Eubacterium rectale-Clostridium Research Articles

Freshwater microalgae biomasses exert a prebiotic effect on human colonic microbiota

The effects of digested microalgae biomass of Chlorella vulgaris, Desmodesmus maximus, Chlorococcum sp. cf hypnosporum, and Spirulina platensis or fructooligosaccharides on human colonic microbiota during in vitro fermentation (48 h) were evaluated. Fructooligosaccharides and digested microalgae biomass increased Lactobacillus-Enterococcus and Bifidobacterium relative abundances and decreased Prevotellaceae-Bacteroidaceae, Clostridium histolyticum, and Eubacterium rectale-Clostridium coccoides. The digested microalgae biomass s showed a higher prebiotic index than fructooligosaccharides, evidenced through the higher selective stimulation of beneficial microorganisms and inhibition of undesirable bacteria on colonic microbiota. S. platensis biomass contained nystose, raffinose, and phenolics, increased the relative abundance of Lactobacillus-Enterococcus and Bifidobacterium, and decreased Eubacterium rectale-Clostridium coccoides, Clostridium histolyticum, and Prevotellaceae-Bacteroidaceae. After 24 h of incubation, the medium fermented with S. platensis promoted an increase in the concentration of acetic and propionic acids. D. maximus biomass showed higher insoluble fibers and kestose, the greatest Lactobacillus-Enterococcus relative abundance, and increases in the butyric and propionic acids after 48 h. C. vulgaris biomass showed the highest glucose and fructose concentrations, some phenolics, and the highest Bifidobacterium relative abundance after 48 h.

Impact of 2'-Fucosyllactose on Gut Microbiota Composition in Adults with Chronic Gastrointestinal Conditions: Batch Culture Fermentation Model and Pilot Clinical Trial Findings.

Intestinal dysbiosis has been described in patients with certain gastrointestinal conditions including irritable bowel syndrome (IBS) and ulcerative colitis. 2′-fucosyllactose (2′-FL), a prebiotic human milk oligosaccharide, is considered bifidogenic and butyrogenic. To assess prebiotic effects of 2′-FL, alone or in combination with probiotic strains (potential synbiotics), in vitro experiments were conducted on stool from healthy, IBS, and ulcerative colitis adult donors. In anaerobic batch culture fermenters, Bifidobacterium and Eubacterium rectale-Clostridium coccoides counts, and short-chain fatty acids (SCFAs) including butyrate increased during fermentation with 2′-FL and some of the 2′-FL/probiotic combinations. In a subsequent open-label pilot trial, the effect of a 2′-FL-containing nutritional formula was evaluated in twelve adults with IBS or ulcerative colitis. Gastrointestinal Quality of Life Index (GIQLI) total and gastrointestinal symptoms domain scores, stool counts of Bifidobacterium and Faecalibacterium prausnitzii, and stool SCFAs including butyrate, increased after six weeks of intervention. Consistent with documented effects of 2′-FL, the batch culture fermentation experiments demonstrated bifidogenic and butyrogenic effects of 2′-FL during fermentation with human stool samples. Consumption of the 2′-FL-containing nutritional formula by adults with IBS or ulcerative colitis was associated with improvements in intra- and extra-intestinal symptoms, and bifidogenic and butyrogenic effects.

The Association between Cardiorespiratory Fitness and Gut Microbiota Composition in Premenopausal Women

The aim of this study was to investigate the association between cardiorespiratory fitness and gut microbiota composition in premenopausal women. The participants consisted of 71 premenopausal Finnish women (aged 19–49 years). Gut microbiota were analyzed using flow cytometry, 16S rRNA gene hybridization and DNA-staining. Maximum oxygen uptake (VO2max) was assessed by respiratory gas analyzer and body composition by Bioimpdance. We found that participants with low VO2max had lower Bacteroides, but higher Eubacterium rectale-Clostridium coccoides than the high VO2max group (p < 0.05 for all). VO2max was inversely associated with EreC (r = −0.309, p = 0.01) but not with other bacteria. VO2max also negatively correlated with fat% (r = −0.755, p < 0.001), triglycerides (r = −0.274, p = 0.021) and leptin (r = −0.574, p < 0.001). By contrast, EreC was positively associated with fat% (r = 0.382, p = 0.002), dietary fat intake (r = 0.258, p = 0.034), triglycerides (r = 0.390, p = 0.002) and leptin (r = 0.424, p = 0.001), but negatively with carbohydrate intake (r = −0.252, p = 0.034) and HDL (r = −0.26, p = 0.028). After adjusting for age and dietary intake, all the significant associations remained. However, after adjusting for fat%, the associations between VO2max and EreC disappeared. Our results suggest that cardiorespiratory fitness is associated with gut microbiota composition, independent of age and carbohydrate or fat intake. The association between VO2max and EreC, however, appears to be mediated by body fatness.

