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ABSTRACT: The human gut represents a highly complex ecosystem, which is densely colonized by a myriad of microorganisms that influence the physiology, immune function and health status of the host. Among the many members of the human gut microbiota, there are microorganisms that have co-evolved with their host and that are believed to exert health-promoting or probiotic effects. Probiotic bacteria isolated from the gut and other environments are commercially exploited, and although there is a growing list of health benefits provided by the consumption of such probiotics, their precise mechanisms of action have essentially remained elusive. Genomics approaches have provided exciting new opportunities for the identification of probiotic effector molecules that elicit specific responses to influence the physiology and immune function of their human host. In this review, we describe the current understanding of the intriguing relationships that exist between the human gut and key members of the gut microbiota such as bifidobacteria and lactobacilli, discussed here as prototypical groups of probiotic microorganisms.
Cellular and Molecular Life Sciences CMLS 03/2013; · 6.57 Impact Factor
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ABSTRACT: The composition of the microbiota associated with the human ileum and colon in the early weeks of life of two preterm infants was examined, with particular emphasis on the Lactobacillus and Bifidobacterium members. Culturing work showed that bifidobacteria and lactobacilli in the ileostomy changed over time, compared with the colostomy effluent where there was far less variation. The colostomy infant was dominated by two phyla, Actinobacteria and Firmicutes, while in the ileostomy samples, Proteobacteria emerged at the expense of Actinobacteria. Bacteroidetes were only detected following the reversal of the ileostomy in the final fecal sample and were not detected in any colonic fluid samples. Clostridia levels were unstable in the colostomy fluid, suggesting that the ileostomy/colostomy itself influenced the gut microbiota, in particular the strict anaerobes. Pyrosequencing analysis of microbiota composition indicated that bifidobacteria and lactobacilli are among the dominant genera in both the ileal and colonic fluids. Bifidobacteria and lactobacilli levels were unstable in the ileostomy fluid, with large reductions in numbers and relative proportions of both observed. These decreases were characterized by an increase in proportions of Streptococcus and Enterobacteriaceae. Clostridium was detected only in the colonic effluent, with large changes in the relative proportions over time.
MicrobiologyOpen. 01/2013;
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ABSTRACT: Next-generation sequencing technologies have had a dramatic impact in the field of genomic research through the provision of a low cost, high-throughput alternative to traditional capillary sequencers. These new sequencing methods have surpassed their original scope and now provide a range of utility-based applications, which allow for a more comprehensive analysis of the structure and content of microbial genomes than was previously possible. With the commercialization of a third generation of sequencing technologies imminent, we discuss the applications of current next-generation sequencing methods and explore their impact on and contribution to microbial genome research.
Briefings in functional genomics 01/2013; · 4.13 Impact Factor
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Eoin Barrett,
Colm Kerr,
Kiera Murphy,
Orla O'Sullivan,
C Anthony Ryan,
Eugene M Dempsey,
Brendan P Murphy, Paul W O'Toole,
Paul D Cotter,
Gerald F Fitzgerald,
R Paul Ross,
Catherine Stanton
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ABSTRACT: OBJECTIVE: To examine the composition of the evolving microbiota of preterm infants at weeks 2 and 4 of life. SETTINGS: The paediatric intensive care unit of the Cork University Maternity Hospital. METHODS: The microbial diversity of faecal samples from 10 preterm infants was determined using 16S rRNA amplicon pyrosequencing technology. RESULTS: In total, 452 863 sequences were obtained from 20 faecal samples collected from 10 preterm infants, allowing a level of analysis not previously reported. The preterm infant microbiota samples were dominated by Proteobacteria (46%), followed by Firmicutes (45%), while the phyla Actinobacteria (2%) and Bacteroidetes (7%) were detected at much lower levels at week 2 of life. This colonisation pattern was similar at week 4 of life. At the family level, Enterobacteriaceae were detected at 50% and 58% at weeks 2 and 4, respectively. The preterm infants were characterised by a lack of detectable Bifidobacterium and Lactobacillus genera commonly associated with the infant gut. In addition to the dominance of the Proteobacteria, a high level of interindividual variation was observed, indeed the relative proportions of different phyla, families and genera in different infants ranged from <1% to >90%. CONCLUSIONS: The results indicate that in addition to an uncharacteristic microbiota relative to that reported for healthy term infants, there was a large interindividual variation in the faecal microbiota diversity of preterm infants suggesting that the preterm microbiota is individual-specific and does not display a uniformity among infants.
