Retrospective analysis of a listeria monocytogenes contamination episode in raw milk goat cheese using quantitative microbial risk assessment tools

in Journal of Food Protection (2012), 75(12), 2122-2135

In 2005, the Belgian authorities reported a Listeria monocytogenes contamination episode in cheese made from raw goat's milk. The presence of an asymptomatic shedder goat in the herd caused this ... [more ▼]

In 2005, the Belgian authorities reported a Listeria monocytogenes contamination episode in cheese made from raw goat's milk. The presence of an asymptomatic shedder goat in the herd caused this contamination. On the basis of data collected at the time of the episode, a retrospective study was performed using an exposure assessment model covering the production chain from the milking of goats up to delivery of cheese to the market. Predictive microbiology models were used to simulate the growth of L. monocytogenes during the cheese process in relation with temperature, pH, and water activity. The model showed significant growth of L. monocytogenes during chilling and storage of the milk collected the day before the cheese production (median increase of 2.2 log CFU/ml) and during the addition of starter and rennet to milk (median increase of 1.2 log CFU/ml). The L. monocytogenes concentration in the fresh unripened cheese was estimated to be 3.8 log CFU/g (median). This result is consistent with the number of L. monocytogenes in the fresh cheese (3.6 log CFU/g) reported during the cheese contamination episode. A variance-based method sensitivity analysis identified the most important factors impacting the cheese contamination, and a scenario analysis then evaluated several options for risk mitigation. Thus, by using quantitative microbial risk assessment tools, this study provides reliable information to identify and control critical steps in a local production chain of cheese made from raw goat's milk. © International Association for Food Protection. [less ▲]

Validation of a method for simultaneous isolation of shiga toxin-producing Escherichia coli O26, O103, O111, and O145 from minced beef by an international ring-trial

in Foodborne Pathogens and Disease (2012), 9(5), 412-417

An isolation method described by Possé et al. (FEMS Microbiol Lett 2008;282:124-131) was satisfactorily validated in an international ring-trial using artificially contaminated minced beef samples. Until ... [more ▼]

An isolation method described by Possé et al. (FEMS Microbiol Lett 2008;282:124-131) was satisfactorily validated in an international ring-trial using artificially contaminated minced beef samples. Until now, no validated method existed for the simultaneous isolation of Shiga toxin-producing Escherichia coli serogroups O26, O103, O111, and O145 in food. Twelve laboratories from five European countries participated and received 16 inoculated beef samples contaminated with cold-stressed cells of the four serogroups O26, O103, O111, and O145 in two levels (approximately 30 and 300 CFU 25g-1) in duplicate. In addition, they received four non-inoculated samples. The isolation protocol comprised a selective enrichment step, a selective isolation step on a non-O157 agar plate differentiating the serogroups by color, followed by confirmation by plating on confirmation agar media and agglutination. All laboratories were able to isolate the inoculated serogroups from the samples, both for the high and the low inoculation level. Results did not differ whether in-house-prepared or ready-to-use non-O157 agar plates were used, demonstrating that by following the instructions laboratories managed to perform the complete protocol with success. © Copyright 2012, Mary Ann Liebert, Inc. [less ▲]

Caractérisation des viandes bovines à très longue durée de conservation sous vide

Poster (2011, December 09)

Le but de cette étude a été d’évaluer la conservabilité de viandes bovines de différentes origines (Royaume-Uni et Irlande, Australie et Brésil) et l’influence sur celle-ci de la température de ... [more ▼]

Le but de cette étude a été d’évaluer la conservabilité de viandes bovines de différentes origines (Royaume-Uni et Irlande, Australie et Brésil) et l’influence sur celle-ci de la température de conservation (1 °C vs. +4 °C). Des paramètres physico-chimiques (pH, couleur, proportion des différentes formes redox de la myoglobine (FRMb), indice TBARS et acides organiques) et microbiologiques (flore aérobie totale, flore lactique, Enterobacteriaceae, Pseudomonas spp. et Brochothrix thermosphacta) ont été mesurés sur sept lots de contre-filet conditionnés sous vide : aux ⅔ de la DLC et à la fin de la DLC. La diversité bactérienne a été évaluée par galeries API50 CHL et par métagénomique. Le pH a diminué au cours de la conservation dans deux lots. La couleur et la proportion des FRMb sont restées stables. Une augmentation de l’indice TBARS, plus prononcée à +4 °C, a été observée. Les viandes australiennes et brésilienne ont présenté des taux en acides acétique et citrique plus élevés. Tous les lots conservés à 1 °C ont présenté une qualité microbiologique satisfaisante à la fin de leur DLC (viandes britanniques et irlandaises = 35 ~ 45 jours; australiennes = 140 jours et brésilienne = 120 jours). La conservation à +4 °C a favorisé la croissance d’entérobactéries, facteur limitant de la conservation de plusieurs lots. L’identification bactérienne a révélé la présence de bactéries connues pour leur effet bioprotecteur. La phase ultérieure de ce travail consistera à étudier la dynamique de la flore microbienne endogène en fonction des conditions environnementales appliquées (température, atmosphère). [less ▲]

