Bayesian global regression model relating product characteristics of intermediate moisture food products to heat inactivation parameters for Salmonella Napoli and Eurotium herbariorum mould spores.

Thermal inactivation of pathogenic and spoilage organisms in low and intermediate moisture foods is of critical importance for guaranteeing microbiological safety and stability of these products. Producers tendentially reduce salt in low and intermediate moisture foods because of nutritional health considerations, but it is unclear how this affects microbial inactivation rates during pasteurization. In this study we predict the time to achieve a pre-defined 6-log reduction for Salmonella enterica subsp. enterica serovar Napoli (hereafter: S. Napoli) and Eurotium herbariorum mould spores (hereafter: E. herbariorum spores) and the relationship with product characteristics. We tested 31 design products for heat inactivation of S. Napoli and 29 design products for heat inactivation of E. herbariorum spores. We used a global Bayesian regression combining primary Weibull models with a secondary regression model to relate pasteurization temperature and product characteristics (water activity (aw), pH, and fractions of sodium chloride, sucrose and oil on product) to microbial counts. With this model, we predict the time to 6-log reduction. Thermal inactivation rates were much higher for vegetative S. Napoli than for E. herbariorum spores. Also, inactivation curves were non-linear for many experiments. There were significant associations between the Weibull model parameters and temperature, and pH and aw for S. Napoli and E. herbariorum spores, respectively. We parameterized an inactivation model for S. Napoli and E. herbariorum spores using design products with a broad range of characteristics and showed how the simplified approach of using D-values does not accurately describe the non-linearity of thermal inactivation for both types of organism. Results of our model can be used to produce accurate heat inactivation predictions as input for the pasteurization process in factories where intermediate moisture foods are manufactured.

Mode of antimicrobial action of vanillin against Escherichia coli, Lactobacillus plantarum and Listeria innocua.

AIMS: To investigate the mode of action of vanillin, the principle flavour component of vanilla, with regard to its antimicrobial activity against Escherichia coli, Lactobacillus plantarum and Listeria innocua. METHODS AND RESULTS: In laboratory media, MICs of 15, 75 and 35 mmol l(-1) vanillin were established for E. coli, Lact. plantarum and L. innocua, respectively. The observed inhibition was found to be bacteriostatic. Exposure to 10-40 mmol l(-1) vanillin inhibited respiration of E. coli and L. innocua. Addition of 50-70 mmol l(-1) vanillin to bacterial cell suspensions of the three organisms led to an increase in the uptake of the nucleic acid stain propidium iodide; however a significant proportion of cells still remained unstained indicating their cytoplasmic membranes were largely intact. Exposure to 50 mmol l(-1) vanillin completely dissipated potassium ion gradients in cultures of Lact. plantarum within 40 min, while partial potassium gradients remained in cultures of E. coli and L. innocua. Furthermore, the addition of 100 mmol l(-1) vanillin to cultures of Lact. plantarum resulted in the loss of pH homeostasis. However, intracellular ATP pools were largely unaffected in E. coli and L. innocua cultures upon exposure to 50 mmol l(-1) vanillin, while ATP production was stimulated in Lact. plantarum cultures. In contrast to the more potent activity of carvacrol, a well studied phenolic flavour compound, the extent of membrane damage caused by vanillin is less severe. CONCLUSIONS: Vanillin is primarily a membrane-active compound, resulting in the dissipation of ion gradients and the inhibition of respiration, the extent to which is species-specific. These effects initially do not halt the production of ATP. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding the mode of action of natural antimicrobials may facilitate their application as natural food preservatives, particularly for their potential use in preservation systems employing multiple hurdles.

Debating the biological reality of modelling preservation.

Predictive food microbiology is a rapidly developing science and has made great advances. The aim is to debate a number of issues in modelling preservation: (1) inoculum and prehistory effects on lag times and process susceptibility; (2) mechanistic vs. empirical modelling; and (3) concluding remarks (the Species concept, methodology and biovariability). Increasing the awareness in these issues may bridge the gap between the complex reality in food microbial physiology and the application potential of predictive models. The challenge of bringing integrated preservation or risk analysis further and developing ways to truly model and link biological susceptibility distributions from raw ingredients via process survival to outgrowth probabilities in the final product remains.

