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Biofilm formation on abiotic surfaces has become a major public health concern because of the serious problems they can cause in various fields. Biofilm cells are extremely resistant to stressful conditions, because of their complex structure impedes antimicrobial penetration to deep-seated cells. The increased resistance of biofilm to currently applied control strategies underscores the urgent need for new alternative and/or supplemental eradication approaches. The combination of two or more methods, known as Hurdle technology, offers an excellent option for the highly effective control of biofilms. In this perspective, the use of functional enzymes combined with biosourced antimicrobial such as essential oil (EO) is a promising alternative anti-biofilm approach. However, these natural antibiofilm agents can be damaged by severe environmental conditions and lose their activity. The microencapsulation of enzymes and EOs is a promising new technology for enhancing their stability and improving their biological activity. This review article highlights the problems related to biofilm in various fields, and the use of encapsulated enzymes with essential oils as antibiofilm agents. KEY POINTS: • Problems associated with biofilms in the food and medical sectors and their subsequent risks on health and food quality. • Hurdle technology using enzymes and essential oils is a promising strategy for an efficient biofilms control. • The microencapsulation of enzymes and essential oils ensures their stability and improves their biological activities.

Biofilms consist of microbial communities enclosed in a self-produced extracellular matrix which is mainly responsible of biofilm virulence. Targeting this matrix could be an effective strategy to control biofilms. In this work, we examined the efficacy of two proteolytic enzymes, pepsin and trypsin, to degrade P. aeruginosa and E. faecalis biofilms and their synergistic effect when combined with carvacrol. The minimum dispersive concentrations (MDCs) and the contact times of enzymes, as well as the minimal inhibitory concentrations (MICs) and contact times of carvacrol, were determined against biofilms grown on polystyrene surfaces. For biofilms grown on stainless steel surfaces, the combined pepsin or trypsin with carvacrol treatment showed more significant reduction of both biofilms compared with carvacrol treatment alone. This reduction was more substantial after sequential treatment of both enzymes, followed by carvacrol with the greatest reduction of 4.7 log CFU mL−1 (p < 0.05) for P. aeruginosa biofilm and 3.3 log CFU mL−1 (p < 0.05) for E. faecalis biofilm. Such improved efficiency was also obvious in the epifluorescence microscopy analysis. These findings demonstrate that the combined effect of the protease-dispersing activity and the carvacrol antimicrobial activity could be a prospective approach for controlling P. aeruginosa and E. faecalis biofilms. This work was supported by the University of Lille, CNRS, INRAE, Centrale Lille, UMET-Unité Matériaux et Transformations under Grant [UMR 8207].

Environmental conditions in food and medical fields enable the bacteria to attach and grow on surfaces leading to resistant bacterial biofilm formation. Indeed, the first step in biofilm formation is the bacterial irreversible adhesion. Controlling and inhibiting this adhesion is a passive approach to fight against biofilm development. This strategy is an interesting path in the inhibition of biofilm formation since it targets the first step of biofilm development. Those pathogenic structures are responsible for several foodborne diseases and nosocomial infections. Therefore, to face this public health threat, researchers employed cold plasma technologies in coating development. In this review, the different factors influencing the bacterial adhesion to a substrate are outlined. The goal is to present the passive coating strategies aiming to prevent biofilm formation via cold plasma treatments, highlighting antiadhesive elaborated surfaces. General aspects of surface treatment, including physico-chemical modification and application of cold plasma technologies, were also presented. Key points • Factors surrounding pathogenic bacteria influence biofilm development. • Controlling bacterial adhesion prevents biofilm formation. • Materials can be coated via cold plasma to inhibit bacterial adhesion.

The biofilm lifestyle plays a major role in the resistance and virulence of

In food industries, microbial contaminations are difficult to control due to the recurrent formation of biofilms that hinders antimicrobials penetration and efficiency. An understanding of Salmonella Enteritidis biofilms behavior under flow conditions is a key to develop efficient preventive and control strategies. S. Enteritidis biofilms displayed 5.96, 6.28 and 6.80 log CFU cm

Made available in DSpace on 2021-06-25T10:56:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-04-01 In this study, we evaluated the effect of complexation and microencapsulation with pea protein on the antioxidant activity of protein hydrolysates from bycatch in Brazil. The zeta potential values of complexes changed from negative to positive with the increase of pea protein as a result of positively charged complexes formation. The increase in the ratio of pea protein/hydrolysates also resulted in increased turbidity in all samples. Particle size measurements indicated that the complexes tended to form larger aggregates (ranged from 61.5 ± 1.7 μm to 183 ± 2.8 μm). The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of the pea protein/fish protein hydrolysate complexes was higher than that of the protein hydrolysates alone. Moreover, increasing levels of pea protein did not affect the antioxidant activity of fish protein hydrolysates. The complexes of the Paralonchurus brasiliensis were chosen for the microencapsulation process by spray-drying. The results revealed that spray-drying did not have a significant effect (P > 0.05) on the protein hydrolysate antioxidant activity when they were complexed with pea protein. Thus, this work suggests that the complexation with pea protein and subsequent microencapsulation by spray-drying is an efficient way to protect the biological activity of protein hydrolysates obtained from bycatch. This study provides evidence for the potential use of bycatch from shrimp fisheries as functional ingredients or nutraceuticals. São Paulo State University (UNESP), Aquaculture Center, Campus Jaboticabal UMR 8207 - UMET - Unité Matériaux Et Transformations Univ Lille CNRS INRAE Centrale Lille LAGEPP UMR 5007 Univ Lyon Université Claude Bernard Lyon 1 CNRS, 43 Bd 11 Novembre 1918 São Paulo State University (UNESP), Aquaculture Center, Campus Jaboticabal