Allergic diseases of the skin and drug allergies – 2013. Longitudinal analysis of fecal microbiota of infants followed-up for eczema till 2 years of age

Methods Children with eczema at 2 years old (n=26) and their matched (for gender, mode of delivery and feeding in first 6 months) healthy controls (n=26) were selected from the placebo group of a cohort of at-risk infants participating in an randomized double-blind placebo controlled trial on the protective effects of supplemental probiotics (first 6 months) on eczema and allergies. Children with eczema were subclassified into atopic eczema (n=12) and non-atopic eczema (n=14). Molecular evaluation of fecal microbiota were conducted using Fluorescence In Situ Hybridization-Flow Cytometry (FISH-FC) for fecal samples collected at 3 days, 1, 3, and 12 months. Probes were selected to target Eubacterium rectale-Clostridium coccoides group (Erec482), Clostridium leptum subgroup (Clep866 and the corresponding competitor probes), Bacteroides-Prevotella group (Bac303), Bifidobacterium genus (Bif164), Atopobium group (Ato291), LactobacilliEnterococci group (Lab158), Enterobacteriaceae family (Enter1432) and Clostridium perfringens (Cperf191). Linear mixed model was used to evaluate the longitudinal differences (i.e. 4 time points) of bacterial targets while adjusting for gender, mode of delivery, feeding up to 6 months, and allergic rhinitis and wheezing within the eczema group at 2 years of age. Results Longitudinal analyses over four time points showed that higher relative abundance of Enterobacteriaceae [coefficient (B): 1.104, 95% confidence interval (CI):0.175-2.033, adj p=0.022] in children with eczema by 2 years of age. Similar observations were made when eczema group was subanalyzed into non-atopic and atopic eczema, where higher relative abundance of Enterobacteriaceae [B:1.357, 95%CI; 0.382-2.332, adj p=0.008] and [B:1.165, 95%CI; 0.221-2.109, adj p=0.019] were observed respectively as compared to healthy controls. Relative abundance of Clostridium perfringens were also higher when subanalyzed for non-atopic [B:0.572, 95%CI; 0.0009-1.144, adj p=0.050] and atopic eczema [B:0.000451, 95%CI: 0.0001-0.0007, adj p=0.012] compared to healthy controls.

Preliminary investigation of intestinal microflora in patients with hepatic cirrhosis

To examine the differential levels of fecal Bifidobacterium, Bacteroides, Eubacterium rectale-Clostridium, Escherichia coli, Enterococcus, and Clostridium difficile between patients with hepatic cirrhosis and healthy controls. Fecal samples were collected from 29 patients with hepatic cirrhosis treated in the Department of Digestive Diseases at Zunyi Hospital between March and December of 2010. Fecal samples were collected from 13 healthy college students for use as controls. All samples were assessed by pH measurement, bacterial culture for turbidity, fluorescence in situ hybridization, and laser scanning confocal microscopy. The t-test and rank correlation test were used to determine statistical significance of intergroup differences in each tested parameter. The feces of patients with hepatic cirrhosis had higher pH than that of healthy controls (6.79+/-0.64 vs. 6.18+/-0.74, P less than 0.05). The bacterial turbidity was not significantly different between the feces of hepatic cirrhosis patients and healthy controls (1.15+/-0.59 vs. 1.39+/-1.01, P more than 0.05). The numbers of Bifidobacterium, Bacteroides, Eubacterium rectale-Clostridium, Escherichia coli, Enterococcus, and Clostridium difficile in feces of patients with hepatic cirrhosis were significantly lower than those of the controls (all P less than 0.01). No significant correlation was found between the number or ratio of bacteria species and the severity of hepatic cirrhosis (Child-Pugh scores; P more than 0.05). The total quantity of intestinal bacteria in patients with hepatic cirrhosis is not significantly different from that in healthy patients. However, the profile of intestinal bacteria is different, which may explain the increased pH of fecal samples from patients with hepatic cirrhosis, but the differential profile is not correlated to cirrhosis pathogenesis.

