Design of challenge testing experiments to assess the variability of Listeria monocytogenes growth in foods

The assessment of the evolution of micro-organisms naturally contaminating food must take into account the variability of biological factors, food characteristics and storage conditions. A research project involving eight French laboratories was conducted to quantify the variability of growth parameters of Listeria monocytogenes obtained by challenge testing in five food products. The residual variability corresponded to a coefficient of variation (CV) of approximately 20% for the growth rate (μ_max) and 130% for the parameter K = μ_max × lag. The between-batch and between-manufacturer variability of μ_max was very dependent on the food tested and mean CV of approximately 20 and 35% were observed for these two sources of variability, respectively. The initial physiological state variability led to a CV of 100% for the parameter K. It appeared that repeating a limited number of three challenge tests with three different batches (or manufacturers) and with different initial physiological states seems often necessary and adequate to accurately assess the variability of the behavior of L. monocytogenes in a specific food produced by a given manufacturer (or for a more general food designation).     

Predictive microbiology models vs. modeling microbial growth within Listeria monocytogenes risk assessment: what parameters matter and why

Predictive microbiology models are essential tools to model bacterial growth in quantitative microbial risk assessments. Various predictive microbiology models and sets of parameters are available: it is of interest to understand the consequences of the choice of the growth model on the risk assessment outputs. Thus, an exercise was conducted to explore the impact of the use of several published models to predict Listeria monocytogenes growth during food storage in a product that permits growth. Results underline a gap between the most studied factors in predictive microbiology modeling (lag, growth rate) and the most influential parameters on the estimated risk of listeriosis in this scenario (maximum population density, bacterial competition). The mathematical properties of an exponential dose-response model for Listeria accounts for the fact that the mean number of bacteria per serving and, as a consequence, the highest achievable concentrations in the product under study, has a strong influence on the estimated expected number of listeriosis cases in this context.     

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.     

Critical evaluation of the EU-technical guidance on shelf-life studies for L. monocytogenes on RTE-foods: a case study for smoked salmon

In November 2008, a technical guidance document on the challenge test protocol was published by the EU CRL (Community of Reference Laboratory) for L. monocytogenes. This document describes the practical aspects on the execution of a challenge test in order to comply to the EU Commission regulation N° 2073/2005 on microbiological criteria for foodstuff. In this guideline two approaches are specified. On the one hand challenge tests, based on actual data measurements at the beginning and end of the shelf-life of products stored under reasonably foreseen T-profile, are described. On the other hand, growth potential is calculated by predictive models using a validated maximum specific growth rate.The present study evaluates the two above mentioned approaches on cold smoked salmon, a typical risk product for L. monocytogenes. The focus is on: (i) the relative importance of intrabatch versus interbatch variability, (ii) the concept of a simple challenge test based on actual data at start and end of shelf life versus a modelling approach and (iii) the interpretation of challenge tests. Next to this, available tertiary models were used to estimate the growth potential of these products based on their initial physicochemical characteristics.From the results it could be concluded that in some batches considerable intrabatch variability was obtained. In general, however, the interbatch variability was significantly higher than intrabatch variability. Concerning the two above mentioned methods for challenge tests, it can be stated that the first approach (simple challenge test) can be set up rather rapidly and is cost-effective for SMEs (small and medium enterprises) but provides only a single isolated outcome. This implies that challenge tests should be redone if changes occur in composition or production process. The second (modelling) approach, using extended challenge tests to establish growth parameters needs larger set ups and more complicated data analysis, which makes them more expensive. Using available tertiary models has the major advantage that the most important intrinsic and extrinsic factors can be included for the prediction of the growth parameter. It was clear that product specific models, taking into account the interaction effects with background flora, performed the best. Regarding the challenge tests, it can be concluded that the best approach to choose will depend on the particular context as in the end both approaches will lead to the same conclusion.     

Assessing biofilm formation by Listeria monocytogenes strains

When a microtitre plate assay was used to quantify biofilm production by Listeria monocytogenes strains following growth in Tryptone Soy Broth (TSB) for 48 h at 20 degrees C, 127 of 138 strains (92.0%) were classified as weak, 9 of 138 strains (6.5%) as moderate and only 2 of 138 strains (1.5%) as strong biofilm formers. The strains included environmental, animal, food (persistent and sporadic strains) and clinical isolates previously typed using esterase electrophoresis (ESE) and multi-locus enzyme electrophoresis (MEE). Strains from different sources produced similar quantities of biofilm, whereas biofilm production by ESE type II strains, irrespective of source, was greater than that observed for other ESE types. No correlation between MEE type and biofilm production was observed. A Petri dish assay which allowed parallel quantification and microscopic examination of biofilms was used to examine biofilm formation by selected L. monocytogenes strains during growth in TSB for 14 days at 20 degrees C. Results from these assays showed that following prolonged incubation, some L. monocytogenes strains categorized as weak biofilm formers by the 48 h microtitre assay, were able to form biofilms similar in terms of quantity and structure to those produced by strains classified as strong or medium biofilm formers. Results from 14-day Petri dish assays confirmed 48 h microtitre assays regarding greater biofilm production by ESE type II strains compared to other ESE types of L. monocytogenes. Biofilm production was similar for ESE type II persistent and sporadic food isolates but reduced for ESE type II clinical strains.     

