Impact of multiple hurdles on Listeria monocytogenes dispersion of survivors.

Pathogen exposure to multiple hurdles could result in variation in the number of survivors, which needs to be carefully considered using appropriate regression models for dealing with survivor dispersion. The aim of this study was to evaluate the impact of the hurdles on the random component of the measured variation and on its unexplained part (over or under-dispersion) representing the departure from randomness, i.e. non-randomness, in survivors of a multi-strain mixture of L. monocytogenes. The pathogen inactivation curves were fitted to the Weibull model within the Conway-Maxwell-Poisson process. In all the 20 hurdle combinations, the surviving cells, whether they showed an upward curvature or linear kinetics, displayed the randomness revealed by the degree of dispersion of the inactivation parameters (-b and p). In 15 combinations, a significant dispersion coefficient (c0), which reflected the non-random component of variation was evident, denoting either over-dispersion (c0 > 0 in 13 combinations) or under-dispersion (c0 < 0 in 2 combinations). The observed dependence of the under- and over-dispersion conditions on the inactivation rate was confirmed by a Monte Carlo simulation based on the inactivation parameter -b. Including both randomness and non-randomness provides a more accurate estimation of survivors, which certainly impacts on intervention practices.

The COM-Poisson Process for Stochastic Modeling of Osmotic Inactivation Dynamics of Listeria monocytogenes.

Controlling harmful microorganisms, such as Listeria monocytogenes, can require reliable inactivation steps, including those providing conditions (e.g., using high salt content) in which the pathogen could be progressively inactivated. Exposure to osmotic stress could result, however, in variation in the number of survivors, which needs to be carefully considered through appropriate dispersion measures for its impact on intervention practices. Variation in the experimental observations is due to uncertainty and biological variability in the microbial response. The Poisson distribution is suitable for modeling the variation of equi-dispersed count data when the naturally occurring randomness in bacterial numbers it is assumed. However, violation of equi-dispersion is quite often evident, leading to over-dispersion, i.e., non-randomness. This article proposes a statistical modeling approach for describing variation in osmotic inactivation of L. monocytogenes Scott A at different initial cell levels. The change of survivors over inactivation time was described as an exponential function in both the Poisson and in the Conway-Maxwell Poisson (COM-Poisson) processes, with the latter dealing with over-dispersion through a dispersion parameter. This parameter was modeled to describe the occurrence of non-randomness in the population distribution, even the one emerging with the osmotic treatment. The results revealed that the contribution of randomness to the total variance was dominant only on the lower-count survivors, while at higher counts the non-randomness contribution to the variance was shown to increase the total variance above the Poisson distribution. When the inactivation model was compared with random numbers generated in computer simulation, a good concordance between the experimental and the modeled data was obtained in the COM-Poisson process.

Praedicere Possumus: An Italian web-based application for predictive microbiology to ensure food safety.

The use of predictive modelling tools, which mainly describe the response of microorganisms to a particular set of environmental conditions, may contribute to a better understanding of microbial behaviour in foods. In this paper, a tertiary model, in the form of a readily available and userfriendly web-based application Praedicere Possumus (PP) is presented with research examples from our laboratories. Through the PP application, users have access to different modules, which apply a set of published models considered reliable for determining the compliance of a food product with EU safety criteria and for optimising processing throughout the identification of critical control points. The application pivots around a growth/no-growth boundary model, coupled with a growth model, and includes thermal and non-thermal inactivation models. Integrated functionalities, such as the fractional contribution of each inhibitory factor to growth probability (f) and the time evolution of the growth probability (Pt), have also been included. The PP application is expected to assist food industry and food safety authorities in their common commitment towards the improvement of food safety.

Survival strategies of Bacillus spores in food.

Control of bacterial spores is one of the major problem in the food preservation. Spores of Bacillus genus are commonly present in different environments, including soil and the gut of insects and animals and, as a result, they can be spread to all kind of foods. Due to their high resistance properties, their complete inactivation in food is often impossible without changing the product characteristics. Surviving spores can germinate and grow out to vegetative cells, with the consequent great risk of food spoilage and food poisoning after consumption. Spores have evolved various mechanisms, including phenotypic variability, to protect themselves from a wide range of damage resulting from food preservation treatments. Even if the phenotypic heterogeneity contributes to increase the chances of survival of Bacillus spore to conventional preservation treatments, in some specific instances, an homogeneous response could be the result of a strategy adopted by the spores to increase resistance to those treatments.

A simplified approach for modelling the bacterial growth/no growth boundary.

