Bioprotective yeasts: Potential to limit postharvest spoilage and to extend shelf life or improve microbial safety of processed foods.

Yeasts are a widespread group of microorganisms that are receiving increasing attention from scientists and industry. Their diverse biological activities and broad-spectrum antifungal activity make them promising candidates for application, especially in postharvest biocontrol of fruits and vegetables and food biopreservation. The present review focuses on recent knowledge of the mechanisms by which yeasts inhibit pathogenic fungi and/or spoilage fungi and bacteria. The main mechanisms of action of bioprotective yeasts include competition for nutrients and space, synthesis and secretion of antibacterial compounds, mycoparasitism and the secretion of lytic enzymes, biofilm formation, quorum sensing, induced systemic resistance of fruit host, as well as the production of reactive oxygen species. Preadaptation of yeasts to abiotic stresses such as cold acclimatization and sublethal oxidative stress can improve the effectiveness of antagonistic yeasts and thus more effectively play biocontrol roles under a wider range of environmental conditions, thereby reducing economic losses. Combined application with other antimicrobial substances can effectively improve the efficacy of yeasts as biocontrol agents. Yeasts show great potential as substitute for chemical additives in various food fields, but their commercialization is still limited. Hence, additional investigation is required to explore the prospective advancements of yeasts in the field of biopreservation for food.

Minimal Processing Technologies for Production and Preservation of Tailor-Made Foods§.

Tailor-made foods, also known as foods with programmable properties, are specialised systems with unique composition prepared by different methods, using the known mechanisms of action of their bioactive ingredients. The development of tailor-made foods involves the evaluation of individual components, including bioactive substances derived from waste products of other productions, such as essential oils. These components are evaluated both individually and in combination within food compositions to achieve specific functionalities. This review focuses on the application of minimal processing technologies for the production and preservation of tailor-made foods. It examines a range of approaches, including traditional and emerging technologies, as well as novel ingredients such as biomolecules from various sources and microorganisms. These approaches are combined according to the principles of hurdle technology to achieve effective synergistic effects that enhance food safety and extend the shelf life of tailor-made foods, while maintaining their functional properties.

Volatilome Analysis and Evolution in the Headspace of Packed Refrigerated Fish.

Fresh fish is a perishable food in which chemical (namely oxidation) and microbiological degradation result in undesirable odor. Non-processed fish (i.e., raw fish) is increasingly commercialized in packaging systems which are convenient for its retailing and/or which can promote an extension of its shelf-life. Compared to fish sent to its retail unpackaged, fish packaging results in a modification of the gaseous composition of the atmosphere surrounding it. These modifications of atmosphere composition may affect both chemical and microbiological degradation pathways of fish constituents and thereby the volatile organic compounds produced. In addition to monitoring Total Volatile Basic Nitrogen (TVB-N), which is a common indicator to estimate non-processed fish freshness, analytical techniques such as gas chromatography coupled to mass spectrometry or techniques referred to as "electronic nose" allow either the identification of the entire set of these volatile compounds (the volatilome) and/or to selectively monitor some of them, respectively. Interestingly, monitoring these volatile organic compounds along fish storage might allow the identification of early-stage markers of fish alteration. In this context, to provide relevant information for the identification of volatile markers of non-processed packaged fish quality evolution during its storage, the following items have been successively reviewed: (1) inner atmosphere gaseous composition and evolution as a function of fish packaging systems; (2) fish constituents degradation pathways and analytical methods to monitor fish degradation with a focus on volatilome analysis; and (3) the effect of different factors affecting fish preservation (temperature, inner atmosphere composition, application of hurdle technology) on volatilome composition.

Structural characterization of exopolysaccharide from Leuconostoc mesenteroides P35 and its binding interaction with bovine serum albumin using surface plasmon resonance biosensor.