Growth/no growth model of Listeria monocytogenes as a function of temperature, pH, citric acid and ascorbic acid

A linear logistic regression model was built to describe the growth/no growth boundaries of Listeria monocytogenes as a function of temperature (4–30 °C), pH (4.5–6), citric acid (0–0.4%) and ascorbic acid (0–0.4%). A fractional factorial design was followed among the factors considered and an inoculum size of 10⁵ CFU/ml was used. Evaluation of growth was performed by optical density measurements in Bioscreen C (Labsystems, Finland), during 21 days. Data of optical density were transformed to log CFU/ml by using a calibration curve. Among the 232 combinations of factors tested, growth was observed in 133 and no growth in 99. The degree of agreement between predictions and observations was 97.8% concordant and 2.2% discordant. An internal validation with additional data within the interpolation region of the model was performed. The predictions were concordant in 94% of the cases, and all the wrong cases failed to the safe side of the boundary region. The probability of growth was strongly influenced at low temperatures (<15 °C). The effect of pH was more notorious below 5.3, since the minimum temperature that inhibited growth was higher. Citric acid was more effective than ascorbic acid when the analysis was based on the undisssociated acid concentration (u.a.c.). However, when dealing with the undissociated fraction (u.a.f.), ascorbic acid presented more inhibitory effect. Organic acids also accentuate the temperature and pH inhibition of bacterial growth limits, and increased the minimum pH at which growth was detected. These results have a practical implication for stakeholders and risk managers in order to identify the treatments that can be applied to food and ensure that no growth of this pathogenic microorganism will occur.     

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.     

Effect of combining nisin and/or lysozyme with in-package pasteurization for control of Listeria monocytogenes in ready-to-eat turkey bologna during refrigerated storage

This study investigated the efficacy of in-package pasteurization combined with pre-surface application of nisin and/or lysozyme to reduce and prevent the subsequent recovery and growth of Listeria monocytogenes during refrigerated storage on the surface of low-fat turkey bologna. Sterile bologna samples were treated with solutions of nisin (2 mg/ml=5000 AU/ml), lysozyme (10 mg/ml=80 AU/ml) and a mixture of nisin and lysozyme (2 mg nisin+10mg lysozyme/ml) before in-package pasteurization at 65 degrees C for 32s. In-package pasteurization resulted in an immediate 3.5-4.2 log CFU/cm(2) reduction in L. monocytogenes population for all treatments. All pasteurized treatments also resulted in a significant reduction of L. monocytogenes by 12 weeks compared to un-pasteurized bologna. In-package pasteurization in combination with nisin or nisin-lysozyme treatments was effective in reducing the population below detectable levels by 2-3 weeks of storage. Results from this study could have a significant impact for the industry since a reduction in bacterial population was achieved by a relatively short pasteurization time and antimicrobials reduced populations further during refrigerated storage.     

Review--Persistence of Listeria monocytogenes in food industry equipment and premises

To understand why Listeria monocytogenes may persist in food industry equipment and premises, notably at low temperature, scientific studies have so far focused on adhesion potential, biofilm forming ability, resistance to desiccation, acid and heat, tolerance to increased sublethal concentration of disinfectants or resistance to lethal concentrations. Evidence from studies in processing plants shows that the factors associated with the presence of L. monocytogenes are those that favor growth. Interestingly, most conditions promoting bacterial growth were shown, in laboratory assays, to decrease adhesion of L. monocytogenes cells. Good growth conditions can be found in so-called harborage sites, i.e. shelters due to unhygienic design of equipment and premises or unhygienic or damaged materials. These sites are hard to eliminate. A conceptual model of persistence/no persistence based on the relative weight of growth vs. outcome of cleaning and disinfection is suggested. It shows that a minimum initial bacterial load is necessary for bacteria to persist in a harborage site and that when a low initial bacterial charge is applied, early cleaning and disinfection is the only way to avoid persistence. We conclude by proposing that there are no strains of L. monocytogenes with unique properties that lead to persistence, but harborage sites in food industry premises and equipment where L. monocytogenes can persist.     

