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.     

Radiosensitization of Salmonella spp. and Listeria spp. in ready-to-eat baby spinach leaves

The FDA recently approved irradiation treatment of leafy greens such as spinach up to 1 kGy; however, it is important to reduce the dose required to decontaminate the produce while maintaining its quality. Thus, the objectives of this study were: (1) to assess the radiation sensitivities of Salmonella spp. and Listeria spp. inoculated in ready-to-eat baby spinach leaves under modified atmosphere packaging (MAP) and irradiated using a 1.35-MeV Van de Graff accelerator (the leaves were irradiated both at room temperature and at -5 °C); and (2) to understand and optimize the synergistic effect of MAP and irradiation by studying the radiolysis of ozone formation under different temperatures, the effect of dose rate on its formation, and its decomposition. Results showed that increased concentrations of oxygen in the packaging significantly increased the radiation sensitivity of the test organisms, ranging from 7% up to 25% reduction in D(10)-values. In particular, radiosensitization could be effected (P < 0.05) by production of ozone, which increases with increasing dose-rate and oxygen concentration, and reducing temperatures. Radiosensitization was demonstrated for both microorganisms with irradiation of either fresh or frozen (-5 °C) baby spinach. These results suggest that low-dose (below 1 kGy) e-beam radiation under modified atmosphere packaging (100% O(2) and N(2):O(2)[1:1]) may be a viable tool for reducing microbial populations or eliminating Salmonella spp. and Listeria spp. from baby spinach. A suggested treatment to achieve a 5-log reduction of the test organisms would be irradiation at room temperature under 100% O(2) atmosphere at a dose level of 0.7 kGy. Practical Application: Decontamination of minimally processed fruits and vegetables from food-borne pathogens presents technical and economical challenges to the produce industry. Internalized microorganisms cannot be eliminated by the current procedure (water-washed or treated with 200-ppm chlorine). The only technology available commercially is ionizing radiation; however, the actual radiation dose required to inactivate pathogens is too high to be tolerated by the product without unwanted changes. This study shows a new approach in using MAP with 100% O(2), which is converted to ozone to radiosensitize pathogens while improving the shelf life of minimally processed fruits and vegetables. The process results in a high level of microorganism inactivation using lower doses than the conventional irradiation treatments.     

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.     

Growth potential of Salmonella enterica in thirty-four different RTE vegetable salads during shelf-life

Thirty-four different ready-to eat (RTE) vegetable salads were inoculated with a cocktail of three Salmonella enterica strains, and stored under a modified atmosphere for up to 168 h at 4, 7, 12 and 16°C. Eighteen (18) of the salad samples comprised of two or more vegetable ingredients (also referred to as MV RTE salads), and 16 were made up of single vegetable ingredients (SV RTE salads). Generally, the growth potential of inoculated S. enterica varied depending on temperature and type of RTE vegetable salad. The higher temperature was generally more favourable for the growth of S. enterica. Among all 34 salad samples, 5, 11, 18 and 24 salad samples supported the growth of Salmonella at 4, 7, 12 and 16°C, respectively. All salads consisting of multiple vegetable ingredients except two: one comprised of carrots, lettuce and beetroot and another comprised of white cabbage and purple cabbage, supported the growth of Salmonella at high temperatures (either 12 or 16 or both 12 and 16°C). Although the growth of Salmonella was variable in the different types of RTE salads, and growth was generally low at 4°C, Salmonella exhibited consistently minimal growth in some vegetable salads such as those comprised of carrots, lettuce and beetroot, carrots, beetroots, cabbage and cucumber, as well as one comprised of beetroot and corn at all temperature conditions tested.     

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.     

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.     

Population dynamics of Salmonella spp. and Shigella spp. in ready-to-eat Mediterranean vegetable salads

This study evaluated the behavior of Salmonella and Shigella (5–6 log CFU/g) in tomato–cucumber (TC) salad without additives (control), TC with 1.0% lemon juice and 0.5% salt, TC with 10% wt/wt tahini, coleslaw, and toum sauce at 4, 10, or 24°C for 5 days. At 4°C, both pathogens survived well in all salads, with a 0.2–1.6 log CFU/g reduction after 5 days (except for toum sauce with >3.5 log CFU/g reduction after 4 days). At 10°C, Salmonella in the different TC salads remained constant, whereas Shigella numbers significantly increased by 1.0–1.7 log CFU/g after 5 days. Yet, both pathogens significantly decreased by 1.2–1.4 log CFU/g in coleslaw after 5 days and by >3.5 log CFU/g in toum sauce after 3 days. At 24°C, Salmonella significantly increased in TC salad without additives by 1.4 log CFU/g after 5 days and were below the detection level in the other types of salad after 5 days. However, Shigella numbers significantly increased by 1.0 log CFU/g in TC with tahini, but they significantly declined by 1.9–2.9 log CFU/g in TC salads after 5 days, and the pathogen was not detected in coleslaw and toum sauce after 4 days.     

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.     

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.     

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.     

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.     

我国零售食品单增李斯特菌污染的健康风险分级研究

目的对我国零售阶段食品中单增李斯特菌污染的健康风险进行评估和分级。方法利用全国污染物监测网中2010—2013年食品中单增李斯特菌污染水平监测数据、2002年中国居民营养与健康状况调查中的膳食消费量数据以及2011年中国统计年鉴资料,对李斯特菌病敏感人群通过5大类17小类食品暴露于单增李斯特菌的风险进行评估和分级。结果生食水产品是单增李斯特菌污染水平较高的食品类别,其次是散装熟肉制品。豆腐皮/丝是导致李斯特菌病每年发病风险最高的食品,散装熟肉食品导致的单增李斯特菌健康风险是定型包装的4~10倍。结论生食水产品和散装熟肉制品导致的每餐单增李斯特菌病发病风险最高,即食非发酵豆制品和熟肉制品是可能导致我国每年发生单增李斯特菌病的最主要食品。     

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.     

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.