Prolongation of the shelf-life of minimally processed vegetables by mild treatments.
Explore the source record for details and available documents.
Biomedical subjects
Publications and source records attributed to J Debevere.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
This study investigated the usefulness of two selected lactic acid bacteria, Lactobacillus sakei subsp. carnosus (10A) and the lactocin S producing L. sakei 148 (LS5), to extend the shelf-life of cooked meat products. The interaction between these potential protective cultures and the spoilage organisms, Leuconostoc mesenteroides (LM4) and Brochothrix thermosphacta (BT1), were examined in co-culture studies on a model cooked ham product at 7 degrees C under vacuum packaged conditions. Furthermore, the influence of the glucose content of the model cooked ham on the interaction phenomena was investigated. When artificially contaminating the model cooked ham with BT1 at 10(2) cfu/g in combination with 10A at 10(5) cfu/g, the growth of BT1 was significantly slower compared to a simultaneous mono-culture experiment. In a similar experiment with LM4, LM4 reached a level of 10(7) cfu/g +/-14 days later when LM4 grew together with 10A compared to its growth in mono-culture. The lactocin S producing LS5 did not demonstrate an inhibitory action towards LM4 or BT1 and is therefore not useful as protective culture on cooked meat products. The glucose level of the model cooked ham had no influence on the observed antagonistic interactions of 10A towards LM4 or BT1, indicating that the action of the biopreservative 10A in cooked meat products is independent of the substrate glucose.
Two lactic acid bacteria, Lactobacillus sakei subsp. carnosus (10A) and lactocin S producing Lactobacillus sakei 148 (LS5), were examined for their usefulness as protective culture in the biopreservation of cooked meat products. Co-culture experiments on a model cooked ham (MCH) between 10A or LS5 and a cocktail of three Listeria monocytogenes strains were performed to examine the influence of inoculum level (10(5) vs. 10(6)cfu/g), storage temperature (4 vs. 7 degrees C) and packaging type (vacuum-packaging vs. modified atmosphere-packaging). At 7 degrees C, applying Lactobacillus sakei 10A at 10(6) cfu/g limited the growth of Listeria monocytogenes to <1 log(10) cfu/g during 27 days, whilst an application level of 10(5) cfu/g failed to prevent growth to unacceptable levels. Lactobacillus sakei LS5 did not demonstrate an antagonistic effect towards Listeria monocytogenes. Lowering the temperature to 4 degrees C or switching from vacuum-packaging to modified atmosphere packaging (MAP) did not influence the ability of strain 10A to grow on the MCH, as its dominance did not change. A combination of strain 10A and 4 degrees C or a MAP containing 50% CO(2) completely inhibited the growth of Listeria monocytogenes. Sensory assessments and pH measurements confirmed that 10A, even when present at a high level for prolonged storage times, did not acidify the cooked ham to a point of sensory rejection.
N-butanoyl-L-homoserine lactone (C4-HSL) production by Aeromonas hydrophila 519 has been established. C4-HSL production at 22 and 30 degrees C was found after 20-24 h of incubation corresponding to a population density of ca. 10(8)-10(9) cfu/ml, respectively. Reduced C4-HSL production was noted after 72 h of incubation at 12 degrees C when the culture reached ca. 10(9) cfu/ml. No C4-HSL production was detected at 37 degrees C, although a dense population was obtained. In LB broth supplemented with 0.1% and 0.5% glucose, C4-HSL production was noted whereas with 1% glucose no C4-HSL was detected although a high colony count was obtained. In the latter culture residual levels of glucose (0.65%) were found after 43 h whereas in the 0.1% and 0.5% supplemented LB, glucose was quickly consumed which may have stimulated C4-HSL production. In conclusion, the present study shows an effect of environmental conditions (temperature, glucose concentration) on the C4-HSL production and warrants further investigation to elucidate the effect of external conditions on production of AHL signal molecules to reveal the relevance of quorum sensing in, e.g. foods.
