Interactions between Kluyveromyces marxianus from cheese origin and the intestinal symbiont Bacteroides thetaiotaomicron: Impressive antioxidative effects

The effects of yeast Kluyveromyces marxianus S-2-05, of cheese origin, were assessed on the intestine anaerobe symbiont Bacteroides thetaiotaomicron ATCC 29741 to unveil any changes in its antioxidant properties. To this end, these microorganisms were grown and incubated either separately, or co-incubated, under anaerobic atmosphere. Afterwards, the microbial cells were recovered and washed, and extracts were prepared using a sterile detergent solution to mimic the intestine detergent content. The extracts prepared from K. marxianus S-2-05 and reference strain K. marxianus MUCL 29917, grown under different conditions, were assessed for their antioxidant properties against superoxide anion and hydrogen peroxide. Extracts from both yeasts showed antioxidative effects, which were particularly important for K. marxianus S-02-5 after anaerobic incubation. Moreover, K. marxianus S-02-5 displayed a high level of activity against the aforementioned reactive oxygen species, enhancing that of B. thetaiotaomicron ATCC 29741, after the co-incubation process. Two-dimensional polyacrylamide gel electrophoresis was used to separate the proteins extracted. Superoxide dismutase, thiol peroxidase, rubrerythrin -intensively produced by B. thetaiotaomicron induced by the yeast-were identified by mass spectrometry. The antioxidative potential evidenced for K. marxianus S-02-5 is another advantage which could justify the utilization of this strain as a probiotic for countering intestinal inflammatory processes. © 2017 Elsevier Lt

Edible films and coatings as carriers of living microorganisms: a new strategy towards biopreservation and healthier foods

Edible films and coatings have been extensively studied in recent years due to their unique properties and advantages over more traditional conservation techniques. Edible films and coatings improve shelf life and food quality, by providing a protective barrier against physical and mechanical damage, and by creating a controlled atmosphere and acting as a semipermeable barrier for gases, vapor, and water. Edible films and coatings are produced using naturally derived materials, such as polysaccharides, proteins, and lipids, or a mixture of these materials. These films and coatings also offer the possibility of incorporating different functional ingredients such as nutraceuticals, antioxidants, antimicrobials, flavoring, and coloring agents. Films and coatings are also able to incorporate living microorganisms. In the last decade, several works reported the incorporation of bacteria to confer probiotic or antimicrobial properties to these films and coatings. The incorporation of probiotic bacteria in films and coatings allows them to reach the consumers gut in adequate amounts to confer health benefits to the host, thus creating an added value to the food product. Also, other microorganisms, either bacteria or yeast, can be incorporated into edible films in a biocontrol approach to extend the shelf life of food products. The incorporation of yeasts in films and coatings has been suggested primarily for the control of the postharvest disease. This work provides a comprehensive review of the use of edible films and coatings for the incorporation of living microorganisms, aiming at the biopreservation and probiotic ability of food products.Ana Guimaraes received support through grant SFRH/BD/ 103245/2014 from the Portuguese Foundation for Science and Technology (FCT). Luís Abrunhosa was supported by grant UMINHO/BPD/51/2015 from project UID/BIO/04469/2013 financed by FCT/MEC (OE). This study was supported by FCT under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and of BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Vectors used in Figure were designed by Freepik.info:eu-repo/semantics/publishedVersio

Research of antagonistic yeasts, with probiotic potential, in artisanal products of the North of France Study of Kluyveromyces marxianus and K. lactis isolated from an artisanal cheese, the « Tomme d’Orchies »

