J.F. Van Impe

Concentration of carbon dioxide in the water-phase as a parameter to model the effect of a modified atmosphere on microorganisms

The effect of modified atmosphere packaging can mainly be attributed to the bacteriostatic action of CO2. The dissolved CO2 in the water-phase of a food product is strongly dependent on several intrinsic and extrinsic parameters and will determine the effectiveness of a modified atmosphere packaging configuration. The effect of pH, gas/product ratio, initial %CO2 in the gas-phase, lard content and storage temperature on the amount of dissolved CO2 was screened in a preliminary experiment. The initial CO2-concentration in the gas-phase and the gas/product ratio turned out to be the two major factors determining the amount of dissolved CO2. The initial pH also determined significantly the final CO2-concentration in the broth. Temperature and lard content were shown to have only a minor effect on the amount of dissolved CO2 compared to the above mentioned parameters. This demonstrates the importance of the packaging configuration in the effectiveness of a modified atmosphere. In a second step, a model was constructed to predict the amount of dissolved carbon dioxide in modified BHI-broth as a function of the gas/product ratio, the initial CO2-concentration and the temperature by means of Response Surface Methodology (RSM). A second equation was also derived based on Henrys law and was shown to be a powerful tool in the quantification of the effect of intrinsic and extrinsic parameters on the CO2-solubility in food products. The possibility of the use of the concentration of dissolved CO2 in the water-phase as a determinative factor for the inhibitory effect of modified atmospheres was examined on Pseudomonas fluorescens. Growth curves at 7 degrees C of P. fluorescens in different packaging configurations (initial %CO2 and gas/product ratio) resulting in equal amounts of dissolved CO2 were compared. P. fluorescens was shown to be similarly inhibited by equal amounts of dissolved CO2-concentrations, independent of the packaging configuration. This demonstrates the potential of the application of the concentration of dissolved CO2 in the water-phase as a parameter to characterise a modified atmosphere and its inhibition of certain microorganisms.

Using survival analysis to investigate the effect of UV-C and heat treatment on storage rot of strawberry and sweet cherry.

Ultraviolet light and heat treatment are proposed as alternative techniques for the use of chemicals to reduce the development of the spoilage fungi Botrytis cinerea and Monilinia fructigena on strawberry and sweet cherry, respectively, during storage. In order to investigate the effect of both physical techniques on microbial inactivation and on fruit quality, inoculated berries were subjected to different temperatures (40-48 degrees C) and UV-C doses (0.05-1.50 J/cm2). For each condition, 20 berries were used. After the treatment, fungal growth, visual damage (holes, stains) and fruit firmness were evaluated during a period of 10 days. The experimental data were analysed statistically using survival analysis techniques. Fungal growth on strawberries was significantly retarded using UV-C doses of 0.05 J/cm2 and higher. The same treatment had no significant effect when applied to cherries. The highest doses (1.00 and 1.50 J/cm2) had a negative effect on the calyx of the strawberry, causing browning and drying of the leaves. No beneficial effect of a low temperature treatment (40-48 degrees C) on the shelf life of strawberries was observed, but fungal development on cherries was retarded at temperatures of 45 and 48 degrees C. These temperatures caused severe damage on strawberries (soft stains, holes, decreased firmness), but had no influence on the quality of sweet cherries.

Effect of dissolved carbon dioxide and temperature on the growth of Lactobacillus sake in modified atmospheres

Carbon dioxide is known to be the active gas in modified atmosphere packaging. However, only the dissolved part of the CO2 will be active in the inhibition of microorganisms. When the effect of a modified atmosphere on the growth of microorganisms is modeled, the amount of dissolved CO2 should be the parameter to evaluate the effect of CO2 in modified atmospheres. Lactobacillus sake was isolated from cooked ham, identified and was chosen as a representative organism for the spoilage of modified atmosphere packed cooked meat products. In a first step a suitable liquid simulation medium for cooked meat products was selected. The growth rate of L. sake was significantly higher in modified MRS than in cooked ham, while no significant difference was observed between the growth parameters of L. sake in modified BHI and in cooked ham. Careful evaluation of the applied simulation medium was shown to be essential before application to the development of predictive models for nutrient demanding microorganisms such as lactic acid bacteria. In a second step, the influence of dissolved CO2 and the storage temperature on the growth parameters of L. sake was modeled. Temperature was shown to have a major effect on the growth rate and lag phase of L. sake. Carbon dioxide negatively influenced the growth rate of L. sake. A negative, linear relationship was observed between the growth rate and the amount of dissolved carbon dioxide. Carbon dioxide was shown, however, to have no significant effect on the lag phase of L. sake. A significant interaction term between temperature and dissolved CO2 on the growth rate of L. sake was established.