Archives of Disease in Childhood - Fetal and Neonatal Edition 01/2013; · 3.05 Impact Factor
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ABSTRACT: It is well established that diet influences the health of an individual and that a diet rich in plant-based foods has many advantages in relation to the health and well-being of an individual. What has been unclear until recently is the large contribution of the gut microbiota to this effect. As well as providing basic nutritional requirements, the long-term diet of an animal modifies its gut microbiota. In adults, diets that have a high proportion of fruit and vegetables and a low consumption of meat are associated with a highly diverse microbiota and are defined by a greater abundance of Prevotella compared to Bacteroides, while the reverse is associated with a diet that contains a low proportion of plant-based foods. Furthermore, it is becoming increasingly clear that the effect of the microbial ecology of the gut goes beyond the local gut immune system and is implicated in immune-related disorders, such as IBS, diabetes and inflamm-ageing. In this review, we investigate the evidence that a balanced diet leads to a balanced, diverse microbiota with significant consequences for healthy ageing by focusing on conditions of interest.
Nutrients 01/2013; 5(1):234-52. · 0.68 Impact Factor
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ABSTRACT: Links between the gut microbiota and host metabolism have provided new perspectives on obesity. We previously showed that the link between the microbiota and fat deposition is age- and time-dependent subject to microbial adaptation to diet over time. We also demonstrated reduced weight gain in diet-induced obese (DIO) mice through manipulation of the gut microbiota with vancomycin or with the bacteriocin-producing probiotic Lactobacillus salivarius UCC118 (Bac(+)), with metabolic improvement achieved in DIO mice in receipt of vancomycin. However, two phases of weight gain were observed with effects most marked early in the intervention phase. Here, we compare the gut microbial populations at the early relative to the late stages of intervention using a high throughput sequencing-based analysis to understand the temporal relationship between the gut microbiota and obesity. This reveals several differences in microbiota composition over the intervening period. Vancomycin dramatically altered the gut microbiota composition, relative to controls, at the early stages of intervention after which time some recovery was evident. It was also revealed that Bac(+) treatment initially resulted in the presence of significantly higher proportions of Peptococcaceae and significantly lower proportions of Rikenellaceae and Porphyromonadaceae relative to the gut microbiota of L. salivarius UCC118 bacteriocin negative (Bac(-)) administered controls. These differences were no longer evident at the later time. The results highlight the resilience of the gut microbiota and suggest that interventions may need to be monitored and continually adjusted to ensure sustained modification of the gut microbiota.
PLoS ONE 01/2013; 8(6):e65790. · 4.09 Impact Factor
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ABSTRACT: The aim of this study was to investigate the diversity and composition of the intestinal microbiota of elderly subjects using a combination of culture dependent techniques and 16S rRNA gene amplicon sequencing. The study was performed as part of the ELDERMET project, in which 368 faecal samples were assessed for viable numbers of Bifidobacterium spp., Lactobacillus spp. and Enterobacteriaceae on selective agar. However, the Bifidobacterium selective medium (BSM) also supported the growth of Clostridium perfringens, which appeared as distinct colonies and were subsequently characterised phenotypically and genotypically. All the isolates were confirmed as toxin biotype A producers. In addition, three isolates tested also had the genetic determinants for the beta-2 (β2) toxin. Of the 368 faecal samples assessed, C. perfringens was detected in 28 samples (7.6%). Moreover, C. perfringens was observed in samples from subjects in all the residence locations assessed; but was most prevalent in subjects from longstay residential care, with 71.4% of the samples (63.2% of the subjects) being from this residence location, and with a shedding level in excess of 106 c.f.u. g-1 faeces. Microbiota profiling revealed some significant compositional changes across both the family and genus taxonomic levels between the C. perfringens positive and negative data-sets. Levels of culturable Bifidobacterium spp. and Lactobacillus spp. were significantly (P<0.05) lower in the C. perfringens positive samples. Sequence based methods also confirmed a significant difference in the Bifidobacterium spp. level (P<0.05) between both the datasets. Taken together, these data suggest that a high viable count (in excess of 106 c.f.u. g-1 faeces) of C. perfringens in stool samples may be indicative of a less healthy microbiota in the intestine of elderly people in longstay residential care.