This paper describes the structural elucidation of Leuconostoc mesenteroides P35 exopolysaccharide (EPS-LM). Ln. mesenteroides P35 strain was isolated from a French goat cheese for its capacity to produce EPS increasing the viscosity of a whey-based fermentation medium. The chemical structure of EPS-LM analysis was elucidated by determination of optical rotation degree, macromolecular characterization, sugar units and methylation analyses, FT-IR, 1D NMR spectroscopy (1H and 13C NMR), 2D NMR spectroscopy (1H1H COSY, HSQC and HMBC). EPS-LM was a high molecular weight (ranging from 6.7 × 106 Da to 9.9 × 106 Da) dextran that is composed of only d-glucose units containing α (1 â†’ 6) linkages and paltry α (1 â†’ 3) branches. Since polysaccharide-protein interactions can be exploited to control and design food matrices, EPS-LM interactions with bovine serum albumin (the main constituent of bovine plasma) were investigated by surface plasmon resonance (SPR). Kinetic properties of EPS-LM binding with immobilized BSA via showed an increase of EPS-LM affinity (equilibrium constant (Kd)) for BSA from (2.50 ± 0.01) × 10-5 M-1 at 298 K to (9.21 ± 0.05) × 10-6 M-1 at to 310 K. The thermodynamic parameters revealed that van der Waals and hydrogen binding forces play a major role in the interaction of EPS-LM with BSA. However, EPS-LM-BSA interaction was non-spontaneous, entropy driven and an EPS-LM - BSA binding process was endothermic (ΔG > 0). The structural findings suggested that Ln. mesenteroides P35 α-D-glucan might find widespread technological applications in the biopolymer, medical and food industries.

Chitosan-Sodium Caseinate Composite Edible Film Incorporated with Probiotic Limosilactobacillus fermentum: Physical Properties, Viability, and Antibacterial Properties.

Single-use synthetic plastics that are used as food packaging is one of the major contributors to environmental pollution. Hence, this study aimed to develop a biodegradable edible film incorporated with Limosilactobacillus fermentum. Investigation of the physical and mechanical properties of chitosan (CS), sodium caseinate (NaCas), and chitosan/sodium caseinate (CS/NaCas) composite films allowed us to determine that CS/NaCas composite films displayed higher opacity (7.40 A/mm), lower water solubility (27.6%), and higher Young's modulus (0.27 MPa) compared with pure CS and NaCas films. Therefore, Lb. fermentum bacteria were only incorporated in CS/NaCas composite films. Comparison of the physical and mechanical properties of CS/NaCas composite films incorporated with bacteria with those of control CS/NaCas composite films allowed us to observe that they were not affected by the addition of probiotics, except for the flexibility of films, which was improved. The Lb. fermentum incorporated composite films had a 0.11 mm thickness, 17.9% moisture content, 30.8% water solubility, 8.69 A/mm opacity, 25 MPa tensile strength, and 88.80% elongation at break. The viability of Lb. fermentum after drying the films and the antibacterial properties of films against Escherichia coli O157:H7 and Staphylococcus aureus ATCC 29213 were also evaluated after the addition of Lb. fermentum in the composite films. Dried Lb. fermentum composite films with 6.65 log10 CFU/g showed an inhibitory effect against E. coli and S. aureus (0.67 mm and 0.80 mm inhibition zone diameters, respectively). This shows that the Lb.-fermentum-incorporated CS/NaCas composite film is a potential bioactive packaging material for perishable food product preservation.

Biopreservation of Chocolate Mousse with Lactobacillus helveticus 2/20: Microbial Challenge Test

Probiotic bacteria are used for food biopreservation because their metabolic products might contribute to ensuring food microbiological safety and/or increase its shelf life without the addition of chemical preservatives. Moreover, biopreserved foods are excellent vehicles for the delivery of probiotic bacteria. The aim of the study was to investigate the potential of chocolate mousse food matrix for the delivery of the probiotic strain Lactobacillus helveticus 2/20 (Lb. helveticus 2/20) and to investigate its capacity to inhibit the growth of two foodborne pathogenic bacteria (Staphylococcus aureus and Escherichia coli). Therefore, the populations of free or encapsulated in calcium alginate Lb. helveticus 2/20 cells and/or of each pathogen (used to voluntarily contaminate each sample) were monitored both in complex nutrient medium (MRS broth) and in chocolate mousse under refrigeration conditions and at room temperature. Lb. helveticus 2/20 alone in free or encapsulated state effectively inhibited the growth of Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923 in chocolate mousse when stored at 20 ± 2 °C. Practically no viable unwanted bacteria were identified on the 7th day from the beginning of the process. High viable Lb. helveticus 2/20 cell populations were maintained during storage under refrigerated conditions (4 ± 2 °C) and at room temperature. Chocolate mousse is thus a promising food matrix to deliver probiotic Lb. helveticus 2/20 cells, which could also protect it from contamination by unwanted bacteria.