La métagénomique au service de la microbiologie alimentaire : étude de l’évolution des populations microbiennes lors du vieillissement de deux matrices alimentaires

Conference (2011, November 17)

La viande et les préparations à base de viande représentent des biotopes de choix pour les bactéries. L’optimisation de la conservation de ces denrées, importante tant du point de vue économique que de ... [more ▼]

La viande et les préparations à base de viande représentent des biotopes de choix pour les bactéries. L’optimisation de la conservation de ces denrées, importante tant du point de vue économique que de santé publique passe par une meilleure connaissance de ces biotopes et des processus de détérioration de leurs qualités. Les microbiologistes ont depuis longtemps abordé ce problème en utilisant différentes approches. Ainsi, les études basées sur les méthodes de culture ont été complétées par des stratégies axées sur des approches indépendantes de la culture microbiologique. Les techniques actuelles de séquençage de nouvelle génération ont permis de donner une nouvelle dimension à l’écologie microbienne à travers l’analyse métagénomique d’un grand nombre d’individus au sein d’une population microbienne mixte. Notre propos est de démontrer que cette méthodologie peut être appliquée avec succès à l’étude des flores microbiennes des denrées alimentaires et pourra être adaptée, à court terme, aux exigences spécifiques de la microbiologie alimentaire. Dans cette étude, un produit de viande crue et une préparation de viande cuite, la viande hachée de porc et le boudin blanc, ont subi un test de vieillissement dans différentes conditions de conservation (Température et packaging). L’ADN ribosomal 16S a été extrait des produits originaux et des échantillons à la date limite de consommation et, après normalisation, les régions hypervariables V5 et V6 de l’ADN 16S ont été séquencées. Un total d’environ 130.000 séquences ont été obtenues et une analyse métagénomique a abouti à la classification taxonomique au genre pour 80% de cette population. L’analyse subséquente des populations microbiennes montre que les populations microbiennes majoritaires à la date limite de conservation sont bien celles qui sont généralement observées lors d’analyse microbiologiques de ces produits de viande. Toutefois, les populations sous-dominantes et surtout plusieurs populations de germes non cultivables ont pu être identifiée. Ces groupes de bactéries, plus difficile à obtenir par d’autres méthodes (culture-dépendante et indépendante confondues), doivent être étudiées car elles participent aux processus de détériorations de ces matrices alimentaires. Enfin, la sensibilité de cette technologie rend possible l’analyse des denrées alimentaires présentant un taux microbien très faible et permet donc l’identification des contaminants microbiens avant leur développement. [less ▲]

Development of a quantitative risk assessment for cheese made from raw goat milk contaminated by Listeria monocytogenes

Poster (2011, September)

A retrospective study was performed to assess the potential risk of human listeriosis following a contamination by L. monocytogenes of cheeses made from goat raw milk reported by the Belgian Federal ... [more ▼]

