Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas

Villaviciosa, Spain

Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas

Villaviciosa, Spain
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Zivkovic M.,University of Belgrade | Miljkovic M.,University of Belgrade | Ruas-Madiedo P.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Strahinic I.,University of Belgrade | And 3 more authors.
Applied and Environmental Microbiology | Year: 2015

Lactobacillus paraplantarum BGCG11, a putative probiotic strain isolated from a soft, white, artisanal cheese, produces a highmolecular-weight heteropolysaccharide, exopolysaccharide (EPS)-CG11, responsible for the ropy phenotype and immunomodulatory activity of the strain. In this study, a 26.4-kb region originating from the pCG1 plasmid, previously shown to be responsible for the production of EPS-CG11 and a ropy phenotype, was cloned, sequenced, and functionally characterized. In this region 16 putative open reading frames (ORFs), encoding enzymes for the production of EPS-CG11, were organized in specific loci involved in the biosynthesis of the repeat unit, polymerization, export, regulation, and chain length determination. Interestingly, downstream of the eps gene cluster, a putative transposase gene was identified, followed by an additional rfb gene cluster containing the rfbACBD genes, the ones most probably responsible for dTDP-L-rhamnose biosynthesis. The functional analysis showed that the production of the high-molecular-weight fraction of EPS-CG11 was absent in two knockout mutants, one in the eps and the other in the rfb gene cluster, as confirmed by size exclusion chromatography analysis. Therefore, both eps and rfb genes clusters are prerequisites for the production of high-molecular-weight EPS-CG11 and for the ropy phenotype of strain L. paraplantarum BGCG11. © 2015, American Society for Microbiology.


Tojo R.,Hospital Of Cabuenes | Suarez A.,Hospital Universitario Central Of Asturias Huca | Clemente M.G.,Hospital Universitario Central Of Asturias Huca | De Los Reyes-Gavilan C.G.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | And 3 more authors.
World Journal of Gastroenterology | Year: 2014

The pool of microbes inhabiting our body is known as "microbiota" and their collective genomes as "microbiome". The colon is the most densely populated organ in the human body, although other parts, such as the skin, vaginal mucosa, or respiratory tract, also harbour specific microbiota. This microbial community regulates some important metabolic and physiological functions of the host, and drives the maturation of the immune system in early life, contributing to its homeostasis during life. Alterations of the intestinal microbiota can occur by changes in composition (dysbiosis), function, or microbiota-host interactions and they can be directly correlated with several diseases. The only disease in which a clear causal role of a dysbiotic microbiota has been demonstrated is the case of Clostridium difficile infections. Nonetheless, alterations in composition and function of the microbiota have been associated with several gastrointestinal diseases (inflammatory bowel disease, colorectal cancer, or irritable bowel syndrome), as well as extra-intestinal pathologies, such as those affecting the liver, or the respiratory tract (e.g. , allergy, bronchial asthma, and cystic fibrosis), among others. Species of Bifidobacterium genus are the normal inhabitants of a healthy human gut and alterations in number and composition of their populations is one of the most frequent features present in these diseases. The use of probiotics, including bifidobacteria strains, in preventive medicine to maintain a healthy intestinal function is well documented. Probiotics are also proposed as therapeutic agents for gastrointestinal disorders and other pathologies. The World Gastroenterology Organization recently published potential clinical applications for several probiotic formulations, in which species of lactobacilli are predominant. This review is focused on probiotic preparations containing Bifidobacterium strains, alone or in combination with other bacteria, which have been tested in human clinical studies. In spite of extensive literature on and research into this topic, the degree of scientific evidence of the effectiveness of probiotics is still insufficient in most cases. More effort need to be made to design and conduct accurate human studies demonstrating the efficacy of probiotics in the prevention, alleviation, or treatment of different pathologies. © 2014 Baishideng Publishing Group Inc. All rights reserved.