International audience; This study aimed to improve the physical, mechanical, and biological properties of a monolayer pectin (P) film containing nanoemulsified trans-Cinnamaldehyde (TC) by incorporating it between inner and outer layers of ethylcellulose (EC). The nanoemulsion had an average size of 103.93 nm and a zeta potential of −46 mV. The addition of the nanoemulsion increased the opacity of the film, reduced its moisture absorption capacity, and improved its antimicrobial activity. However, the tensile strength and elongation at break of the pectin films decreased after the incorporation of nanoemulsions. Multilayer films (EC/P/EC) showed a higher resistance to breaking and better extensibility compared to monolayer films. The antimicrobial activity of both mono and multilayer films was effective in inhibiting the growth of foodborne bacteria during storage of ground beef patties at 8 °C for 10 days. This study suggests that biodegradable antimicrobial multilayer packaging films can be effectively designed and applied in the food packaging industry.; Cette étude visait à améliorer les propriétés physiques, mécaniques et biologiques d'un film monocouche de pectine (P) contenant du trans-cinnamaldéhyde (TC) nanoémulsifié en l'incorporant entre les couches internes et externes d'éthylcellulose (EC). La nanoémulsion avait une taille moyenne de 103,93 nm et un potentiel zêta de -46 mV. L'ajout de la nanoémulsion a augmenté l'opacité du film, réduit sa capacité d'absorption de l'humidité et amélioré son activité antimicrobienne. Cependant, la résistance à la traction et l'élongation à la rupture des films de pectine ont diminué après l'incorporation des nanoémulsions. Les films multicouches (EC/P/EC) ont montré une plus grande résistance à la rupture et une meilleure extensibilité que les films monocouches. L'activité antimicrobienne des films monocouches et multicouches a permis d'inhiber la croissance des bactéries d'origine alimentaire pendant le stockage de galettes de bœuf haché à 8 °C pendant 10 jours. Cette étude suggère que les films d'emballage multicouches biodégradables et antimicrobiens peuvent être conçus et appliqués efficacement dans l'industrie de l'emballage alimentaire.

International audience; Nisin is a small peptide produced byLactococcus lactisssplactisthat is currently industrially produced. This preservative is often used for growth prevention of pathogenic bacteria contaminating the food products. However, the use of nisin as a food preservative is limited by its low production during fermentation. This low production is mainly attributed to the multitude of parameters influencing the fermentation progress such as bacterial cells activity, growth medium composition (namely carbon and nitrogen sources), pH, ionic strength, temperature, and aeration. This review article focuses on the main parameters that affect nisin production byLactococcus lactisbacteria. Moreover, nisin applications as a food preservative and the main strategies generally used are also discussed.

Essential oils active components emerged as captivating antimicrobials to control biofilms developed on food contact surfaces. Free and nanoencapsulated carvacrol (CAR) and thymol (THY) were assessed as antimicrobials against Salmonella Enteritidis biofilms adhered to stainless steel. The developed spherical nanocapsules using the spray-drying technique showed an average size ranged between 159.25 and 234.76 nm and zeta potential values ranged between 23.60 and 24.66 mV. The minimal inhibitory concentrations (MIC) of free THY and CAR were both 1.25 mg L-1. Nanoencapsulation reduced MIC values to 0.62 mg L-1 (THY) and 0.31 mg L-1 (CAR). Furthermore, the exposure to free and nanoencapsulated CAR and THY induced a destabilization of bacterial membranes with obvisous morphological deformations and a pronounced leakage of potassium ions and green fluorescent proteins. Eradication of S. Enteritidis biofilms developed on stainless steel was achieved following a 15 min treatment with nanoencapsulated CAR and THY at 2 MIC. Free antimicrobial exposures induced up to 4.27 log CFU cm(-2) reductions. Additionally, the ecotoxicity tests against Daphnia magna crustaceans reported a non-toxicity of both free and nanoencapsulated CAR and THY after 48 h exposure. Thereby, both CAR and THY antimicrobials proved to be promising natural surface disinfectants that require further exploration and incorporation in food industries.

Mycosubtilin belongs to the family of lipopeptides. Different isoforms with various antifungal activities can be obtained according to the length and the isomery of the fatty acid. In this work, the activities of the mycosubtilin isoforms were first studied against the pathogen

International audience; Pathogenic bacterial biofilms invading surfaces in food and medical fields are a challenge to overcome. Despite all the strategies applied to fight their formation, the microbiological risk associated to bacterial biofilms remains an important threat for at risk population and for food and healthcare sectors. The prevention of biofilm formation might be an effective approach to confront this problem. In this study, stainless steel surfaces were functionalized by nisin, a natural antimicrobial peptide. The mechanism of action of immobilized nisin against sensitive bacteria is not fully understood. Therefore, nisin was grafted onto the surface by either its carboxylic group or its amino group. The generated coating's chemical, topographical and antibacterial properties were studied to understand the nisin mode of action, when immobilized, and identify the section of the bacteriocin responsible for the antimicrobial activity. The antimicrobial activity of the elaborated coatings was tested against Listeria monocytogenes. Indeed, the surfaces coated with nisin linked by its amino group showed an efficient antibacterial activity while the surface with nisin linked by its carboxylic group showed less antimicrobial effect. The antimicrobial results showed almost 2 log reduction of colony forming units for efficient antibacterial coatings while the other showed no bacterial reduction. The surface properties analysis permitted to understand the chemical and topographical characteristics of treated surfaces including nisin conformation and quantification. A tight relation was concluded between the surface topography, the nisin conformation, and the antibacterial activity of the bacteriocin-coated surfaces.

