria spp. in three types of ready-to-eat chicken meat products

This study investigated contamination sources of Listeria spp. in frozen, ready-to-eat, roasted, steamed, and fried chicken meat products from a plant in Thailand, as well as the correlation between Listeria contamination in the production environment and the finished product. The cooking processes used in this factory (with a product core temperature of 80 degrees C for 1 min) were confirmed as adequate for eliminating Listeria spp. However, Listeria spp. were detected at the packing stage of roasted and steamed chicken products. An environmental swab test was conducted by means of the zone concept, whereby surfaces in the production area were divided into three zones. Zone 1 was made up of the equipment surfaces that came into direct contact with the products. Zone 2 consisted of equipment surfaces that were not in direct contact with the products, including surfaces that were difficult to be cleaned. Zone 3 included surfaces that did not come in direct contact with the products and were located far from the products. The results showed that the prevalence of Listeria spp. in roasted and steamed products was affected by the prevalence of Listeria contamination in all zones, especially zone 1, which demonstrated the highest correlation. In addition, the prevalence of Listeria contamination in zones 2 and 3 affected the prevalence of Listeria in zone 1. A correlation between Listeria on roasted chicken products and the surfaces of zone 1 at the start of production was also established.     

Prevalence of Salmonella spp. and Listeria monocytogenes at small-scale spanish factories producing traditional fermented sausages

The manufacturing of fermented sausages is subject to natural contamination processes that can potentially carry foodborne pathogens along the process chain and result in contamination of the final product. The aim of this study was to evaluate the occurrence of Salmonella spp. and Listeria monocytogenes at different sampling points during the manufacturing process of fuet, a type of traditional fermented sausage, at 10 small-scale Spanish factories. The presence of both pathogens was studied in the raw materials (19 casings and 19 meat batters), the final products (19 fermented sausages), and the factory equipment (12 mincing, 12 mixing, and 19 stuffing machines, 19 cutting tables, 11 knives, and 12 cold rooms) by using classical microbiological techniques and real-time PCR. Salmonella was not detected in the equipment analyzed or in the final products, but it was detected in the raw materials (23.7% of samples). L. monocytogenes showed higher incidence than Salmonella and was detected in the equipment (11.8% of samples), the raw materials (28.9%), and the final products (15.8%), confirming its ubiquity throughout the manufacturing process of fermented sausages. Five factories were further investigated to study the changes in the distribution of pathogens in the fuet production process over a period of either 2 or 3 years. There was considerable variation in the incidence of both pathogens at different sampling periods, and there was no relation between seasonal variations or geographic location of the factories.     

Effectiveness of irradiation treatments in inactivating Listeria monocytogenes on fresh vegetables at refrigeration temperature

Ionizing radiation can be effective in controlling the growth of food spoilage and foodborne pathogenic bacteria. This study reports on an investigation of the effectiveness of irradiation treatment to eliminate Listeria monocytogenes on laboratory-inoculated broccoli, cabbage, tomatoes, and mung bean sprouts. Irradiation of broccoli and mung bean sprouts at 1.0 kGy resulted in reductions of approximately 4.88 and 4.57 log CFU/g, respectively, of a five-strain cocktail of L. monocytogenes. Reductions of approximately 5.25 and 4.14 log CFU/g were found with cabbage and tomato, respectively, at a similar dose. The appearance, color, texture, taste, and overall acceptability did not undergo significant changes after 7 days of postirradiation storage at 4 degrees C, in comparison with control samples. Therefore, low-dose ionizing radiation treatment could be an effective method for eliminating L. monocytogenes on fresh and fresh-cut produce.     

Measurements and predictions of growth for Listeria monocytogenes and Salmonella during fluctuating temperature II. Rapidly changing temperatures

Growth of Listeria monocytogenes and Salmonella was examined during various rates of increase and decrease in temperature from and to the minimum for growth. Growth was little affected by even the most rapid changes and injury or lag was not observed. Subsequent investigations of growth during periods of rapid variation in temperature from and to temperatures below the growth minimum again had little effect and growth was satisfactorily predicted using the dynamic model of Baranyi and Roberts [Int. J. Food Microbiol. 23 (1994) 277] in conjunction with the data of Food Micromodel.     