A retrospective study was performed to assess the potential risk of human listeriosis following a contamination by L. monocytogenes of cheeses made from goat raw milk reported by the Belgian Federal Agency for the Safety of the Food Chain in 2005. The source of the contamination was related to a shedder goat, excreting 2.6 log cfu (colonies forming units) L. monocytogenes / ml without any clinical symptom. On the basis of the collected data, a quantitative microbial risk assessment model was developed covering the production chain from the milking of goats until the consumed products. Predictive microbiology models were used to simulate the growth of L. monocytogenes during the process of cheeses made from goat raw milk. The modular exposure assessment model showed a significant growth of L. monocytogenes during chilling and storage of the milk collected the day before the cheese production (increase of 1.7 log cfu/ml for the median) and during the step of starter and rennet adjunction to milk (increase of 0.8 log cfu/ml for the median). The median estimated final result (in the fresh cheese) was equal to 3.5 log cfu/g. The model estimates (expressed as median final result issued from the exposure assessment) were realistic compared to the number of L. monocytogenes measured in the fresh cheese (3.6 log cfu/g) reported during the cheese contamination period. The average number of expected cases of human listeriosis was between 0 and 1 for a high-risk sub-population and 0 for a low-risk healthy sub-population. Scenario analysis was finally performed to identify the most significant factors and aid in developing priorities for risk mitigation. Thus, by using quantitative risk assessment and predictive microbiology models, this study provided valuable information to identify and to control critical steps in a local production chain of goat cheese made from raw milk. [less ▲]

Influence of temperature on conservability of chilled vacuum packed beef from different origins

Poster (2011, August 07)

The objective of this experiment was to study the conservability of chilled vacuum-packed meat depending on storage temperature (–1 °C vs. +4 °C) during the last third of their shelf life. Physicochemical ... [more ▼]

The objective of this experiment was to study the conservability of chilled vacuum-packed meat depending on storage temperature (–1 °C vs. +4 °C) during the last third of their shelf life. Physicochemical parameters (pH and colour) and microbiological growth (total aerobic bacteria, lactic acid bacteria, Enterobacteriaceae, Pseudomonas spp. and Brochothrix thermosphacta) of Longissimus dorsi samples from different origins (United Kingdom and Ireland, Australia and Brazil) were measured at: i) 2/3 of their shelf life and ii) the end of their shelf life. Sample bacteria population growing on MRS was identified by API 50 CHL strips. Unlike Irish and British samples, pH of some Australian and Brazilian samples decreased during conservation. The colour of the samples remained stable and it did not seem to be influenced by temperature. All samples conserved at –1 °C presented a satisfactory microbiological quality at the end of their shelf life (British and Irish meat = 35~45 days; Australian meat = 140 days and Brazilian meat = 120 days). On Australian and Brazilian samples, temperature did not influence total aerobic bacteria growth, but conservation at +4 °C favoured lactic acid bacteria and Enterobacteriaceae growth. API 50 CHL strip identifications revealed the presence of bacteria like Lactobacillus brevis, Carnobacterium maltaromaticum and Lactobacillus fermentum, which occur naturally in fresh meat and are known for their bioprotective effect against other microorganisms. Further analyses are being carried out using molecular methods in order to study the initial bacteria population diversity and it evolution during storage. [less ▲]

A new tool to control meat products safety: a web based application of predictive microbiology models

Conference (2011)

Predictive microbiology is considered by the European legislation as a tool to control food safety. Meat and meat products are particularly sensitive to contamination with pathogens. However, development ... [more ▼]

Predictive microbiology is considered by the European legislation as a tool to control food safety. Meat and meat products are particularly sensitive to contamination with pathogens. However, development of predictive microbiology models and interpretation of results require specific knowledge. A free web based model has been developed for an easy use by people who are not experts in this field as industries and public authorities. The model can simulate the growth of Salmonella spp, Listeria monocytogenes and Escherichia coli O157 in minced pork meat and on pork meat product (white pudding) under different environmental conditions. The model provides simulations under static or dynamic conditions over time. The user also has the opportunity to import the specific growth rate and cardinal parameters of a bacterium. Unlike polynomial models currently available, this free web access model is distinguished by the use of secondary square roots and primary logistic model with delay. This model permits to have a real time process management, to prospect new formulation for safer products or to design safer processes, to estimate the shelf life of a food product, etc [less ▲]

Bifidobacterium pseudolongum are efficient indicators of animal fecal contamination in raw milk cheese industry

in BMC Microbiology (2011), 11(178),

Background: The contamination of raw milk cheeses (St-Marcellin and Brie) from two plants in France was studied at several steps of production (raw milk, after addition of rennet - St-Marcellin - or after ... [more ▼]

