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Biomedical subjects

Miguel Gueimonde

Publications and source records attributed to Miguel Gueimonde.

At least 19 recordsLinked to original sources
PubMed2006

Induction of alpha-L-arabinofuranosidase activity by monomeric carbohydrates in Bifidobacterium longum and ubiquity of encoding genes.

Bifidobacterium longum can be isolated from human faeces, some strains being considered probiotics. B. longum NIZO B667 produces an exo-acting alpha-L-arabinofuranosidase, AbfB, previously purified by us, that releases L-arabinose from arabinan and arabinoxylan. This activity was subjected to two-seven-fold induction by L-arabinose, D-xylose, L-arabitol and xylitol and to repression by glucose. Maximum activity was obtained at 48 h incubation except for D-xylose that was at 24 h. High concentrations (200 mM) of L-arabitol also caused repression of the arabinofuranosidase. A unique band of activity showing the same migration pattern as the purified AbfB was found in zymograms of cell free extracts, indicating that the activity was likely due to this sole enzyme. The assessment of the influence of inducers and repressors on the activity of AbfB and on the expression of the abfB gene by real time PCR indicated that regulation was transcriptional. DNA amplifications using a pair of degenerated primers flanking an internal fragment within alpha-L-arabinofuranosidase genes of the family 51 of glycoside hydrolases evidenced that these enzymes are widespread in Bifidobacterium. The aminoacidic sequences of bifidobacteria included a fragment of four to six residues in the position 136-141 that was absent in other microorganisms.

Arabinose↗
PubMed2006

Competitive exclusion of enteropathogens from human intestinal mucus by Bifidobacterium strains with acquired resistance to bile--a preliminary study.

The ability to inhibit the adhesion and to displace selected pathogens from human intestinal mucus of two Bifidobacterium strains with acquired resistance to bile, were assessed and compared with those of their bile sensitive original strains. A preliminary characterization of the macromolecules involved in the adhesion was also carried out. The inhibition of adhesion and the displacement of enteropathogens previously adhered were found to be specific, depending on the strains used. The cholate-resistant strain Bifidobacterium bifidum M6dCo, that adhered more to mucus than its original, was able to inhibit the adhesion and to displace pathogens from mucus significantly more than its original cholate-sensitive strain B. bifidum M6. Contrary to this, two strains showing similar adhesion levels, B. bifidum A1 and its bile resistant derivative B. bifidum A1dOx, did not display any differences. Different molecules appear to be involved in the adhesion of the strains B. bifidum M6 and B. bifidum M6dCo. These differences in the cellular surface may explain the differences in competitive exclusion observed between both strains.

Adaptation, Physiological↗
PubMed2006

Effect of maternal consumption of lactobacillus GG on transfer and establishment of fecal bifidobacterial microbiota in neonates.

BACKGROUND: Establishment of the gut microbiota at birth provides a substantial source of microbial stimuli for the maturation of the immune system. Deviations in this process precede the development of specific diseases providing the rationale for the use of probiotics to counteract them. OBJECTIVE: This study was designed to characterize both the mother-infant bifidobacteria transfer at birth and the development of bifidobacteria microbiota during the first weeks of life in infants whose mothers received Lactobacillus rhamnosus GG or placebo. METHODS: Species-specific PCR was used to assess the fecal bifidobacterial composition of mothers before and after delivery and in infants at 5 days and 3 weeks of age. RESULTS: Bifidobacterium longum was the species most commonly found in the mothers. Bifidobacterium catenulatum was the most prevalent group in infants at 5 days of age and B. longum the predominant species at 3 weeks. At 5 days of age, infants whose mothers received L. rhamnosus GG showed a significantly higher occurrence of B. breve and lower of B. adolescentis than those from the placebo group. In addition, L. rhamnosus GG consumption increased the bifidobacterial diversity in infants and reduced the Bifidobacterium microbiota similarity between mother and infant. CONCLUSIONS: These results indicate that specific changes in the transfer and initial establishment of bifidobacteria in neonates take place as consequence of the consumption of L. rhamnosus GG by the mothers.

Adult↗
PubMed2006

Probiotic intervention in neonates--will permanent colonization ensue?

The presence of Lactobacillus rhamnosus GG at 6 and 12 months in the feces of 128 infants receiving the strain or placebo for 6 months after birth was monitored. No permanent establishment of the strain was found, but transient colonization was observed at 6 months of age. The presence of the strain at the end of the administration period was correlated with a reduced prevalence of atopic eczema later in life.

Colony Count, Microbial↗
PubMed2006

Presence of specific antibiotic (tet) resistance genes in infant faecal microbiota.