Reduction and survival of Listeria monocytogenes in ready-to-eat meats after irradiation

A five-strain Listeria monocytogenes culture was inoculated onto six different types of ready-to-eat (RTE) meats (frankfurters, ham, roast beef, bologna, smoked turkey with lactate, and smoked turkey without lactate). The meats were vacuum packed and stored at 4 degrees C for 24 h prior to irradiation. Populations of L. monocytogenes were recovered by surface plating on nonselective and selective media. The margins of safety studied include 3-log (3D) and 5-log (5D) reduction of pathogenic bacteria to achieve an optimal level of reduction while retaining organoleptic qualities of the meats. A 3-log reduction of L. monocytogenes was obtained at 1.5 kGy when nonselective plating medium was used. The dosages for 3-log reduction were 1.5 kGy for bologna, roast beef, and both types of turkey and 2.0 kGy for frankfurters and ham on the basis of use of selective medium. The D10-values ranged from 0.42 to 0.44 kGy. A 5-log reduction of L. monocytogenes was obtained at 2.5 kGy with nonselective medium. With selective medium, the dosages were 2.5 kGy for bologna, roast beef, and both types of turkey and 3.0 kGy for frankfurters and ham. Survival of L. monocytogenes in the same RTE meat types after irradiation was also studied. Meats were inoculated with 5 log L. monocytogenes per g and irradiated at doses of 2.0 and 4.0 kGy. Recovery of the surviving organisms was observed during storage at temperatures of 4 and 10 degrees C for 12 weeks. Preliminary results showed no growth in meats irradiated at 4.0 kGy. Survivors were observed for irradiated meats at 2.0 kGy stored at 10 degrees C after the second week. No growth was observed in samples irradiated at 2.0 kGy stored at 4 degrees C until the fifth week.     

Inhibition of Listeria monocytogenes by Carnobacterium spp. strains in a simulated cold smoked fish system stored at 4 degrees C.

Preservation of smoked salmon from bacterial spoilage, and especially from Listeria monocytogenes by bacteriocin producers is a promising challenge. Over a hundred lactic acid bacteria, isolated from commercial vacuum packaged cold smoked salmon, were screened for their antagonistic activity against L. innocua. Twenty-two strains were able to produce bacteriocin-like proteinaceous substances. These strains were characterized physiologically and biochemically as Carnobacterium strains. Three different groups were determined by pulsed-field gel electrophoresis after Sma I and Apa I DNA digestion. Peptidoglycan hydrolases patterns completed the characterization of these strains. All were confirmed as being Carnobacterium piscicola. Growth and bacteriocin production of three strains of each group and two well known bacteriocin producers (C. divergens V41 and C. piscicola V1) were tested in a simulated cold smoked fish system at 4 degrees C. These strains were able to reach 10(8) cfu ml(-1) in 21 days and to produce as much bacteriocin activities in the cold smoked fish system as in the rich media. Carnobacterium divergens V41 and C. piscicola V1 were the most effective strains in co-culture experiments, inhibiting L. monocytogenes as early as day 4, whereas C. piscicola SF668 inhibiting effect was observed at day 13. The potential for using such biopreservation treatments on whole smoked salmon is discussed.     

Combined effect of packaging atmosphere and storage temperature on growth of Listeria monocytogenes on ready-to-eat shrimp

Cooked, peeled, and deveined shrimp were inoculated with a 5 strain mixture of Listeria monocytogenes and packaged in air, vacuum, and a 100% carbon dioxide modified atmosphere. The packaged shrimp were then stored at 3, 7, and 12 degrees C for 15 days to monitor the growth of L. monocytogenes and psychrotrophic bacteria. Uninoculated shrimp were also subjected to sensory evaluation by a trained panel to measure odor and appearance over the storage period. Results demonstrated that shrimp packaged in CO(2) and stored at 3 degrees C did not permit growth of L. monocytogenes during the 15-day storage period, while all other packaging/temperature combinations allowed for multiplication of the bacterium. Carbon dioxide packaging also resulted in the slowest growth of psychrotrophic bacteria and resulted in shrimp having acceptable sensory odor and appearance scores at the end of storage. When strict temperature control is difficult, such as during processing, transportation, retail display, or home use, additional antimicrobial hurdles may be necessary to ensure safety.     

Multiplex Real-time PCR for the Simultaneous Detection of Salmonella spp. and Listeria monocytogenes in Food Samples

In this study, we have developed a rapid method for the simultaneous detection of Listeria monocytogenes and Salmonella spp. in foods, combining culture enrichment and a multiplex real-time polymerase chain reaction (PCR). The assay used two pre-existing primer-probe sets, labelled with different reporter dyes to enable the direct distinction of the original contaminating agent. Amplification efficiency and inclusivity/exclusivity of the combined assay was successfully assessed. The overall process included the culture enrichment based on the ISO standard, consisting of 24 h incubation in appropriate media (Half Fraser Broth for Listeria and buffered peptone water (BPW) for Salmonella), followed by a single DNA extraction of mixed enrichment aliquots, and real-time PCR detection of the hly and bipA genes of L. monocytogenes and Salmonella spp., respectively. An internal amplification control, co-amplified during the PCR run, was included in the assay to verify the results. The tool was evaluated with a variety of artificially inoculated samples of fresh products and ready to eat and cooked dishes, allowing the identification of the target pathogens down to 5 CFU/25 g of food sample. Moreover, the analysis saved a considerable amount of time compared to the ISO standard, being performed in less than 2 working days. Specificity, sensitivity and accuracy were satisfactorily tested by comparison to the standard methods ISO 11290-2:1998 and ISO 6579:2002, suggesting that the tool has a great potential as a reliable alternative for food safety assurance providing rapid detection of both pathogens in food samples.     