Intense light pulses (ILP) is a new method intended for decontamination of food surfaces by killing microorganisms using short time high frequency pulses of an intense broad spectrum, rich in UV-C light. This work studied in a first step the effect of food components on the killing efficiency of ILP. In a second step, the decontamination of eight minimally processed (MP) vegetables by ILP was evaluated, and thirdly, the effect of this treatment on the shelf-life of MP cabbage and lettuce stored at 7 degrees C in equilibrium modified atmosphere packages was assessed by monitoring headspace gas concentrations, microbial populations and sensory attributes. Proteins and oil decreased the decontamination effect of ILP, whilst carbohydrates and water showed variable results depending on the microorganism. For this reason, high protein and fat containing food products have little potential to be efficiently treated by ILP. Vegetables, on the other hand, do not contain high concentrations of both compounds and could therefore be suitable for ILP treatment. For the eight tested MP vegetables, log reductions up to 2.04 were achieved on aerobic mesophilic counts. For the shelf-life studies, respiration rates at 3% O2 and 7 degrees C were 14.63, 17.89, 9.17 and 16.83 ml O2/h kg produce for control and treated cabbage, and control and treated lettuce respectively; used packaging configurations prevented anoxic conditions during the storage times. Log reductions of 0.54 and 0.46 for aerobic psychrothrophic count (APC) were achieved after flashing MP cabbage and lettuce respectively. APC of treated cabbage became equal than that from control at day 2, and higher at day 7, when the tolerance limit (8 log) was reached and the panel detected the presence of unacceptable levels of off-odours. Control never reached 8 log in APC and were sensory acceptable until the end of the experiment (9 days). In MP lettuce, APC of controls reached rejectable levels at day 2, whilst that of treated samples did after 3 days. Both samples were sensory unacceptable at day 3, controls because of bad overall visual quality (OVQ), off-odour and leaf edge browning and treated samples due to bad OVQ; browning inhibitors might be proposed to preserve OVQ. Yeasts and lactic acid bacteria counts were low in all the samples. It seems that ILP treatment alone under the conditions used in this work does not increase MP vegetables shelf-life in spite of the reduction in the initial microbial load.
Solid phase cytometry (SPC) in conjunction with fluorescent viability staining has been investigated as a tool to detect viable but non-culturable Campylobacter jejuni in drinking water. Inoculated water samples were filtered over a polyester membrane filter and the retained cells were stained using a carboxyfluorescein ester as a substrate for intracellular esterases. The number of green fluorescent bacteria was automatically counted by an Ar laser scanning device (ChemScan) in 3 min. In parallel, the plate count was determined on Columbia Blood Agar. The number of culturable cells decreased below the detection limit of plate counting in less than 50 days. In contrast, the number of fluorescent bacteria remained at its initial level for at least 85 days. The discrepancy between the two results can be attributed to the transition of culturable C. jejuni cells into VBNC C. jejuni cells. Furthermore, as SPC can distinguish between low numbers of dividing and non-dividing cells of Campylobacter it has the potential to monitor attempts to resuscitate VBNC cells.
The influence of housing system on the initial bacterial contamination of the eggshell was studied. Two long-term experiments were performed. Bacterial eggshell contamination, as expressed by total count of aerobic and Gram-negative bacteria, was periodically analysed for eggs from a conventional cage, a furnished cage with nest boxes containing artificial turf or grids as nest-floor material and an aviary housing system. Results were log-transformed prior to statistical analyses. For both experiments no systematic differences were found between the conventional cage and furnished cage. The type of nest-floor material in the nest boxes of the furnished cages also did not systematically influence the bacterial contamination. A possible seasonal influence on contamination with a decrease in the winter period (up to > 0.5 log cfu/eggshell) of total count of aerobic and Gram-negative bacteria was observed in the first experiment. The contamination with total aerobic flora was higher (more than 1.0 log) on eggs from the aviary housing system compared to the conventional and the furnished cage systems. For Gram-negative bacteria this was not the case. During the entire period of both experiments, independent of housing system, shell contamination was not influenced by age of hens or period since placing the birds in the houses. For the total count of aerobic bacteria a restricted positive correlation (r2 = 0.66) was found between the concentration of total bacteria in the air of the poultry houses and initial shell contamination.