Association between the ABO blood group and the human intestinal microbiota composition

BackgroundThe mucus layer covering the human intestinal epithelium forms a dynamic surface for host-microbial interactions. In addition to the environmental factors affecting the intestinal equilibrium, such as diet, it is well established that the microbiota composition is individually driven, but the host factors determining the composition have remained unresolved.ResultsIn this study, we show that ABO blood group is involved in differences in relative proportion and overall profiles of intestinal microbiota. Specifically, the microbiota from the individuals harbouring the B antigen (secretor B and AB) differed from the non-B antigen groups and also showed higher diversity of the Eubacterium rectale-Clostridium coccoides (EREC) and Clostridium leptum (CLEPT) -groups in comparison with other blood groups.ConclusionsOur novel finding indicates that the ABO blood group is one of the genetically determined host factors modulating the composition of the human intestinal microbiota, thus enabling new applications in the field of personalized nutrition and medicine.

Women With and Without Metabolic Disorder Differ in Their Gut Microbiota Composition

The aim of this study was to investigate whether overweight/obese women in metabolic disorder group (MDG, n = 27) differ in their gut microbiota composition from overweight/obese women in non-metabolic disorder group (NMDG, n = 47) and normal weight women group (NWG, n = 11). Gut microbiota was profiled from fecal samples by 16S rRNA fluorescence in situ hybridization and flow cytometry in 85 premenopausal women. Body composition was measured by bioimpedance, and dietary intakes were collected via food diaries. Standard procedures were used to assess plasma glucose, serum insulin, lipids, and inflammatory status. We found that the proportion of bacteria belonging to Eubacterium rectale-Clostridium coccoides group, indicating efficient energy harvest from nutrients in gut, was higher in MDG compared to NMDG and NWG, while no difference was found between NMDG and NWG. The proportion of Eubacterium rectale-Clostridium coccoides group correlated positively with weight, BMI, total fat, fat mass percentage (FM%), visceral fat area, and serum triglycerides, and negatively with high-density lipoprotein (HDL). Our results indicate that certain members of Eubacterium rectale-Clostridium coccoides group are associated with obesity-related MDs not obesity per se.

Distinct microbial populations exist in the mucosa‐associated microbiota of sub‐groups of irritable bowel syndrome

There is increasing evidence to support a role for the gastrointestinal microbiota in the etiology of irritable bowel syndrome (IBS). Given the evidence of an inflammatory component to IBS, the mucosa-associated microbiota potentially play a key role in its pathogenesis. The objectives were to compare the mucosa-associated microbiota between patients with diarrhea predominant IBS (IBS-D), constipation predominant IBS (IBS-C) and controls using fluorescent in situ hybridization and to correlate specific bacteria groups with individual IBS symptoms. Forty-seven patients with IBS (27 IBS-D and 20 IBS-C) and 26 healthy controls were recruited to the study. Snap-frozen rectal biopsies were taken at colonoscopy and bacterial quantification performed by hybridizing frozen sections with bacterial-group specific oligonucleotide probes. Patients with IBS had significantly greater numbers of total mucosa-associated bacteria per mm of rectal epithelium than controls [median 218 (IQR - 209) vs 128 (121) P = 0.007], and this was chiefly comprised of bacteroides IBS [69 (67) vs 14 (41) P = 0.001] and Eubacterium rectale-Clostridium coccoides [52 (58) vs 25 (35) P = 0.03]. Analysis of IBS sub-groups demonstrated that bifidobacteria were lower in the IBS-D group than in the IBS-C group and controls [24 (32) vs 54 (88) vs 32 (35) P = 0.011]. Finally, amongst patients with IBS, the maximum number of stools per day negatively correlated with the number of mucosa-associated bifidobacteria (P < 0.001) and lactobacilli (P = 0.002). The mucosa-associated microbiota in patients with IBS is significantly different from healthy controls with increases in bacteroides and clostridia and a reduction in bifidobacteria in patients with IBS-D.