Journal of Medical Microbiology 12/2012; · 2.50 Impact Factor
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ABSTRACT: AIMS: To isolate and characterize bacteriocins produced by predominant species of lactic acid bacteria (LAB) from faeces of elderly subjects. METHODS AND RESULTS: Screening over 70,000 colonies, from faecal samples collected from 266 subjects, using the indicator organisms Lactobacillus bulgaricus LMG 6901 and Listeria innocua DPC 3572, identified 55 antimicrobial-producing bacteria. Genomic fingerprinting following ApaI digestion revealed 15 distinct strains. The antimicrobial activities associated with 13 of the 15 strains were sensitive to protease treatment. The predominant antimicrobial-producing species were identified as Lactobacillus salivarius, Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus crispatus and Enterococcus spp. A number of previously characterized bacteriocins, including ABP-118 and salivaricin B (from Lb. salivarius), Enterocin B (Ent. faecium), Lactacin B (Lb. acidophilus), Gassericin T and a variant of Gassericin A (Lb. gasseri), were identified. Interestingly, two antimicrobial-producing species, not generally associated with intestinally-derived microorganisms were also isolated: Lactococcus lactis producing Nisin Z and Streptococcus mutans producing Mutacin II. CONCLUSION: These data suggest that bacteriocin production by intestinal isolates against our chosen targets under the screening conditions used was not frequent (0.08%). © 2012The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.
Journal of Applied Microbiology 11/2012; · 2.34 Impact Factor
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ABSTRACT: Older people constitute a growing proportion of the worldwide population. Despite this, older people are often excluded from clinical research and are generally underrepresented in intervention studies. This severely restricts the ability to generalise outcomes from research involving younger populations, which may impact on the progression of knowledge and the development of best practice guidelines for the care of older people. This opinion piece outlines the challenges and practical difficulties experienced and overcome by the ELDERMET project. The ELDERMET project has recruited almost 500 subjects, aged 65 years and older, across a range of health states from the very frail to the very fit, half of whom have been studied at multiple time points. All ELDERMET subjects have participated in an extensive protocol and supplied multiple biological sample types. The challenges and obstacles faced by both researchers in recruiting older subjects and older people engaging with research and intervention studies are set out. Strategies are discussed for: the recruitment and retention of older subjects; recruiting subjects with physical or cognitive impairment; recruitment from specific locations; collecting accurate and robust data, particularly from subjects with mild to severe cognitive impairment; intervention product design, delivery and compliance. Practical and realistic solutions for maximising the engagement of older people with research and intervention studies are offered. The increased benefit brought by the generalisation and application of research and intervention outcomes to older populations is discussed.
Gerontology 11/2012; · 2.78 Impact Factor
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ABSTRACT: Irritable Bowel Syndrome (IBS) is a clinically heterogeneous disorder which is likely to involve a number of causative factors. The contribution of altered intestinal microbiota composition or function to this disorder is controversial, and is the subject of much current research. Until recently, the technical limitations of the methodologies available have not permitted an adequate survey of low-abundance microbial species. Recent technological developments have enabled the analysis of the global population of the microbiome using high through-put, culture independent, 16S rRNA amplicon pyrosequencing. Using these new methodologies, we are able to gain important biological insights into the link between functional bowel disorders and the microbiome. This addendum contextualizes and summarizes the results of these studies, and defines the future challenges and opportunities in the field.
Gut Microbes 11/2012; 3(6).
