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J Debevere

Publications and source records attributed to J Debevere.

At least 19 recordsLinked to original sources

Individual and combined effects of ph and lactic acid concentration on Listeria innocua inactivation: development of a predictive model and assessment of experimental variability.

In food technology, organic acids (e.g., lactic acid, acetic acid, and citric acid) are popular preservatives. The purpose of this study was to separate the individual effects of the influencing factors pH and undissociated lactic acid on Listeria innocua inactivation. Therefore, the inactivation process was investigated under controlled, initial conditions of pH (pH0) and undissociated lactic acid ([LaH]0). The resulting inactivation curves consisted of a (sometimes negligible) shoulder period followed by a descent phase. In a few cases, a tailing phase was observed. Depending on the conditions, the descent phase contained one or two log-linear parts or had a convex or concave shape. In addition, the inactivation process was characterized by a certain variability, dependent on the severity of the conditions. Furthermore, in the neighborhood of the growth/no growth interface sometimes contradictory observations occurred. Overall, the individual effects of the influencing factors pH and undissociated lactic acid could clearly be distinguished and were also apparent based on fluorescence microscopy. Appropriate model types were developed and enabled prediction of which conditions of pH0 and [LaH]0 are necessary to obtain a predetermined inactivation (number of decimal reductions) within a predetermined time range.

Food Preservatives↗

Evaluation of two viral extraction methods for the detection of human noroviruses in shellfish with conventional and real-time reverse transcriptase PCR.

AIMS: Comparison of two viral extraction methods in order to establish a sensitive and simple detection method for human noroviruses (NV) in bivalve shellfish. METHODS AND RESULTS: A direct RNA extraction method and an alkaline virus elution-concentration method were tested on artificially contaminated mussels. The latter used an alkaline buffer and polyethylene glycol (PEG) to isolate and concentrate the virus particles from shellfish. In both methods Trizol was used to release RNA. The final RNA extracts were amplified and detected with conventional and real-time reverse transcriptase PCR. The direct RNA extraction method was not able to detect low inoculation levels. However, the virus elution-concentration method was more sensitive. CONCLUSIONS: The alkaline elution-PEG concentration method followed by Trizol effectively removed inhibitory components and fulfilled the demands to be a useful tool for routine testing of shellfish for NV detection. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of the lack of standardized methods to detect NV in shellfish, many 'in-house' extraction methods are used in practice. A comparison of these methods aims to determine a simple, rapid and sensitive method that could be a candidate method for screening suspected shellfish.

Animals↗

Influence of pH, water activity and acetic acid concentration on Listeria monocytogenes at 7 degrees C: data collection for the development of a growth/no growth model.

Growth/no growth models can be used to determine the chance that microorganisms will grow in specific environmental conditions. As a consequence, these models are of interest in the assessment of the safety of foods which can be contaminated with food pathogens. In this paper, growth/no growth data for Listeria monocytogenes (in a monoculture and in a mixed strain culture) are presented. The data were gathered at 7 degrees C in Nutrient Broth with different combinations of environmental factors pH (5.0-6.0, six levels), water activity (0.960-0.990, six levels) and acetic acid concentration (0-0.8% (w/w), five levels). This combination of environmental factors for the development of a growth/no growth model was based on the characteristics of sauces and mayonnaise based salads. The strains used were chosen from screening experiments in which the pH, water activity and acetic acid resistance of 26 L. monocytogenes strains (LFMFP culture collection) was determined at 30 degrees C in Brain Heart Infusion broth. The screening showed that most L. monocytogenes strains were not able to grow at a(w)<0.930, pH<4.3 or a total acetic acid concentration >0.4% (w/w). Among these strains, the ones chosen were the most resistant to one of these factors in the hope that, if the resulting model predicted no growth at certain conditions for those more resistant strains, then these predictions would also be valid for the less resistant strains. A mixed strain culture was also examined to combine the strains that were most resistant to one of the factors. A full factorial design with the selected strains was tested. The experiments were performed in microtiter plates and the growth was followed by optical density measurements at 380 nm. The plates were inoculated with 6 log CFU/ml and twenty replicates were made for each treatment combination. These data were used (1) to determine the growth/no growth boundary and (2) to estimate the influence of the environmental conditions on the time to detection. From the monoculture and mixed strain data, the growth boundary of L. monocytogenes is shown not to be a straight cut-off but a rather narrow transition zone. The experiments also showed that in the studied region, a(w) did not have a pronounced influence on the position of the growth/no growth boundary while a low concentration of acetic acid (0.2% (w/w)) and a pH decrease from 6.0 to 5.8 was sufficient to significantly reduce the possibility of growth. The determination of the time to detection showed a significant increase at the combinations of environmental conditions near the 'no growth zone'. For example, at 0.2% (w/w) acetic acid, there was an increase from +/-10 days to 30 days by lowering pH from 5.8 to 5.6 at a(w) values of 0.985 and 0.979, while at pH 5.4 less than 50% growth occurred for all a(w) values.