Microbial contaminations are responsible for many chronic, healthcare, persistent microbial infections and illnesses in the food sector, therefore their control is an important public health challenge. Over the past few years, essential oils (EOs) have emerged as interesting alternatives to synthetic antimicrobials as they are biodegradable, extracted from natural sources and potent antimicrobials. Through their multiple mechanisms of actions and target sites, no microbial resistance has been developed against them till present. Although extensive documentation has been reported on the antimicrobial activity of EOs, comparisons between the use of whole EOs or their active components alone for an antimicrobial treatment are less abundant. It is also essential to have a good knowledge about EOs to be used as alternatives to the conventional antimicrobial products such as chemical disinfectants. Moreover, it is important to focus not only on planktonic vegetative microorganisms, but to study also the effect on more resistant forms like spores and biofilms. The present article reviews the current knowledge on the mechanisms of antimicrobial activities of EOs and their active components on microorganisms in different forms. Additionally, in this review, the ultimate advantages of encapsulating EOs or combining them with other hurdles for enhanced antimicrobial treatments are discussed. This work was supported by the Partenariat Hubert Curien (PHC)- Cèdre program [42281SD] and the open access funding was provided by the Qatar National Library .

Bioactive aminooxime ligands based on optically pure (R)-limonene have been synthesized in two steps. Their ruthenium (II) cationic water-soluble complex was prepared by a reaction between dichloro (para-cymene) ruthenium (II) dimers and aminooxime ligands in a 1:2 molar ratio. Antibacterial and antibiofilm activities of the synthetized complex were assessed against Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis. The results revealed that the ruthenium (II) complex has higher antibacterial and antibiofilm activities in comparison with free ligands or the enantiopure (R)-limonene. Moreover, microencapsulation of this complex reduced its cytotoxicity and improved their minimum inhibitory concentration and antibiofilm activity toward the considered bacteria. The ruthenium (II) complex targets the bacterial cell membrane, which leads to rapid leakage of intracellular potassium. Our study suggests that the developed ruthenium (II) complexes could be useful as an alternative to conventional disinfectants.

International audience; Emerging technology regarding antimicrobial coatings contributes to fighting the challenge of pathogenic bacterial biofilms in medical and agri-food environments. Stainless steel is a material widely used in those fields since it has satisfying mechanical properties, but it, unfortunately, lacks the required bio-functionality, rendering it vulnerable to bacterial adhesion and biofilm formation. Therefore, this review aims to present the coatings developed by employing biocides grafted on stainless steel. It also highlights antimicrobial peptides (AMPs)used to coat stainless steel, particularly nisin, which is commonly accepted as a safe alternative to prevent pathogenic biofilm development.

International audience; The aim of this work is to characterize the potential probiotic properties of 14 antifungal Lactobacillus strains isolated from traditional fermenting Moroccan green olives. The molecular identification of strains indicated that they are composed of five Lactobacillus brevis, two Lactobacillus pentosus, and seven Lactobacillus plantarum. In combination with bile (0.3%), all the strains showed survival rates (SRs) of 83.19–56.51% at pH 3, while 10 strains showed SRs of 31.67–64.44% at pH 2.5. All the strains demonstrated high tolerance to phenol (0.6%) and produced exopolysaccharides. The autoaggregation, hydrophobicity, antioxidant activities, and surface tension value ranges of the strains were 10.29–41.34%, 15.07–34.67%, 43.11–52.99%, and 36.23–40.27 mN/m, respectively. Bacterial cultures exhibited high antifungal activity against Penicillium sp. The cell-free supernatant (CFS) of the cultures showed important inhibition zones against Candida pelliculosa (18.2–24.85 mm), as well as an antibacterial effect against some gram-positive and gram-negative bacteria (10.1–14.1 mm). The neutralized cell-free supernatant of the cultures displayed considerable inhibitory activity against C. pelliculosa (11.2–16.4 mm). None of the strains showed acquired or horizontally transferable antibiotic resistance or mucin degradation or DNase, hemolytic, or gelatinase activities. Lactobacillus brevis S82, Lactobacillus pentosus S75, and Lactobacillus plantarum S62 showed aminopeptidase, β-galactosidase, and β-glucosidase activities, while the other enzymes of API-ZYM were not detected. The results obtained revealed that the selected antifungal Lactobacillus strains are considered suitable candidates for use both as probiotic cultures for human consumption and for starters and as biopreservative cultures in agriculture, food, and pharmaceutical industries.

Biofilms are involved in serious problems in medical and food sectors due to their contribution to numerous severe chronic infections and foodborne diseases. The high resistance of biofilms to antimicrobial agents makes their removal as a big challenge. In this study, spray-drying was used to develop microcapsules containing carvacrol, a natural antimicrobial agent, to enhance its activity against P. aeruginosa and E. faecalis biofilms. The physicochemical properties and microscopic morphology of the realized capsules and cells were characterized. The minimum inhibitory concentration of encapsulated carvacrol (E-CARV) (1.25 mg mL-1) was 4-times lower than that of free carvacrol (F-CARV) (5 mg mL-1) against P. aeruginosa, while it remained the same against E. faecalis (0.625 mg mL-1). E-CARV was able to reduce biofilm below the detection limit for P. aeruginosa and by 5.5 log CFU ml-1 for E. faecalis after 15 min of treatment. Results also showed that F-CARV and E-CARV destabilize the bacterial cell membrane leading to cell death. These results indicate that carvacrol exhibited a strong antimicrobial effect against both bacterial biofilms. In addition, spray-drying could be used as an effective tool to enhance the antibiofilm activity of carvacrol, while reducing the concentrations required for disinfection of abiotic surfaces.

Bioactive aminooxime ligands based on optically pure (R)-limonene have been synthesized in two steps. Their ruthenium (II) cationic water-soluble complex was prepared by a reaction between dichloro (para-cymene) ruthenium (II) dimers and aminooxime ligands in a 1:2 molar ratio. Antibacterial and antibiofilm activities of the synthetized complex were assessed against Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis. The results revealed that the ruthenium (II) complex has higher antibacterial and antibiofilm activities in comparison with free ligands or the enantiopure (R)-limonene. Moreover, microencapsulation of this complex reduced its cytotoxicity and improved their minimum inhibitory concentration and antibiofilm activity toward the considered bacteria. The ruthenium (II) complex targets the bacterial cell membrane, which leads to rapid leakage of intracellular potassium. Our study suggests that the developed ruthenium (II) complexes could be useful as an alternative to conventional disinfectants.