Polysaccharide-Based Edible Films Incorporated with Essential Oil Nanoemulsions: Physico-Chemical, Mechanical Properties and Its Application in Food Preservation-A Review.

Edible films with essential oils (EOs) are becoming increasingly popular as an alternative to synthetic packaging due to their environmentally friendly properties and ability as carriers of active compounds. However, the required amounts of EOs to impart effective antimicrobial properties generally exceed the organoleptic acceptance levels. However, by nanoemulsifying EOs, it is possible to increase their antimicrobial activity while reducing the amount required. This review provides an overview of the physico-chemical and mechanical properties of polysaccharide-based edible films incorporated with EOs nanoemulsions and of their application to the preservation of different food types. By incorporating EOs nanoemulsions into the packaging matrix, these edible films can help to extend the shelf-life of food products while also improving the quality and safety of the food product during storage. It can be concluded that these edible films have the potential to be used in the food industry as a green, sustainable, and biodegradable method for perishable foods preservation.

A critical review on approaches to regulate the release rate of bioactive compounds from biopolymeric matrices.

Recently, there has been a growing interest in novel food formulations and packaging materials using controlled release systems in order to enhance the functional performance of food bioactive ingredients and to prolong the shelf-life of food products. Correspondingly, numerous types of matrices (polymers, biopolymers, synthetic materials) could be implemented as carriers based on their compatibility with the bioactive ingredients and the type of releasing medium. A plethora of applications of controlled release can be identified in the pharmaceutical, agriculture, cosmetics, and particularly the food industries. Moreover, several physical and chemical stimuli are applied to regulate the release behavior of a bioactive molecule in the desired medium, which is under development. This review highlights the strategies applied to regulate the release rate of biopolymeric networks mainly including mixing of biopolymers, physical and chemical crosslinkers along with encapsulation systems, furthermore, the future prospects of release concept are discussed.

Staphylococcus aureus membrane-damaging activities of four phenolics.

The membrane-damaging activities of four phenolics chosen for their bactericidal activity against Staphylococcus aureus CNRZ3 were investigated: 5,7-dihydroxy-4-phenylcoumarin (DHPC), 5,8-dihydroxy-1,4-naphthoquinone (DHNQ), epigallocatechin gallate (EGCG) and isobutyl 4-hydroxybenzoate (IBHB). Staphylococcus aureus CNRZ3 cells, as well as model liposomes mimicking its membrane phospholipids composition, were treated with each phenolic at its minimal bactericidal concentration. Membrane integrity, intracellular pH and intracellular esterase activity were examined by flow cytometric analysis of S. aureus cells stained with propidium iodide and SYTO® 9, 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein acetoxymethyl ester, and 5(6)-carboxyfluorescein diacetate, respectively. While intracellular pH was affected by the foyr phenolics, only DHNQ and to a lesser extent EGCG, caused a loss of membrane integrity. Flow cytometric analysis of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and DPPC/POPG (2-oleoyl-1-palmitoyl-sn-glycero-3-phosphoglycerol) liposomes stained with Coumarin 6 (which penetrates the lipid bilayer) or 5-N(octadecanoyl)-amino-fluorescein (which binds to the liposome shell) suggested that only EGCG and DHNQ penetrated the bilayer of phospholipids of liposomes. Taken together, these findings support the hypothesis that EGCG and DHNQ bactericidal activity results from their accumulation in the phospholipid bilayer of S. aureus cells membrane causing its disruption.

Phenolic-Rich Plant Extracts With Antimicrobial Activity: An Alternative to Food Preservatives and Biocides?