Ruiz L.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Margolles A.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Sanchez B.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas
Frontiers in Microbiology | Year: 2013

Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. Most of the probiotic bacteria currently available in the market belong to the genera Lactobacillus and Bifidobacterium, and specific health-promoting activities, such as treatment of diarrhea or amelioration of gastrointestinal discomfort, have been attributed to them. In order to be able to survive the gastrointestinal transit and transiently colonize our gut, these bacteria must be able to counteract the deleterious action of bile salts, which are the main components of bile. Bile salts are detergent-like biological substances synthesized in the liver from cholesterol. Host enzymes conjugate the newly synthesized free bile acids in the liver with the amino acids glycine or taurine, generating conjugated bile salts. These compounds are stored in the gall bladder and they are released into the duodenum during digestion to perform their physiological function, which is the solubilization of fat coming from diet. These bile salts possess strong antimicrobial activity, since they are able to disorganize the structure of the cell membrane, as well as trigger DNA damage. This means that bacteria inhabiting our intestinal tract must have intrinsic resistance mechanisms to cope with bile salts. To do that, Lactobacillus and Bifidobacterium display a variety of proteins devoted to the efflux of bile salts or protons, to modify sugar metabolism or to prevent protein misfolding. In this manuscript, we review and discuss specific bile resistance mechanisms, as well as the processes responsible for the adaptation of bifidobacteria and lactobacilli to bile. © 2013 Ruiz, Margolles and Sánchez.


Sanchez B.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Ruiz L.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Gueimonde M.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Ruas-Madiedo P.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Margolles A.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas
Pharmacological Research | Year: 2013

Members of the genus Bifidobacterium are considered to be important constituents of the microbiota of animals, from insects to mammals. They are gut commensals extensively used by the food industry as probiotic microorganisms, since some strains have been shown to have specific beneficial effects. However, the molecular processes underlying their functional capacities to promote a healthy status in the host, as well as those involved in survival, colonization and persistence of bifidobacteria in the gut, are far from being completely understood. This review summarizes the current knowledge on the mechanisms used by bifidobacteria to cope with gastrointestinal factors and to adapt to them, and discusses the advantages of the adaptive traits acquired by these microorganisms as a consequence of their interactions with the gastrointestinal tract environment, as well as the impact of such adaptations in the functional characteristics of bifidobacteria. © 2012 Elsevier Ltd. All Rights Reserved.


Hidalgo-Cantabrana C.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Hidalgo-Cantabrana C.,University of Parma | Sanchez B.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Milani C.,University of Parma | And 3 more authors.
Applied and Environmental Microbiology | Year: 2014

For many years, bacterial exopolysaccharides (EPS) have received considerable scientific attention, mainly due to their contribution to biofilm formation and, above all, because EPS are potential virulence factors. In recent times, interest in EPS research has enjoyed a welcome boost thanks to the discovery of their ability to mediate communication processes with their surrounding environment and to their contribution to host health maintenance. In this review, we provide a fresh perspective on the genetics and activity of these polymers in members of the Bifidobacterium genus, a common gut inhabitant of humans and animals that has been associated with several health-promoting effects. Bifidobacteria can use EPS to protect themselves against the harsh conditions of the gastrointestinal tract, thus improving their persistence in the host. Indeed, the relevant function of EPS for bifidobacteria is underlined by the fact that most genomes sequenced until now contain genes related to EPS biosynthesis. A high interspecies variability in the number of genes and structural organization is denoted among species/subspecies; thus, eps clusters in this genus do not display a consensus genetic architecture. Their different G+C content compared to that of the whole genome suggests that eps genes have been acquired by horizontal transfer. From the host perspective, EPS-producing bifidobacteria are able to trigger both innate and adaptive immune responses, and they are able to modulate the composition and activity of the gut microbiota. Thus, these polymers seem to be critical in understanding the physiology of bifidobacteria and their interaction with the host. © 2014, American Society for Microbiology.


