Reducing viral contamination from finger pads: handwashing is more effective than alcohol-based hand disinfectants.

BACKGROUND: Hand hygiene is important for interrupting transmission of viruses through hands. Effectiveness of alcohol-based hand disinfectant has been shown for bacteria but their effectiveness in reducing transmission of viruses is ambiguous. AIM: To test efficacy of alcohol hand disinfectant against human enteric and respiratory viruses and to compare efficacy of an alcohol-based hand disinfectant and handwashing with soap and water against norovirus. METHODS: Efficacies of a propanol and an ethanol-based hand disinfectant against human enteric and respiratory viruses were tested in carrier tests. Efficacy of an alcohol-based hand disinfectant and handwashing with soap and water against noroviruses GI.4, GII.4, and MNV1 were tested using finger pad tests. FINDINGS: The alcohol-based hand disinfectant reduced the infectivity of rotavirus and influenza A virus completely within 30s whereas poliovirus Sabin 1, adenovirus type 5, parechovirus 1, and MNV1 infectivity were reduced <3 log10 within 3 min. MNV1 infectivity reduction by washing hands with soap and water for 30s (>3.0 ± 0.4 log10) was significantly higher than treating hands with alcohol (2.8 ± 1.5 log10). Washing with soap and water for 30s removed genomic copies of MNV1 (>5 log10), noroviruses GI.4 (>6 log10), and GII.4 (4 log10) completely from all finger pads. Treating hands with propanol-based hand disinfectant showed little or no reduction to complete reduction with mean genomic copy reduction of noroviruses GI.4, GII.4, and MNV1 being >2.6, >3.3, and >1.2 log10 polymerase chain reaction units respectively. CONCLUSIONS: Washing hands with soap and water is better than using alcohol-based hand disinfectants in removing noroviruses from hands.

Inactivation of Listeria monocytogenes by disinfectants and bacteriophages in suspension and stainless steel carrier tests.

To simulate food contact surfaces with pits or cracks, stainless steel plates with grooves (depths between 0.2 and 5 mm) were constructed. These plates were artificially contaminated with Listeria monocytogenes in clean conditions, with organic soiling, or after 14 days of biofilm formation after which inactivation of the pathogen by Suma Tab D4 (sodium dichloroisocyanurate, 240 and 300 mg/liter), Suma Bac D10 (quaternary ammonium compound, 740 mg/liter), and bacteriophage suspension (Listex P100) was determined. Both chemical disinfectants performed well in suspension tests and in clean carrier tests according to the European standard with a reduction of more than 5 and 4 log units, respectively, of Listeria cells after 5 min of contact time. However, for the plates with grooves, the reduction could not meet the standard requirement, although a higher reduction of L. monocytogenes was observed in the shallow grooves compared with the deeper grooves. Furthermore, presence of food residues and biofilm reduced the effect of the disinfectants especially in the deep grooves, which was dependent on type of food substrate. Bacteriophages showed the best antimicrobial effect compared with the chemical disinfectants (sodium dichloroisocyanurate and quaternary ammonium compound) in most cases in the shallow grooves, but not in the deep grooves. The chlorine based disinfectants were usually less effective than quaternary ammonium compound. The results clearly demonstrate that surfaces with grooves influenced the antimicrobial effect of the chemical disinfectants and bacteriophages because the pathogen is protected in the deep grooves. The use of bacteriophages to inactivate pathogens on surfaces could be helpful in limited cases; however, use of large quantities in practice may be costly and phage-resistant strains may develop.

Risk evaluation and management to reaching a suggested FSO in a steam meal.

Steam meals are ready-to-eat meals composed of raw and semi-cooked ingredients, which get cooked while microwave heating. In this study, an Indian style meal was selected, Chicken Tandoori, from two different producers. These meals were first evaluated with the Risk Ranger® to identify the main foodborne pathogens risks, which were Listeria monocytogenes, Salmonella Typhimurium and Bacillus cereus. Thereafter, quantitative microbiology was applied using different models and verified with growth and inactivation challenge tests. It was observed that the gamma model and the ComBase program® showed very similar results. However, in some cases the results obtained with the challenge tests showed different results. The information gathered was used to create different scenarios which indicate how to manage the risks by setting Performance Objectives during the different stages of the food chain of this product and hence reaching a suggested Food Safety Objective.

