Bioingénierie et Dynamique Microbienne aux Interfaces Alimentaires

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 towards 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.

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.

Nisin is a natural preservative for many food products. This bacteriocin is mainly used in dairy and meat products. Nisin inhibits pathogenic food borne bacteria such as Listeria monocytogenes and many other Gram-positive food spoilage microorganisms. Nisin can be used alone or in combination with other preservatives or also with several physical treatments. This paper reviews physicochemical and biological properties of nisin, the main factors affecting its antimicrobial effectiveness, and its food applications as an additive directly incorporated into food matrices.

In recent years, the use of lytic bacteriophages as antimicrobial agents controlling pathogenic bacteria has appeared as a promising new alternative strategy in the face of growing antibiotic resistance which has caused problems in many fields including medicine, veterinary medicine, and aquaculture. The use of bacteriophages has numerous advantages over traditional antimicrobials. The effectiveness of phage applications in fighting against pathogenic bacteria depends on several factors such as the bacteriophages/target bacteria ratio, the mode and moment of treatment, environmental conditions (pH, temperature...), the neutralization of phage and accessibility to target bacteria, amongst others. This report presents these factors and the challenges involved in developing phage therapy applications.

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.

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.

Nisin is the only bacteriocin approved as a food preservative because of its antibacterial effectiveness and its negligible toxicity for humans. Typical problems encountered when nisin is directly added to foods are mainly fat adsorption leading to activity loss, heterogeneous distribution in the food matrix, inactivation by proteolytic enzymes, and emergence of resistance in normally sensitive bacteria strains. To overcome these problems, nisin can be immobilized in solid matrices that must act as diffusional barriers and allow controlling its release rate. This strategy allows maintaining a just sufficient nisin concentration at the food surface. The design of such antimicrobial materials must consider both bacterial growth kinetics but also nisin release kinetics. In this review, nisin incorporation in polymer-based materials will be discussed and special emphasis will be on the applications and properties of antimicrobial food packaging containing this bacteriocin.

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.

Three different methods to standardize biofilm removal for in situ sanitary control of closed surfaces in the food industry have been developed and compared, i.e. sonication, enzymatic treatment and a combined treatment which involved the application of ultrasound to enzyme preparations. The biofilm studied was an Escherichia coli model biofilm, made with milk on stainless steel sheets. Plate counting and epifluorescence microscopy were used to assess the efficiency of each treatment. The results are expressed in percentages, 100% denoting total removal, obtained with a flat ultrasonic transducer (T1) developed and presented in a previous study. The application of ultrasound by a patented curved transducer, T2 (10 s, 40 kHz), specifically devised for closed surfaces, was not sufficient to completely remove the biofilm (30 +/- 7%). This biofilm was dislodged by two proteolytic enzyme preparations tested by immersion, viz. a 15-min application of protease (84 +/- 1%) and a 30-min trypsin application (95 +/- 8%). Using a combined treatment, the results showed a synergism between ultrasonic waves and proteolytic or glycolytic enzyme preparations, with removal of a significant amount of biofilm, i.e. 61-96% depending on the conditions tested, i.e. two to three times greater compared to sonication alone (30%). This application was in agreement with an industrial control, i.e. a good reproducible recovery of the biofilm in 10 s compared with 30 or 15 min with the enzyme alone.

The citrate metabolism has been extensively studied in lactic acid bacteria (LAB) for its aroma compound production. Among the 4-carbon (C4) by-products obtained from citrate fermentation, diacetyl is one of the better known products for its contribution to the buttery aroma of dairy products. A lot of documents deal with ways to improve diacetyl concentration in food matrices. Apart from these organoleptic advantages, in a microbial ecosystem, the citrate metabolism gives selective advantages to citrate positive microorganisms. Citrate metabolism allows the LAB to use another carbon source for their growth, withstand acidic conditions and generate a “proton motive force” (PMF). Moreover, the citrate/glucid co-metabolism leads to the fast release of organic compounds known for having bacteriostatic effects. Under specific conditions, the C4 pathway liberates diacetyl which is bacteriostatic. In this review we first describe the citrate metabolism and the enzymes involved in the two homo- and heterofermentative LABLc diacetylactisandLeuconostocspp. Moreover, the way to shift the metabolic pathway toward the production of aromatic compounds is discussed for both of these fermentative types of bacteria. Finally, the selective advantages of citrate metabolism for LAB in complex microbial ecosystems are delineated.

The antibacterial mechanism of a Cinnamomum cassia essential oil from Vietnam and of its main component (trans-cinnamaldehyde, 90% (m/m) of C. cassia essential oil) against a Listeria innocua strain was investigated to estimate their potential for food preservation. In the presence of C. cassia essential oil or trans-cinnamaldehyde at their minimal bactericidal concentration (2700 μg·mL(-1)), L. innocua cells fluoresced green after staining with Syto9® and propidium iodide, as observed by epifluorescence microscopy, suggesting that the perturbation of membrane did not cause large pore formation and cell lysis but may have introduced the presence of viable but nonculturable bacteria. Moreover, the fluidity, potential, and intracellular pH of the cytoplasmic membrane were perturbed in the presence of the essential oil or trans-cinnamaldehyde. However, these membrane perturbations were less severe in the presence of trans-cinnamaldehyde than in the presence of multicomponent C. cassia essential oil. This indicates that in addition to trans-cinnamaldehyde, other minor C. cassia essential oil components play a major role in its antibacterial activity against L. innocua cells.

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.

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.