Acetic Acid↗

The influence of modified atmospheres and their interaction with water activity on the radial growth and fumonisin B(1) production of Fusarium verticillioides and F. proliferatum on corn. Part I: the effect of initial headspace carbon dioxide concentration.

The effect of modified atmospheres on the growth and fumonisin B(1) production of Fusarium verticillioides and Fusarium proliferatum on corn is presented in a series of two papers. In this, the first part, the effect of initial headspace (IH) carbon dioxide concentration and its interaction with water activity (a(w)) on growth and fumonisin B(1) production was evaluated. It was observed that at all a(w) values studied, increase in the IH CO(2) concentration generally resulted in a decrease in the colony growth rate (g, mm day(-1)) and maximum colony diameter (D(max), mm) and an increase in the lag phase duration (lambda, day). Although both a(w) and IH CO(2) concentration had significant and synergistic effects on g, a(w) had the largest effect. As little as 10% IH CO(2) completely inhibited the production of fumonisin B(1) by F. verticillioides. F. proliferatum was more resistant and required 40, 30 and 10% IH CO(2) at a(w) 0.984, 0.951 and 0.930, respectively, to completely inhibit fumonisin B(1) production. These results demonstrate that modified atmospheres containing high CO(2) levels could potentially be employed for the protection of corn from fungal spoilage and mycotoxin contamination during the post-harvest period.

Carbon Dioxide↗

The influence of modified atmospheres and their interaction with water activity on the radial growth and fumonisin B(1) production of Fusarium verticillioides and F. proliferatum on corn. Part II: The effect of initial headspace oxygen concentration.

This paper is the second in a series of two that describe the effect of modified atmospheres on the growth and mycotoxin production of Fusarium verticillioides and Fusarium proliferatum on corn. In this part, the effect of initial headspace (IH) oxygen concentration and its interaction with water activity (a(w)) on growth and fumonisin B(1) production was investigated. In addition, the impact of vacuum packaging and in-cooperation of O(2) scrubbing sachets was also studied. It was observed that at all a(w) values studied, reduction of IH O(2) concentration from 20 to 2% had no significant effect on the colony growth rate (g, mm d(-1)) and lag phase duration (lambda, d). However, g and lambda were positively and negatively correlated to a(w). The IH O(2) concentration was determined to have a a(w) dependent effect on the oxygen consumption rate. Although the maximum colony diameter (D(max), mm) decreased with the reduction of the IH O(2) level, the greatest mycelial density occurred at 10% IH O(2) for both isolates. This observation was accompanied by a trend of a decrease in the value of the IH O(2) level at which the most fumonisin B(1) was produced from 15 to 5% when the a(w) was decreased from 0.976 to 0.930 for F. verticillioides. For F. proliferatum the optimum conditions for fumonisin B(1) production shifted from 20% at a(w) 0.976 to 10% at both 0.951 and 0.930. Vacuum packaging and the in-cooperation of O(2) absorbing sachets completely inhibited the growth of both isolates. These results together with those reported in Part I of the study indicate that O(2) should preferably be completely excluded from modified atmospheres that are employed to protect stored corn from fungal growth and mycotoxin production.