The widespread use of antibiotics for medical and veterinary purposes has led to an increase of microbial resistance. The antibiotic resistance of pathogenic bacteria has been studied extensively. However, antibiotics are not only selective for pathogens: they also affect all members of the gut microbiota. These microorganisms may constitute a reservoir of genes carrying resistance to specific antibiotics. This study was designed to characterize the gut microbiota with regard to the presence of genes encoding tetracycline resistance proteins (tet) in the gut of healthy exclusively breast-fed infants and their mothers. For this purpose we determined the prevalence of genes encoding ribosomal protection proteins (tet M, tet W, tet O, tet S, tet T and tet B) by PCR and characterized the gut microbiota by FISH in stools of infants and their mothers. The gene tet M was found in all the breast-fed infants and their mothers. tet O was found in all of the mothers' samples, whilst only 35% of the infants harboured this gene. tet W was less frequently found (85% of the mothers and 13% of the infants). None of the other genes analysed was found in any sample. Our results suggest that genes carrying antibiotic resistance are common in the environment, as even healthy breast-fed infants with no direct or indirect previous exposure to antibiotics harbour these genes.

Adult↗
PubMed2006

Adhesion properties and competitive pathogen exclusion ability of bifidobacteria with acquired acid resistance.

The adhesion properties of Bifidobacterium longum and Bifidobacterium catenulatum strains with an acquired resistance to acid and their ability to competitively exclude Salmonella enterica serovar Typhimurium, Escherichia coli, Listeria monocytogenes, Enterobacter sakazakii, and Clostridium difficile from adhering to human intestinal mucus were evaluated and compared with the results when the same experiments were run with the original acid-sensitive strains. In half of the four studied cases, the acid-resistant derivative showed a greater ability to adhere to human intestinal mucus than the original strain. The ability of bifidobacteria to inhibit pathogen adhesion to mucus was not generally improved by the acquisition of acid resistance. In contrast, three of the four acid-resistant strains showed a greater ability to displace preadhered pathogens than the original strains, especially preadhered Salmonella Typhimurium and C. difficile. Overall, the induction of acid resistance in bifidobacteria could be a strategy when selecting strains with enhanced stability and improved surface properties that favor their potential functionality as probiotics against specific pathogens.

Acids↗
PubMed2006

Exopolysaccharides produced by probiotic strains modify the adhesion of probiotics and enteropathogens to human intestinal mucus.

Exopolysaccharides (EPSs) are exocellular polymers present in the surface of many bacteria, including Lactobacillus and Bifidobacterium. The genome sequence of several strains revealed the presence of EPS-encoding genes. However, the physiological role that EPSs play in the bacterial ecology still remains uncertain. In this study, we have assessed the effect of EPSs produced by Lactobacillus rhamnosus GG, Bifidobacterium longum NB667, and Bifidobacterium animalis IPLA-R1 on the adhesion of probiotic and enteropathogen strains to human intestinal mucus. The EPS fraction GG had no significant effect on the adhesion of L. rhamnosus GG and B. animalis IPLA-R1. However, the EPS fractions NB667 and IPLA-R1 significantly reduced the adherence of both probiotic strains. In contrast, the three EPS fractions increased the adhesion of Enterobacter sakazakii ATCC 29544 and Escherichia coli NCTC 8603. Higher adherence of Salmonella enterica serovar Typhimurium ATCC 29631 and Clostridium difficile ATCC 9689 was detected in the presence of the EPS fractions GG and NB667. In general, these effects were obtained at EPS concentrations of up to 5 mg/ml, and they were EPS dose dependent. The competitive exclusion of probiotics in the presence of EPS could suggest the involvement of these biopolymers in the adhesion to mucus. The increase in the adherence of enteropathogens could be explained if components of the pathogen surface are able to bind to specific EPSs and the bound EPSs are able to adhere to mucus. To the best of our knowledge, this is the first work reporting the effect of EPSs from probiotics on bacterial adhesion properties.

Antibiosis↗
PubMed2005

Comparison of four methods to enumerate probiotic bifidobacteria in a fermented food product.

Four methods of enumeration were compared by monitoring levels of probiotic bifidobacteria in fermented oat drink during storage. Strains of Bifidobacterium longum and B. lactis were quantified by plate counts, fluorescent in situ hybridization (FISH), quantitative real-time PCR and commercial LIVE/DEAD BacLight bacterial viability kit, and the methods were further developed to suit the enumeration of bifidobacteria in fermented foods. Plate counts of both B. lactis and B. longum were lower than the PCR and FISH counts. The LIVE/DEAD counts of B. lactis were comparable to PCR and FISH counts. The plate counts of B. lactis were slightly but significantly lower than LIVE/DEAD counts, suggesting that the cells that were not able to grow on plates may have become dormant. The plate counts of B. longum were several log units lower than LIVE/DEAD counts, suggesting that a remarkable part of the cells were dormant. Real-time PCR and FISH were shown to suit the quantification of the total amount of probiotic bifidobacteria in foods. Plate counts and LIVE/DEAD counts provided conflicting information on viability especially in the case of B. longum. We conclude that the choice of enumeration method for probiotic bacteria may have significant effect on the results of the analysis. The strain-specific properties and the objects of the analysis should be taken into account when enumeration methods for different probiotic strains are chosen.