Multiple-locus variable number of tandem repeat analysis (MLVA) of Listeria monocytogenes directly in food samples

Listeria monocytogenes is the etiologic agent of listeriosis responsible for severe and fatal infections in humans. Listeria contamination occurs quite often in a wide range of foods due to its ubiquitous nature. Isolates need to be characterized to a strain level for accurate diagnosis of Listeria infection, epidemiological studies, investigation of outbreaks and effective prevention and control of food-borne listeriosis. The purpose of this research was to evaluate the multiple-locus variable number of tandem repeat analysis (MLVA) for sub-typing L. monocytogenes isolates in pure cultures and in food matrices. Two multiplex PCR assays were formulated to amplify six specific loci using fluorescently-labeled primers; and the amplicons were analyzed by capillary electrophoresis. The MLVA method resulted in 34 unique DNA fingerprint patterns from 46 L. monocytogenes isolates of 10 serotypes which had 29 or 30 PFGE patterns with a single restriction enzyme and 34 AFLP patterns. The MLVA patterns of the 46 isolates remained unchanged in the presence of pre-enriched food matrices including sausage, ham, chicken, milk and lettuce. The MLVA method successfully typed L. monocytogenes strains spiked in cheese, roast beef, egg salad and vegetable samples after 48 h enrichment at the initial inoculation levels of 1-5 CFU per 25 g of food or higher. The limits of detection (typing) of the MLVA method were 10³-10⁴ CFU/mL of pre-enriched food broth when evaluated using post-spiked sausage, ham, chicken, milk and lettuce samples. The MLVA method was simple, highly discriminatory, and easy to perform with portable (numerical) results. To our knowledge, this is the first report that describes the application of the MLVA method directly to food samples and demonstrates the possibility to obtain rapid and accurate subtyping results before an isolate is obtained.     

Control of Listeria monocytogenes growth in a ready-to-eat poultry product using a bacteriophage

A bacteriophage (phage) that infected strains of the species Listeria monocytogenes as well as Listeria ivanovii and Listeria welshimeri, but not Listeria grayi or Listeria innocua, was isolated from sheep faeces. The phage had a contractile tail and an icosohedral head indicating that it was a myovirus, and was morphologically similar to phage A511. At 30 °C, phages added at 5.2 × 10⁷ PFU ml⁻¹ prevented the growth in broth of L. monocytogenes present at approximately twice this concentration for 7 h, but re-growth occurred such that the concentration after 24 h incubation was similar in both control and phage-treated cultures. At the same temperature, but on the surface of vacuum-packed ready-to-eat chicken breast roll, there was an immediate 2.5 log₁₀ CFU cm⁻² reduction in pathogen concentration following addition of phages and then re-growth. However, at a temperature reflecting that at which a chilled food might be held (5°C), this re-growth was prevented over 21 days incubation. The data suggest a dose-dependent rapid reduction in pathogen concentration followed by no continued phage-mediated effect. These results, alongside other published data, indicate that a high concentration of phages per unit area is required to ensure significant inactivation of target pathogens on food surfaces.     

Effect of preinoculation growth media and fat levels on thermal inactivation of a serotype 4b strain of Listeria monocytogenes in frankfurter slurries

Preinoculation growth conditions and fat levels were evaluated for effects on the heat resistance of Listeria monocytogenes strain MFS 102 in formulated frankfurter slurries and on frankfurter surfaces. Comparison of linear inactivation rates (D-values) for cells heated in frankfurter slurry showed that growth conditions were significant (P<0.05) factors affecting subsequent thermal resistance. The average D(60 degrees C)-values for the five preinoculation growth media tested from most resistant to least heat resistant were: tryptic soy broth with 0.6% yeast extract (TSBYE) (2.2 min) and 8.5% fat slurry (2.2 min), followed by 23% fat slurry (1.7 min) and 11% fat slurry (1.7 min), and then TSYBE with quaternary ammonium compounds added (TSBYE+Q) (1 min). The fat level in the frankfurter heating media also had a significant (P<0.05) effect on the thermal death rate of L. monocytogenes. Cells heated in 8.5% fat slurry had a significantly higher (P<0.05) D(60 degrees C)-value (2.2 min) than those heated in 11% fat (1.0 min) and 23% fat slurry (0.9 min). Growth media (TSBYE, 8.5% fat slurry, and TSBYE+Q), and fat level (15% and 20%), however, were not significant factors (P>0.05) affecting thermal inactivation rates on frankfurter surfaces. Heat inactivation rates were consistently higher on frankfurter surfaces compared to similar treatments done in frankfurter slurry. On frankfurter surfaces, a 2.3- to 5.1-log(10) reduction was achieved after 15 min depending on frankfurter surface type. The time necessary to achieve a 3-log(10) reduction using post-processing pasteurization of frankfurters in a hot water-bath at 60 degrees C almost doubled for cells grown in TSBYE and heated in 23% fat frankfurter slurry (19.6 min) versus cells grown and heated in 8.5% fat frankfurter slurry (10.8 min).     