Combinations of pulsed white light and UV-C or mild heat treatment to inactivate conidia of Botrytis cinerea and Monilia fructigena.

The use of pulses of intense white light to inactivate conidia of the fungi Botrytis cinerea and Monilia fructigena, responsible for important economical losses during postharvest storage and transport of strawberries and sweet cherries, was investigated in this study. In the first stage, a light treatment applying pulses of 30 micros at a frequency of 15 Hz was investigated, resulting in a treatment duration varying from 1 to 250 s. The conidia of both fungi showed similar behaviour to pulsed light, with a maximal inactivation of 3 and 4 log units for B. cinerea and M. fructigena, respectively. The inactivation of the conidia increased with increasing treatment intensity, but no complete inactivation was achieved. The sigmoidal inactivation pattern obtained by the pulsed light treatment was described using a modification of the model of Geeraerd et al. [Int. J. Food Microbiol. 59 (2000) 185]. Hereto, the shoulder length was incorporated explicitly and relative values for the microbial populations were used. In the second stage, combinations of light pulses and ultraviolet-C or heat were applied. The UV light used in the experiments is the short-wave band or UV-C, running from 180 to 280 nm with a peak at 254 nm (UV-B runs from 280 to 320 nm and UV-A from 320 to 380 nm). The UV-C doses were 0.025, 0.05 and 0.10 J/cm(2), and the temperatures for the thermal treatment ranged from 35 to 45 degrees C during 3-15 min. When combining UV-C and light pulses, there was an increase in inactivation for both B. cinerea and M. fructigena, and synergism was observed. There was no effect of the order of the treatments. For the heat-light pulses combination, there was a difference between both fungi. The order of the treatments was highly significant for B. cinerea, but not for M. fructigena. Combining heat and light treatments improved the inactivation, and synergism between both methods was again observed. Complete inactivation of M. fructigena conidia was obtained after, e.g., a 40-s pulsed light treatment and 15 min at 41 degrees C, or after an 80-s light treatment and 10 min at 41 degrees C.

Application of artificial neural networks as a non-linear modular modeling technique to describe bacterial growth in chilled food products

In many chilled, prepared food products, the effects of temperature, pH and %NaCl on microbial activity interact and this should be taken into account. A grey box model for prediction of microbial growth is developed. The time dependence is modeled by a Gompertz model-based, non-linear differential equation. The influence of temperature, pH and %NaCl reflected in the model parameters is described by using low-complexity, black box artificial neural networks (ANNs). The use of this non-linear modeling technique makes it possible to describe more accurately interacting effects of environmental factors when compared with classical predictive microbiology models. When experimental results on the influence of other environmental factors become available, the ANN models can be extended simply by adding more neurons and/or layers.

Prevalence and challenge tests of Listeria monocytogenes in Belgian produced and retailed mayonnaise-based deli-salads, cooked meat products and smoked fish between 2005 and 2007