Modeling the growth rate and lag time of different strains of Salmonella enterica and Listeria monocytogenes in ready-to-eat lettuce

The growth parameters (growth rate, μ and lag time, λ) of three different strains each of Salmonella enterica and Listeria monocytogenes in minimally processed lettuce (MPL) and their changes as a function of temperature were modeled. MPL were packed under modified atmosphere (5% O₂, 15% CO₂ and 80% N₂), stored at 7–30 °C and samples collected at different time intervals were enumerated for S. enterica and L. monocytogenes. Growth curves and equations describing the relationship between μ and λ as a function of temperature were constructed using the DMFit Excel add-in and through linear regression, respectively. The predicted growth parameters for the pathogens observed in this study were compared to ComBase, Pathogen modeling program (PMP) and data from the literature. High R² values (0.97 and 0.93) were observed for average growth curves of different strains of pathogens grown on MPL. Secondary models of μ and λ for both pathogens followed a linear trend with high R² values (>0.90). Root mean square error (RMSE) showed that the models obtained are accurate and suitable for modeling the growth of S. enterica and L. monocytogenes in MP lettuce. The current study provides growth models for these foodborne pathogens that can be used in microbial risk assessment.     

Incidence of Listeria spp. and Salmonella spp. in horsemeat for human consumption

The incidence of Salmonella spp., Listeria spp. and Listeria monocytogenes in horsemeat for human consumption was investigated. One-hundred and twenty-one samples of frozen horsemeat collected from two Brazilian abattoirs were analysed over a period of 1 year. Twenty-two samples (18.2%) were positive for Listeria spp. with nine (7.4%) containing L. monocytogenes. None of the samples harbored Salmonella spp.     

Comparison of a cold enrichment and the FDA method for isolating Listeria monocytogenes and other Listeria spp. from ready-to-eat food on retail sale in the U.K

A cold enrichment and the modified FDA selective enrichment method were compared for their ability to detect Listeria monocytogenes and other Listeria species from various ready-to-eat foods on sale in the UK. Of 57 food samples examined using cold enrichment, five yielded L. monocytogenes, and two L. innocua. The FDA enrichment method yielded three samples positive for L. monocytogenes only. Foods examined included soft cheeses, fermented meat sausages, pates and salads.     

Enhanced antimicrobial effects of combination of lactate and diacetate on Listeria monocytogenes and Salmonella spp. in beef bologna

The antimicrobial activities of salts of organic acids such as lactate and acetate are well documented, but there is limited information on their effect when used in combination. We previously reported enhanced inhibition of Listeria monocvtogenes and Salmonella enterica serovar Enteritidis in sterile comminuted beef at 5 and 10 degrees C by combinations of sodium lactate (SL) (2.5%) and sodium diacetate (SDA) (0.2%). The present study was undertaken to evaluate the inhibitory effect of these salts, alone and in combination, in ready-to-eat (RTE) meat. Single strains and six-strain mixtures of each of the pathogens ( approximately 3 log CFU/g) were tested in beef bologna during aerobic storage at 5 and 10 degrees C for up to 60 days. The growth rate of the six-strain mixture of Listeria was faster than that of the single strain (Scott A) in the lactate/diacetate-free product. While each of the salts delayed growth of the listeriae at 5 degrees C, the effect of their combination was listericidal for the single strain and listeriostatic for the six-strain mixture. Enhanced inhibition by the salt combination was also observed at 10 degrees C. Salmonella numbers declined to undetectable levels in the untreated meat product and in each of the treatments after 20-30 days. However, the decline was more rapid in meat with the combination of the salts during storage at both 5 and 10 degrees C. Each of the salts further delayed the growth of the background microflora during storage at 5 degrees C, with their combinations showing the most effect.     

Growth of Listeria monocytogenes in vacuum-packed, smoked salmon, during storage at 4 degrees C.

Samples of smoked salmon of different hygienic quality were inoculated with low (6 cfu/g) and high (600 cfu/g) levels of a mixture of three strains of Listeria monocytogenes, after which they were vacuum-packed and stored at 4 degrees C for up to 5 weeks. L. monocytogenes grew well during storage in all the inoculated sample groups. Growth was, however, slightly faster in the fish with the better hygienic quality. The smoked salmon was still sensorically acceptable after 4 weeks. All three strains were found after 4 weeks in the fish with the better quality, while only two strains were recovered after the same time from the poorer quality salmon.     