Colony Count, Microbial↗

Quantitative risk assessment of Campylobacter spp. in poultry based meat preparations as one of the factors to support the development of risk-based microbiological criteria in Belgium.

The objective of this study was to do an exercise in risk assessment on Campylobacter spp. for poultry based meat preparations in Belgium. This risk assessment was undertaken on the demand of the competent national authorities as one of the supportive factors to define risk-based microbiological criteria. The quantitative risk assessment model follows a retail to table approach and is divided in different modules. The contamination of raw chicken meat products (CMPs) was represented by a normal distribution of the natural logarithm of the concentration of Campylobacter spp. (ln[Camp]) in raw CMPs based on data from surveillance programs in Belgium. To analyse the relative impact of reducing the risk of campylobacteriosis associated with a decrease in the Campylobacter contamination level in these types of food products, the model was run for different means and standard deviations of the normal distribution of the ln[Camp] in raw CMPs. The limitation in data for the local situation in Belgium and on this particular product and more precisely the semi-quantitative nature of concentration of Campylobacter spp. due to presence/absence testing, was identified as an important information gap. Also the knowledge on the dose-response relationship of Campylobacter spp. was limited, and therefore three different approaches of dose-response modelling were compared. Two approaches (1 and 2), derived from the same study, showed that the reduction of the mean of the distribution representing the ln[Camp] in raw CMPs is the best approach to reduce the risk of Campylobacter spp. in CMPs. However, for the simulated exposure and approach 3 it was observed that the reduction of the standard deviation is the most appropriate technique to lower the risk of campylobacteriosis. Since the dose-response models used in approach 1 and 2 are based on limited data and the reduction of the mean corresponds with a complete shift of the contamination level of raw CMPs, demanding high efforts from the poultry industry, it is proposed to lower the standard deviation of the concentration of Campylobacter spp. in raw CMPs. This proposal corresponds with the elimination of the products that are highly contaminated. Simulation showed that eating raw chicken meat products can give rise to exposures that are 10(10) times higher than when the product is heated, indicating that campaigns are important to inform consumers about the necessity of an appropriate heat treatment of these type of food products.

Animals↗

Modelling Yersinia enterocolitica inactivation in coculture experiments with Lactobacillus sakei as based on pH and lactic acid profiles.

In food processing and preservation technology, models describing microbial proliferation in food products are a helpful tool to predict the microbial food safety and shelf life. In general, the available models consider microorganisms in pure culture. Thus, microbial interactions are ignored, which may lead to a discrepancy between model predictions and the actual microbial evolution, particularly for fermented and minimally processed food products in which a background flora is often present. In this study, the lactic acid mediated negative microbial interaction between the lactic acid bacterium Lactobacillus sakei and the psychrotrophic food pathogen Yersinia enterocolitica was examined. A model describing the lactic acid induced inhibition (i.e., early induction of the stationary phase) of the pathogen [Vereecken, K.M., Devlieghere, F., Bockstaele, A., Debevere, J., Van Impe, J.F., 2003. A model for lactic acid induced inhibition of Yersinia enterocolitica in mono- and coculture with Lactobacillus sakei. Food Microbiology 20, 701-713.] was extended to describe the subsequent inactivation (i.e., decrease of the cell concentration to values below the detection limit). In the development of a suitable model structure to describe the inactivation process, critical points in the variation of the specific evolution rate mu [1/h] with the dynamic (time-varying) pH and undissociated lactic acid profiles were taken into account. Thus, biological knowledge, namely, both pH and undissociated lactic acid have an influence on the microbial evolution, was incorporated. The extended model was carefully validated on new data. As a result, the newly developed model is able to accurately predict the growth, inhibition and subsequent inactivation of Y. enterocolitica in coculture as based on the dynamic pH and lactic acid profiles of the medium.