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ABSTRACT: Enhanced barrier function is one mechanism whereby commensals and probiotic bacteria limit translocation of foreign antigens or pathogens in the gut. However, barrier protection is not exhibited by all probiotic or commensals and the strain-specific molecules involved remain to be clarified. We evaluated the effects of 33 individual Lactobacillus salivarius strains on the hydrogen peroxide (H(2)O(2))-induced barrier impairment in human epithelial Caco-2 cells. These strains showed markedly different effects on H(2)O(2)-induced reduction in transepithelial resistance (TER). The effective strains such as UCC118 and CCUG38008 attenuated H(2)O(2)-induced disassembly and relocalization of tight junction proteins, but the ineffective strain AH43324 did not. Strains UCC118 and CCUG38008 induced phosphorylation of extracellular signal-regulated kinase (ERK) in Caco-2 cells, and the ERK inhibitor U0126 attenuated the barrier-protecting effect of these strains. In contrast, the AH43324 strain induced phosphorylation of Akt and p38, which was associated with an absence of a protective effect. Global transcriptome analysis of UCC118 and AH43324 revealed that some genes in a bacteriocin gene cluster were up-regulated in AH43324 under TER assay conditions. A bacteriocin-negative UCC118 mutant displayed significantly greater suppressive effect on H(2)O(2)-induced reduction in TER compared to wild type UCC118. The wild type strain augmented H(2)O(2)-induced phosphorylation of Akt and p38, while a bacteriocin-negative UCC118 mutant did not. These observations indicate that L. salivarius strains are widely divergent in their capacity for barrier protection and this is underpinned by differences in the activation of intracellular signaling pathways. Furthermore, bacteriocin production appears to have an attenuating influence on lactobacillus-mediated barrier protection.
AJP Gastrointestinal and Liver Physiology 09/2012; · 3.43 Impact Factor
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ABSTRACT: OBJECTIVES: The human intestinal microbiota composition alters naturally with age, but is unusually perturbed by antibiotic therapy. The impact of antibiotic therapy on the composition of the intestinal microbiota of a cross-section of elderly Irish subjects (n = 185, ≥65 years) was investigated, taking into consideration their residence location. METHODS: Forty-two of the 185 elderly subjects were treated with at least one antibiotic within 1 month prior to faecal microbiota profiling. The residence locations of the subjects varied from long-term nursing care and rehabilitation wards to day hospitals and the community. RESULTS: Culture-dependent methods indicated that faecal Bifidobacterium spp. numbers were significantly reduced following antibiotic treatment (P = 0.004, 7-fold reduction), while levels of Lactobacillus spp. and Enterobacteriaceae were unaffected. The largest decrease in Bifidobacterium spp. numbers was linked to the administration of nucleic acid synthesis inhibitors (P = 0.004, 23-fold reduction). Microbiota profiling revealed a significant compositional change across nine genera following antibiotic therapy, including a relative increase in Lactobacillus spp. (P = 0.031), as well as a decrease in the number of genera identified in the antibiotic-treated subjects (n = 58), when compared with untreated subjects (n = 79). More alterations in the intestinal microbiota were observed post-nucleic acid synthesis inhibitor therapy, most notably a decrease in relative Faecalibacterium spp. numbers (P < 0.001). CONCLUSIONS: The impact of antibiotic therapy on the intestinal microbiota in the elderly should be considered for long-term health effects, and differential susceptibility may require the development of products (e.g. prebiotics and probiotics) for at-risk subjects.
Journal of Antimicrobial Chemotherapy 09/2012; · 5.07 Impact Factor
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ABSTRACT: Bacteroides fragilis and Bacteroides thetaiotaomicron are members of the normal human intestinal microbiota. However, both organisms are capable of causing opportunistic infections, during which the environmental conditions to which the bacteria are exposed change dramatically. To further explore their potential for contributing to infection, we have characterized the expression in B. thetaiotaomicron of four homologues of the gene encoding the C10 cysteine protease SpeB, a potent extracellular virulence factor produced by Streptococcus pyogenes.
We identified a paralogous set of genes (btp genes) in the B. thetaiotaomicron genome, that were related to C10 protease genes we recently identified in B. fragilis. Similar to C10 proteases found in B. fragilis, three of the B. thetaiotaomicron homologues were transcriptionally coupled to genes encoding small proteins that are similar in structural architecture to Staphostatins, protease inhibitors associated with Staphopains in Staphylococcus aureus. The expression of genes for these C10 proteases in both B. fragilis and B. thetaiotaomicron was found to be regulated by environmental stimuli, in particular by exposure to oxygen, which may be important for their contribution to the development of opportunistic infections.