Membrane permeabilization in relation to inactivation kinetics of Lactobacillus species due to pulsed electric fields.

Membrane permeabilization due to pulsed electric field (PEF) treatment of gram-positive Lactobacillus cells was investigated by using propidium iodide uptake and single-cell analysis with flow cytometry. Electric field strength, energy input, treatment time, and growth phase affected membrane permeabilization of Lactobacillus plantarum during PEF treatment. A correlation between PEF inactivation and membrane permeabilization of L. plantarum cells was demonstrated, whereas no relationship was observed between membrane permeabilization and heat inactivation. The same results were obtained with a Lactobacillus fermentum strain, but the latter organism was more PEF resistant and exhibited less membrane permeabilization, indicating that various bacteria have different responses to PEF treatment. While membrane permeabilization was the main factor involved in the mechanism of inactivation, the growth phase and the acidity of the environment also influenced inactivation. By using flow cytometry it was possible to sort cells in the L. plantarum population based on different cell sizes and shapes, and the results were confirmed by image analysis. An apparent effect of morphology on membrane permeabilization was observed, and larger cells were more easily permeabilized than smaller cells. In conclusion, our results indicate that the ability of PEF treatment to cause membrane permeabilization is an important factor in determining inactivation. This finding should have an effect on the final choice of the processing parameters used so that all microorganisms can be inactivated and, consequently, on the use of PEF treatment as an alternative method for preserving food products.

Synergistic antibacterial action of heat in combination with nisin and magainin II amide.

Lactobacillus plantarum has been exposed to mild heat at temperatures between 48 and 56 degrees C in combination with low concentrations of the lantobiotic nisin in different sequential set-ups. Exposure to heat and nisin caused synergistic reductions of Lact. Plantarum viability. Efficient antimicrobial action was dependent on the growth state of the culture as well as on levels and sequences of treatment applications. Listeria monocytogenes and Escherichia coli were treated at 55 degrees C in the presence of magainin II amide. Synergistic reductions in viable counts could be observed for L. monocytogenes and, after prolonged exposure, also for E. coli. the bacterial membrane could be identified by fluorometry and flow cytometry as an important target of applied treatment combinations.

Noroleanane saponins from Celmisia spectabilis.

Two new saponins were isolated as the major components of the deacylated saponin extract from the underground parts of Celmisia spectabilis. Their structures were established by NMR and mass spectral data and derivative formation as 2 beta,3 beta,17,23- tetrahydroxy-28-norolean-12-en-16-one-3-O-alpha-L-arabinopyrano syl (1-->2)-alpha-L-arabinopyranosyl(1-->6)-beta-D-glucopyranoside and 2 beta,3 beta,17,23- tetrahydroxy-28-norolean-12-en-16-one-3-O-alpha-L-arabinopyrano syl (1-->2)-alpha-L-arabinopyranosyl(1-->6)-[alpha-L-rhamnopyranosyl (1-->2)]-beta-D-glucopyranoside.

Flow cytometric analysis of Lactobacillus plantarum to monitor lag times, cell division and injury.

Flow cytometry in combination with fluorescent molecular markers 5- (and 6-) carboxyfluorescein succinimidylester (CFSE) and propidium iodide (PI) have been applied to determine lag times, numbers of cell divisions and injury after mild heat (50 degrees C, 5 min) and nisin treatments (0.1 and 1.0 microgram ml-1) of Lactobacillus plantarum. Initial labelling with covalently bound dye CFSE (20 and 100 micrograms ml-1) allowed determination of lag times and cell proliferation for up to eight generations. Double-labelling with CFSE and PI (5 micrograms ml-1) provided additional information about damage levels and distributions within populations. Subpopulations surviving treatment could be identified easily and selectively sorted.