The persistence of

International audience; The persistence of Salmonella enterica on abiotic surfaces in hospitals and the agri-food industries leads to several infections worldwide. In this context, this work aimed to study the adhesion of S. Enteritidis on plasma-modified stainless steel to prevent biofilm-associated-infections. Surface modification was achieved by the elaboration of organosilicon coatings from the monomer 1,1,3,3-tetramethyldisiloxane, mixed with oxygen, using a flowing nitrogen microwave post-discharge plasma polymerization technique. The effect of cold plasma parameters on the properties of the coatings, the coated surface topography and S. Enteritidis cell adhesion was studied. The results showed that the surface topography influenced the bacterial adhesion rate. Indeed, rough surfaces did not repel S. Enteritidis since the number of attached cells on these coatings was between 30 ± 4 to 65 ± 4 bacteria per microscopic field. Otherwise, smoother surfaces demonstrated an anti-adhesive character since the number of attached cells was almost nil on these coatings.

Dodecyltrimethylammonium chloride (DTAC) was trapped into maltodextrins/pectin spray dried microcapsules to improve its activity against

International audience; The biofilm formation on food-contact-surfaces is a serious threat for public health and often results in huge human and economic losses. Thus, it is of importance to develop new strategies that allow the monitoring of the biofilm formation on food-contact-surfaces. Such strategies permit effective and timely countermeasures and, therefore, to avoid the microbiological risk and associated environmental impact due to cleaning processes. Therefore, this work investigated a non-invasive method for monitoring and detection of the biofilm formation using Coda Wave Interferometry (CWI) method. The principle of this method and the calculation of decorrelation coefficient based on the ultrasonic measurements were explained. The results underlined that our developed method is able to detect the early stage of the biofilm formation of Staphylococcus aureus on the stainless steel. Namely, it is shown that the bacterial cell kinetic is well captured by the evolution of the decorrelation coefficient as a function of incubation times. Overall, this work showed that this cost effective CWI method is a promising tool for the detection of early stage biofilm formation on food-contact-surfaces.

International audience; Dodecyltrimethylammonium chloride (DTAC) was trapped into maltodextrins/pectin spray dried microcapsules to improve its activity against Salmonella enteritidis and Staphylococcus aureus biofilms. Two different microcapsules were prepared: uncomplexed DTAC-microcapsules (UDM), containing DTAC and maltodextrins; and complexed DTAC-microcapsules (CDM) containing DTAC complexed with pectin and maltodextrins. The minimum inhibitory concentrations (MIC) of both free and microencapsulated DTAC were investigated against S. Enteritidis and S. aureus. The MICs of DTAC were significantly lower when encapsulated. CDM treatment resulted in a 2 and 3.2 log reduction in S. aureus and S. Enteritidis biofilm culturable biomass, respectively. Microencapsulation reduced the cytotoxicity of DTAC by up to 32-fold. Free DTAC and CDM targeted the cell membrane resulting in the leakage of the intracellular molecules and subsequent cell death. The development of DTAC microcapsules reduced the amount of DTAC required to maintain the high standards of cleanliness and hygiene required in the food processing industries.

International audience; Aims: In this study, benzalkonium chloride (BAC) microcapsules were developed for surface disinfection purpose and were evaluated against Listeria monocytogenes and Escherichia coli biofilms.Methods and Results: Microcapsules were prepared with two different strategies: uncomplexed BAC‐microcapsules (UBM), containing BAC and maltodextrins and complexed BAC‐microcapsules (CBM) containing BAC complexed by pectin and maltodextrins. The minimum inhibitory concentrations (MICs) of free and microencapsulated BAC was investigated against two food pathogens: L. monocytogenes and E. coli. The antibiofilm activities of UBM and CBM against L. monocytogenes and E. coli biofilms formed on stainless steel at 37°C were evaluated and compared to BAC used under its free form. MICs of encapsulated BAC were up to 4‐fold lower than those of free BAC. The UBM and CBM showed higher antibiofilm effect when compared to the free BAC.Conclusions: Overall, results demonstrated that microencapsulation enhanced the antibacterial activity of BAC against L. monocytogenes and E. coli biofilms.Significance and Impact of the Study: The application of such BAC microcapsule‐based delivery systems can improve surface disinfection procedures and reduce the required BAC concentrations and the related cytotoxicity of this antimicrobial compound.

International audience; Front-face fluorescence spectroscopy technique coupled with chemometric tools was used for predicting the freshness state of a Lebanese dairy product called Qishta, stored up to 20 days. Acid, peroxide and thiobarbituric acid reactive substances (TBARS) values reached no more than 0.93 mg NaOH g−1 fat, 6.22 meq O2 Kg−1 fat, and 0.0313 mg malonaldehyde (MA) kg−1 sample, respectively, throughout the investigated storage time. In parallel, fluorescence emission spectra of tryptophan and riboflavin, and fluorescence excitation spectra of vitamin A were recorded and showed the highest fluorescence intensity for the Qishta samples aged of 20 days and the lowest intensity for the fresh ones. The primary and secondary indicators of lipid oxidation showed that Qishta can be stored for 20 days without any alteration despite the increase in the TBARS after 16 days of storage. Principal component analysis (PCA) applied on riboflavin emission spectra allowed better discrimination between Qishta samples with a clear distinction of those aged 20 days while some overlapping was noticed between samples aged below 16 days. A high correlation (R2 = 0.923) was observed between the peroxide value and the intensity of the riboflavin fluorescence recorded at 460 nm.