Les fromages au lait cru présentent un écosystème microbien complexe où chaque acteur interagit avec ses voisins et avec le milieu. Parmi ces interactions, l’une d’elle est particulièrement intéressante, l’antagonisme, autrement dit la capacité d’un individu à inhiber la croissance d’un autre. Ces interactions antagonistes sont particulièrement étudiées dans les écosystèmes bactériens fromagers et ont conduit à l’identification de bactéries lactiques productrices de bactériocines. Parmi elles, certaines souches ont révélé des propriétés probiotiques et présentent ainsi un fort potentiel dans la lutte contre les résistances aux antibiotiques. D’un autre côté, l’écosystème fongique des fromages est encore peu étudié. Ce dernier est cependant prometteur, notamment par la possibilité d’abriter des levures productrices de mycocines et/ou possédant des propriétés probiotiques. Dans ce cadre, les présents travaux ont porté sur l’étude d’un fromage du terroir des Hauts-de-France, la Tomme d’Orchies. Ils ont permis la mise en évidence de deux levures antagonistes non-Saccharomyces, présentant un potentiel probiotique. L’écosystème microbien de la Tomme d’Orchies sera décrit et la recherche et la caractérisation de levures antagonistes seront effectuées au sein de son écosystème fongique. Deux souches de Kluyveromyces (K. marxianus S-2-05 et K. lactis S-3-05) seront mises en évidence. Ces dernières présentant un pouvoir antagoniste large, contre des bactéries à Gram positifs et négatifs et contre une levure pathogène. De plus, un haut potentiel d’utilisation comme agents probiotiques a été observé, complété par une activité antioxydante d’intérêt médical, observé chez K. marxianus.Raw milk cheeses show a complex microbial ecosystem, where each actor interacting with its neighbor and with its environment. Among these interactions, one is particularly interesting, the antagonism, in other words the capacity of an individual to inhibit the growth of another. Antagonistic interactions are massively studied in bacterial cheese ecosystems and lead to the identification of bacteriocin-producing lactic acid bacteria. Among these strains, some revealed probiotics properties and a high potential in the fight against antibiotics resistances. Additionally, cheese fungal ecosystems were poorly studied. However, this ecosystem is promising as it might contain mycocin-producing yeasts and/or showing probiotic properties. In this frame, our work focused on a local French cheese, the “Tomme d’Orchies”. Two non Saccharomyces antagonistic yeasts, with a probiotic potential, were identified in this cheese. Microbial ecosystems of the “Tomme d’Orchies” will be described before presenting the research and the characterization of antagonistic yeast in the fungal ecosystem. Two strains of Kluyveromyces (K. marxianus S-2-05 and K. lactis S-3-05) will be highlighted, with broad antagonistic properties, against Gram positive and negative bacteria, but also against pathogenic yeast. A high potential to be used as probiotic agents could be observed, in addition to antioxidative properties for K. marxianus, with medical relevance

Anti-Salmonella activity and probiotic trends of Kluyveromyces marxianus S-2-05 and Kluyveromyces lactis S-3-05 isolated from a French cheese named "Tomme d'Orchies"

Kluyveromyces marxianus S-2-05 and Kluyveromyces lactis S-3-05 were recently isolated from a traditional French cheese, Tomme d'Orchies, and characterized here for their advantages using a different application perspective. First, we established their anti-Salmonella activity and downregulation of the virulence sopD gene of Salmonella enterica subsp. enterica serovar Typhimurium, mainly in the presence of K. marxianus S-2-05. In addition to their antagonism, these non-Saccharomyces yeasts were able to survive under conditions mimicking the gastrointestinal environment and to form biofilms on an abiotic device such as polystyrene. These strains also displayed highly hydrophilic cell wall surfaces properties and capacity for adhesion to intestinal Caco-2 cells, thus enhancing their potential as probiotic strains

Use of a metagenetic approach to monitor the bacterial microbiota of “Tomme d’Orchies” cheese during the ripening process

The study of microbial ecosystems in artisanal foodstuffs is important to complete in order to unveil its diversity. The number of studies performed on dairy products has increased during the last decade, particularly those performed on milk and cheese derivative products. In this work, we investigated the bacterial content of "Tomme d'Orchies" cheese, an artisanal pressed and uncooked French cheese. To this end, a metagenetic analysis, using Illumina technology, was utilized on samples taken from the surface and core of the cheese at 0, 1, 3, 14 and 21 days of ripening process. In addition to the classical microbiota found in cheese, various strains likely from environmental origin were identified. A large difference between the surface and the core content was observed within samples withdrawn during the ripening process. The main species encountered in the core of the cheese were Lactococcus spp. and Streptococcus spp., with an inversion of this ratio during the ripening process. Less than 2.5% of the whole population was composed of strains issued from environmental origin, as Lactobacillales, Corynebacterium and Brevibacterium. In the core, about 85% of the microbiota was attributed to the starters used for the cheese making. In turn, the microbiota of the surface contained less than 30% of these starters and interestingly displayed more diversity. The predominant genus was Corynebacterium sp., likely originating from the environment. The less abundant microbiota of the surface was composed of Bifidobacteria, Brevibacterium and Micrococcales. To summarize, the “Tomme d’Orchies” cheese displayed a high diversity of bacterial species, especially on the surface, and this diversity is assumed to arise from the production environment and subsequent ripening process

Yeast diversity in a traditional French cheese “Tomme d'orchies” reveals infrequent and frequent species with associated benefits

peer reviewe

Fungal diversity of “Tomme d'Orchies” cheese during the ripening process as revealed by a metagenomic study

Tomme d'Orchies is an artisanal pressed and uncooked cheese produced and marketed in the north of France. This study aimed at showing the fungal microbiota evolution of this cheese using a metagenetic based Illumina technology targeting the ITS2 domain of 5.8S fungal rDNAs. To this end, samples were taken from the rind and the core of different cheeses, after 0, 1, 3, 7, 14 and 21 days of ripening. The data underpinned the prevalence of Yarrowia lipolytica and Galactomyces geotrichum for both microbiotas. Unusual species including Clavispora lusitaniae, Kazachstania unispora and Cladosporium cladosporioides were also detected, but their origins remain to be ascertained. The metagenomic revealed also the presence of Kluyveromyces and Debaryomyces species. © 2017 Elsevier B.V