Survival of Listeria monocytogenes in vanilla-flavored soy and dairy products stored at 8 degrees C

The survival of Listeria monocytogenes V37 in vanilla-flavored yogurt (low-fat and nonfat) and soy milk (low-fat and Plus) stored at 8 degrees C for 31 days was investigated. Commercial samples of yogurt and soy milk were used. These samples were inoculated with either 10(4) or 10(7) CFU of L. monocytogenes per ml. Sampling was carried out every 3 to 4 days initially and was then carried out weekly, for a total storage time of 31 days. Each time a sample was collected, the pH of the sample was measured. After 31 days, low-fat plain, low-fat vanilla, and nonfat plain yogurt samples inoculated with 10(4) CFU/ml showed 2.5-log reductions in viable cell populations, and nonfat vanilla yogurt showed a 3.5-log reduction. For yogurt inoculated with 10(7) CFU/ml, reductions of 2.5 log CFU/ml were observed for plain low-fat and nonfat yogurts, and reductions of 5 log CFU/ml were observed for vanilla-flavored low-fat and nonfat yogurts. In vanilla-flavored and plain low-fat and Plus soy milk samples, cell counts increased from 10(4) and 10(7) CFU/ml to 10(9) CFU/ml at 7 and 3 days of storage, respectively, at 8 degrees C. Coagulation in soy milk samples was observed when the cell population reached 10(9) CFU/ml. In soy milk, the L. monocytogenes population did not change for up to 31 days. Vanillin had an inhibitory effect on L. monocytogenes in yogurt but not in soy milk.     

Irradiation temperature influences product quality factors of frozen vegetables and radiation sensitivity of inoculated Listeria monocytogenes

Four frozen vegetables (broccoli, corn, lima beans, and peas) were gamma irradiated at subfreezing temperatures ranging from -5 to -20 degrees C to determine (i) the radiation sensitivity of an inoculated outbreak strain of Listeria monocytogenes (ATCC 49594), (ii) the effect of changing irradiation conditions (i.e., temperature) on that sensitivity, and (iii) the effect of the recommended radiation dose on the texture and color of irradiated frozen vegetables. The amounts of radiation necessary to reduce the bacterial population by 90% (D10-values) for L. monocytogenes differed significantly among vegetables at each irradiation temperature. D10 increased significantly with decreasing temperature for all vegetables, with each vegetable showing a different response pattern. At an irradiation temperature of -5 degrees C, D10 ranged from 0.505 kGy for broccoli to 0.613 kGy for corn. At -20 degrees C, D10 ranged from 0.767 kGy for lima beans to 0.916 kGy for peas. At -20 degrees C, radiation doses sufficient to achieve a 5-log10 kill (3.9 to 4.6 kGy) caused significant softening of peas and broccoli stems but not of corn or lima beans. Lower doses of comparable antimicrobial efficacy delivered at -5 degrees C (2.5 to 3.1 kGy) did not cause significant changes in texture in any vegetable. Color varied significantly among the dose-temperature combinations only for broccoli florets; this variation did not demonstrate a clear pattern of quality changes in response to irradiation.     

Molecular ecology of Listeria monocytogenes and other Listeria species in small and very small ready-to-eat meat processing plants

A longitudinal study was conducted to track Listeria contamination patterns in ready-to-eat meats from six small or very small meat processing plants located in three states over 1 year. A total of 688 environmental sponge samples were collected from nonfood contact surfaces during bimonthly visits to each plant. Overall, L. monocytogenes was isolated from 42 (6.1%) environmental samples, and its prevalence ranged from 1.7 to 10.8% across different plants. Listeria spp., other than L. monocytogenes, were isolated from 9.5% of samples overall, with the prevalence ranging from 1.5 to 18.3% across different plants. The prevalence of L. monocytogenes correlated well with that of other Listeria spp. for some but not all plants. One L. monocytogenes isolate representing each positive sample was characterized by molecular serotyping, EcoRI ribotyping, and pulsed-field gel electrophoresis typing. Seven sample sites tested positive for L. monocytogenes on more than one occasion, and the same ribotype was detected more than once at five of these sites. Partial sigB sequencing was used to speciate other Listeria spp. isolates and assign an allelic type to each isolate. Other Listeria spp. were isolated more than once from 14 sample sites, and the same sigB allelic type was recovered at least twice from seven of these sites. One plant was colonized by an atypical hemolytic L. innocua strain. Our findings indicate that small and very small meat processing plants that produce ready-to-eat meat products are characterized by a varied prevalence of Listeria, inconsistent correlation between contamination by L. monocytogenes and other Listeria spp., and a unique Listeria molecular ecology.