Antibiosis↗

Volatile metabolite production of spoilage micro-organisms on a mixed-lettuce agar during storage at 7 degrees C in air and low oxygen atmosphere.

This paper describes the volatile metabolite production of spoilage bacteria (Pantoea agglomerans and Rahnella aquatilis) and spoilage yeasts (Pichia fermentans and Cryptococcus laurentii), previously isolated from mixed lettuce, on a simulation medium of shredded mixed lettuce (mixed-lettuce agar) both under air conditions and modified atmosphere (MA)-conditions at 7 degrees C. These latter conditions simulated the equilibrium modified atmosphere packaging, which is used to extend the shelf-life of shredded mixed lettuce. Besides volatile metabolites, organic acid metabolites and consumption of sugars were measured. Microbiological growth on the mixed-lettuce agar resulted in metabolite production and consumption of sugars. Bacteria and yeasts produced a range of volatile organic compounds both under air conditions and MA-conditions: ethanol, ethyl acetate, 2-methyl-1-propanol, 2-methyl-1-butanol, 3-methyl-1-butanol, 2,3-butanedione, 3-methyl-1-pentanol, 1-butanol and 1-hexanol. Under MA-conditions, 2-methyl-1-butanol, 3-methyl-1-butanol and ethanol were the first compounds that were detected in the headspace as being produced by the inoculated micro-organisms. In the case of the yeast P. fermentans, production of these compounds was detected from a count of 5.0+/-0.1 log cfu/cm(2) with a fast increase when exceeding 6.0-6.5 log cfu/cm(2). Unlike P. fermentans, the yeast C. laurentii showed a slow metabolism under MA-conditions, compared to air conditions. In the case of the bacteria, production of 2-methyl-1-butanol and 3-methyl-1-butanol was detected starting from a count of 6.7+/-0.1 log cfu/cm(2) in the case of R. aquatilis and from a count of 7.1+/-0.4 log cfu/cm(2) in the case of P. agglomerans with a fast increase when exceeding 8 log cfu/cm(2). No production of ethanol by the bacteria under MA-conditions was detected in contradiction to air conditions. It could be concluded that, if these counts are reached on the cut surfaces of shredded mixed lettuce which are simulated by the mixed-lettuce agar, sensorial quality of shredded mixed lettuce could be influenced by the microbiological production of metabolites.

Bacteria↗

Eggshell factors influencing eggshell penetration and whole egg contamination by different bacteria, including Salmonella enteritidis.

Trans-shell infection routes and whole egg contamination of 7 selected bacterial strains; Staphylococcus warneri, Acinetobacter baumannii, Alcaligenes sp., Serratia marcescens, Carnobacterium sp., Pseudomonas sp. and Salmonella enteritidis, recovered from egg contents, were studied. The first objective was to correlate bacterial eggshell penetration with various eggshell characteristics and bacterial strains. An agar approach was used to assess the eggshell penetration. The second objective was to assess the contamination of whole eggs with the bacterial strains; whole intact eggs were used in this case. The intact shells of agar-filled and whole eggs were inoculated with 10(3) -10(4) cfu of the selected strains. During 3 weeks storage at 20 degrees C and 60% relative humidity, the bacterial eggshell penetration was regularly monitored. The whole egg contamination was only analyzed after 3 weeks. The eggshell characteristics such as area eggshell, shell thickness and number of pores did not influence the bacterial eggshell penetration. For each individual bacterial strain the mean cuticle deposition was lower for penetrated compared to non-penetrated eggshells. For the individual strain Carnobacterium sp. and for the global results of all strains this difference was statistical significantly. The whole egg contamination was not influenced by neither the area of the eggshell nor the porosity of the eggshell. The results of the agar approach indicate that the Gram-negative, motile and non-clustering bacteria penetrated the eggshell most frequently; Pseudomonas sp. (60%) and Alcaligenes sp. (58%) were primary invaders followed by S. enteritidis (43%). All selected strains were able to penetrate; penetration was observed most frequently after ca. 4-5 days. Particularly S. enteritidis was a primary invader of whole eggs: the membranes and/or the content of 32% of the whole eggs was contaminated. The remaining bacterial eggshell contamination with the selected strain was determined after 3 weeks storage. Penetrated eggshells and contaminated whole eggs showed a significantly higher bacterial contamination on the eggshell compared to non-penetrated eggshells and non-contaminated whole eggs respectively (global results of all strains). The influence of hen age on bacterial eggshell penetration and egg content contamination was not significant. While the agar approach is suitable to study the influence of the eggshell characteristics on the bacterial eggshell penetration, the intact egg approach gives an estimation of the penetration of the shell followed by the probability of survival and migration in whole eggs.