Rodriguez-Rubio L.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Martinez B.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Rodriguez A.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Donovan D.M.,Animal and Natural Resources Institute | And 2 more authors.
PLoS ONE | Year: 2013

The increase in antibiotic resistance world-wide revitalized the interest in the use of phage lysins to combat pathogenic bacteria. In this work, we analyzed the specific cleavage sites on the staphylococcal peptidoglycan produced by three phage lytic proteins. The investigated cell wall lytic enzymes were the endolysin LysH5 derived from the S. aureus bacteriophage vB_SauS-phi-IPLA88 (phi-IPLA88) and two fusion proteins between lysostaphin and the virion-associated peptidoglycan hydrolase HydH5 (HydH5SH3b and HydH5Lyso). We determined that all catalytic domains present in these proteins were active. Additionally, we tested for the emergence of resistant Staphylococcus aureus to any of the three phage lytic proteins constructs. Resistant S. aureus could not be identified after 10 cycles of bacterial exposure to phage lytic proteins either in liquid or plate cultures. However, a quick increase in lysostaphin resistance (up to 1000-fold in liquid culture) was observed. The lack of resistant development supports the use of phage lytic proteins as future therapeutics to treat staphylococcal infections.


Cuervo A.,University of Oviedo | Valdes L.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Salazar N.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | De Los Reyes-Gavilan C.G.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | And 3 more authors.
Journal of Agricultural and Food Chemistry | Year: 2014

Several studies have addressed the use of dietary fibers in the modulation of intestinal microbiota; however, information about other highly correlated components in foods, such as polyphenols, is scarce. The aim of this work was to explore the association between the intake of fibers and polyphenols from a regular diet and fecal microbiota composition in 38 healthy adults. Food intake was recorded using an annual food frequency questionnaire (FFQ). Quantification of microbial populations in feces was performed by quantitative PCR. A negative association was found between the intake of pectins and flavanones from oranges and the levels of Blautia coccoides and Clostridium leptum. By contrast, white bread, providing hemicellulose and resistant starch, was directly associated with Lactobacillus. Because some effects on intestinal microbiota attributed to isolated fibers or polyphenols might be modified by other components present in the same food, future research should be focused on diet rather than individual compounds. © 2014 American Chemical Society.


Salazar N.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Ruas-Madiedo P.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Prieto A.,CSIC - Biological Research Center | Calle L.P.,CSIC - Biological Research Center | De Los Reyes-Gavilan C.G.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas
Journal of Agricultural and Food Chemistry | Year: 2012

Bifidobacteria are natural members of the human intestinal microbiota and some strains are being used as probiotics. Adaptation to bile can allow them to increase survival in gastrointestinal conditions, thus improving their viability. Bifidobacterium longum NB667 and the cholate-resistant strain B. longum IPLA B667dCo produced exopolysaccharides (EPS) that were partially characterized. Analysis by size exclusion chromatography-multiangle laser light scattering indicated that the EPS crude fractions of both strains contained two polymer peaks of different molar mass. On the basis of chromatographic techniques both peaks appeared to be heteropolysaccharides. The smaller peak was mainly composed of glucose, galactose and rhamnose whose molar ratios and linkage types showed slight variations between the EPS fractions of both strains. The bigger peak consisted of glucose and galactose; the monosaccharide composition was identical in the EPS fractions of the two microorganisms, but their infrared spectra presented some differences regarding compounds other than carbohydrates that seem to be associated to the polymer. Differences in the composition of EPS fractions did not affect the capability of crude EPS from B. longum to be fermented by the human intestinal microbiota in fecal batch cultures. © 2012 American Chemical Society.