This study aims at exploring the chemical composition of a traditional Lebanese dairy product known as Qishta, describing the process of how to prepare it and understanding the mechanisms leading to its formation. The process of making Qishta can be divided into two phases: a hot phase during which milk is heated in a stainless-steel large shallow vessel, and a cold phase consisting of draining, cooling and packaging. According to milk temperature, two reaction zones were identified: zone A with an average temperature of 100 °C, and zone B with an average temperature of 60 °C. The results showed that Qishta had a moisture, fat, protein, lactose and ash content of 68%, 11.7%, 12.1%, 5.4% and 1.6%, respectively. Our findings showed that Qishta is a lipoprotein product having an equal amount of fat and proteins (≈12%); this composition is almost similar to that of Ricotta cheese made from whole milk. In addition, our results assert that the interactions between caseins and whey proteins lead to gel formation. Milk initial fat percentage had a significant effect on Qishta production. The highest yields were obtained when the initial fat percentage was 3.6% (182.5 g of Qishta).

International audience; The objective of this work is to study the oleuropein (OLP) biodegradation by a strain of Lactobacillus plantarum FSO175 under conditions (pH 4.5, NaCl 5% and glucose 1%), during 7 days of incubation at 30 °C, in modified MRS broth containing OLP as carbon source. The results obtained, by HPLC analyses, showed that the biodegradation of OLP by L. plantarum FSO175, is accompanied with increase of hydroxytyrosol (HT) content and acidity values. The yields of OLP degradation and HT accumulation are depending on carbon sources, stress conditions and time of incubation. So that, the drastic reduction in OLP biodegradation, obtained at pH 6.7 (trial B), decreased significantly (p

International audience; Mycosubtilin belongs to the family of lipopeptides. Different isoforms with various antifungal activities can be obtained according to the length and the isomery of the fatty acid. In this work, the activities of the mycosubtilin isoforms were first studied against the pathogen Aspergillus niger, revealing the high activity of the anteiso-C17 isoform. Modification of the mycosubtilin isoform patterns during cultures of the natural strain Bacillus subtilis ATCC 6633 was then investigated through amino acids feeding experiments. In parallel, single gene knockouts and single gene overexpression leading to the overproduction of the anteiso-C15 fatty acid chains were predicted using informatics tools which provide logical reasoning with formal models of reaction networks. In this way, it was in silico predicted that the single overexpression of the ilvA gene as well as the single knockout of the codY gene may lead to the overproduction of anteiso-C15 fatty acid chains. For the first time, it has been demonstrated that overexpression of ilvA allows an enhancement of furniture of odd anteiso fatty acids leading to a favoured mycosubtilin anteiso-C17 production pattern (+41%). Alternatively, a knock-out codY mutant led to a higher furniture of even iso fatty acids, leading to a favoured mycosubtilin iso-C16 production pattern (+180%). These results showed that increased selective synthesis of particular isoforms of mycosubtilin through metabolic engineering is feasible, disclosing the interest of these approaches for future development of lipopeptide producing strains.

International audience; Protein–protein cross-linking and protein-fat interactions in the traditional Lebanese dairy product, Qishta, were investigated. Liquid chromatography coupled to mass spectrometry was used for the detection of lysinoalanine (LAL) and lanthionine (LAN) during the production of Qishta, which is made by heating whole milk in an open shallow vessel for more than 2 h and harvesting the surface aggregates formed. LAL and LAN cross-links were seen in Qishta at concentrations higher than those in milk, indicating their importance in the gel formation and therefore their impact on Qishta texture. Disulphide bridges were also involved in Qishta formation. The amino acid residues (cysteine, serine or threonine) involved in the β-elimination and in dehydroalanine formation were identified. Confocal scanning laser microscopy showed the role of fat globules in the gel structure.

Salmonella is a pathogen transmitted by foods and it is one of the most important target bacteria in food irradiation studies. Few works were carried out on the effectiveness of gamma radiation against biofilms formed by this bacterium. Salmonella can form a biofilm on different material surfaces. The physicochemical properties of surfaces and environmental factors influence the adhesion of this pathogen. The present study investigated the effect of gamma radiation (1 and 2 kGy) and temperature (28°C and 37°C) on the development of Salmonella Hadar biofilm on polyvinyl chloride (PVC), glass, cellophane paper (CELLO), and polystyrene (POLY). The obtained results indicated that biofilm production is surface and temperature dependent. In addition, biofilm formation decreased significantly after gamma irradiation at either 1 or 2 kGy doses. However, the agfD and adrA genes expression did not demonstrate significant decrease. This work highlighted that gamma radiation treatment could reduce the biofilm formation of Salmonella enterica serovar Hadar on different food contact surfaces.

International audience; Objective: This work aims to evaluate the impact of the use of ultrasound during essential oils extraction from garlic bulbs. Material and methods: The essential oils were extracted by Conventional Hydrodistillation method (HD), Ultrasound-Assisted hydro-distillation (US-HD) and Sono-hydrodistillation (SHD) and compared using yield profile, chemical composition and antioxidant activity. Results: Higher performances were obtained when the direct application of ultrasound were used. Indeed, SHD allowed a higher yield after only 30 min of extraction time against 70 min and 90 min for the US-HD and the HD respectively. Among the total identified compounds by GC/MS, Sulphur compounds were found to be the most abundant with qualitative similarities for the three processes. Essential oils showed a potential DPPH radical scavenging activity with IC50 values of 0.96, 1.17 and 1.23 respectively for the three HD, US-HD and SHD processes. Conclusion: The use of ultrasound for the extraction of essential oil from garlic reduced extraction time without causing significant changes in volatile composition and antioxidant activity.

Citral-in-water emulsions were prepared with two different essential oil concentrations of 2.5 and 5.0% (w/w), then spray-dried in the presence of the same amount of maltodextrins (20%). The microcapsules were prepared with two different emulsifier compositions: monolayer microcapsules (ML) stabilized by sodium caseinate alone and layer-by-layer microcapsules (LBL) stabilized by sodium caseinate and pectin. The encapsulation efficiency was higher for LBL microcapsules (e.g. 99.6 ± 0.4% for 2.5% citral) than that for ML ones (e.g. 78.6 ± 0.6% for 2.5% citral) which confirm that the additional pectin layer was able to protect citral during the spray-drying process whatever citral concentration. Furthermore, our results showed that the antibacterial activity of the obtained microcapsules significantly depends on both citral concentration and interfacial membrane composition. The presence of two layers surrounding the citral droplets may result in a progressive and controlled release of the encapsulated citral.