Animals↗

Influence of food system conditions on N-acyl-L-homoserine lactones production by Aeromonas spp.

Eleven of 13 Aeromonas strains were shown to produce AHLs. Results of TLC showed that N-butanoyl-L-homoserine lactone (C4-HSL) was the main AHL produced in LB medium at 30 degrees C. The influence of different carbon sources, temperature, pH values and salt concentrations on AHL production was determined in eight A. hydrophila and one A. caviae strain. Additionally a quantitative study of C4-HSL production by A. hydrophila strain 519 under different conditions was performed. Positive results were found in the AHL induction assay for some Aeromonas strains in cultures in LB agar incubated at 12 degrees C after 72-96 h. The induction of the sensor strains by Aeromonas spp. occurred in LB medium supplemented with all carbon sources in a concentration of 0.5%. The production of C4-HSL by A. hydrophila 519 was found until 3.5% (w/v) of NaCl. For pHs close to the neutrality the C4-HSL production by A. hydrophila was evident after 24-48 h of incubation. A. hydrophila 519 produced C4-HSL under anaerobic conditions. Also, the AHL production by Aeromonas strains was studied in simulate agar of shrimp, fish and some vegetables. The production of AHLs was evident by almost all the test strains in shrimp simulated agar. In fish agar only for one of three fish species tested, positive results were found. Induction assay in vegetables simulated agar showed principally negative results, probably because of the presence of inhibitory compounds in these vegetables.

4-Butyrolactone↗

Effect of preincubation temperature and pH on the individual cell lag phase of Listeria monocytogenes, cultured at refrigeration temperatures.

The impact of precultural temperature and pH on the distribution of the lag phase of individual Listeria monocytogenes cells was assessed during preincubation at 7 degrees C, using a dilution protocol to obtain single cells, and optical density measurements to estimate the individual lag phase. Firstly, the pure temperature effect (37, 15, 10, 7, 4 and 2 degrees C) was investigated on a subsequent growth at 7 degrees C and pH 7.4. Secondly, low precultural temperatures (10, 7 and 4 degrees C) were combined with a controlled pH at 7.4 and 5.7 with a subsequent growth at 7 degrees C and at different pH values (7.4, 6.0 and 5.5). For all temperature-pH combinations, the individual cell lag phase was determined using a three-phase linear growth model. It was observed that at low precultural temperatures (2, 4 and 7 degrees C), a high proportion of L. monocytogenes cells were able to grow at 7 degrees C with almost no lag phase, consequently, the resulting distributions were positively skewed. Beside this, the variability observed was lower than at higher precultural temperatures. Regarding the precultural pH effect, at pH 7.4 the mean values of the lag phases were shorter at lower preincubation temperatures; while at pH 5.7 small pH transitions produced shorter individual lag phases at all precultural temperatures. The quantification of the effect of precultural conditions on the individual cell lag phase duration would improve the accuracy of the existing growth models, especially when a series of processing and storage steps are linked together in a process model or exposure assessment. Distributions will be fitted to the data for every set of conditions, generating useful tools for further risk assessment purposes.