In recent years, the search for natural plant-based antimicrobial compounds as alternatives to some synthetic food preservatives or biocides has been stimulated by sanitary, environmental, regulatory, and marketing concerns. In this context, besides their established antioxidant activity, the antimicrobial activity of many plant phenolics deserved increased attention. Indeed, industries processing agricultural plants generate considerable quantities of phenolic-rich products and by-products, which could be valuable natural sources of natural antimicrobial molecules. Plant extracts containing volatile (e.g., essential oils) and non-volatile antimicrobial molecules can be distinguished. Plant essential oils are outside the scope of this review. This review will thus provide an overview of current knowledge regarding the promises and the limits of phenolic-rich plant extracts for food preservation and biofilm control on food-contacting surfaces. After a presentation of the major groups of antimicrobial plant phenolics, of their antimicrobial activity spectrum, and of the diversity of their mechanisms of action, their most promising sources will be reviewed. Since antimicrobial activity reduction often observed when comparing in vitro and in situ activities of plant phenolics has often been reported as a limit for their application, the effects of the composition and the microstructure of the matrices in which unwanted microorganisms are present (e.g., food and/or microbial biofilms) on their activity will be discussed. Then, the different strategies of delivery of antimicrobial phenolics to promote their activity in such matrices, such as their encapsulation or their association with edible coatings or food packaging materials are presented. The possibilities offered by encapsulation or association with polymers of packaging materials or coatings to increase the stability and ease of use of plant phenolics before their application, as well as to get systems for their controlled release are presented and discussed. Finally, the necessity to consider phenolic-rich antimicrobial plant extracts in combination with other factors consistently with hurdle technology principles will be discussed. For instance, several authors recently suggested that natural phenolic-rich extracts could not only extend the shelf-life of foods by controlling bacterial contamination, but could also coexist with probiotic lactic acid bacteria in food systems to provide enhanced health benefits to human.

Biological preservation of mayonnaise with Lactobacillus plantarum LBRZ12, dill, and basil essential oils.

In this study, different variants of egg-free mayonnaise containing free and immobilized Lactobacillus plantarum LBRZ12 cells and essential oils taken from basil and dill were prepared. The composition and antimicrobial properties of essential oils were investigated. The main constituents of basil oil were methyl chavicol (36.81%), methyl eugenol (20.40%), ß-linanool (14.35%), eugenol (10.55%), and L(-)-carvone (39.05%), whereas dill oil contained mostly d-limonene (21.11%) and α-phellandrene (22.68%). The essential oils exhibited strong antimicrobial activity against all test-microorganisms. The mayonnaise variants were kept refrigerated for 40 days and changes in pH, concentration of viable cells of lactobacilli, microbiological, and organoleptic characteristics were monitored. The pH decreased from 6.5 to 4.5 over the period of storage. The number of undesired microflora in mayonnaise preserved with lactobacilli and essential oils decreased significantly (0 after the 20th day) indicating their effectiveness as biological preservatives. The mayonnaise variants demonstrated pleasant organoleptic characteristics, thus meet customers' requirements.

Antibacterial Properties of Polyphenols: Characterization and QSAR (Quantitative Structure-Activity Relationship) Models.