Background: The contamination of raw milk cheeses (St-Marcellin and Brie) from two plants in France was studied at several steps of production (raw milk, after addition of rennet - St-Marcellin - or after second maturation - Brie -, after removal from the mold and during ripening) using bifidobacteria as indicators of fecal contamination. Results: Bifidobacterium semi-quantitative counts were compared using PCR-RFLP and real-time PCR. B. pseudolongum were detected in 77% (PCR-RFLP; 1.75 to 2.29 log cfu ml-1) at the different production steps) and 68% (real-time PCR; 2.19 to 2.73 log cfu ml-1) of St-Marcellin samples and in 87% (PCR-RFLP; 1.17 to 2.40 log cfu ml-1) of Brie cheeses samples. Mean counts of B. pseudolongum remained stable along both processes. Two other populations of bifidobacteria were detected during the ripening stage of St-Marcellin, respectively in 61% and 18% of the samples (PCR-RFLP). The presence of these populations explains the increase in total bifidobacteria observed during ripening. Further characterization of these populations is currently under process. Forty-eight percents (St-Marcellin) and 70 % (Brie) of the samples were B. pseudolongum positive / E. coli negative while only 10 % (St-Marcellin) and 3 % (Brie) were B. pseudolongum negative / E. coli positive. Conclusions: The increase of total bifidobacteria during ripening in Marcellin’s process does not allow their use as fecal indicator. The presence of B. pseudolongum along the processes defined a contamination from animal origin since this species is predominant in cow dung and has never been isolated in human feces. B. pseudolongum was more sensitive as an indicator than E. coli along the two different cheese processes. B. pseudolongum should be used as fecal indicator rather than E. coli to assess the quality of raw milk and raw milk cheeses. Results: Bifidobacterium semi-quantitative counts were compared using PCR-RFLP and real-time PCR. Bif. pseudolongum were detected in 77% (PCR-RFLP; 1.75 to 2.29 log cfu ml-1) at the different production steps) and 68% (real-time PCR; 2.19 to 2.73 log cfu ml-1) of St-Marcellin samples and in 87% (PCR-RFLP; 1.17 to 2. 40 log cfu ml-1) of Brie cheeses samples. Mean counts of Bif. pseudolongum remained stable along both processes. Two other populations of bifidobacteria were detected during the ripening stage of St-Marcellin, respectively in 61% and 18% of the samples (PCR-RFLP). The presence of these populations explains the increase in total bifidobacteria observed during ripening. Further identification of these species is currently under process. Forty-eight percents (St-Marcellin) and 70 % (Brie) of the samples were Bif. pseudolongum positive / E. coli negative while only 10 % (St-Marcellin) and 3 % (Brie) were Bif. pseudolongum negative / E. coli positive. Conclusions: The increase of total bifidobacteria during ripening in Marcellin’s process does not allow their use as fecal indicator. The presence of Bif. pseudolongum along the processes defined a contamination from animal origin since this species is predominant in cow dung and has never been isolated in human feces. Bif. pseudolongum was more sensitive as an indicator than E. coli along the two different cheese processes. Bif. pseudolongum should be used as fecal indicator rather than E. coli to assess the quality of raw milk and raw milk cheeses. [less ▲]

A New Method To Enumerate Low Level Of Listeria Monocytogenes And Salmonella Spp. In Meat Products

Poster (2010, September)

Modeling Microbial Cross-contamination in Quick Service Restaurants by Means of Experimental Simulations With Bacillus Spores

Poster (2010)

Cross contamination has been frequently mentioned as being in the origin of a wide range of food borne outbreaks. Handling of food is one of the ways through which cross contamination may occur. For many ... [more ▼]

Cross contamination has been frequently mentioned as being in the origin of a wide range of food borne outbreaks. Handling of food is one of the ways through which cross contamination may occur. For many different reasons, quick service restaurants are particularly at risk. Due to its importance, cross contamination via the hands should be taken into consideration when carrying out a quantitative risk assessment. The main goal of this study was to determine transfer rates of bacteria to and via the hands, reduction rates of two hand sanitizing procedures and to apply the results to a quantitative microbial risk assessment model. According to our results, handling of a portion of raw minced meat contaminated at 4.104 cfu leads to the presence of 24 cfu on both hands, 3 cfu on ready-to-eat product (RTE) manipulated with unwashed hands, 1 cfu on RTE manipulated with wiped hands and absence on RTE manipulated with washed hands. This study provides adequate quantitative data for quantitative microbial risk assessment. [less ▲]