Colony Count, Microbial↗

Effect of environmental parameters (temperature, pH and a(w)) on the individual cell lag phase and generation time of Listeria monocytogenes.

The effect of the individual environmental factors temperature (2-30 degrees C), pH (4.4-7.4) and a(w) (0.947-0.995) as well as the combinations of these factors on the individual cell lag phase and the generation time of Listeria monocytogenes was investigated. Individual cells were isolated using a serial dilution protocol in microtiter plates, and subsequent growth was investigated by optical density (OD) measurements at 600 nm. About 100 replicates were made for each set of environmental conditions. Part of the data were previously published in Francois et al. (Francois, K., Devlieghere, F., Smet, K., Standaert, A.R., Geeraerd, A.H., Van Impe, J.F., Debevere, J., 2005a. Modelling the individual cell lag phase: effect of temperature and pH on the individual cell lag distribution of Listeria monocytogenes. Int. J. Food Microbiol. 100, 41-53.), but were recalculated here using the calibration curves for transformation of optical density to colony forming units/ml from Francois et al. (Francois, K., Devlieghere, F., Standaert, A.R., Geeraerd, A.H., Cools, I., Van Impe, J.F., Debevere, J., 2005b. Environmental factors influencing the relationship between optical density and cell count for Listeria monocytogenes. J. Appl. Microbiol. 99, 1503-1515), as this calibration curve appeared to be dependent on the environmental parameters. The previous dataset was also extended with a factor a(w), observed individually and combinations with the above mentioned environmental factors. Individual cell lag phases and subsequent growth rates were calculated assuming an exponential growth model. The results are discussed as mean values to determine the general trends and in addition, histograms are made and statistical distributions are fitted to the different data sets. When stress levels increased, the mean values and the variability observed for the individual cell lag phases increased, resulting in broader histograms and distributions that were shifting to the right. Also the gravity point of the distributions was shifting from a skewed left type to a more symmetrical type. The best description of the data is obtained with an exponential distribution for low stress levels, a gamma distribution for intermediate stress and a Weibull distribution for severe stress levels. When only low stress levels were applied, a significant percentage of the cells showed no lag phase. In those cases, a new approach was used to obtain better fits: cells with a lag phase and those without a lag phase were separated using a binomial distribution while in a second step, a gamma or a Weibull distribution is fitted to the fraction of cells showing a lag phase. A normal distribution is used to describe the variability of the generation times. These distributions can be applied to refine the exposure assessment part of the risk assessment concerning L. monocytogenes by incorporating intercellular variability.

Colony Count, Microbial↗

Role of yeast proliferation in the quality degradation of strawberries during refrigerated storage.

Quality changes of strawberries during storage can be caused both by microbiological and physiological processes. There is little known about the possible contribution of microbiological processes to the quality degradation of strawberries. In this study, quality of strawberries during storage was evaluated by analytical and sensorial analyses. It was the aim to investigate the influence of microbiological activity on the changes of different quality factors of strawberries during storage. During storage at 7 degrees C, quality was mainly determined by the odor and by visual defects. Regarding the odor, highly microbiologically contaminated late-season strawberries packaged in air at 7 degrees C became sensorially unacceptable due to the presence of high amounts of ethyl acetate. This could be attributed to the yeast proliferation: at yeast concentrations above 5.0 log cfu/g, an increase in ethanol was detected in the headspace of the strawberries. It was shown that ethanol was converted to ethyl acetate by strawberries resulting in an unacceptable odor. In an experiment with low microbiologically contaminated early-season strawberries, not reaching the above mentioned yeast counts, less ethyl acetate was detected which resulted in strawberries that were sensorially acceptable during the whole storage period (12 days). Strawberries packaged in modified atmosphere conditions showed a different quality pattern due to the effect of decreased O2-concentrations on both microbiological and physiological processes. This paper demonstrates that also microbiological processes on strawberries should be considered as they could play an important role in the sensorial quality when interacting with physiological processes.