Consumption of raw vegetables and fruits: a risk factor for Campylobacter infections.

The purpose of this study was to determine the prevalence of Campylobacter in fresh vegetables and fruits at retail level in the Netherlands, and to estimate its implications on the importance of vegetables and fruits as risk factor for campylobacteriosis. Thirteen of the 5640 vegetable and fruit samples were Campylobacter positive, resulting in a prevalence of 0.23% (95% confidence interval (Cl): 0.12-0.39%). The prevalence of packaged products (0.36%, 95% Cl: 0.17-0.66) was significantly higher than of unpackaged products (0.07; 95% Cl: 0.01-0.27). No statistical differences were found between seasons. Combining the mean prevalence found in this study with data on the consumption of vegetables and fruits, an exposure of 0.0048 campylobacters ingested per person per day in the Netherlands by transmission via vegetables and fruits, was calculated. This exposure, as input in a Beta-Poisson dose-response model, resulted in an estimated number of 5.3×105 cases of infection with Campylobacter per year for the whole Dutch population. This constitutes the consumption of raw vegetables and fruits, especially when packaged, to be a risk factor for Campylobacter infections.

Survival of Listeria monocytogenes on a conveyor belt material with or without antimicrobial additives.

Survival of Listeria monocytogenes on a conveyor belt material with or without antimicrobial additives, in the absence or presence of food debris from meat, fish and vegetables and at temperatures of 10, 25 and 37 degrees C was investigated. The pathogen survived best at 10 degrees C, and better at 25 degrees C than at 37 degrees C on both conveyor belt materials. The reduction in the numbers of the pathogen on belt material with antimicrobial additives in the first 6h at 10 degrees C was 0.6 log unit, which was significantly higher (P<0.05) than the reduction of 0.2 log unit on belt material without additives. Reductions were significantly less (P<0.05) in the presence of food residue. At 37 degrees C and 20% relative humidity, large decreases in the numbers of the pathogen on both conveyor belt materials during the first 6h were observed. Under these conditions, there was no obvious effect of the antimicrobial substances. However, at 25 degrees C and 10 degrees C and high humidity (60-75% rh), a rapid decrease in bacterial numbers on the belt material with antimicrobial substances was observed. Apparently the reduction in numbers of L. monocytogenes on belt material with antimicrobial additives was greater than on belt material without additives only when the surfaces were wet. Moreover, the presence of food debris neutralized the effect of the antimicrobials. The results suggest that the antimicrobial additives in conveyor belt material could help to reduce numbers of microorganisms on belts at low temperatures when food residues are absent and belts are not rapidly dried.

Fermented soya bean (tempe) extracts reduce adhesion of enterotoxigenic Escherichia coli to intestinal epithelial cells.

AIMS: This study aimed to investigate the effect of processed soya bean, during the successive stages of tempe fermentation and different fermentation times, on adhesion of enterotoxigenic Escherichia coli (ETEC) K88 to intestinal brush border cells as well as Caco-2 intestinal epithelial cells; and to clarify the mechanism of action. METHODS AND RESULTS: Tempe was prepared at controlled laboratory scale using Rhizopus microsporus var. microsporus as the inoculum. Extracts of raw, soaked and cooked soya beans reduced ETEC adhesion to brush border cells by 40%. Tempe extracts reduced adhesion by 80% or more. ETEC adhesion to Caco-2 cells reduced by 50% in the presence of tempe extracts. ETEC K88 bacteria were found to interact with soya bean extracts, and this may contribute to the observed decrease of ETEC adhesion to intestinal epithelial cells. CONCLUSIONS: Fermented soya beans (tempe) reduce the adhesion of ETEC to intestinal epithelial cells of pig and human origin. This reduced adhesion is caused by an interaction between ETEC K88 bacteria and soya bean compounds. SIGNIFICANCE AND IMPACT OF THE STUDY: The results strengthen previous observations on the anti-diarrhoeal effect of tempe. This effect indicates that soya-derived compounds may reduce adhesion of ETEC to intestinal cells in pigs as well as in humans and prevent against diarrhoeal diseases.

