Home style frying of steak and meat products: Survival of Escherichia coli related to dynamic temperature profiles.

Microbial survival of heating and cross-contamination are the two transmission routes during food preparation in the consumers' kitchen that are relevant for QMRA (Quantitative Microbial Risk Assessment). The aim of the present study was to extend the limited amount of data on microbial survival during real-life preparation of meat and meat products and to obtain accessory temperature data that allow for a more general (product unspecific) approach. Therefore survival data were combined with extensive measurements of time- and location dependent temperature using an infrared camera for the surface and buttons for the inside of the product, supplemented with interpolation modelling. We investigated the survival of heating of Escherichia coli O111:H2 in beefsteak, hamburgers (beef and 50% beef 50% pork (HH)), meatballs (beef and HH) and crumbs (HH). For beefsteak, survival as a whole is dominated by the sides, giving a log reduction of 1-2 (rare), 3-4 (medium) and 6-7 (done). Limited measurements indicated that done preparation gave 5-6 log reduction for crumbs and at least 8-9 log for the other products. Medium preparation gave a higher reduction in hamburgers (2-4 log) than in meatballs (1-2 log) and in beef (3-4) than in HH (2-3) hamburgers. In general, our 'done' results give larger inactivation than found in literature, whereas 'rare' and 'medium' results are similar. The experiments resulted in two types of curves of D70/z-values, dependent on product, doneness and for beefsteaks sides vs. top/bottom. One type of curve agrees reasonably with literature D70/z estimates from isothermal temperature experiments, which supports using these estimates for home style cooking QMRA calculations. In case of the other type of curve, which is mainly found for (near) surface contamination in close contact with the pan, these literature estimates cannot be applied. We also applied a simplified approach, assuming thermal inactivation is dominated by the highest temperatures reached. The time duration of this highest temperature gives accessory D-values which prove to fit with isothermal temperature literature data, thus suggesting application of such data for QMRA is possible by this approach also, which is less labor intensive both in terms of measurements and modelling. In real life, variability in product properties and preparation styles is large. Further studies are needed to analyze the effect on survival, preferably focusing on determining the essential variables. More variation in heating time will allow for estimating D70/z point estimates rather than curves representing possible sets of D70/z-values.

Modelling the number of viable vegetative cells of Bacillus cereus passing through the stomach.

AIMS: Model the number of viable vegetative cells of B. cereus surviving the gastric passage after experiments in simulated gastric conditions. MATERIALS AND METHODS: The inactivation of stationary and exponential phase vegetative cells of twelve different strains of Bacillus cereus, both mesophilic and psychrotrophic strains isolated from food and faeces from healthy and ill individuals, in simulated gastric conditions was determined using decimal reduction times at low pH (D(pH)). Subsequently inactivation rates were calculated. Inclusion of the inactivation rates into models describing the course of the gastric pH after the consumption of meal of solid food and the transfer of food from the stomach to the small intestine resulted in numbers of viable Bacillus cereus vegetative cells able to pass the stomach. CONCLUSIONS: According to the model, 3-26% of the ingested vegetative cells from Bacillus cereus may survive the gastric passage, dependent on the growth phase of the vegetative cells, the type of strains, and the age of the consumer. SIGNIFICANCE AND IMPACT OF THE STUDY: Vegetative cells of Bacillus cereus may be involved in the onset of diarrhoeal disease to a greater extent than expected since up to 26% of the ingested cells survive simulated gastric conditions.

Germination of Bacillus cereus spores is induced by germinants from differentiated Caco-2 Cells, a human cell line mimicking the epithelial cells of the small intestine.

Spores of 11 enterotoxigenic strains of Bacillus cereus isolated from foods and humans adhered with similar efficiencies to Caco-2 cells, whereas subsequent germination triggering was observed with only 8 of these strains. Notably, Hep-2 cells did not trigger germination, while spores of all strains displayed similar germination efficiencies in brain heart infusion broth.

Prevalence of potentially pathogenic Bacillus cereus in food commodities in The Netherlands.

Randomly selected food commodities, categorized in product groups, were investigated for the presence and number of Bacillus cereus bacteria. If positive, and when possible, five separate colonies were isolated and investigated for the presence of four virulence factors: presence of genes encoding three enterotoxins (hemolysin BL [HBL], nonhemolytic enterotoxin [NHE], and cytotoxin K) and the ability to produce cereulide. In addition, the presence of psychrotrophic and mesophilic signatures was determined. The genes for NHE are found in more than 97% of the isolates, those for HBL in approximately 66% of the isolates, and the gene for cytotoxin K in nearly 50% of the isolates. Significant associations between product groups and (combinations of) virulence factors were the relatively low percentage of isolates from the "flavorings" group containing genes encoding NHE and the higher-than-average occurrence of both the genes encoding HBL and NHE in the "pastry" group. Cereulide was produced by 8.2% of the isolates but only in combination with the presence of genes for one or more other virulence factors. Most isolates (89.9%) were mesophilic; minorities of the isolates were psychrotrophic (4.4%) or of intermediate signature (5.7%). In the product group "milk and milk products," the incidence of strains with psychrotrophic or intermediate signatures is significantly higher than in the other product groups. In the product groups "flavorings," "milk and milk products," "vegetable(s) and vegetable products," "pastry," and "ready-to-eat foods," a relatively high number of samples contain high numbers of B. cereus bacteria. Within the product group "ready-to-eat foods," the products containing rice and pasta show a relatively high incidence of high numbers of B. cereus bacteria.