Edible chitosan films on ready-to-eat roast beef for the control of Listeria monocytogenes

The use of chitosan as an edible film was evaluated for its antimicrobial activity against Listeria monocytogenes (LM) on the surface of ready-to-eat (RTE) roast beef. L. monocytogenes, decimally diluted to give an initial inoculation of >6.50logCFU/g, was inoculated onto the surface of RTE roast beef cubes, and air-dried. The samples were dipped into chitosan (high or low molecular weights) solutions dissolved with acetic or lactic acid at 0.5% (w/v) or 1% (w/v) then bagged and refrigerated at 4 degrees C. The bacterial counts were determined on days 0, 7, 14, 21, and 28. The samples were spread plated onto modified Oxford agar plates and incubated at 37 degrees C for 48h. An initial 6.50logCFU/g of L. monocytogenes inoculated onto the surface of the non-coated RTE roast beef increased too >10logCFU/g by day 28. On day 14, L. monocytogenes counts were significantly different for all the chitosan-coated samples from the control counts by 2-3logCFU/g and remained significantly different on day 28. Our results have shown that the acetic acid chitosan coating were more effective in reducing L. monocytogenes counts than the lactic acid chitosan coating. Our study indicated that chitosan coatings could be used to control L. monocytogenes on the surface of RTE roast beef.     

The antimicrobial effect of thyme essential oil, nisin, and their combination against Listeria monocytogenes in minced beef during refrigerated storage

The antimicrobial effect of thyme essential oil (EO) at 0.3%, 0.6%, or 0.9%, nisin at 500 or 1000IU/g, and their combination against Listeria monocytogenes was examined in both tryptic soy broth (TSB) and minced beef meat. Thyme EO at 0.3% possessed a weak antibacterial activity against the pathogen in TSB, whereas at 0.9% showed unacceptable organoleptic properties in minced meat. Thus, only the level of 0.6% of EO was further examined against the pathogen in minced meat. Treatment of minced beef meat with nisin at 500 or 1000IU/g showed antibacterial activity against L. monocytogenes, which was dependent on the concentration level of nisin and the strains used. Treatment of minced beef meat with EO at 0.6% showed stronger inhibitory activity against L. monocytogenes than treatment with nisin at 500 or 1000IU/g. All treatments showed stronger inhibitory activity against the pathogens at 10 degrees C than at 4 degrees C. The combined addition of EO at 0.6% and nisin at 500 or 1000IU/g showed a synergistic activity against the pathogen. Most efficient among treatments was the combination of EO at 0.6% with nisin at 1000IU/g, which decreased the population of L. monocytogenes below the official limit of the European Union recently set at 2logcfu/g, during storage at 4 degrees C.     

Longitudinal monitoring of Listeria monocytogenes contamination patterns in a farmstead dairy processing facility

Contamination of dairy products with Listeria monocytogenes is a concern because multiple human listeriosis outbreaks have been linked to contaminated cheese and dairy products. Dairy production on farmstead operations may be a particular concern because L. monocytogenes is also an animal pathogen that can be shed by ruminants with and without clinical symptoms; physical proximity between production animal and dairy processing facilities may thus provide a higher risk for introduction of L. monocytogenes into the dairy production process. To better understand the risks of L. monocytogenes contamination associated with farmstead dairy production, samples from a farmstead dairy processing operation and the milking barn of the directly adjacent dairy sheep operation were tested for L. monocytogenes over a 3-yr period. Prevalence of L. monocytogenes for samples collected on the farm (n = 85) and the dairy production facility (n = 674) was 9.4 and 2.7%, respectively. Molecular subtyping using automated EcoRI ribotyping of L. monocytogenes isolates revealed that distinct subtypes were associated with the dairy production facility and the farm's milking parlor. Although a total of 5 and 4 different ribotypes were identified among isolates obtained from the dairy production facility and the milking parlor, respectively, only 1 ribotype (DUP-1030A) was isolated from both. Different ribotypes were predominant among isolates from the dairy production facility (ribotype DUP-1052A, representing 15 of 18 isolates) and the farm's milking parlor (ribotype DUP-1039A, representing 4 of 8 isolates); each of these ribotypes appeared to persist over time in the respective area. Our data support that i) in farmstead dairy processing facilities, L. monocytogenes present on the farm can largely be prevented from being introduced into the processing facility; and ii) L. monocytogenes can persist on farm and in processing areas, providing a potential high-risk source for contamination. Preventing cross contamination between dairy production and processing facilities and control of persistent L. monocytogenes are thus critical to assuring the microbial safety of farmstead dairy products.     