Genes encoding C10 proteases are increasingly identified in operons which also contain genes encoding proteins homologous to protease inhibitors. The Bacteroides C10 protease gene expression levels are responsive to different environmental stimuli suggesting they may have distinct roles in the bacterial-host interaction.
BMC Microbiology 09/2012; 12:190. · 3.04 Impact Factor
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ABSTRACT: A recent comparative genomic hybridization study in our laboratory revealed considerable plasticity within the bacteriocin locus of gastrointestinal strains of Lactobacillus salivarius. Most notably, these analyses led to the identification of two novel unmodified bacteriocins, salivaricin L and salivaricin T, produced by the neonatal isolate L. salivarius DPC6488 with immunity, regulatory and export systems analogous to those of abp118, a two-component bacteriocin produced by the well characterized reference strain L. salivarius UCC118. In this addendum we discuss the intraspecific diversity of our seven bacteriocin-producing L. salivarius isolates on a genome-wide level, and more specifically, with respect to their salivaricin loci.
Gut Microbes 09/2012; 3(5):468-73.
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Nature Reviews Microbiology 09/2012; 10(9):591-2. · 21.18 Impact Factor
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Marcus J Claesson,
Ian B Jeffery,
Susana Conde,
Susan E Power,
Eibhlís M O'Connor,
Siobhán Cusack,
Hugh M B Harris,
Mairead Coakley,
Bhuvaneswari Lakshminarayanan,
Orla O'Sullivan, [......],
Douwe van Sinderen,
Martina Wallace,
Lorraine Brennan,
Catherine Stanton,
Julian R Marchesi,
Anthony P Fitzgerald,
Fergus Shanahan,
Colin Hill,
R Paul Ross, Paul W O'Toole
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ABSTRACT: Alterations in intestinal microbiota composition are associated with several chronic conditions, including obesity and inflammatory diseases. The microbiota of older people displays greater inter-individual variation than that of younger adults. Here we show that the faecal microbiota composition from 178 elderly subjects formed groups, correlating with residence location in the community, day-hospital, rehabilitation or in long-term residential care. However, clustering of subjects by diet separated them by the same residence location and microbiota groupings. The separation of microbiota composition significantly correlated with measures of frailty, co-morbidity, nutritional status, markers of inflammation and with metabolites in faecal water. The individual microbiota of people in long-stay care was significantly less diverse than that of community dwellers. Loss of community-associated microbiota correlated with increased frailty. Collectively, the data support a relationship between diet, microbiota and health status, and indicate a role for diet-driven microbiota alterations in varying rates of health decline upon ageing.
Nature 07/2012; 488(7410):178-84. · 36.28 Impact Factor
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James Collins,
Jan-Peter van Pijkeren,
Lisbeth Svensson,
Marcus J Claesson,
Mark Sturme,
Yin Li,
Jakki C Cooney,
Douwe van Sinderen,
Alan W Walker,
Julian Parkhill,
Oonagh Shannon, Paul W O'Toole
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ABSTRACT: The marketplace for probiotic foods is burgeoning, measured in billions of euro per annum. It is imperative, however, that all bacterial strains are fully assessed for human safety. The ability to bind fibrinogen is considered a potential pathogenicity trait that can lead to platelet aggregation, serious medical complications, and in some instances, death. Here we examined strains from species frequently used as probiotics for their ability to bind human fibrinogen. Only one strain (CCUG 47825), a Lactobacillus salivarius isolate from a case of septicaemia, was found to strongly adhere to fibrinogen. Furthermore, this strain was found to aggregate human platelets at a level comparable to the human pathogen Staphylococcus aureus. By sequencing the genome of CCUG 47825, we were able to identify candidate genes responsible for fibrinogen binding. Complementing the genetic analysis with traditional molecular microbiological techniques enabled the identification of the novel fibrinogen receptor, CCUG_2371. Although only strain CCUG 47825 bound fibrinogen under laboratory conditions, homologues of the novel fibrinogen binding gene CCUG_2371 are widespread among L. salivarius strains, maintaining their potential to bind fibrinogen if expressed. We highlight the fact that without a full genetic analysis of strains for human consumption, potential pathogenicity traits may go undetected.