AIMS: Understanding spoilage caused by different types of spoilage organisms, associated with vacuum-packaged sliced cooked meat products (CMP). METHODS AND RESULTS: First, strains were characterized in a broth at 7 degrees C under anaerobic conditions to compare their growth rate, acidifying character and metabolite production under conditions simulating refrigerated vacuum-packaged conditions. Brochotrix thermosphacta grew faster than the lactic acid bacteria (LAB). Within the group of the LAB, all strains grew fast except Leuconostoc mesenteroides subsp. dextranicum and Leuconostoc carnosum. Secondly, the organisms were inoculated on a model cooked ham to better understand the relationship between spoilage, microbial growth, pH, metabolite production and accompanying sensory changes. Most rapidly growing strains were Leuc. mesenteroides subsp. mesenteroides followed by B. thermosphacta, while Leuc. mesenteroides subsp. dextranicum and Leuc. carnosum grew very slowly compared with the other LAB. Brochotrix thermosphacta caused sensory deviations at a lower cell number compared with the LAB. The related pH changes, metabolite production and sensory perception are presented. CONCLUSIONS: In this pure culture study, B. thermosphacta and Leuc. mesenteroides subsp. mesenteroides had the highest potential to cause rapid spoilage on CMP. SIGNIFICANCE AND IMPACT OF THE STUDY: A systematic study on the behaviour of spoilage organisms on a model cooked ham to establish the relationship between microbial growth, pH, metabolite formation and organoleptic deviations.
AIM: To determine the influence of several factors on the inactivation of micro-organisms by intense light pulses (ILP). METHODS AND RESULTS: Micro-organisms on agar media were flashed 50 times under different conditions and their inactivation measured. Micro-organisms differed in sensitivity to ILP but no pattern was observed among different groups. Several enumeration methods to quantify the effect of ILP were investigated and showed relevant differences, shading effect and photoreactivation accounted for them, the strike method yielded the most reliable results. Higher decontamination efficiencies were obtained for Petri dishes located close to the strobe and inside the illumination cone. Decontamination efficacy decreased significantly at contamination levels >6.85 log(10). After 13 successive treatments, no resistance to ILP could be demonstrated. Media warming up depended on the distance from the strobe and the number of flashes. CONCLUSIONS: For an industrial implementation: the position and orientation of strobes in an unit will determine the lethality, products should be flashed as soon as possible after contamination occurs, a cooling system should be used for heat-sensitive products and flashed products should be light protected. No resistant flora is expected to develop. SIGNIFICANCE AND IMPACT OF THE STUDY: Conclusions derived from this work will allow a better implementation of this decontamination technique at industrial level.
AIMS: The effect of temperature (2-30 degrees C), pH (4.8-7.4) and water activity (0.946-0.995) on the relationship between optical density (OD) at 600 nm and the plate count (CFU ml(-1)) was investigated for Listeria monocytogenes. METHODS AND RESULTS: Calibration curves, relating OD with plate counts, were collected by measuring the OD of consecutive one-half dilution series, before determining the cell density by classic plate count methods. The calibration curves were observed to be shifting in a parallel way, with increasing stress levels. Especially pH influenced the curve in a great extent, while the other variables were showing more synergetic effects. The reason for the shift was investigated by a microscopic viability test, showing a viability decrease with increasing stress levels, causing the shift of the calibration curve. In a last step a model was made describing the effect of environmental factors on the calibration curve, with different data transformations being tested. A polynomial equation was fitted to the data, taking into account a set of constraints to incorporate microbiological knowledge in the black box model. Hence, illogical interpolation results and overfitting of the data could be avoided. CONCLUSIONS: Different stress factors are affecting the relationship between the OD and the cell count of L. monocytogenes by lowering the cell viability. These effects could be modelled using a constrained polynomial model. SIGNIFICANCE AND IMPACT OF THE STUDY: The observed phenomena are important when calculating growth parameters, like growth rate and lag phase, based on OD data.
AIMS: The objectives of the study were to determine the spread and persistence of Campylobacter in a poultry processing plant and to provide a quantitative estimate of the survival of Campylobacter jejuni on the surface of a cutting board. METHODS AND RESULTS: Several contact surfaces in a poultry processing plant were sampled before the start of processing, after 30 min and after 120 min. Next, the survival of four C. jejuni strains was studied on a beech and polypropylene cutting board during 120 min. CONCLUSIONS: A rapid introduction and spread of Campylobacter in a well cleaned processing plant as well as a significant survival in time on the example of a cutting board is shown. SIGNIFICANCE AND IMPACT OF THE STUDY: The need to prevent cross-contamination in the food processing and preparation area and the importance of an integrated approach throughout the whole food chain to control transmission of Campylobacter is highlighted.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
The research presented in this paper analyses a newly developed experimental protocol for isolating single cells by constructing a simulation model of the process. The protocol involves sequential 50% dilutions of a cell suspension in a microtiter plate, so that eventually, wells are obtained containing exactly one cell. The aim of this modelling study is (i) to gain insight in the governing mechanisms of the dilution process, (ii) to confirm experimental findings and (iii) to enable the prediction of an average outcome for future experiments. The model construction process is presented chronologically. The initial basic model simulates the experiment as a sequence of binomial processes, using Monte Carlo techniques. Statistical analysis of the results shows that aggregational factors need to be taken into account in the form of a lognormal distribution. Several issues involved in this adaptation are discussed. To fully account for cell aggregation in the dilution process, a cell clumping algorithm is built into the simulation model. Simulation data from the resulting model show similar statistical characteristics as the experimental data and yield reliable prediction intervals for the available experimental data. The simulation model is a useful tool to support experimental findings and predict the outcome of future experiments. Even more importantly, this study emphasises the importance of careful statistical analysis in single cell research. The impact of stochastic effects is considerably amplified at the low cell concentrations involved and needs to be taken into account in any modelling effort.