Processed ready-to-eat (RTE) foods with a prolonged shelf-life under refrigeration are at risk products for listeriosis. This manuscript provides an overview of prevalence data (n=1974) and challenge tests (n=299) related to Listeria monocytogenes for three categories of RTE food i) mayonnaise-based deli-salads (1187 presence/absence tests and 182 challenge tests), ii) cooked meat products (639 presence/absence tests and 92 challenge tests), and iii) smoked fish (90 presence/absence tests and 25 challenge tests), based on data records obtained from various food business operators in Belgium in the frame of the validation and verification of their HACCP plans over the period 2005-2007. Overall, the prevalence of L. monocytogenes in these RTE foods in the present study was lower compared to former studies in Belgium. For mayonnaise-based deli-salads, in 80 out of 1187 samples (6.7%) the pathogen was detected in 25 g. L. monocytogenes positive samples were often associated with smoked fish deli-salads. Cooked meat products showed a 1.1% (n=639) prevalence of the pathogen. For both food categories, numbers per gram never exceeded 100 CFU. L. monocytogenes was detected in 27.8% (25/90) smoked fish samples, while 4/25 positive samples failed to comply to the 100 CFU/g limit set out in EU Regulation 2073/2005. Challenge testing showed growth potential in 18/182 (9.9%) deli-salads and 61/92 (66%) cooked meat products. Nevertheless, both for deli-salads and cooked meat products, appropriate product formulation and storage conditions based upon hurdle technology could guarantee no growth of L. monocytogenes throughout the shelf-life as specified by the food business operator. Challenge testing of smoked fish showed growth of L. monocytogenes in 12/25 samples stored for 3-4 weeks at 4 degrees C. Of 45 (non-inoculated) smoked fish samples (13 of which were initially positive in 25 g) which were subjected to shelf-life testing, numbers exceeded 100 CFU/g in only one sample after storage until the end of shelf-life. Predictive models, dedicated to and validated for a particular food category, taking into account the inhibitory effect of various factors in hurdle technology, provided predictions of growth potential of L. monocytogenes corresponding to observed growth in challenge testing. Based on the combined prevalence data and growth potential, mayonnaise-based deli-salads and cooked meat products can be classified as intermediate risk foods, smoked fish as a high risk food.

Nonlinear and Adaptive Control in Biotechnology: A Tutorial

Dynamics of biological reactors. Kinetic modelling and identification. State observers as software sensors. Control of fed-batch biological reactors. Feedback control of continuous biological reactors.

Shelf life of modified atmosphere packed cooked meat products: addition of Na-lactate as a fourth shelf life determinative factor in a model and product validation.

Cooked meat products are often post-contaminated because of a packaging and/or slicing step after the pasteurisation process. The shelf life is therefore limited and can be extended by adding Na-lactate. A previously developed model for the spoilage of gas packed cooked meat products, including temperature, water activity and dissolved CO2 as independent variables, was extended with a fourth factor: the Na-lactate concentration in the aqueous phase of the meat product. Models were developed for the maximum specific growth rate mu(max) and the lag phase lambda of the specific spoilage organism Lactobacillus sake subsp. carnosum. Quadratic response surface equations were compared with extended Ratkowsky models. In general, response surface equations fitted the experimental data best but in the case of mu(max) the response surface model predicted illogical growth behaviour at low water activities and high Na-lactate concentrations. A extensive product validation of the mathematical models was performed by means of inoculated as well as naturally contaminated industrially prepared cooked meat products. The deviations of the experimentally determined versus predicted growth parameters in inoculated cooked meat products were in general small. Both types of models were also able to predict the shelf life of naturally contaminated cooked meat products, except for pâté where an under-estimation of the shelf life was predicted by the response surface equations. The validation studies revealed higher accuracy of the extended Ratkowsky models in comparison to the response surface equations. A significant shelf life extending effect of Na-lactate was predicted, which was more pronounced at low refrigerated temperatures. A synergistic effect has also been noticed between Na-lactate and carbon dioxide which, at least partly, could be explained by the pH-decreasing effect of CO2.

Monte Carlo analysis as a tool to incorporate variation on experimental data in predictive microbiology

Until now, most of the mathematical models used in predictive microbiology are deterministic, i.e. their outcome is a point estimate for the microbial load at a certain time instant. For more advanced exploitation of predictive microbiology in the context of hazard analysis and critical control points and risk analysis studies, stochastic models should be developed. Such models predict a probability mass function for the microbial load at a certain time instant. The objective of this paper is to illustrate methodologically how to generate, starting from the experimental observations and a deterministic growth model, probability density functions for (i) the model parameters and (ii) the predictions as a function of time, by using Monte Carlo analysis. A normal distribution over the experimental data was considered. This probabilistic approach, incorporating experimental variation, is applied to experimental growth data of Escherichia coli K12 and Listeria innocua ATCC 33090.

Modelling the individual cell lag phase. Isolating single cells: protocol development.

Aims: To develop a protocol to isolate single cells in wells of a microtitre plate, having a high certainty of individual cells, combined with a sufficient yield.