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PubMed2002

Influence of resistant starch on the SCFA production and cell counts of butyrate-producing Eubacterium spp. in the human intestine.

AIMS: The genus Eubacterium, which is the second most common genus in the human intestine, includes several known butyrate producers. We hypothesized that Eubacterium species play a role in the intestinal butyrate production and are inducible by resistant starch. METHODS AND RESULTS: In a human pilot study species-specific and group-specific 16S rRNA-targeted, Cy3 (indocarbocyanine)-labelled oligonucleotide probes were used to quantify butyrogenic species of the genera Eubacterium, Clostridium and Ruminococcus. Following the intake of RS type III a significant increase in faecal butyrate but not in total SCFA was observed. However, increase in butyrate was not accompanied by a proliferation in the targeted bacteria. CONCLUSIONS: The tested Eubacterium species have the capacity to produce butyrate but do not appear to play a major role for butyric acid production in the human intestine. SIGNIFICANCE AND IMPACT OF THE STUDY: In view of the fact that the bacteria responsible for butyrate production are largely unknown, it is still difficult to devise a dietary intervention to stimulate butyrogenic bacteria in a targeted way.

Acetates↗

PubMed2002

Reclassification of Eubacterium formicigenerans Holdeman and Moore 1974 as Dorea formicigenerans gen. nov., comb. nov., and description of Dorea longicatena sp. nov., isolated from human faeces.

Two strains of a gram-positively staining, obligately anaerobic, non-spore-forming, rod-shaped bacterium, designated strains 111-13A and 111-35T, were isolated from human faeces. Analysis of the 16S rRNA gene sequences indicated that these strains were members of the Clostridium coccoides rRNA group of organisms. The nearest relatives of the unknown bacterium were Eubacterium formicigenerans (having a sequence similarity of 94%) and an uncultured bacterium (similarity > 99%). Characterization studies indicated that the unidentified faecal bacterium was biochemically distinct from Eubacterium formicigenerans, members of the Clostridium coccoides group and all other described Eubacterium species. On the basis of the data from these studies, it is proposed that the hitherto unknown rod-shaped bacterium be designated a species of a novel genus, namely Dorea longicatena gen. nov., sp. nov., and that Eubacterium formicigenerans be transferred to this genus as Dorea formicigenerans gen. nov., comb. nov.

Eubacterium↗

PubMed2001

Characterisation of Eubacterium-like strains isolated from oral infections.

The genus Eubacterium currently includes a heterogeneous group of gram-positive, non-spore-forming anaerobic bacilli, many of which are slow growing, fastidious and generally unreactive in biochemical tests. As a consequence, cultivation and identification of isolates are difficult and the taxonomy of the group remains indifferent. In this study, 105 isolates from odontogenic infections, infections associated with dental implants or saliva from healthy subjects and provisionally assigned to the genus Eubacterium were subjected to phenotypic and genotypic analysis. Ninety-one of the isolates were identified as belonging to one of 14 previously described species: Atopobium parvulum (5 isolates), A. rimae (29), Bulleidia extructa (2), Cryptobacterium curtum (1), Dialister pneumosintes (1), Eubacterium saburreum (2), E. sulci (8), E. yurii subsp. yurii (1), Filifactor alocis (3), Lactobacillus uli (1), Mogibacterium timidum (13), M. vescum (6), Pseudoramibacter alactolyticus (6) and Slackia exigua (13). The remaining 14 isolates did not correspond to existing species. This study confirms the diversity of organisms provisionally assigned to the genus Eubacterium by conventional identification methods. This group of organisms is frequently isolated from oral infections but their role in the aetiology of these conditions has yet to be determined.

Actinomycetales Infections↗

PubMed1980

Biochemical characterization of cholesterol-reducing Eubacterium.

We characterized two isolates of cholesterol-reducing Eubacterium by conducting conventional biochemical tests and by testing various sterols and glycerolipids as potential growth factors. In media containing cholesterol and plasmenylethanolamine, the tests for nitrate reduction, indole production, and gelatin and starch hydrolyses were negative, and no acid was produced from any of 22 carbohydrates. Both isolates hydrolyzed esculin to esculetin, indicating beta-glycosidase activity. In addition to plasmenylethanolamine, five other lipids which contain an alkenyl ether residue supported growth of Eubacterium strain 403 in a lecithin-cholesterol base medium. Of six steroids tested, cholesterol, cholest-4-en-3-one, cholest-4-en-3 beta-ol (allocholesterol), and androst-5-en-3 beta-ol-17-one supported growth of Eubacterium strain 403. All four steroids were reduced to the 3 beta-ol, 5 beta-H products. The delta 5 steroids cholest-5-en-3 alpha-ol (epicholesterol) and 22,23-bisnor-5-cholenic acid-3-beta-ol were not reduced and did not support growth of the Eubacterium strain.

Carbohydrate Metabolism↗

PubMed1988

Glycyrrhizin stimulates growth of Eubacterium sp. strain GLH, a human intestinal anaerobe.