Bifidobacterium↗
PubMed2005

The genomics of probiotic intestinal microorganisms.

An intestinal population of beneficial commensal microorganisms helps maintain human health, and some of these bacteria have been found to significantly reduce the risk of gut-associated disease and to alleviate disease symptoms. The genomic characterization of probiotic bacteria and other commensal intestinal bacteria that is now under way will help to deepen our understanding of their beneficial effects.

Bacteria↗
PubMed2005

Ability of Bifidobacterium strains with acquired resistance to bile to adhere to human intestinal mucus.

The hydrophobicity and capacity to adhere to human intestinal mucus of Bifidobacterium strains with acquired resistance to bile were assessed and compared with those of their more sensitive original strains. The resistant variants used were previously obtained [Int. J. Food Microbiol. 82 (2003) 191; Int. J. Food Microbiol. 94 (2004) 79] by progressive adaptation of originally more sensitive strains to gradually increasing concentrations of bile. In five out of the seven groups of original and bile-resistant variants tested the resistant strains showed higher adhesion levels to human mucus (range between 1.4- and 4-fold) than their corresponding original strains. However, in the presence of physiologic concentrations of bile (0.3%, w/v) the adhesion level of all Bifidobacterium strains dropped between 7% and 74%, depending on the strain. In spite of this, the adhesion capability of three bile-resistant variants remained higher than that of their originals. Hydrophobicity evidenced considerable variability; in four out of the seven bile-resistant strains it was higher than in the original strains, although no direct correlation between adhesion and hydrophobicity could be established. It was concluded that the acquisition of bile resistance by our Bifidobacterium strains promoted changes in hydrophobicity and in the adhesion of these microorganisms to human intestinal mucus.

Adaptation, Physiological↗
PubMed2005

Similar bifidogenic effects of prebiotic-supplemented partially hydrolyzed infant formula and breastfeeding on infant gut microbiota.

The aim of the study was to assess the quantitative and qualitative differences of the gut microbiota in infants. We evaluated gut microbiota at the age of 6 months in 32 infants who were either exclusively breast-fed, formula-fed, nursed by a formula supplemented with prebiotics (a mixture of fructo- and galacto-oligosaccharides) or breast-fed by mothers who had been given probiotics. The Bifidobacterium, Bacteroides, Clostridium and Lactobacillus/Enterococcus microbiota were assessed by the fluorescence in situ hybridization, and Bifidobacterium species were further characterized by PCR. Total number of bifidobacteria was lower among the formula-fed group than in other groups (P=0.044). Total amounts of the other bacteria were comparable between the groups. The specific Bifidobacterium microbiota composition of the breast-fed infants was achieved in infants receiving prebiotic supplemented formula. This would suggest that early gut Bifidobacterium microbiota can be modified by special diets up to the age of 6 months.

Bacteroides↗
PubMed2005

Removal of the cyanobacterial toxin microcystin-LR by human probiotics.

Three human probiotics, Lactobacillus rhamnosus strains GG and LC-705, and Bifidobacterium lactis strain Bb12, were found to bind the cyanobacterial peptide toxin microcystin-LR from water solutions. The highest removal percentage was 46%, observed with heat-treated L. rhamnosus strain GG (10(10) cells/ml) and a microcystin-LR concentration of 0.5 microg/ml during an incubation of 7 h at 35 degrees C.

Bifidobacterium↗
PubMed2005

Probiotics that modify disease risk.

Probiotics are defined as live bacterial preparations with clinically documented health effects in humans. Probiotics have specific properties and targets in the human intestinal tract and intestinal microbiota. Each probiotic strain, independent of its genus and species is unique and, thus, the properties and the human health effects of each strain have to be assessed in a case-by-case manner. Understanding the mechanisms by which probiotics influence the normal intestinal microbiota and counteract aberrancies in microbiota would facilitate the use of probiotics for both dietary management and reduction in risk of specific diseases. Development of intestinal microbiota is an important factor affecting the health of the newborn. Recent studies suggest that specific bacterial components, especially the bifidobacteria, have a key impact on development of a healthy balanced infant microbiota. The composition of infant and child intestinal microbiota may become aberrant and thus influence the development of diarrheal, inflammatory, and allergic diseases. Based on this understanding, positive health effects of probiotics have been reported in the management of diarrheal, inflammatory, and allergic diseases in infants. Most recently, a reduction in risk of atopic diseases followed early administration of specific probiotics.