Besides their established antioxidant activity, many phenolic compounds may exhibit significant antibacterial activity. Here, the effect of a large dataset of 35 polyphenols on the growth of 6 foodborne pathogenic or food-spoiling bacterial strains, three Gram-positive ones (Staphylococcus aureus, Bacillus subtilis, and Listeria monocytogenes) and three Gram-negative ones (Escherichia coli, Pseudomonas aeruginosa, and Salmonella Enteritidis), have been characterized. As expected, the effects of phenolic compounds were highly heterogeneous ranging from bacterial growth stimulation to antibacterial activity and depended on bacterial strains. The effect on bacterial growth of each of the polyphenols was expressed as relative Bacterial Load Difference (BLD) between a culture with and without (control) polyphenols at a 1 g L-1 concentration after 24 h incubation at 37°C. Reliable Quantitative Structure-Activity Relationship (QSAR) models were developed (regardless of polyphenol class or the mechanism of action involved) to predict BLD for E. coli, S. Enteritidis, S. aureus, and B. subtilis, unlike for L. monocytogenes and P. aeruginosa. L. monocytogenes was generally sensitive to polyphenols whereas P. aeruginosa was not. No satisfactory models predicting the BLD of P. aeruginosa and L. monocytogenes were obtained due to their specific and quite constant behavior toward polyphenols. The main descriptors involved in reliable QSAR models were the lipophilicity and the electronic and charge properties of the polyphenols. The models developed for the two Gram-negative bacteria (E. coli, S. Enteritidis) were comparable suggesting similar mechanisms of toxic action. This was not clearly observed for the two Gram-positive bacteria (S. aureus and B. subtilis). Interestingly, a preliminary evaluation by Microbial Adhesion To Solvents (MATS) measurements of surface properties of the two Gram-negative bacteria for which QSAR models were based on similar physico-chemical descriptors, revealed that MATS results were also quite similar. Moreover, the MATS results of the two Gram-positive bacterial strains S. aureus and B. subtilis for which QSARs were not based on similar physico-chemical descriptors also strongly differed. These observations suggest that the antibacterial activity of most of polyphenols likely depends on interactions between polyphenols and bacterial cells surface, although the surface properties of the bacterial strains should be further investigated with other techniques than MATS.

Potential of Incorporation of Antimicrobial Plant Phenolics Into Polyolefin-Based Food Contact Materials to Produce Active Packaging by Melt-Blending: Proof of Concept With Isobutyl-4-Hydroxybenzoate.

There is an increasing interest for active food packaging incorporated with natural antimicrobial agents rather than synthetic preservatives. However, most of plastics for direct contact with food are made of polyolefins, usually processed by extrusion, injection, or blow-molding methods while most of natural antimicrobial molecules are thermolabile compounds (e.g., essential oils). Therefore, addition of plant phenolics (with low volatility) to different polyolefins might be promising to design active controlled release packaging processed by usual plastic compounding and used for direct contact with food products. Therefore, up to 2% (wt/wt) of isobutyl-4-hydroxybenzoate (IBHB) was mixed with 3 polyolefins: EVA poly(ethylene-co-vinyl acetate), LLDPE (Linear Low Density Polyethylene), and PP (PolyPropylene) by melt-blending from 75 to 170°C and then pelletized in order to prepare heat-pressed films. IBHB was chosen as an antibacterial phenolic active model molecule against Staphylococcus aureus to challenge the entire processing. Antibacterial activity of films against S. aureus (procedure adapted from ISO 22196 standard) were 4, 6, and 1 decimal reductions in 24 h for EVA, LLDPE, and PP films, respectively, demonstrating the preservation of the antibacterial activity after melt processing. For food contact materials, the efficacy of antimicrobial packaging depends on the release of the antimicrobial molecules. Therefore, the three types of films were placed at 23°C in 95% (v/v) ethanol and the release rates of IBHB were monitored: 101 ± 1%, 32 ± 7%, and 72 ± 9% at apparent equilibrium for EVA, LLDPE, and PP films, respectively. The apparent diffusion coefficients of IBHB in EVA and PP films were 2.8 ± 0.3 × 10-12 and 4.0 ± 1.0 × 10-16 m2s-1. For LLDPE films, IBHB crystals were observed on the surface of films by SEM (Scanning Electron Microscopy): this blooming effect was due the partial incompatibility of IBHB in LLDPE and its fast diffusion out of the polymer matrix onto the film surface. In conclusion, none of these three materials was suitable for a relevant controlled release packaging targeting the preservation of fresh food, but a combination of two of them is promising by the design of a multilayer packaging: the release could result from permeation through an inner PE layer combined with an EVA one acting as a reservoir.

Antilisterial activity of dromedary lactoferrin peptic hydrolysates.