Eubacterium sp. strain GLH was isolated from human feces and produced two kinds of beta-D-glucuronidase (EC 3.2.1.31), one new enzyme specific for glycyrrhizin (GL) and the other for phenyl beta-D-glucuronides. GL or p-nitrophenyl-mono-beta-D-glucuronide (pNPG) stimulated the production of GL or pNPG beta-glucuronidases and the growth of strain GLH in a basal medium lacking carbohydrate. D-Glucuronic acid also stimulated the growth of the bacterium, but glycyrrhetic acid did not. The increase of GL beta-glucuronidase paralleled the growth of the Eubacterium strain in pure culture. These results suggest that glucuronides such as GL and pNPG stimulate the growth of the Eubacterium strain in a nutrient-poor medium by providing D-glucuronic acid through the activity of beta-glucuronidases. The increase in GL beta-glucuronidase activity in the presence of GL was observed during the cultivation of human intestinal flora in a general anaerobic medium. During mixed cultivation of the Eubacterium strain with Streptococcus faecalis, which does not produce GL beta-glucuronidase, GL beta-glucuronidase was also increased by GL or pNPG, but not by glycyrrhetic acid and p-nitrophenol. It is suggested that GL stimulates the growth of strain GLH even in the mixed culture.

Anti-Bacterial Agents↗

PubMed2000

Quantification of different Eubacterium spp. in human fecal samples with species-specific 16S rRNA-targeted oligonucleotide probes.

Species-specific 16S rRNA-targeted, Cy3 (indocarbocyanine)-labeled oligonucleotide probes were designed and validated to quantify different Eubacterium species in human fecal samples. Probes were directed at Eubacterium barkeri, E. biforme, E. contortum, E. cylindroides (two probes), E. dolichum, E. hadrum, E. lentum, E. limosum, E. moniliforme, and E. ventriosum. The specificity of the probes was tested with the type strains and a range of common intestinal bacteria. With one exception, none of the probes showed cross-hybridization under stringent conditions. The species-specific probes were applied to fecal samples obtained from 12 healthy volunteers. E. biforme, E. cylindroides, E. hadrum, E. lentum, and E. ventriosum could be determined. All other Eubacterium species for which probes had been designed were under the detection limit of 10(7) cells g (dry weight) of feces(-1). The cell counts obtained are essentially in accordance with the literature data, which are based on colony counts. This shows that whole-cell in situ hybridization with species-specific probes is a valuable tool for the enumeration of Eubacterium species in feces.

Colony Count, Microbial↗

PubMed1990

Chronic arthritis induced in rats by cell wall fragments of Eubacterium species from the human intestinal flora.

To investigate arthritis-inducing properties of Eubacterium species, which are major residents of the human intestinal flora, cell wall fragments (CWF) of several Eubacterium strains were prepared and tested in an animal model. After a single intraperitoneal injection in the rat, CWF of E. aerofaciens, E. contortum, and E. lentum induced a chronic polyarthritis. E. limosum and E. tortuosum CWF induced an acute self-limiting joint inflammation, whereas E. rectale CWF failed to do so. The rhamnose contents of the isolated CWF were not related to their arthritis-inducing properties. Paradoxically, the sensitivity of CWF to lysozyme digestion, which is regarded as a parameter for the clearance of CWF in tissues, appeared to be positively correlated with the ability of Eubacterium CWF to induce chronic joint inflammation. Our findings show the diversity in arthritis-inducing properties among different species of the anaerobic genus Eubacterium and underline the importance of the anaerobic intestinal flora in the induction of joint inflammation.

Animals↗

PubMed1991

Cloning, sequencing, and expression of the gene coding for bile acid 7 alpha-hydroxysteroid dehydrogenase from Eubacterium sp. strain VPI 12708.

Southern blot analysis indicated that the gene encoding the constitutive, NADP-linked bile acid 7 alpha-hydroxysteroid dehydrogenase of Eubacterium sp. strain VPI 12708 was located on a 6.5-kb EcoRI fragment of the chromosomal DNA. This fragment was cloned into bacteriophage lambda gt11, and a 2.9-kb piece of this insert was subcloned into pUC19, yielding the recombinant plasmid pBH51. DNA sequence analysis of the 7 alpha-hydroxysteroid dehydrogenase gene in pBH51 revealed a 798-bp open reading frame, coding for a protein with a calculated molecular weight of 28,500. A putative promoter sequence and ribosome binding site were identified. The 7 alpha-hydroxysteroid dehydrogenase mRNA transcript in Eubacterium sp. strain VPI 12708 was about 0.94 kb in length, suggesting that it is monocistronic. An Escherichia coli DH5 alpha transformant harboring pBH51 had approximately 30-fold greater levels of 7 alpha-hydroxysteroid dehydrogenase mRNA, immunoreactive protein, and specific activity than Eubacterium sp. strain VPI 12708. The 7 alpha-hydroxysteroid dehydrogenase purified from the pBH51 transformant was similar in subunit molecular weight, specific activity, and kinetic properties to that from Eubacterium sp. strain VPI 12708, and it reached with antiserum raised against the authentic enzyme on Western immunoblots. Alignment of the amino acid sequence of the 7 alpha-hydroxysteroid dehydrogenase with those of 10 other pyridine nucleotide-linked alcohol/polyol dehydrogenases revealed six conserved amino acid residues in the N-terminal regions thought to function in coenzyme binding.