Quantification of Campylobacter jejuni cross-contamination via hands, cutlery, and cutting board during preparation of a chicken fruit salad.

Using artificially contaminated chicken, the quantitative overall effect of Campylobacter jejuni cross-contamination, either via cutlery, cutting board, or hands, on the microbiological quality of a chicken salad was tested to identify the most critical transfer route. The end contamination level of salads prepared according to different scenarios, with or without cross-contamination, was compared. It was shown that the mean transfer rate calculated for all salads prepared allowing cross-contamination was 0.12% of the initial number of C. jejuni on the chicken fillet (8.8 +/- 0.2 log CFU). The difference in calculated transfer rates for the tested cross-contamination routes was not significantly different (P > 0.05). The prevention of cross-contamination by replacing cutlery and cutting board after handling raw chicken and the prevention of hand contact resulted in considerably reduced end contamination levels (< 2.4 log CFU) or noncontaminated end products. The results of this study emphasize the importance of preventing cross-contamination during food handling in reducing the risks of foodborne infections, and they provide useful data for quantitative microbiological risk assessment.

Number of Salmonella on chicken breast filet at retail level and its implications for public health risk.

This study aimed to characterize the number of Salmonella on chicken breast filet at the retail level and to evaluate if this number affects the risk of salmonellosis. From October to December 2005, 220 chilled raw filets (without skin) were collected from five local retail outlets in The Netherlands. Filet rinses that were positive after enrichment were enumerated with a three-tube most-probable-number (MPN) assay. Nineteen filets (8.6%) were contaminated above the detection limit of the MPN method (10 Salmonella per filet). The number of Salmonella on positive filets varied from 1 to 3.81 log MPN per filet. The obtained enumeration data were applied in a risk assessment model. The model considered possible growth during domestic storage, cross-contamination from filet via a cutting board to lettuce, and possible illness due to consumption of the prepared lettuce. A screening analysis with expected-case and worst-case estimates for the input values of the model showed that variability in the inputs was of relevance. Therefore, a Monte Carlo simulation with probability distributions for the inputs was carried out to predict the annual number of illnesses. Remarkably, over two-thirds of annual predicted illnesses were caused by the small fraction of filets containing more than 3 log Salmonella at retail (0.8% of all filets). The enumeration results can be used to confirm this hypothesis in a more elaborate risk assessment. Modeling of the supply chain can provide insight for possible intervention strategies to reduce the incidence of rare, but extreme levels. Reduction seems feasible within current practices, because the retail market study indicated a significant difference between suppliers.

Shiga toxin-producing Escherichia coli isolated from patients with diarrhoea in Bangladesh.

The prevalence of Shiga toxin-producing Escherichia coli (STEC) and its characteristics were determined among hospitalized patients with diarrhoea and children with diarrhoea in an urban slum community of Dhaka city using sensitive culture and PCR methods. Stool samples were collected from 410 patients with diarrhoea enrolled in the 2% surveillance system (every 50th patient attending the hospital with diarrhoeal disease is included) at the ICDDR,B hospital and from 160 children of 2-5 years of age with diarrhoea living in an urban slum in Dhaka, between September 2004 and April 2005. Shiga toxin genes (stx) were detected by multiplex PCR in the enrichment broth of nine samples (2.2%) from hospitalized patients and 11 samples (6.9%) from the community patients. STEC was isolated from five stool samples with positive PCR results using a colony patch technique. All five isolates were positive in the Vero cell assay and PCR fragments of stx genes were confirmed by sequencing. Two isolates were positive for the E. coli attaching-and-effacing (eae) gene and four were positive for the enterohaemolysin (hlyEHEC) gene and enterohaemolysin production. The five isolates belonged to five different serotypes:O32:H25, O2:H45, O76:H19, ONT:H25 and ONT:H19. It can be concluded that STEC is not a common pathogen in Bangladesh among hospitalized patients with diarrhoea nor among mild cases of diarrhoea in the community.