The aim of the present work was to study and compare the effect of growth temperature (20, 30, and 37 degrees C) and surface type (stainless steel and polycarbonate) on the production of virulence factors, such as proteases and siderophores, and the risk of surface contamination associated with Pseudomonas aeruginosa biofilm and planktonic cells. The increase of growth temperature from 20 to 37 degrees C increased (approximately twofold) the electronegative charge and the hydrophobicity of the P. aeruginosa biofilm cell surface. P. aeruginosa biofilm cell adhesion to stainless steel and polycarbonate was 5- and 1.5-fold higher than their planktonic counterparts at 20 and 30 degrees C, respectively. The increase of growth temperature from 20 to 37 degrees C increased the production of proteases (twofold) and siderophores (twofold) and the cytotoxicity (up to 30-fold) against the HeLa cell line in the supernatants of P. aeruginosa planktonic and biofilm cultures. This study also highlighted that biofilm and planktonic P. aeruginosa cells exhibited distinct physiological properties with respect to the production of virulence factors and the cytotoxicity against the Hela cell line. Therefore, effective disinfection procedures should be adapted to inactivate bacteria detached from biofilms.

The aim of the present work was to study and compare the effect of growth temperature (20, 30, and 37°C) and surface type (stainless steel and polycarbonate) on the production of virulence factors, such as proteases and siderophores, and the risk of surface contamination associated with

International audience; Bacteriocins from lactic acid bacteria (LAB) are successfully applied as natural alternatives to food preservation and to antibiotics; however, information on these antimicrobial peptides (AMPs) is scattered through the literature and databases. Therefore, we developed the LABiocin database, a specialized database on LAB bacteriocins. The database was stored and compiled using MySQL with NetBeans IDE as the platform. Important data are compiled, including bacteriocin name, class, amino acids and nucleic acid sequences, if available. Target microorganisms, origin, status of the producing strains and their culture conditions and extraction and purification methods are also included in this new database. A phylogenetic tree for the mature peptide bacteriocin sequences has also been created.LABiocin is an interactive database with a user-friendly interface that integrates several tools and services and comprises up to 517 LAB bacteriocins. Besides data searching tools, a BLAST tool was integrated into the database to enable the user to perform a homology search against mature peptide sequences. Users can be linked to other databases that contain additional information, particularly about predicted bacteriocin structure and mechanisms of action.The LABiocin database enables comprehensive functional analysis of this special group of AMPs. This would be useful in food preservation and food safety applications and would also have substantial implications for development of new drugs for medical use. LABiocin database is available at labiocin.net.

International audience; The present study investigated and compared the effect of growth temperature on the susceptibility of biofilm-detached andplanktonic Staphylococcus aureus cells, to benzalkonium chloride (BAC). This study also highlights the impact of BAC on the bacterial physiology and the role of membrane fluidity regulation as a bacterial resistance mechanism. The minimum inhibitory concentration of BAC was characterized with micro-dilution growth inhibition assay. The BAC treatment was performed on S. aureus cultured at 20 degrees C and 37 degrees C, for 24h. The morphology of S. aureus cells was examined using scanning electron microscopy. The loss of bacterial membrane integrity after BAC treatment was studied by monitoring the intracellular potassium ion leakage using the atomic absorption spectroscopy. The bacterial membrane total fatty acid composition, controlling the membrane fluidity, was analyzed by GC/MS. The results showed that the resistance of S. aureus cells to BAC increased with the increase of growth temperature. The planktonic cells were more susceptible to BAC than biofilm-detached ones. The rise of growth temperature resulted in an increase of S. aureus membrane rigidity. Furthermore, a higher membrane fluidity was observed in planktonic cells when compared to that in the biofilm-detached ones. The resistance of S. aureus seems to depend on the growth temperature. Compared to planktonic cells, biofilm-detached cells showed a greater resistance to BAC. The BAC targets and disturbs the bacterial membrane. Membrane fluidity modulation is likely a one of resistance mechanisms for S. aureus to BAC at the cellular scale. Therefore, disinfection procedures, in food sector, should be adapted for bacteria detached from biofilm.

International audience; This study investigated the effect of the growth temperature (20 and 37 degrees C) of Escherichia coli strains isolated from pigs on their adhesion to stainless steel and polycarbonate. This study also evaluated the ability of the DLVO and XDLVO mathematical models to predict this adhesion. The rise of growth temperature from 20 to 37 degrees C significantly influenced the adhesion of studied E. coli strains. The data also underlined that the mathematical prediction did not fully match with the experimental bacterial adhesion to surfaces. Furthermore, results showed that the colistin-resistant and sensitive E. coli strains adhesion depends on the type of abiotic surface. Based on these results, the mathematical models are limited in the prediction of the bacterial adhesion to abiotic surfaces. The surface roughness is a major parameter of the bacterial adhesion and should be included in the future mathematical models predicting the bacterial adhesion.

This study aims to describe interactions that may occur between nisin and bacterial exopolysaccharides (EPS) when they are present together in a model solution. For this purpose, two EPS, namely EPS-GM and EPS-BMS, produced by potential probiotic Bacillus tequilensis-GM and Leuconostoc citreum-BMS, respectively, were used. Results showed that maximal complexation occurred at pH 5.0 between anionic EPS (1.5 g/L) and cationic nisin (1 g/L) mainly through electrostatic interactions. Results provided by scanning electron microscopy, epifluorescence microscopy and turbidity measurements showed that EPS-GM was able to form bigger and higher amounts of complexes with nisin. The nisin antimicrobial activity was interestingly not affected and even improved after its complexation with EPS suggesting that these latter may act as protective agents of the nisin antimicrobial activity when applied in foods. The results of this study should help food technologists to optimize the physicochemical conditions for optimal formulation of food products containing EPS.