Molecular Microbiology 06/2012; 85(5):862-77. · 5.01 Impact Factor
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ABSTRACT: To assess the ability of human intestinally derived strains of Lactobacillus and Bifidobacterium to produce γ-aminobutyric acid (GABA).
Strains of Lactobacillus and Bifidobacterium were grown in medium containing monosodium glutamate (MSG). Growth of the bacteria and conversion of MSG to GABA were measured. Of 91 intestinally derived bacteria assessed, one Lactobacillus strain and four strains of Bifidobacterium produced GABA. Lactobacillus brevis DPC6108 was the most efficient of the strains tested, converting up to 100% of MSG to GABA. The ability of the cultured intestinal strains to produce GABA was investigated using a simple pH-controlled anaerobic faeces-based fermentation, supplemented with 30 mg ml⁻¹ MSG. The addition of Lact. brevis DPC6108 to a faeces-based fermentation significantly increased the GABA concentration (P < 0·001), supporting the notion that this biosynthesis could occur in vivo.
The production of GABA by bifidobacteria exhibited considerable interspecies variation. Lactobacillus brevis and Bifidobacterium dentium were the most efficient GABA producers among the range of strains tested. The addition of Lact. brevis DPC6108 to the culturable gut microbiota increased the GABA concentration in fermented faecal slurry at physiological pH.
Identification of optimal MSG conversion to GABA by particular cultured elements of the commensal intestinal microbiota and the demonstration that this can occur under simulated in vivo conditions offer new prospects for microbiota modulation to promote health.
Journal of Applied Microbiology 05/2012; 113(2):411-7. · 2.34 Impact Factor
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ABSTRACT: Lactobacillus salivarius strain UCC118 is a human intestinal isolate that has been extensively studied for its potential probiotic effects in human and animal models. The objective of this study was to determine the effect of L. salivarius UCC118 on gene expression responses in the Caco-2 cell line to improve understanding of how the strain might modulate intestinal epithelial cell phenotypes. Exposure of Caco-2 cells to UCC118 led to the induction of several human genes (TNFAIP3, NFKBIA, and BIRC3) that are negative regulators of inflammatory signaling pathways. Induction of chemokines (CCL20, CXCL-1, and CXCL-2) with antimicrobial functions was also observed. Disruption of the UCC118 sortase gene srtA causes reduced bacterial adhesion to epithelial cells. Transcription of three mucin genes was reduced significantly when Caco-2 cells were stimulated with the ΔsrtA derivative of UCC118 compared to cells stimulated with the wild type, but there was no significant change in the transcription levels of the anti-inflammatory genes. UCC118 genes that were significantly upregulated upon exposure to Caco-2 cells were identified by bacterial genome microarray and consisted primarily of two groups of genes connected with purine metabolism and the operon for synthesis of the Abp118 bacteriocin. Following incubation with Caco-2 cells, the bacteriocin synthesis genes were transcribed at higher levels in the wild type than in the ΔsrtA derivative. These data indicate that L. salivarius UCC118 influences epithelial cells both through modulation of the inflammatory response and by modulation of intestinal cell mucin production. Sortase-anchored cell surface proteins of L. salivarius UCC118 have a central role in promoting the interaction between the bacterium and epithelial cells.
Applied and environmental microbiology 05/2012; 78(15):5196-203. · 3.69 Impact Factor
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ABSTRACT: Obesity develops from a prolonged imbalance of energy intake and energy expenditure. However, the relatively recent discovery that the composition and function of the gut microbiota impacts on obesity has lead to an explosion of interest in what is now a distinct research field. Here, research relating to the links between the gut microbiota, diet and obesity will be reviewed under five major headings: (1) the gut microbiota of lean and obese animals, (2) the composition of the gut microbiota of lean and obese humans, (3) the impact of diet on the gut microbiota, (4) manipulating the gut microbiota and (5) the mechanisms by which the gut microbiota can impact on weight gain.
Gut Microbes 05/2012; 3(3):186-202.