Behavior of Clostridium perfringens at low temperatures.

Refrigerated storage is an important step in the preparation of foods and inadequate storage is one of the main causes of food poisoning outbreaks of Clostridium perfringens. Therefore, growth and germination characteristics of C. perfringens in a temperature range of 3-42 degrees C were determined in fluid thioglycollate broth (FTG) and Dutch pea soup. To study the effect of adaptation, cells were either inoculated from a 37 degrees C pre-culture or from a temperature-adapted pre-culture. Membrane fatty acid patterns were determined at all temperatures to examine the effect of temperature on membrane composition. Spores were either inoculated with and without heat treatment. Adaptation of cells did not influence growth rate nor lag phase. Growth in pea soup, however, was slower and lag phases tended to be more extended compared to FTG. No growth was observed at temperatures < or =10 degrees C and death rates in pea soup were higher than those in FTG at these low temperatures. Cells preserved the membrane fluidity by reducing the arachidic acid content and increasing the lauric acid content when the temperature dropped. This resulted in a net reduction in chain length. Microscopic analysis of cells grown at 15 degrees C revealed a morphological change: cells were elongated compared to those grown at 37 degrees C. These data demonstrate the ability of C. perfringens to adapt to lower temperatures. However, this did not influence growth characteristics compared to non-adapted cells. Spores of C. perfringens did germinate at all temperatures with and without heat-activation. Combining this fact with the extended survival at low temperatures emphasizes the need for adequate heating of refrigerated foods before consumption to eliminate health risks due to C. perfringens.

A quantitative analysis of cross-contamination of Salmonella and Campylobacter spp. via domestic kitchen surfaces.

Epidemiological data indicate that cross-contamination during food preparation in the home contributes noticeably to the occurrence of foodborne diseases. To help prevent such occurrences, the inclusion of a cross-contamination model in exposure assessments would aid in the development and evaluation of interventions used to control the spread of pathogenic bacteria. A quantitative analysis was carried out to estimate the probability of contamination and the levels of Salmonella and Campylobacter spp. on salads as a result of cross-contamination from contaminated chicken carcasses via kitchen surfaces. Data on the prevalence and numbers of these bacteria on retail chicken carcasses and the use of unwashed surfaces to prepare foods were collected from scientific literature. The rates of bacterial transfer were collected from laboratory experiments and literature. A deterministic approach and Monte Carlo simulations that incorporated input parameter distributions were used to estimate the contamination of the product. The results have shown that the probability of Campylobacter spp. contamination on salads is higher than that of Salmonella spp., since both the prevalence and levels of Campylobacter spp. on chicken carcasses are higher than those of Salmonella spp. It is realistic to expect that a fraction of the human exposure to Campylobacter spp., in particular, originates from cross-contamination in private kitchens during food handling. The number of human campylobacteriosis cases could be reduced either by reducing the degree of Campylobacter spp. contamination on chicken carcasses or by improving the hygiene in private kitchens. To eliminate the cross-contamination route, it is important to use separate surfaces or to properly wash the surfaces during the preparation of raw and cooked foods or ready-to-eat foods.

A new protocol for the detection of Enterobacter sakazakii applied to environmental samples.

Enterobacter sakazakii is a motile, peritrichous, gram-negative rod that was previously known as a yellow pigmented Enterobacter cloacae. It is documented as a rare cause of outbreaks and sporadic cases of life-threatening neonatal meningitis, necrotizing enterocolitis, and sepsis. E. sakazakii has been isolated from milk powder-based formulas, and there is thus a need to investigate whether and where E. sakazakii occurs in these manufacturing environments. For this purpose, a simple detection method was developed based on two features of E. sakazakii: its yellow pigmented colonies when grown on tryptone soy agar and its constitutive alpha-glucosidase, which is detected in a 4-h colorimetric assay. Using this screening method, E. sakazakii strains were isolated from three individual factories from 18 of 152 environmental samples, such as scrapings from dust, vacuum cleaner bags, and spilled product near equipment. The method is useful for routine screening of environmental samples for the presence of E. sakazakii.