International audience; This work aims at studying the impact of silane-based coatings, generated by atmospheric pressure plasma spraying (APPS) of a liquid precursor (hexamethydisiloxane – HMDSO), on dairy antifouling and antibacterial properties of food-grade 316L stainless steel. The influence of the manufacturing parameters (precursor flow rate, nozzle-to-substrate distance and scanning speed) on the coatings properties was investigated using a wide range of characterization techniques (drop shape analysis, X-Ray Mappings, Scanning Electron Microscopy, Fourier-Transformed Infrared Spectroscopy, Atomic Force Microscopy). Coating’s roughness in particular was shown to strongly increase when precursor flow rate increases. A pilot pasteurizer, fed with a model foulant solution (whey protein and calcium), allowed performing two consecutive industrial-like isothermal dairy fouling tests, revealing the promising anti-fouling properties of the HMDSO-coated steel. A fouling reduction of up to 90% compared to bare stainless steel was achieved after first cycle for all samples. The second fouling run allowed to select the best-performing sample, which kept the same antifouling properties as in the first test. Its mechanism of action was investigated, which revealed that a nanostructured, Si-O-Si rich surface was efficient to prevent isothermal dairy fouling. The adhesion of the pathogenic bacterium Staphylococcus aureus also proved to be impacted by this plasma coating, with a significant decrease of adhered cells (−30% compared to native stainless steel).

International audience; Bacteriocins are antimicrobial peptides produced by bacteria Gram-negative and Gram-positive, including lactic acid bacteria (LAB), organisms that are traditionally used in food preservation practices. Bacteriocins have been shown to have an aptitude as biofilm controlling agents in Listeria monocytogenes biofilms, a major risk for consumers and the food industry. Biofilms protect pathogens from sanitization procedures, allowing them to survive and persist in processing facilities, resulting in the cross-contamination of the end products. Studies have been undertaken on bacteriocinogenic LAB, their bacteriocins, and bioengineered bacteriocin derivatives for controlling L. monocytogenes biofilms on different surfaces through inhibition, competition, exclusion, and displacement. These alternative strategies can be considered promising in preventing the development of resistance to conventional sanitizers and disinfectants. Bacteriocins are "friendly" antimicrobial agents, and with high prevalence in nature, they do not have any known associated public health risk. Most trials have been carried out in vitro, on food contact materials such as polystyrene and stainless steel, while there have been few studies performed in situ to consolidate the results observed in vitro. There are strategies that can be employed for prevention and eradication of L. monocytogenes biofilms (such as the establishment of standard cleaning procedures using the available agents at proper concentrations). However, commercial cocktails using alternatives compounds recognized as safe and environmental friendly can be an alternative approach to be applied by the industries in the future.

Background: Mycobacterium spp. are responsible for several diseases, particularly in immunocompromised populations. The spreading of the resistance to antimycobacterial drugs is a significant problem to the public health and requires to find out a new and innovative alternative for the treatment of drug resistant mycobacterial strains. In this study, the antimycobacterial activity of Micromeria barbata, Eucalyptus globulus and Juniperus excelsa essential oils extracted from Lebanese plants was investigated against selected Mycobacterium spp. strains. Methods: Several dilutions of the three aforementioned essential oils were studied for antimycobacterial activity against four Mycobacterium spp. strains: Mycobacterium tuberculosis subsp. tuberculosis (ATCC® 27294™), multidrug-resistant M. tuberculosis (CMUL 157), Mycobacterium kansasii Hauduroy (ATCC® 12478™) and Mycobacterium gordonae Bojalil et al. (ATCC® 14470™). Results: Even with high dilutions, all tested essential oils showed a high antimycobacterial activity against targeted strains. Our data showed that M. barbata, E. globulus and J. excelsa essential oils totally inhibit the mycobacterial growth whatever the tested strains for the dilution 1/250, 1/100 and 1/250, respectively. Conclusion: To our knowledge, this is the first study regarding the antimycobacterial activity of essential oils in Lebanon. Our data show promising results, and encourage to investigate more on these medicinal plants, especially M. barbata. Keywords: Essential oils, Antimycobacterial activity, Micromeria barbata, Eucalyptus globulus, Juniperus excelsa

Introduction: The raw goat milk is considered as a good source of novel bacteriocinogenic lactic acid bacteria (LAB) strains. Method: The bacteriocin, which named enterocin CMUL20-2 was secreted by Enterococcus faecium CMUL20-2. This bacterial strain was originally isolated from raw goat’s milk, was extracted by using a modified adsorption-desorption method and purified via RP-HPLC. antimicrobial activity was tested against several pathogenic and spoilage microbes. Results: The enterocin CMUL20-2 showed a strong adsorption on cell wall of producer strain even in acidic environment which facilitate its extraction in only two simple steps. The recovered purified enterocin has decreased procedure time and diminished the number of undesirable molecules present in Rogosa and Sharpe (MRS) broth. The recovered enterocin showed antimicrobial activity against several foodborne pathogenic and spoilage microbes. Conclusion: The recovered enterocin was able to tolerate a variety of food chain conditions such as high temperature, pH and storage stability, and it can be a good candidate to protect food from spoilage microbes

International audience; Dairy industries are burdened by fouling phenomena that increase costs and environmental impacts of thermal processes. One remedial solution could be to replace stainless-steel equipment by other materials less prone to fouling. This work studied the fouling behaviour and hygienic features of two hydrophobic and non-polar commercial graphite-based composites (Graphilor (R) XC and XTH) that were exposed to isothermal whey protein fouling in an industrial pasteurisation unit and to foodborne pathogenic bacterial strains, namely Staphylococcus aureus, Listeria monocytogenes and Salmonella enterica. Both composites exhibited impressive fouling-release properties: a simple 20 min water rinse was enough to remove all dairy deposit after 1.5 h of pasteurisation, as opposed to usual heavy clean-in-place procedures. The composites also showed a significant effect on bacterial adhesion, exhibiting lower cell counts than stainless-steel surfaces. These results suggest that graphite-based materials might successfully replace stainless steel as equipment material to reduce fouling in dairy fouling industries.