Spores from mesophilic Bacillus cereus strains germinate better and grow faster in simulated gastro-intestinal conditions than spores from psychrotrophic strains.

The species Bacillus cereus, known for its ability to cause food borne disease, consists of a large variety of strains. An important property for discrimination of strains is their growth temperature range. Psychrotrophic strains can grow well at refrigerator temperatures but grow at 37 degrees C with difficulty. Mesophilic strains on the other hand are unable to grow below 10 degrees C, but grow well at 37 degrees C. Spores of six psychrotrophic and six mesophilic strains were investigated for their ability to survive and grow in simulated gastro-intestinal fluids, mimicking the conditions in the gastro-intestinal tract. The germination potential of psychrotrophic and mesophilic spores in simulated intestinal fluid does not differ much. Under conditions simulating the gastro-intestinal passage, 5 out of 6 mesophilic strains showed growth, and only 2 out of 6 psychrotrophic strains. Temperature (37 degrees C) and simulated gastro-intestinal conditions together influenced germination and growth.

Marker antigens to assess exposure to molds and their allergens. II. Alternaria alternata.

The estimation of exposure to molds and their products in the indoor environment, which may lead to the occurrence of allergies or respiratory complaints, by means of enumeration of viable parts is inadequate. Therefore, other methods must be developed. When grown under various circumstances (22 degrees C and 30 degrees C, high and low water activity) under laboratory conditions, Alternaria alternata produces one antigen that can be found under all studied growth conditions in extracts of the water-soluble portion of the mycelium. This common antigen may serve as marker antigen for exposure to A. alternata and its allergens. In extracts of the culture filtrate, three antigens, designated index antigens, have been identified that together may have the function of marker for the exposure to allergens of A. alternata.

Marker antigens to assess exposure to molds and their allergens. I. Aspergillus fumigatus.

BACKGROUND: Exposure to molds and mold products in the indoor environment may lead to allergies, asthma, or respiratory complaints in general. Enumeration of viable parts of molds in the environment is insufficient to estimate exposure. Therefore, other methods have to be developed. METHODS: Aspergillus fumigatus (Af) was grown under various circumstances (22 degrees C and 30 degrees C, high and low water activity) in the laboratory. At various moments during culture, extracts were taken, and antigen and allergen content was examined by acrylamide electrophoresis and immunoblot. RESULTS: In extracts of the culture filtrate, two antigens were found to be produced under all studied growth conditions (common antigens). In the extracts of the water-soluble portion of the mycelium, one common antigen was found. CONCLUSIONS: The three common antigens may serve as marker antigens for exposure to Af and its products. In view of the simultaneous presence of two of these common antigens with Af allergens, these two marker antigens may be used to estimate exposure to allergens of Af.

Immunoblotting technique for the detection of allergens of Aspergillus fumigatus: influence of Nonidet P-40 on the sensitivity.

Immunoblotting provides a useful technique for the study of antigens, antibodies and allergens. To overcome problems regarding the loss of antigenic properties during the blotting and developing procedures, several solutions have been described. The inclusion of Nonidet P-40, recommended to increase the sensitivity of developing procedures for immunoblots, in an existing procedure for the detection of allergens of Aspergillus fumigatus, however, led to decreased sensitivity of the method.

Purification and characterization of a 93 kDa Aspergillus fumigatus antigen with diagnostic potential.

A glycoprotein with an apparent molecular weight of 93 kDa was purified from a water-soluble extract of Aspergillus fumigatus NCPF 2109 by single step affinity chromatography using the mannose-specific snowdrop (Galanthus nivalis) lectin coupled to agarose. The carbohydrate moiety contained only mannose and galactose. Partial sequencing of cyanogen bromide fragments of the antigen yielded two sequences, KQNKP and GEIPMKF?PQL, with no homology to any reported proteins. In a preliminary evaluation of its diagnostic potential the 93 kDa antigen was recognized by the sera of four patients with allergic bronchopulmonary aspergillosis, in addition to a monoclonal antibody raised against a partially purified fraction of the A. fumigatus water-soluble extract.

Human serum antibodies to extracellular polysaccharides (EPS) of moulds.

The presence of antibodies to extracellular polysaccharides (EPS) of moulds in sera of healthy subjects (N = 125) was determined. Antibodies against the EPS of Penicillium digitatum, Mucor racemosus, Cladosporium cladosporioides, Fusarium moniliforme and Botrytis tulipae were found in relatively high amounts in all sera. No effect of age on antibodies present could be demonstrated. Antibodies against each of the EPS tested were only neutralized by the homologous EPS and by EPS of moulds belonging to the same genus or a taxonomically closely related genus. Antibodies against the EPS of P. digitatum were inhibited by methyl-beta-D-galactofuranoside, indicating that the galactofuranose part of this EPS is immunodominant.