Effect of cooling on Clostridium perfringens in pea soup.

Foods associated with Clostridium perfringens outbreaks are usually abused after cooking. Because of their short generation times, C. perfringens spores and cells can grow out to high levels during improper cooling. Therefore, the potential of C. perfringens to multiply in Dutch pea soup during different cooling times was investigated. Tubes of preheated pea soup (50 degrees C) were inoculated with cocktails of cells or heat-activated spores of this pathogen. The tubes were linearly cooled to 15 degrees C in time spans of 3, 5, 7.5, and 10 h and were subsequently stored in a refrigerator at 3 or 7 degrees C for up to 84 h. Cell numbers increased by 1-log cycle during the 3-h cooling period and reached their maximum after 10 h of cooling. Subsequent refrigeration hardly reduced cell numbers. Cooling of 3.75 liters of pea soup in an open pan showed that this amount of pea soup cooled from 50 to 15 degrees C in 5 h, which will allow a more than 10-fold increase in cell numbers. These findings emphasize the need of good hygienic practices and quick cooling of heated foods after preparation.

Tolerance of Salmonella Enteritidis and Staphylococcus aureus to surface cleaning and household bleach.

Effective cleaning and sanitizing of food preparation sites is important because pathogens are readily spread to food contact surfaces after preparation of contaminated raw products. Tolerance of Salmonella Enteritidis and Staphylococcus aureus to surface cleaning by wiping with regular, microfiber, and antibacterial-treated cloths was investigated. Wiping with cleaning cloths resulted in a considerable reduction of microorganisms from surfaces, despite the greater difficulty in removing S. aureus than Salmonella Enteritidis. Depending on the cloth type, S. aureus were reduced on surfaces from initial numbers of approximately 10(5) CFU/100 cm2 to numbers from less than 4 CFU/100 cm2 (below the detection limit) to 100 CFU/100 cm2. Directly after the cloths were used to clean the contaminated surfaces, they contained high numbers of bacteria (10(4) to 10(5) CFU/100 cm2), except for the disposable antibacterial-treated cloths, in which no bacteria could be detected. The tolerance of these pathogens to sodium hypochlorite was studied in the suspension test and in cloths. S. aureus showed a better tolerance for sodium hypochlorite than Salmonella Enteritidis. Inactivation of microorganisms in cloths required a higher concentration of sodium hypochlorite than was needed in the suspension test. Repeated exposure to sodium hypochlorite, however, resulted in an increase in susceptibility to this compound. This study provides essential information about the transfer of bacteria when wiping surfaces and highlights the need for a hygiene procedure with cleaning cloths that sufficiently avoids cross-contamination in the household environment.

Survival of foodborne pathogens on stainless steel surfaces and cross-contamination to foods.

The retention of bacteria on food contact surfaces increases the risk of cross-contamination of these microorganisms to food. The risk has been considered to be lowered when the surfaces are dry, partly because bacterial growth and survival would be reduced. However, some non-spore-forming bacteria might be able to withstand dry conditions on surfaces for an extensive period of time. In this study the survival of Salmonella enteritidis, Staphylococcus aureus and Campylobacter jejuni on stainless steel surfaces at different initial levels was determined at room temperature. The transfer rates of these pathogens from kitchen sponges to stainless steel surfaces and from these surfaces to foods were also investigated. Staph. aureus was recovered from the surfaces for at least 4 days when the contamination level was high (10(5) CFU/cm2) or moderate (10(3) CFU/cm2). At low levels (10 CFU/cm2), the surviving numbers decreased below the detection limit (4 CFU/100 cm2) within 2 days. S. enteritidis was recovered from surfaces for at least 4 days at high contamination levels, but at moderate level, the numbers decreased to the detection limit within 24 h and at low level within 1 h. C. jejuni was the most susceptible to slow-air-drying on surfaces; at high contamination levels, the numbers decreased below the detection limit within 4 h. The test microorganisms were readily transmitted from the wet sponges to the stainless steel surfaces and from these surfaces to the cucumber and chicken fillet slices, with the transfer rates varied from 20% to 100%. This study has highlighted the fact that pathogens remain viable on dry stainless steel surfaces and present a contamination hazard for considerable periods of time, dependent on the contamination levels and type of pathogen. Systematic studies on the risks of pathogen transfer associated with surface cleaning using contaminated sponges provide quantitative data from which a model of risks assessment in domestic setting could lead.