The aim of this study was to explore the antibacterial peptides derived from dromedary lactoferrin (LFc). The LFc was purified from colostrum using a batch procedure with a cation exchange chromatography support and was hydrolyzed with pepsin to generate peptic digest. This peptic digest was fractionated by cation exchange chromatography, and the antilisterial activity of LFc, peptic digest, and obtained fractions was investigated using the bioscreen method. The growth of Listeria innocua ATCC 33090 and LRGIA 01 strains was not inhibited by LFc and its hydrolysates. Two fractions of dromedary lactoferrin peptic hydrolysate were active against both strains. A tandem mass spectroscopy analysis revealed that the 2 active fractions comprised at least 227 different peptides. Among these peptides, 9 found in the first fraction had at least 50% similarity with 10 known antimicrobial peptides (following sequence alignments with the antimicrobial peptide database from the University of Nebraska Medical Center, Omaha). Whereas 9 of these peptides presented homology with honeybee, frog, or amphibian peptides, the 10th peptide, F152SASCVPCVDGKEYPNLCQLCAGTGENKCACSSQEPYFGY192 (specifically found in 1 separated fraction), exibited 54% homology with a synthetic antibacterial peptide (AP00481) derived from human lactoferrin named kaliocin-1. Similarly, the second fraction contained 1 peptide similar to lactoferrampin B, an antibacterial peptide derived from bovine milk. This result suggests that peptic hydrolysis of LFc releases more active antimicrobial peptides than their protein source and thus provides an opportunity for their potential use to improve food safety by inhibiting undesirable and spoilage bacteria.

Plant antimicrobial polyphenols as potential natural food preservatives.

BACKGROUND: The growing demand for natural food preservatives in the last decade has promoted investigations on their application for preserving perishable foods. In this context, the present review is focused on discussing the prospective application of plant extracts containing phenolics or isolated plant phenolics as natural antimicrobials in foods. Plant essential oils are outside the scope of this review since utilization of their antimicrobial activity for food preservation has been extensively reviewed. RESULTS: Although the exact antimicrobial mechanisms of action of phenolic compounds are not yet fully understood, it is commonly acknowledged that they have diverse sites of action at the cellular level. Antimicrobial phenolics can be added directly to the formulation of perishable food products or incorporated into food-contact materials to release them in the immediate zone of perishable foods. Edible coatings or active food packaging materials can thus be used as carriers of plant bioactive compounds. CONCLUSION: These materials could be an interesting delivery system to improve the stability of phenolics in foods and to improve the shelf life of perishable foods. This review will thus provide an overview of current knowledge of the antimicrobial activity of phenolic-rich plant extracts and of the promises and limits of their exploitation for the preservation of perishable foods. © 2018 Society of Chemical Industry.

pH-dependent complexation of lysozyme with low methoxyl (LM) pectin.

In order to understand the effect of pH on the formation of electrostatic complexes between lysozyme and low methoxyl (LM) pectin, mixtures were prepared at a fixed lysozyme concentration (0.714g.L-1) by progressive addition of LM pectin (from 0 to 4g.L-1). Turbidity analysis allowed to determine specific conditions of pH and lysozyme/LM pectin ratio for optimal complex aggregation. The intrinsic fluorescence enhancement observed upon binding of LM pectin to lysozyme was correlated with the formation of intermolecular aggregates. Conversely, the intrinsic fluorescence decrease observed at higher LM pectin amounts was correlated with the dissociation of intermolecular aggregates. UV absorption spectroscopy showed modifications in lysozyme conformation during both the aggregation phase and the dissociation phase. The role of electrostatic interactions in the formation of lysozyme/LM pectin complexes is discussed in relation to the overall structure and the charge density profile of the two biopolymers.

Using complexation for the microencapsulation of nisin in biopolymer matrices by spray-drying.

The aim of this study is to investigate the potential of complexation to encapsulate nisin (5g/L concentration) using spray-drying technique and to evaluate how complexation with pectin or alginate (2g/L concentration) can preserve nisin structure and antimicrobial activity. Spray-drying of nisin-low methoxyl pectin or nisin-alginate electrostatic complexes has led to the microencapsulation of the peptide in different networks that were highly influenced by the polysaccharide type. Turbidity and particle size measurements indicated that while spray-drying promoted the aggregation of nisin-pectin complexes, it favored the dissociation of nisin-alginate aggregates to form individual complexes. Structural changes of nisin induced by complexation with pectin or alginate and spray-drying were studied by using UV-Vis absorption and fluorescence spectroscopy. The results showed that complexation with pectin or alginate preserved nisin structure as well as its antimicrobial activity during spray-drying.