Effect of a Milk Formula Containing Probiotics on the Fecal Microbiota of Asian Infants at Risk of Atopic Diseases

The fecal microbiota of 37 infants with (n = 20) or without (n = 17) probiotic administration was evaluated on D 3, and at 1, 3, and 12 mo by fluorescence in situ hybridization-flow cytometry (FISH-FC), PCR, and bacteriological culture methods. They represent consecutive subjects of an ongoing double-blind, placebo-controlled trial on a probiotic formula (LGG and Bifidobacterium longum) administered during the first 6 mo of life. Despite varying composition in each baby, there was a general bacterial colonization pattern in the first year. Bifidobacteria increased markedly (p = 0.0003) with a parallel decrease in Enterobacteriaceae (p < 0.001) and Bacteroides-Prevotella (p = 0.005) populations. Eubacterium rectale-Clostridium coccoides (p < 0.001) and Atopobium (p = 0.039) groups also gradually increased. This overall pattern was unaffected by probiotic administration (p > 0.05). B. longum (p = 0.005) and Lactobacillus rhamnosus (p < 0.001) were detected more frequently in probiotic group during supplementation, but no difference after supplementation had ceased (p > 0.05). Cultured lactic acid bacteria were also more numerous in the probiotic-administered babies during treatment period (log CFU/g 8.4 versus 7.4; p = 0.035). Our results indicate that supplemented strains could be detected but did not persist in the bowel once probiotic administration had ceased.

Design and evaluation of group-specific oligonucleotide probes for quantitative analysis of intestinal ecosystems: their application to assessment of equine colonic microflora

Nine oligonucleotide probes complementary to conserved regions of small subunit rRNA from phylogenetically defined clusters of intestinal anaerobic bacteria were designed and evaluated for use in quantitative analysis of intestinal microflora. Optimum wash temperatures ( T w) were determined according to the temperature of dissociation ( T d) of each probe and target group specificity was demonstrated by comparing hybridisation to target and non-target rRNA immobilised on nylon membranes. Three probes are targeted to phylogenetic clusters of Clostridiaceae, clusters III, IV and IX, with three probes designed to target previously undefined clusters within the low %G+C Gram-positive phyla. The remaining three probes encompass the Cytophaga-Flexibacter-Bacteroides assemblage, the Bacillus-Lactobacillus-Streptococcus group and the Spirochaetaceae. Application of these probes, alongside available probes targeted to other intestinal bacterial groups, against rRNA extracted from equine colonic samples has provided the first quantitative data on the predominant bacterial populations inhabiting the equine large intestine. Results show the Spirochaetaceae, the Cytophaga-Flexibacter-Bacteroides assemblage, the Eubacterium rectale-Clostridium coccoides group, and ‘Unknown cluster C’ of the Clostridiaceae to be the largest populations in the equine gut, each comprising 10–30% of the total microflora in each horse sampled. Other detected notable populations were the Bacillus-Lactobacillus-Streptococcus group, Fibrobacter and ‘Unknown cluster B’, each comprising 1–10% of the total flora. Average coverage using the suite of probes described here exceeded 75% of the total microbial community.

Effects of alternative dietary substrates on competition between human colonic bacteria in an anaerobic fermentor system.

Duplicate anaerobic fermentor systems were used to examine changes in a community of human fecal bacteria supplied with different carbohydrate energy sources. A panel of group-specific fluorescent in situ hybridization probes targeting 16S rRNA sequences revealed that the fermentors supported growth of a greater proportion of Bacteroides and a lower proportion of gram-positive anaerobes related to Faecalibacterium prausnitzii, Ruminococcus flavefaciens-Ruminococcus bromii, Eubacterium rectale-Clostridium coccoides, and Eubacterium cylindroides than the proportions in the starting fecal inoculum. Nevertheless, certain substrates, such as dahlia inulin, caused a pronounced increase in the number of bacteria related to R. flavefaciens-R. bromii and E. cylindroides. The ability of three strictly anaerobic, gram-positive bacteria to compete with the complete human fecal flora was tested in the same experiment by using selective plating to enumerate the introduced strains. The Roseburia-related strain A2-183(F) was able to grow on all substrates despite the fact that it was unable to utilize complex carbohydrates in pure culture, and it was assumed that this organism survived by cross-feeding. In contrast, Roseburia intestinalis L1-82(R) and Eubacterium sp. strain A2-194(R) survived less well despite the fact that they were able to utilize polysaccharides in pure culture, except that A2-194(R) was stimulated 100-fold by inulin. These results suggest that many low-G+C-content gram-positive obligate anaerobes may be selected against during in vitro incubation, although several groups were stimulated by inulin. Thus, considerable caution is necessary when workers attempt to predict the in vivo effects of probiotics and prebiotics from their effects in vitro.