Comparison of media for enumeration of Clostridium perfringens from foods.

Many media have been developed to enumerate Clostridium perfringens from foods. In this study, six media [iron sulfite (IS) agar, tryptose sulfite cycloserine (TSC) agar, Shahidi Ferguson perfringens (SFP) agar, sulfite cycloserine azide (SCA), differential clostridial agar (DCA), and oleandomycin polymyxin sulfadiazine perfringens (OPSP) agar] were compared in a prestudy, of which four (IS, TSC, SCA, and DCA) were selected for an international collaborative trial. Recovery of 15 pure strains was tested in the prestudy and recovery of one strain from foodstuffs was tested in the collaborative trial. Results from the prestudy did reveal statistical difference of the media but recoveries on all media were within the microbiological limits (+/-30%) of IS, which was set as a reference medium. Recoveries on the media tested in the collaborative trial were statistically different as well, but these differences were of no microbiological-analytical relevance. Food matrices did not affect the recovery of C. perfringens in general. DCA and SCA, in particular, are labor-intensive to prepare and DCA frequently failed to produce black colonies; gray colonies were quite common. Since IS medium is nonselective, it was concluded that TSC was the most favorable medium for the enumeration of C. perfringens from foods.

Optimizing sporulation of Clostridium perfringens.

Many sporulation media have been developed for Clostridium perfringens, but none stimulates sporulation for all strains. The aim of our experiments was to develop a sporulation method using Duncan and Strong (DS) medium, which supports sporulation of a wide variety of strains. Different inoculation levels were tested, and the effects of sporulation-promoting substances and acid shock were evaluated. Furthermore, DS medium was compared with other sporulation media. Highest spore numbers in DS medium were obtained with a 10% 24-h fluid thioglycollate broth inoculum (5.0 x 10(5)/ml). Addition of theophylline and replacement of starch by raffinose increased spore yields for some strains, but most strains were not affected (average increases in log N/ml of 0.2 and 0.3, respectively). One strain was enhanced by the addition of bile, but other strains were strongly inhibited (average decrease in log N/ml of 2.5); agar did not influence sporulation. Neither short-time acid exposure nor addition of culture supernatant fluids of well-sporulating strains resulted in higher spore numbers in DS medium. None of the tested methods enhanced sporulation in general; only strain-dependent effects were obtained. Peptone bile theophylline medium was the most promising sporulation medium tested; peptone bile theophylline starch medium yielded highest spore numbers (2.5 x 10(5)/ml), but some strains failed to sporulate. In conclusion, adding theophylline to DS medium may optimize sporulation of C. perfringens, but peptone bile theophylline medium with or without starch is most suitable.

Effects of antibacterial dishwashing liquid on foodborne pathogens and competitive microorganisms in kitchen sponges.