Role of quantitative risk assessment and food safety objectives in managing Listeria monocytogenes on ready-to-eat meats

Listeria monocytogenes may be found on ready-to-eat (RTE) meats, posing a public health risk. To minimize the public health impact, an appropriate level of protection (ALOP) can be established for a population with respect to L. monocytogenes, and ideally should be based on a scientific assessment of the risk, as well as societal and economic factors. Food safety systems can be based on meeting the ALOP. Food safety objectives (FSO) provide a link between the ALOP and performance objectives that are established to control a foodborne hazard. An FSO can be used as a risk management tool for L. monocytogenes in RTE meats, as the FSO establishes the stringency of the measures being used to control the hazard, by specifying the frequency and/or cell number of the pathogen in the food that should not be exceeded at the time of consumption. Typically, this requires setting performance objectives or performance criteria at an earlier point in the food chain, to ensure that the product will meet the FSO. Establishing an FSO requires an assessment of the risk of the hazard to the population of interest. Risk management strategies such as use of HACCP systems and Good Manufacturing Practices can then be used to ensure that the FSO is met.     

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 egg salad and pasta salad formulated with mayonnaise of various pH and stored at refrigerated and abuse temperatures

This study investigated and modeled the behavior of Listeria monocytogenes in egg salad and pasta salad as affected by mayonnaise pH (3.8, 4.2, 4.6, and 5.0) and storage temperature (4, 8, and 12 degrees C). At each storage temperature, L. monocytogenes was able to grow in both salads regardless of the mayonnaise pH. The lag-phase durations (LPD) of L. monocytogenes in egg salad ranged from 33 to 85, 15 to 50, and 0 to 19 h, and the growth rates (GR) ranged from 0.0187 to 0.0318, 0.0387 to 0.0512, and 0.0694 to 0.1003 log(10)cfu/h at 4, 8, and 12 degrees C, respectively. The LPD of L. monocytogenes in pasta salad ranged from 210 to 430, 49 to 131, and 21 to 103 h, and GR ranged from 0.0118 to 0.0350, 0.0153 to 0.0418, and 0.0453 to 0.0718 log(10)cfu/h at 4, 8, and 12 degrees C, respectively. The growth of L. monocytogenes was more rapid in egg salad than in pasta salad, indicating that a better growth environment for L. monocytogenes existed in egg salad. In both salads, the LPD decreased and the GR increased as the storage temperature increased. Mathematical models and response surface plots describing the LPD and GR of L. monocytogenes in both salads as affected by the mayonnaise pH and storage temperature were developed. The models confirmed that the growth of L. monocytogenes in egg salad and pasta salad was primarily promoted by higher storage temperatures and, secondarily, by higher mayonnaise pH. The conditions under which the models may be applied to estimate the growth of L. monocytogenes in both salads were identified.     

ADSA Foundation Scholar Award--An integrated science-based approach to dairy food safety: Listeria monocytogenes as a model system

Transmission of food- and milkborne pathogens often involves complex interactions among the pathogen, the environment, and one or multiple host species. A complete understanding of these interactions is critical to allow the development of science-based, effective intervention strategies for foodborne infectious diseases. This article summarizes our studies on the transmission, ecology, pathogenesis and population genetics of Listeria monocytogenes, which we have used as model for a food- and milkborne pathogen that infects multiple hosts and also has considerable ability to survive and multiply in nonhost environments. Application of molecular subtyping tools in conjunction with phenotypic characterization of selected strains has allowed us to define distinct L. monocytogenes subtypes and clonal groups that appear to differ in relevant phenotypic characteristics that may affect their abilities to be transmitted through food systems. For example, a genetic group designated as lineage I has been shown to be not only more common among human listeriosis cases than among animal cases, but lineage I strains also appear to show an increased in vitro ability to spread intracellularly from host cell to host cell. These findings are consistent with the fact that while genetically diverse strains may be classified to one bacterial species, these strains often differ from one another in important genetic and phenotypic characteristics. I thus propose that evolutionary- and molecular subtyping-based definitions of bacterial subtypes and clonal groups will provide critical insight into the microbial ecology of dairy food systems, including not only foodborne pathogens, but also organisms important for dairy fermentation and spoilage.