Acetates↗

Metabolite production of yeasts on a strawberry-agar during storage at 7 degrees C in air and low oxygen atmosphere.

Changes of different quality factors of strawberries have been described until now from a physiological point of view. Possible effects on quality caused by yeast proliferation have not been described. To elucidate the metabolic activity of yeasts (i.e. Debaryomyces melissophilus, Rhodotorula glutinis, Cryptococcus laurentii), isolated from strawberries, they were inoculated on a simulation medium of strawberries (strawberry-agar). Their activity was measured by analysing and correlating microbiological counts, metabolite concentration in the headspace as well as in the medium, and sugar consumption. The isolated yeasts from strawberries could grow on the strawberry-agar, both under air conditions and modified atmosphere (MA) conditions. The maximum count for the yeasts reached 7.5-8.5 log cfu cm(-2) (air conditions) and 5.6-6.4 log cfu cm(-2) (MA conditions). Production or consumption of a number of compounds could be detected when microbial counts reached levels between 4.7 log cfu cm(-2) and 8.5 log cfu cm(-2) depending on species and atmospheric conditions. A range of volatile organic compounds, produced by the yeasts, was detected: acetone, ethyl acetate, ethanol, isopropyl acetate, ethyl butyrate, 1-propanol, 2-methyl-1-propanol, 1-butanol, 2-methyl-1-butanol, 3-methyl-1-butanol, 1-hexanol and hexyl acetate. These compounds are able to influence the sensory properties of strawberries. A simultaneous decrease in sugar concentrations (sucrose, glucose, fructose) was observed. When ethanol reached high concentrations, ethyl acetate and ethyl butyrate were produced. This production can be attributed to a detoxification of ethanol by yeasts. The fermentative metabolism of yeasts during aerobic conditions could be explained by the Crabtree effect. As the detected volatile organic compounds produced by yeasts are also found in fresh strawberries, it can be concluded that these compounds are produced both by microbiological and physiological processes.

Colony Count, Microbial↗

Bacterial shell contamination in the egg collection chains of different housing systems for laying hens.

The bacterial eggshell contamination of eating eggs in different commercial housing systems; two conventional cages, one organic aviary system and one barn production, were compared. The total counts of aerobic bacteria and the total counts of Gram-negative bacteria on the shell were used to detect key points where contamination occurred and to study the progress of contamination in the egg collection and transportation chains. The key points in the chain were those where eggs accumulated on a short conveyor belt, initial shell contamination in the alternative housing systems and extra nest-boxes placed on the ground. The high bacterial load of floor eggs (>6.3 log CFU total aerobic flora/eggshell) explains why they cannot be used for eating. On average higher initial shell contamination with total counts of aerobic bacteria was found for eggs from the alternative housing systems compared to the conventional systems; respectively 5.46 compared to 5.08 log CFU/eggshell. However, initial contamination with total counts of Gram-negative bacteria on the shells was less in the alternative systems: 3.31 compared to 3.85 log CFU/shell. Initial bacterial shell contamination tended to correlate positively with the concentration of bacteria in the air of the poultry houses. Storing shell eggs, whether temporarily refrigerated or not, for 9 d or more, resulted in a decrease in bacterial eggshell contamination for both bacterial variables.

Air Microbiology↗

Single cell variability of L. monocytogenes grown on liver pâté and cooked ham at 7 degrees C: comparing challenge test data to predictive simulations.