In response to increasing concern about home hygiene, the use of antibacterial products to reduce microorganisms in kitchen sponges and cleaning cloths is strongly promoted by some producers of detergent for domestic use. The effects of an antibacterial dishwashing liquid on Escherichia coli, Salmonella Enteritidis, Staphylococcus aureus, and Bacillus cereus were investigated in a modified suspension test and in used sponges with and without food residues under laboratory conditions. A limited study was conducted in households to assess the efficacy of antibacterial dishwashing liquid as used by the consumer. In the suspension tests, S. aureus and B. cereus were shown to be susceptible to low concentrations of antibacterial dishwashing liquid (0.5%), whereas E. coli and Salmonella Enteritidis maintained their initial numbers for at least 24 h at 25 degrees C. At higher concentrations (2 to 4%), all test organisms decreased to below the detection limit after 24 h. Over a 24-h period, the antibacterial dishwashing liquid did not significantly reduce these organisms in used sponges in which food residues were present. The antibacterial product did not reduce the competitive microorganisms either. Similar results were found for sponges involved in daily household use. The results of this study demonstrate that the antibacterial dishwashing liquid was effective in reducing pathogens in the suspension test but not in the used sponges. This finding indicates that to determine the efficacy of antibacterial products, their use in a household setting must be considered.

Stepwise quantitative risk assessment as a tool for characterization of microbiological food safety.

This paper describes a system for the microbiological quantitative risk assessment for food products and their production processes. The system applies a stepwise risk assessment, allowing the main problems to be addressed before focusing on less important problems. First, risks are assessed broadly, using order of magnitude estimates. Characteristic numbers are used to quantitatively characterize microbial behaviour during the production process. These numbers help to highlight the major risk-determining phenomena, and to find negligible aspects. Second, the risk-determining phenomena are studied in more detail. Both general and/or specific models can be used for this and varying situations can be simulated to quantitatively describe the risk-determining phenomena. Third, even more detailed studies can be performed where necessary, for instance by using stochastic variables. The system for quantitative risk assessment has been implemented as a decision supporting expert system called SIEFE: Stepwise and Interactive Evaluation of Food safety by an Expert System. SIEFE performs bacterial risk assessments in a structured manner, using various information sources. Because all steps are transparent, every step can easily be scrutinized. In the current study the effectiveness of SIEFE is shown for a cheese spread. With this product, quantitative data concerning the major risk-determining factors were not completely available to carry out a full detailed assessment. However, this did not necessarily hamper adequate risk estimation. Using ranges of values instead helped identifying the quantitatively most important parameters and the magnitude of their impact. This example shows that SIEFE provides quantitative insights into production processes and their risk-determining factors to both risk assessors and decision makers, and highlights critical gaps in knowledge.

Occurrence and survival of verocytotoxin-producing Escherichia coli O157 in meats obtained from retail outlets in The Netherlands.

In 1996 and 1997, 2,941 fresh and processed meat products obtained from supermarkets and butcher shops in The Netherlands were examined for the presence of verocytotoxin-producing Escherichia coli of serogroup O157 (O157 VTEC). Additionally, the fate of O157 VTEC in raw meat products stored at low temperatures and the effect of different additives were evaluated. O157 VTEC strains were isolated from 6 (1.1%) of 571 samples of raw minced beef, 2 (0.5%) of 402 samples of raw minced mixed beef and pork, 1 (1.3%) of 76 samples of raw minced pork, 1 (0.3%) of 393 samples of other raw pork products, and 1 (0.3%) of 328 samples of cooked or fermented ready-to-eat meats. Other raw beef products (n = 223) and meat samples originating from poultry (n = 819), sheep or lamb (n = 46), or wild animals (n = 83) were all found to be negative for O157 VTEC. For the survival experiments we used tartaar (minced beef with a fat content of less than 10%) and filet americain (tartaar mixed with a mayonnaise-based sauce [80 to 20%]). The O157 VTEC strain tested was able to survive in tartaar and filet americain stored at -20, 0, 5, or 7 degrees C for 3 days. At both 7 and at 15 degrees C, O157 VTEC counts in tartaar and filet americain remained virtually unchanged throughout a storage period of 5 days. Addition of acetic acid (to pH 4.0), sodium lactate (1 and 2% [wt/wt]), or components of the lactoperoxidase-thiocyanate-hydrogen peroxide system to filet americain did not result in a reduction of viable O157 VTEC cells during storage at 7 or 15 degrees C. It was concluded that raw meat contaminated with O157 VTEC will remain a hazard even if the meat is held at low or freezing temperatures.