Antimicrobials are therapeutic substances used to prevent or treat infections. Disinfectants are antimicrobial agents applied to non-living surfaces. Every year, several thousand tonnes of antimicrobials and their by-products are released into the environment and in particular into the aquatic environment. This type of xenobiotic has ecological consequences in the natural environment but also in technological environments such as wastewater treatment plants and methane fermentation sewage sludge treatment plants. The constant exposure of microbial communities not only to high concentrations but also to sub-inhibitory concentrations of antibiotics is a key element in the development of antibiotic resistance in aquatic environments and in soils. The future of antimicrobials lies in the development of biosourced or bioinspired molecules. The observation and deciphering of interactions between living organisms is the key to this development.

The aim of this study was to evaluate the antimicrobial activity of camel caseins and their hydrolysates by gastrointestinal proteolytic enzymes against 3 Gram-positive and 2 Gram-negative bacterial strains. Camel caseins (CN) were hydrolysed by successive action of pepsin and pancreatin. Hydrolysis of CN was checked by electrophoresis and gel filtration chromatography (GFC). Both techniques showed that CN was hydrolysed into peptides. Among the tested bacteria, a decrease of 19.3%±0.02 of E. coli XL1 blue cells growth was observed in the presence of undigested camel casein at a concentration of 20 mg ml −1 . After successive hydrolyses by pepsin and pancreatin, camel milk casein hydrolysates still exhibited anti-bacterial activity against E. coli XL1 blue strain (19.73±0.01% growth inhibition under the same conditions). Gram-positive strain growth was not affected by intact camel CN, while, at the same concentration (20 mg ml –1 ), their hydrolysates slightly inhibited the growth of these bacteria. This suggests that antibacterial peptidic fragments of caseins were generated by pepsin and pancreatin.

ABSTRACT Antimicrobial textiles (65% cotton – 35% polyester) were functionalized using a patented technology that combines an antimicrobial molecule – polyhexamethylene biguanide (PHMB) and a precipitating agent – sodium laurylsulphate. Surface characterization was performed by x‐ray photoelectron spectroscopy and time‐of‐flight secondary ion mass spectrometry, and both techniques made clear signatures of PHMB at the top surface of treated textiles. Washing led to a strong decrease of PHMB concentration at the surface. Comparison of textile surface analysis and antimicrobial tests indicated that the amount of PHMB at its extreme surface of textiles after five or 10 industrial washings was sufficient to inhibit Listeria innocua but not Pseudomonas aeruginosa growth. The viability of L. innocua cells after contact with PHMB‐treated textile after one industrial washing was estimated using the Live/Dead BacLight kit (Molecular Probes, Eugene, OR): the combination of epifluorescence microscopy observations coupled with classic enumeration allowed detection of the presence of viable but nonculturable cells. PRACTICAL APPLICATIONS Protective clothing is required in the food‐processing industry to protect products from being contaminated by microorganisms carried by workers' clothes or filtration systems. Consequently, there is an increasing interest in the use of antimicrobial functionalized textiles in the food industry to avoid that textiles could be vectors for pathogenic or food spoilage microorganisms. In the present study, the correlation between PHMB (the antimicrobial agent) at the surface of textiles (monitored by surface analysis characterization methods) and their antibacterial activity was assessed. After contact with antimicrobial textiles, the enumeration of bacteria was performed either by plate counting or by direct observation by epifluorescence microscopy in the presence of fluorescent viability markers in order to determine whether viable but nonculturable bacterial cells were present.

Natural starters have been extensively used for many centuries to make many different fermented food products from different raw materials: Milk, meat, roots, vegetables, etc. The industrialisation of food production at the end of the 19th century necessitated the use of regular selected starters to standardize the organoleptic characteristics of the final product. As a consequence, during the 20th century, there was a decline in the use of natural starters in Western countries except in the production of local cheeses or sourdough breads. The beginning of this new millennium has witnessed a deep change in consumer demand, in pursuit of quality, safety and pleasure. In this context, natural starters could, in the future, play an important role in the development of fermented products. However, food producers and researchers have first to cope with fundamental problems in the understanding of these complex ecosystems. The dynamic evolution of the microbial population inside the natural starter (its resilience, its genetic and physiological aptitudes) and the consequences on the product are still partially unknown. This document reviews a broad range of articles concerning the use of natural starters with a specific focus on cheeses and breads, and discusses the major stakes for local food production and the consumption of typical products.

The purpose of the present study was to investigate the effect of increasing sodium caseinate (CAS) of low methoxyl pectin (LMP) based-film formulations as well as pH on the release kinetics of incorporated natamycin (45 mg/L of film-forming suspension) and the antimicrobial activity of films against two fungal strains. The diffusion coefficient values of natamycin from composite films to imidazole-acetate buffer (pH 7.0) were shown to depend on both protein/polysaccharide ratio and pH of formed films. Diffusion agar tests showed that films containing natamycin were more effective in inhibiting the growth of Penicillium chrysogenum than Aspergillus brasiliensis incubated at 30°C during 4 days. Also, active films were able to reduce the growth of P. chrysogenum voluntarily inoculated on a cured meat model with the highest spore count reduction obtained for composite film with CAS/LMP ratio of 1.00 prepared at pH 3.0. Practical applications Natamycin has been safely used as a natural mold inhibitor for the surface treatment of cheese, meat, and other food products due to its low solubility in water and most organic solvents. The application of natamycin in some food products can prevent the production of mycotoxins produced by molds which are known to be harmful for human health. However, the antifungal effects of natamycin can be lowered in the case of direct application in the food matrix. Thus, incorporation of natamycin in various protein/polysaccharide-based films has been investigated as an alternative in order to reduce the disadvantages of its direct addition into food products. The controlled release of antifungal agent from the film plays an important role in the surface treatment of food.

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.

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°C 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 3 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±0.3x10-12 and 4.0±1.0x10-16 m2.s-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 3 materials was suitable for a relevant controlled release packaging targeting the preservation of fresh food, but a combination of 2 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.