Valdes-Varela L.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Hernandez-Barranco A.M.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Ruas-Madiedo P.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Gueimonde M.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas
Frontiers in Microbiology | Year: 2016

The intestinal overgrowth of Clostridium difficile, often after disturbance of the gut microbiota by antibiotic treatment, leads to C. difficile infection (CDI) which manifestation ranges from mild diarrhea to life-threatening conditions. The increasing CDI incidence, not only in compromised subjects but also in traditionally considered low-risk populations, together with the frequent relapses of the disease, has attracted the interest for prevention/therapeutic options. Among these, probiotics, prebiotics, or synbiotics constitute a promising approach. In this study we determined the potential of selected Bifidobacterium strains for the inhibition of C. difficile growth and toxicity in different carbon sources. We conducted co-cultures of the toxigenic strain C. difficile LMG21717 with four Bifidobacterium strains (Bifidobacterium longum IPLA20022, Bifidobacterium breve IPLA20006, Bifidobacterium bifidum IPLA20015, and Bifidobacterium animalis subsp. lactis Bb12) in the presence of various prebiotic substrates (Inulin, Synergy, and Actilight) or glucose, and compared the results with those obtained for the corresponding mono-cultures. C. difficile and bifidobacteria levels were quantified by qPCR; the pH and the production of short chain fatty acids was also determined. Moreover, supernatants of the cultures were collected to evaluate their toxicity using a recently developed model. Results showed that co-culture with B. longum IPLA20022 and B. breve IPLA20006 in the presence of short-chain fructooligosaccharides, but not of Inulin, as carbon source significantly reduced the growth of the pathogen. With the sole exception of B. animalis Bb12, whose growth was enhanced, the presence of C. difficile did not show major effects upon the growth of the bifidobacteria. In accordance with the growth data, B. longum and B. breve were the strains showing higher reduction in the toxicity of the co-culture supernatants. © 2016 Valdés-Varela, Hernández-Barranco, Ruas-Madiedo and Gueimonde.


Valdes-Varela L.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Alonso-Guervos M.,University of Oviedo | Garcia-Suarez O.,University of Oviedo | Gueimonde M.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas | Ruas-Madiedo P.,Institute Productos Lacteos Of Asturias Consejo Superior Of Investigaciones Cientificas
Frontiers in Microbiology | Year: 2016

Clostridium difficile is an opportunistic pathogen inhabiting the human gut, often being the aetiological agent of infections after a microbiota dysbiosis following, for example, an antibiotic treatment. C. difficile infections (CDI) constitute a growing health problem with increasing rates of morbidity and mortality at groups of risk, such as elderly and hospitalized patients, but also in populations traditionally considered low-risk. This could be related to the occurrence of virulent strains which, among other factors, have high-level of resistance to fluoroquinolones, more efficient sporulation and markedly high toxin production. Several novel intervention strategies against CDI are currently under study, such as the use of probiotics to counteract the growth and/or toxigenic activity of C. difficile. In this work, we have analyzed the capability of twenty Bifidobacterium and Lactobacillus strains, from human intestinal origin, to counteract the toxic effect of C. difficile LMG21717 upon the human intestinal epithelial cell line HT29. For this purpose, we incubated the bacteria together with toxigenic supernatants obtained from C. difficile. After this co-incubation new supernatants were collected in order to quantify the remnant A and B toxins, as well as to determine their residual toxic effect upon HT29 monolayers. To this end, the real time cell analyser (RTCA) model, recently developed in our group to monitor C. difficile toxic effect, was used. Results obtained showed that strains of Bifidobacterium longum and B. breve were able to reduce the toxic effect of the pathogen upon HT29, the RTCA normalized cell-index values being inversely correlated with the amount of remnant toxin in the supernatant. The strain B. longum IPLA20022 showed the highest ability to counteract the cytotoxic effect of C. difficile acting directly against the toxin, also having the highest capability for removing the toxins from the clostridial toxigenic supernatant. Image analysis showed that this strain prevents HT29 cell rounding; this was achieved by preserving the F-actin microstructure and tight-junctions between adjacent cells, thus keeping the typical epithelium-like morphology. Besides, preliminary evidence showed that the viability of B. longum IPLA20022 is needed to exert the protective effect and that secreted factors seems to have anti-toxin activity. © 2016 Valdés-Varela, Alonso-Guervos, García-Suárez, Gueimonde and Ruas-Madiedo.

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