Recent Advances on Multi-Parameter Flow Cytometry to Characterize Antimicrobial Treatments.

The investigation on antimicrobial mechanisms is a challenging and crucial issue in the fields of food or clinical microbiology, as it constitutes a prerequisite to the development of new antimicrobial processes or compounds, as well as to anticipate phenomenon of microbial resistance. Nowadays it is accepted that a cells population exposed to a stress can cause the appearance of different cell populations and in particular sub-lethally compromised cells which could be defined as viable but non-culturable (VBNC). Recent advances on flow cytometry (FCM) and especially on multi-parameter flow cytometry (MP-FCM) provide the opportunity to obtain high-speed information at real time on damage at single-cell level. This review gathers MP-FCM methodologies based on individual and simultaneous staining of microbial cells employed to investigate their physiological state following different physical and chemical antimicrobial treatments. Special attention will be paid to recent studies exploiting the possibility to corroborate MP-FCM results with additional techniques (plate counting, microscopy, spectroscopy, molecular biology techniques, membrane modeling) in order to elucidate the antimicrobial mechanism of action of a given antimicrobial treatment or compound. The combination of MP-FCM methodologies with these additional methods is namely a promising and increasingly used approach to give further insight in differences in microbial sub-population evolutions in response to antimicrobial treatments.

Complex coacervation for the development of composite edible films based on LM pectin and sodium caseinate.

Coacervation between sodium caseinate (CAS) and low methoxyl pectin (LMP) at pH 3 was investigated as a function of protein/polysaccharide ratio. The highest amount of complex coacervates was formed at a CAS/LMP ratio of 2 at which the ζ-potential value was zero and the turbidity reached its highest value. Then, the properties of films based on these complex coacervates were studied. Coacervation resulted in decreasing water content and water sorption of films as the protein concentration increased. The mechanical properties of films were highly influenced by the formation of electrostatic complexes. The highest values of Young's modulus (182.97± 6.48MPa) and tensile strength (15.64±1.74MPa) with a slight increase of elongation at break (9.35±0.10%) were obtained for films prepared at a CAS/LMP ratio equal to 0.05. These findings show that interactions between LMP and CAS can be used to develop innovative packaging containing active molecules.

Antimicrobial Activity of Nisin and Natamycin Incorporated Sodium Caseinate Extrusion-Blown Films: A Comparative Study with Heat-Pressed/Solution Cast Films.

Antimicrobial edible films based on sodium caseinate, glycerol, and 2 food preservatives (nisin or natamycin) were prepared by classical thermomechanical processes. Food preservatives were compounded (at 65 °C for 2.5 min) with sodium caseinate in a twin-screw extruder. Anti-Listeria activity assays revealed a partial inactivation of nisin following compounding. Thermoplastic pellets containing food preservatives were then used to manufacture films either by blown-film extrusion process or by heat-press. After 24 h of incubation on agar plates, the diameters of K. rhizophila growth inhibition zones around nisin-incorporated films prepared by solution casting (control), extrusion blowing or heat pressing at 80 °C for 7 min of nisin-containing pellets were 15.5 ± 0.9, 9.8 ± 0.2, and 8.6 ± 1.0 mm, respectively. Since heat-pressing for 7 min at 80 °C of nisin-incorporated pellets did not further inactivate nisin, this indicates that nisin inactivation during extrusion-blowing was limited. Moreover, the lower diameter of the K. rhizophila growth inhibition zone around films prepared with nisin-containing pellets compared to that observed around films directly prepared by solution casting confirms that nisin inactivation mainly occurred during the compounding step. Natamycin-containing thermoplastic films inhibited Aspergillus niger growth; however, by contrast with nisin-containing films, heat-pressed films had higher inhibition zone diameters than blown films, therefore suggesting a partial inactivation of natamycin during extrusion-blowing.