Khelissa, O., Jama, Abdallah, Boukherroub, R., Faille, C., Chihib, N.-E. (2017). Effect of incubation duration, growth temperature, and abiotic surface type on cell surface properties, adhesion and pathogenicity of biofilm-detached Staphylococcus aureus cells. AMB Express, 7 (191). , DOI : 10.1186/s13568-017-0492-0 https://prodinra.inra.fr/record/417260; International audience; Following publication of the original article (Khelissa et al. 2017), the authors reported that the legend for Fig. 3 contained an error. Instead of “Adhesion of planktonic and biofilm-detached Staphylococcus aureus cells on stainless steel and polycarbonate. Cell cultures were grown at 20, 30 and 37 °C, during 24 h (white square) and 48 h (white square). Planktonic cells adhesion on stainless steel (a) and polycarbonate (b). Adhesion of stainless steel-biofilm-detached-cells on stainless steel 24 (c) and polycarbonate-biofilm-detached-cells on polycarbonate (d)”, the legend should read “Adhesion of planktonic and biofilm-detached Staphylococcus aureus cells on stainless steel and polycarbonate. Cell cultures were grown at 20, 30 and 37 °C, during 24 h (black square) and 48 h (white square). Planktonic cells adhesion on stainless steel (a) and polycarbonate (b). Adhesion of stainless steel-biofilm-detached-cells on stainless steel 24 (c) and polycarbonate-biofilm-detached-cells on polycarbonate (d)”. The original article has been corrected.

International audience; The goal of this study was to investigate the effect of growth conditions such as the temperature (20, 30 and 37 °C), incubation duration (24 and 48 h) and surface type (stainless steel and polycarbonate) on the cell surface physicochemical properties and adhesion to abiotic surfaces of biofilm-detached and planktonic Staphylococcus aureus cells. This study tested also the hypothesis that S. aureus planktonic cells exhibit distinct pathogenic properties compared with their sessile counterparts. The results showed that the changes of the growth conditions promoted changes in the zeta potential, hydrophobicity, electron donor/acceptor character of the studied cell populations. Biofilm-detached cells showed a greater adhesion to stainless steel and polycarbonate compared with planktonic cells. Compared with planktonic cells, sessile ones showed higher cytotoxic effect against HeLa cells, DNase activity, and siderophore levels. The higher cytotoxic effect and production of DNase and siderophore increased with the increase of temperature and duration of incubations. Based on the obtained data, the S. aureusbiofilm-detached cells were found to be distinct in many physiological properties compared with their planktonic counterparts.

This study investigated the effect of growth temperature changes (20, 30, and 37°C) on the adhesion behavior of Pseudomonas aeruginosa and Staphylococcus aureus to stainless steel and polycarbonate. Adhesion assays were performed under static conditions at 20°C. In addition, the validity of the thermodynamic and extended Derjaguin, Landau, Verwey, and Overbeek theories as predictive tools of bacterial adhesion were studied. The surface properties of the bacterial cells and the substrates of attachment were characterized, and atomic force microscopy was used to analyze the surface topography. The results indicated that the highest adhesion rate of P. aeruginosa and S. aureus on both surfaces was observed when the cells were grown at 37°C. The bacterial adhesion to stainless steel was found to be two times higher than to polycarbonate for both bacteria, whatever the condition used. The present study underlined that the thermodynamic and the extended Derjaguin, Landau, Verwey, and Overbeek theories were able to partially predict the empirical results of P. aeruginosa adhesion. However, these theories failed to predict the adhesion behavior of S. aureus to both surfaces when the growth temperature was changed. The results of the microbial adhesion to solvent indicated that the adhesion rate to abiotic surfaces may correlate with the hydrophobicity of bacterial surfaces. The effect of surface topography on bacterial adhesion showed that surface roughness, even on the very low nanometer scale, has a significant effect on bacterial adhesion behavior.

Emerging resistance to antiviral agents is a growing public health concern worldwide as it was reported for respiratory, sexually transmitted and enteric viruses. Therefore, there is a growing demand for new, unconventional antiviral agents which may serve as an alternative to the currently used drugs. Meanwhile, published literature continues shedding the light on the potency of lactic acid bacteria (LAB) and their bacteriocins as antiviral agents. Health-promoting LAB probiotics may exert their antiviral activity by (1) direct probiotic-virus interaction; (2) production of antiviral inhibitory metabolites; and/or (3) via stimulation of the immune system. The aim of this review was to highlight the antiviral activity of LAB and substances they produce with antiviral activity.

The goal of this study was to investigate the effect of the environmental conditions such as the temperature change, incubation time and surface type on the resistance of Staphylococcus aureus biofilms to disinfectants. The antibiofilm assays were performed against biofilms grown at 20 °C, 30 °C and 37 °C, on the stainless steel and polycarbonate, during 24 and 48 h. The involvement of the biofilm matrix and the bacterial membrane fluidity in the resistance of sessile cells were investigated. Our results show that the efficiency of disinfectants was dependent on the growth temperature, the surface type and the disinfectant product. The increase of growth temperature from 20 °C to 37 °C, with an incubation time of 24 h, increased the resistance of biofilms to cationic antimicrobials. This change of growth temperature did not affect the major content of the biofilm matrix, but it decreased the membrane fluidity of sessile cells through the increase of the anteiso-C19 relative amount. The increase of the biofilm resistance to disinfectants, with the rise of the incubation time, was dependent on both growth temperature and disinfectant product. The increase of the biofilm age also promoted increases in the matrix production and the membrane fluidity of sessile cells. The resistance of S. aureus biofilm seems to depend on the environment of the biofilm formation and involves both extracellular matrix and membrane fluidity of sessile cells. Our study represents the first report describing the impact of environmental conditions on the matrix production, sessile cells membrane fluidity and resistance of S. aureus biofilms to disinfectants.