Bacteria↗
PubMed2005

Probiotic bacteria may become dormant during storage.

The determination of bacterial viability in probiotic products is of economic, technological, and clinical significance. We compared four methods to enumerate three Bifidobacterium strains in fermented oat products during storage. A subpopulation of nonculturable cells retained a functional cell membrane typical of viable cells, indicating that probiotic bacteria become dormant during storage.

Avena↗
PubMed2005

Adhesion of selected Bifidobacterium strains to human intestinal mucus and the role of adhesion in enteropathogen exclusion.

The ability of potential probiotic strains to adhere to the intestinal mucosa and exclude and displace pathogens is of utmost importance for therapeutic manipulation of the enteric microbiota. The ability of seven selected human bifidobacterial strains and five human enteropathogenic strains to adhere to human intestinal mucus was analyzed and compared with that of four strains isolated from chicken intestines. The adhesion of the bifidobacterial strains ranged from 3 to 16% depending on the strain. Bifidobacterium strains of animal origin adhered significantly better than did strains of human origin. Of the pathogenic bacteria, Escherichia coli NCTC 8603 had the highest adhesion value (20%), Salmonella Typhimurium ATCC 29631, Enterobacter sakazakii ATCC 29544, and Clostridium difficile ATCC 9689 had adhesion values ranging from 10 to 15%, and Listeria monocytogenes ATCC 15313 had the lowest adhesive value (3%). The ability of these bifidobacteria to inhibit pathogen adhesion and to displace pathogens previously adhering to mucus was also tested. The inhibition of pathogens adhesion by these bifidobacterial strains was variable and clearly strain dependent. In general, bifidobacterial strains of animal origin were better able to inhibit and displace pathogens than were human strains. Preliminary characterization of bacterial adhesion was accomplished using different pretreatments to explore adhesion mechanisms. The results indicate that different molecules are implicated in the adhesion of bifidobacteria to the human intestinal mucus, constituting a multifactorial process.

Analysis of Variance↗
PubMed2004

Effect of the adaptation to high bile salts concentrations on glycosidic activity, survival at low PH and cross-resistance to bile salts in Bifidobacterium.

Six derivatives with increased resistance to ox gall (MIC: > or = 1% w/v) and one derivative resistant to sodium cholate (MIC: 0.8% w/v) were obtained from more sensitive original Bifidobacterium strains. These microorganisms, and two additional cholate resistant derivatives obtained in a previous study (Int. J. Food Microbiol. 82 (2003) 191), were partially characterised in this study. Acquisition of resistance against a given bile salt, also conferred cross-resistance to other bile salts, and promoted an increase in the survival of these microorganisms at low pH. Bile resistance levels of derivatives were dependent on the external pH so that the resistance was lower at neutral pH values than in acidic environments. In addition, the acquisition of bile resistance induced changes on glycoside-hydrolysing activities of derivatives obtained from five out of eight original strains, with certain activities such as beta-glucosidase showing more than tenfold increases in some of these microorganisms. These data suggest that the exposure to high bile salts concentrations may have induced a synergic response on Bifidobacterium for the adaptation to the conditions of the gastrointestinal tract. This could have improved the survival at low pH in these microorganisms, the resistance to high bile salts concentrations, and the assimilation of non-digestible carbohydrates by the enhancement of some glycoside-hydrolysing activities.

Adaptation, Physiological↗
PubMed2004

New real-time quantitative PCR procedure for quantification of bifidobacteria in human fecal samples.

The application of a real-time quantitative PCR method (5' nuclease assay), based on the use of a probe labeled at its 5' end with a stable, fluorescent lanthanide chelate, for the quantification of human fecal bifidobacteria was evaluated. The specificities of the primers and the primer-probe combination were evaluated by conventional PCR and real-time PCR, respectively. The results obtained by real-time PCR were compared with those obtained by fluorescent in situ hybridization, the current gold standard for intestinal microbiota quantification. In general, a good correlation between the two methods was observed. In order to determine the detection limit and the accuracy of the real-time PCR procedure, germfree rat feces were spiked with known amounts of bifidobacteria and analyzed by both methods. The detection limit of the method used in this study was found to be about 5 x 10(4) cells per g of feces. Both methods, real-time PCR and fluorescent in situ hybridization, led to an accurate quantification of the spiked samples with high levels of bifidobacteria, but real-time PCR was more accurate for samples with low levels. We conclude that the real-time PCR procedure described here is a specific, accurate, rapid, and easy method for the quantification of bifidobacteria in feces.

Adult↗