AIMS: The variability in growth between individual Listeria monocytogenes cells was investigated on liver pâté and cooked ham. These results were compared to Monte Carlo simulations based on data collected previously in broths (Francois et al., submitted for publication). METHODS AND RESULTS: Single cells were isolated by a dilution protocol and inoculated on 15 g samples of liver pâté and cooked ham, pasteurized in the packaging. Of each product, 250 samples were inoculated, of which 50 samples were analysed for L. monocytogenes on each analysis day. Results were compared to simulations, based on distributions that describe the variability of the individual cell lag phases and generation times of L. monocytogenes cultivated in broths. Based on the same simulation techniques, the variability effect was investigated for different inoculum levels (10, 100, 10,00 and 10,000 cells). It was demonstrated that the expected variability of the outgrowth of L. monocytogenes in a challenge test is very high for low inoculum levels. CONCLUSIONS: The variability in growth characteristics observed between different single L. monocytogenes cells on foods is very large. The simulations based on the previously collected optical density data in broths, could be confirmed by foods inoculated with single L. monocytogenes cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The large variability between different individual L. monocytogenes cells has serious consequences for the experimental design of a challenge test. One thousand cells per portion are necessary in order to reduce the variability to acceptable levels and quantify the behaviour of the pathogen consistently with a reasonable number of challenge tests.

Animals↗

The effect of a commercial UV disinfection system on the bacterial load of shell eggs.

AIMS: To study the effect of UV irradiation on the bacterial load of shell eggs and of a roller conveyor belt. METHODS AND RESULTS: The natural bacterial load on the eggshell of clean eggs was significantly reduced by a standard UV treatment of 4.7 s; from 4.47 to 3.57 log CFU per eggshell. For very dirty eggs no significant reduction was observed. Eggs inoculated with Escherichia coli and Staphylococcus aureus (4.74 and 4.64 log CFU per eggshell respectively) passed the conveyor belt and were exposed to UV for 4.7 and 18.8 s. The reduction of both inoculated bacteria on the eggshell was comparable and significant for both exposure times (3 and 4 log CFU per eggshell). Escherichia coli was reduced but still detectable on the conveyor rollers. The internal bacterial contamination of eggs filled up with diluent containing E. coli or S. aureus was not influenced by UV irradiation. CONCLUSIONS: There is a significant lethal effect of UV irradiation on the bacterial contamination of clean eggshells and recent shell contamination, contamination of rollers can be controlled and the internal contamination of eggs is not reduced. SIGNIFICANCE AND IMPACT OF THE STUDY: The penetration of UV into organic material appears to be poor and UV disinfection can be used as an alternative for egg washing.

Animals↗

Risk assessment of Listeria monocytogenes: impact of individual cell variability on the exposure assessment step.

Recently, the lag phase research in predictive microbiology is focusing more on the individual cell variability, especially for pathogenic microorganisms that typically occur in very low contamination levels, like Listeria monocytogenes. In this study, the effect of this individual cell lag phase variability was introduced in an exposure assessment study for L. monocytogenes in a liver pâté. A basic framework was designed to estimate the contamination level of pâté at the time of consumption, taking into account the frequency of contamination and the initial contamination levels of pâté at retail. Growth was calculated on pâté units of 150 g, comparing an individual-based approach with a classical population-based approach. The two different protocols were compared using simulations. If only the individual cell lag variability was taken into account, important differences were observed in cell density at the time of consumption between the individual-based approach and the classical approach, especially at low inoculum levels, resulting in high variability when using the individual-based approach. Although, when all variable factors were taken into account, no significant differences were observed between the different approaches, allowing the conclusion that the individual cell lag phase variability was overruled by the global variability of the exposure assessment framework. Even in more extreme conditions like a low inoculum level or a low water activity, no differences were created in cell density at the time of consumption between the individual-based approach and the classical approach. This means that the individual cell lag phase variability of L. monocytogenes has important consequences when studying specific growth cases, especially when the applied inoculum levels are low, but when performing more general exposure assessment studies, the variability between the individual cell lag phases is too limited to have a major impact on the total exposure assessment.

Animals↗