A simplified growth/no growth (G/NG) model, conceptually derived from the Gamma model and making direct and explicit use of growth limits of bacteria through a normalization constant (η), was proposed. The η value, which quantifies the product of the cardinal optimal distances for growth probability, is a species-independent constant. This is of importance when experimental data is missing or insufficient. The simplified G/NG model was developed including the effect of temperature, pH and water activity, and was expanded incorporating the preservative effects. As a practical application, the model was investigated for its ability to describe published data. The successful validation of the simplified G/NG model is discussed in regard to its potential applicability as a first estimate method for the development of safe food products.

A novel multipurpose Excel tool for equilibrium speciation based on Newton-Raphson method and on a hybrid genetic algorithm.

A new algorithm for simulation of chemical equilibria is developed, based on classical Newton-Raphson method applied to mass balance. This tool, named EST (Equilibrium Speciation Tool), is improved by using a robust Genetic Algorithm. In addition, EST works by using Excel spreadsheets and therefore offers the innovation of a great simplicity and versatility. In fact, it allows the users to simulate, or to obtain from experimental data, desired chemical-physical parameters as well as to interact with other available or freely created Excel tools. The reliability of this utility is here proved by comparison with some published data by other authors, concerning both complicated homogeneous and heterogeneous equilibria. In addiction its flexibility is tested computing thermodynamic parameters by using experimental calorimetric data referred to the complex formation of cobalt(II) with a macrocyclic ligand. A brief review and comparison of the relative robustness and quickness of main numerical methods are also reported.

Mixed nitrogen/oxygen ligand affinities for bipositive metal ions and dioxygen binding to cobalt(II) complexes.

The complex formation of Co(II) and Cd(II) with mixed N/O ligands in dimethylsulfoxide (dmso) at 298 K is investigated by means of potentiometric, UV-Vis, calorimetric, FT-IR, NMR and electrochemical techniques. The linear and cyclic ligands investigated are: 2,2'-oxydiethylamine (NON), N-(2-hydroxyethyl)-ethylenediamine (NNO), 1,4,10-trioxa-7,13-diaza-cyclopentadecane (2,1) and 1,4,7,10-tetraoxa-13-azacyclopentadecane (2,2). The results are discussed by taking into account the donor strength of donor atoms, strain effects in the formation of chelate rings, steric and inductive effects and the size of the cavity when macrocyclic ligands are concerned. DFT studies are also performed to gain an insight into the stabilization of the lower oxidation state of the CoL2(2+/3+) couple in order to understand the different reactivity of the Co(II) complexes towards dioxygen. The kinetics of dioxygen uptake has been studied by UV-Vis measurements and the results reveal an interesting strong solvent effect, which is active in lowering the rate constant for formation of the initial superoxo complex.

Cobalt(II) dioxygen carriers based on simple diamino ligands: kinetic and ab initio studies.

The kinetics of the oxygenation reaction of CoL2(2+) complexes (L=ethylenediamine (en), N,N'-dimethylethylenediamnine (dmen)) have been investigated in dimethyl sulfoxide (dmso) at 298 K and in a medium adjusted to 0.1 mol dm(-3) with Et4NClO4 by means of a UV-vis spectrophotometric technique. The reaction mechanisms are consistent with the fast formation of superoxo 1:1 initial CoL2-O2 species (L=en, dmen), whereas the dimeric mu-peroxo (CoL2)2O2 adduct is formed only when L=en, in the rate determining step. The kinetic results are discussed taking into account the effects of ligand/solvent substitution. EPR results give information on the electronic structure and the coordination geometry of the Co(II) complexes and further confirm the stoichiometry of the species formed. Ab initio calculations provide insights on the geometrical parameters of all the complexes investigated and allow us to draw some hypotheses about the influence of H.H nonbonded interactions in the eventual formation of the dimeric mu-peroxo (CoL2)2O2 complexes. Solvational effects are also considered. The formation of the (CoL3)2O2 adduct is also proved when L=en by means of O2 volumetric absorption.

SOLVERSTAT: a new utility for multipurpose analysis. An application to the investigation of dioxygenated Co(II) complex formation in dimethylsulfoxide solution.

A new utility for multipurpose analysis, SOLVERSTAT, taking advantage of the versatility of spreadsheets is here described. By means of this tool advanced statistical tests have introduced in Microsoft Excel Solver thus allowing regression diagnostic and discrimination between different models. The utility is here applied to the determination, by UV-Vis spectroscopy, of the stability constant for the uptake of molecular dioxygen by the 1:2 complex of Co(II) with N,N'-dimethylethylenediamine (dmen) in the aprotic solvent dimethylsulfoxide (dmso) at 298 K and in a medium adjusted to 0.1 mol dm(-3) with Et(4)NClO(4). The reliability of the model and parameters obtained are discussed and the results compared with those obtained by Dynafit, a different software package, and by independent voltammetric measurements. The validity of SOLVERSTAT has been also examined applying it to the discrimination between different models already discussed in the literature.