The Effect of the Prebiotic Inulin and the ProbioticBifidobacterium longumon the Fecal Microflora of Healthy Volunteers Measured by FISH and DGGE

The effect of pre- and probiotics on the human intestinal microflora was investigated by fluorescent in situ hybridization (FISH) with specific 16S rRNA-targeted oligonucleotide probes, and by denaturing gradient gel electrophoresis (DGGE) of specifically PCR-amplified 16S rRNA genes. The prebiotic inulin was orally administered in the form of Raftiline ® HP (Orafti, Belgium). Daily doses of 9 g were taken for 14 days by 10 healthy adult volunteers. The probiotic Bifidobacterium longum , encapsulated in gelatin/pectin capsules, was orally administered in the form of Bifina ® (Morishita Jintan, Japan). Nine capsules were taken each day for 14 days by 14 healthy adult volunteers. For the prebiotic study, FISH probes were used to enumerate all bacteria, bifidobacteria, the Eubacterium rectale–Clostridium coccoides group (Erec group), Bacteroides , and eubacteria of the low G+C group. No significant changes were found in the total population of bacteria, the Bacteroides nor in the low G+C group during the study. The population of bifidobacteria increased significantly, while there was a significant decrease in numbers of the Erec group. The FISH analysis of the probiotic study focused on the enumeration of bifidobacteria only, which did not increase significantly during the study. Samples collected in both studies were analyzed by Bifidobacterium -specific PCR and DGGE. The results showed that a stable bifidobacterial population was present throughout each study. Using both FISH and DGGE, we have shown that for modulation of the bifidobacterial populations in the gut, prebiotics are better candidates than probiotics, since inulin increased bifidobacterial numbers without changing the species composition, whilst the probiotic had almost no effect. Keywords: inulin, non-digestible oligosaccharides, prebiotics, probiotics, intestinal microflora, bifidobacteria, fluorescent in situ hybridization, probes, DGGE.

Intestinal anaerobic bacteria in early rheumatoid arthritis (RA)

Increasing attention has recently been paid to the normal intestinal flora as a potential source of etiological agents in RA. Changes in the intestinal flora, due to fasting or diet, have been shown to reflect improvement of the patients when they are divided into high and low-responders. Previously, evidence has also been presented that intestinal flora in the early RA is different from that of non-RA controls, due primarily to anaerobic bacteria. The present study was designed to compare the fecal microbiota of the patients with early RA with the microbiota of the control patients using 16SrRNA oligonucleotide probes, detecting a variety of anaerobic bacteria in the normal intestinal flora. Fecal samples of 25 early, disease modifying antirheumatic drugs, naive RA patients and 23 control patients suffering from noninflammatory pain were investigated. The contribution of five bacterial groups was determined by using whole cell hybridization with seven fluorescently labeled 16SrRNA-targeted oligonucleotide probes. These probes cover one third to a half of the total bacteria in the human intestine. Patients with early RA had significantly less bacteria belonging to the Bacteroides, Prevotella and Porphyromonas genera than the controls (4.7% vs. 9.5%, P = 0.00005). The finding was confirmed with a probe specific for bacteria of the Bacteroides fragilis group (1.6% vs. 2.6%, P = 0.02). The samples of RA patients and the controls did not differ significantly when five other oligonucleotide probes were applied. They were detecting bacteria in the genera Atobium, Coriobacterium, Collinsella, Bifidobacterium and Fusobacterium, and in the Eubacterium rectale-Clostridium coccoides group. We conclude that the content of anaerobic bacteria in the intestinal flora of the patients with early RA is significantly different than that of the controls. The number of bacteria belonging to the Bacteroides-Prevotella-Porphyromonas group was, on average, in RA patients only half that of the controls. If this finding can be confirmed, together with a recent suggestion that certain Bacteroides species are required for fortification of the barrier function in the intestinal epithelium, it adds further evidence to the hypothesis that intestinal bacterial flora plays a role in the etiopathogenesis of RA.