Annie Beaufort

Modeling microbial competition in food: Application to the behavior of Listeria monocytogenes and lactic acid flora in pork meat products

Competition between background microflora and microbial pathogens raises questions about the application of predictive microbiology in situ, i.e., in non-sterile naturally contaminated foods. In this article, we present a review of the models developed in predictive microbiology to describe interactions between microflora in foods, with a special focus on two approaches: one based on the Jameson effect (simultaneous deceleration of all microbial populations) and one based on the Lotka-Volterra competition model. As an illustration of the potential of these models, we propose various modeling examples in estimation and in prediction of microbial growth curves, all related to the behavior of Listeria monocytogenes with lactic acid bacteria in three pork meat products (fresh pork meat and two types of diced bacon).

A contribution to the improvement of Listeria monocytogenes enumeration in cold-smoked salmon

For the enumeration of Listeria monocytogenes in food, a sensitive enumeration method based on membrane filtration followed by transfer of the filter to a selective medium has been developed. This study was carried out with cold-smoked salmon, a product likely to be contaminated with L. monocytogenes. The operating protocol utilizes three filtration runs in parallel (5, 15 and 30 ml) of a 1 in 10 dilution of the salmon suspension through 0.45-microm pore-size cellulose ester membranes, and then culture of the filters on Aloa agar (AES Laboratoires, Combourg, France). The results obtained with the technique were compared with those from the reference EN ISO 11290-2 method and found to provide more precise results in the enumeration of L. monocytogenes from both artificially and naturally contaminated cold-smoked salmon. Moreover, for several samples contaminated at low levels, L. monocytogenes could be recovered only by the filtration method. The examination of increasing volumes of salmon suspension enabled readable results to be obtained for all levels of L. monocytogenes and competitive microflora investigated. In most cases, the optimised operating protocol enabled 5.1 g of salmon to be examined, instead of 0.01-0.1 g with the reference EN ISO 11290-2 method, thus improving the sensitivity of the method.

Uncertainty Distribution Associated with Estimating a Proportion in Microbial Risk Assessment

The uncertainty associated with estimates should be taken into account in quantitative risk assessment. Each inputs uncertainty can be characterized through a probabilistic distribution for use under Monte Carlo simulations. In this study, the sampling uncertainty associated with estimating a low proportion on the basis of a small sample size was considered. A common application in microbial risk assessment is the estimation of a prevalence, proportion of contaminated food products, on the basis of few tested units. Three Bayesian approaches (based on beta(0, 0), beta(1/2, 1/2), and beta(l, 1)) and one frequentist approach (based on the frequentist confidence distribution) were compared and evaluated on the basis of simulations. For small samples, we demonstrated some differences between the four tested methods. We concluded that the better method depends on the true proportion of contaminated products, which is by definition unknown in common practice. When no prior information is available, we recommend the beta (1/2, 1/2) prior or the confidence distribution. To illustrate the importance of these differences, the four methods were used in an applied example. We performed two-dimensional Monte Carlo simulations to estimate the proportion of cold smoked salmon packs contaminated by Listeria monocytogenes, one dimension representing within-factory uncertainty, modeled by each of the four studied methods, and the other dimension representing variability between companies.

Use of quantitative microbial risk assessment when investigating foodborne illness outbreaks: The example of a monophasic Salmonella Typhimurium 4,5,12:i: — outbreak implicating beef burgers

A major community outbreak of salmonellosis occurred in France in October 2010. Classical epidemiological investigations led to the identification of beef burgers as the cause of the outbreak and the presence of the emerging monophasic Salmonella Typhimurium 4,5,12:i:-. The objective of this study was to understand the events that led to this large outbreak, that is to say, what are the contributing factors associated with consumer exposure to Salmonella. To this end, intensive microbiological investigations on several beef burgers were conducted and a risk assessment model was built. The microbiological results confirm the presence of Salmonella in all analysed frozen burgers at high levels of contamination above 1000 MPN/g. These results in frozen burgers combined with a model of thermal destruction were used to estimate the dose ingested by the exposed persons. Most people that consumed cooked beef burgers were exposed from 1.6 to 3.1 log₁₀ (MPN). The number of sick people predicted with a dose-response relationship for Salmonella is consistent with the observed number of salmonellosis cases. The very high initial contamination level in frozen beef burgers is the primary cause of this large outbreak rather than bad cooking practices. Intensive investigations, modelling of the initial contamination and quantitative exposure and risk assessments are complementary to epidemiological investigation. They can be valuable elements for the assessment of missing information or the identification of the primary causes of outbreaks.

Evaluation of an enumeration method for Listeria monocytogenes at low contamination levels in cold-smoked salmon.

For the enumeration of Listeria monocytogenes in cold-smoked salmon, a sensitive enumeration method, based on membrane filtration followed by transfer of the filter on a selective medium has been recently developed (Gnanou Besse et al., 2004, A contribution to the improvement of L. monocytogenes enumeration in cold-smoked salmon. International Journal of Food Microbiology, 91, 119-127). The aim of the study was to assess the performance of this enumeration method through an inter-laboratory study, using cold-smoked salmon artificially contaminated at 2 different levels (approximately 0.6 and 1.6 log10 CFU g(-1)). A reproducibility standard deviation of 0.23 log10 CFU g(-1)and 0.15 log10 CFU g(-1) was obtained for the method respectively at the lower level and the higher level. Under certain conditions, the uncertainty of measurement can be derived from the method reproducibility standard deviation and was calculated to be 0.46 log10 CFU g(-1) for the lower contamination level and 0.30 log10 CFU g(-1) for the higher contamination level. These values can be considered as satisfactory for such low contamination levels.

Impact of−2°C Superchilling before Refrigerated Storage (4 and 8°C) on the Microbiological and Sensory Qualities of Cold-Smoked Salmon

Detection and enumeration of Listeria monocytogenes and total spoilage bacteria in 40 batches of cold-smoked salmon (one batch = 42 products from the same day of manufacture) straight from the factory were carried out. If L. monocytogenes was detected in at least one of the nine samples analyzed on receipt at the laboratory, 9 products of the same batch were stored for 10 days at 4°C, which was followed by 18 days at 8°C (control), 12 products were superchilled for 14 days at −2°C, and 12 other products were superchilled for 28 days at −2°C and then stored under the same conditions as the control was stored. L. monocytogenes was detected in 7% of the 40 batches analyzed immediately after receipt at the laboratory. L. monocytogenes prevalence was similar (approximately 25%) throughout the storage at 4 and 8°C, both in control and super-chilled products at −2°C for 14 days. After superchilling for 28 days at −2°C, L. monocytogenes was found in 9% of products, and in 39% at the end of the storage above 0°C. ...