The individual-based approach of the lag phase is gaining interest, especially for pathogens that initially contaminate food products in low amounts. In this paper, the effect of temperature (30, 10, 7, 4 and 2 degrees C) and pH (7.4, 6.1, 5.5, 5.0, 4.7 and 4.4) on the individual cell lag phase of Listeria monocytogenes was examined in a factorial design, using OD measurements. Individual lag phases of about 100 individual cells per condition were examined and calculated using a linear extrapolation method. Generation times were calculated out of the slope. The obtained data were analyzed at three different levels: in a first approach, the mean values were calculated for each set of environmental conditions and compared to predictions made by the USDA's Pathogen Modeling Program (PMP) for analogous growth conditions. The PMP predictions of the generation times were in the same order of magnitude as the obtained data, although a persistent underestimation could be observed. The observed individual cell lag data differed from lag phase predictions by PMP. Possible reasons for this discrepancy are discussed. Secondly, histograms of individual lag phase measurements were constructed for the different temperature-pH combinations. In this way, the influence of both factors on the variability of individual lag phases could be estimated. At low stress levels, most individual cells showed a short lag phase resulting in a compression of the histograms at the zero-lag level, while, at high stress levels, the histograms shifted to longer lag phases with a significant increase in variability. Thirdly, 37 different distribution types were fitted to the datasets to reveal the distributions that fitted best the obtained data. The gamma distribution was preferred at moderate stress levels, while the Weibull distribution was chosen for harsher growth conditions.
In this study, 91 strains, originating from meat products, were subjected to a step-by-step screening and characterisation to search for potential protective cultures to be used in the cooked cured meat industry. Strains were first tested on their homofermentative and psychrotrophic character and salt tolerance. Secondly, the antibacterial capacities towards Listeria monocytogenes, Leuconostoc mesenteroides, Leuconostoc carnosum and Brochotrix thermosphacta were determined in an agar spot test. In total, 38% of the tested strains were inhibitory towards all indicator strains. However, 91%, 88% and 74% of the strains could inhibit, respectively, L. monocytogenes, B. thermosphacta and Leuc. mesenteroides. Finally, 12 strains, with the highest antibacterial capacities, were evaluated on their competitive nature by comparing their growth rate, acidifying character and lactic acid production at 7 degrees C under anaerobic conditions in a liquid broth. All 12 strains, except for a bacteriocin producing Lactobacillus plantarum strain and the lactocin S producing Lactobacillus sakei 148, combined a fast growth rate with a deep and rapid acidification caused by the production of high levels of lactic acid. The 12 selected strains were then further investigated for their growth capacity on a model cooked ham product to establish whether the presence of these cultures on the ham did not negatively influence the sensory properties of the ham. All strains grew in 6 days at 7 degrees C from a level of 10(5)-10(6) to 10(7)-10(8) cfu/g and again the bacteriocin producing L. plantarum strain was the slowest growing strain. As the glucose level of the model cooked ham product was low (0.09+/-0.03%), growth of the putative protective cultures resulted in glucose depletion and a limited lactic acid production and accompanying pH decrease. Cooked ham inoculated with isolates 13E, 10A, 14A (all three identified as L. sakei subsp. carnosus by SDS-PAGE) and with strains L. sakei 148 (LS5) and L. sakei subsp. carnosus SAGA 777 (LS8) were not rejected by the sensory panel at the 34th day of the vacuum packaged storage at 7 degrees C. Therefore, these strains could have potential for the use as protective culture in cooked meat products.