Amino Acid Sequence↗

PubMed2000

Competition in the metabolism of glycyrrhizin with glycyrrhetic acid mono-glucuronide by mixed Eubacterium sp. GLH and Ruminococcus sp. PO1-3.

Eubacterium sp. GLH possessing glycyrrhizin (GL) and glycyrrhetic acid mono-glucuronide (GAMG) beta-D-glucuronidases, Ruminococcus sp. PO1-3 possessing GL and GAMG beta-D-glucuronidases and 3beta-hydroxysteroid dehydrogenase and these mixed bacteria were cultured in GAM medium with and without GL, GAMG or both. GL added to Eubacterium sp. GLH accelerated the peaks of enhanced GL beta-D-glucuronidase activity and suppressed GAMG beta-D-glucuronidase activity, and GAMG delayed the peaks of the enhanced growth with GL and GAMG beta-D-glucuronidase activities. GL added to Ruminococcus sp. PO1-3 enhanced gradually the growth with GL and GAMG beta-D-glucuronidase activities, and GAMG enhanced slowly GL beta-D-glucuronidase activity and rapidly the growth with GAMG beta-D-glucuronidase activity. The metabolite glycyrrhetic acid (GA) was produced by Eubacterium sp. GLH and Ruminococcus sp. PO1-3 in larger amounts and faster from GAMG than from GL. GL (1.0 mM) and 1.0 mM GAMG added to these mixed bacteria enhanced the growth with GL and GAMG beta-D-glucuronidase activities and were metabolized almost completely to GA in culture of 2 d and 1 d, respectively. It was found that the metabolism of GAMG was faster than that of GL. GL with GAMG added to mixed Eubacterium sp. GLH and Ruminococcus sp. PO1-3 cultured for 0 and 1 d led to a lower level of these enzyme activities and the consumption of GAMG more quickly, not GL. Low GAMG beta-D-glucuronidase had the ability to hydrolyze GAMG well.

Carbohydrate Sequence↗

PubMed1999

Increased growth of Bifidobacterium and Eubacterium by germinated barley foodstuff, accompanied by enhanced butyrate production in healthy volunteers.

Germinated barley foodstuff (GBF) derived from the aleurone and scutellum fractions of germinated barley mainly consists of low-lignified hemicellulose and glutamine-rich protein. GBF improves the proliferation of intestinal epithelial cells and defecation, through the bacterial production of short chain fatty acids (SCFA), especially butyrate. In this study we investigated the mechanism of production of butyrate by microflora in humans and in vitro. Daily administration of 9 g GBF for 14 successive days significantly increased fecal butyrate content. Fecal Bifidobacterium and Eubacterium were also significantly increased by GBF administration in healthy volunteers. Ten anaerobic micro-organisms selected from intestinal microflora were cultured in vitro in the medium containing GBF as a sole carbon source (GBF medium). After a 3-day incubation, 7 strains (Bifidobacterium breve, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus casei subsp. casei, Bacteroides ovatus, Clostridium butyricum, and Eubacterium limosum) lowered the medium pH producing SCFA. Eubacterium grown together with Bifidobacterium in GBF medium efficiently produced butyrate. On the other hand, GBF changed the intestinal microflora and increased probiotics such as Bifidobacterium in the intestinal tract. As a result, butyrate was produced by the mutual action of Eubacterium and Bifidobacterium. This butyrate is considered to enhance the proliferation of colonic epithelial cells.

Adult↗

PubMed1996

Identification and discrimination of oral asaccharolytic Eubacterium spp. by pyrolysis mass spectrometry and artificial neural networks.

Curie-point pyrolysis mass spectra were obtained from 29 oral asaccharolytic Eubacterium strains and 6 abscess isolates previously identified as Peptostreptococcus heliotrinreducens. Pyrolysis mass spectrometry (PyMS) with cluster analysis was able to clarify the taxonomic position of this group of organisms. Artificial neural networks (ANNS) were then trained by supervised learning (with the back-propagation algorithm) to recognize the strains from their pyrolysis mass spectra; all Eubacterium strains were correctly identified, and the abscess isolates were identified as un-named Eubacterium taxon C2 and were distinct from the type strain of P. heliotrinreducens. These results demonstrate that the combination of PyMS and ANNs provides a rapid and accurate identification technique.

Bacterial Typing Techniques↗

PubMed1989

Antimicrobial susceptibilities of Eubacterium, Peptostreptococcus, and Bacteroides isolated from root canals of teeth with periapical pathosis.

Eubacterium, Peptostreptococcus, and Bacteroides were isolated in high frequency from root canals with acute periapical inflammation. The antimicrobial susceptibilities of these strains were studied by determining minimum inhibitory concentrations of different agents. Although all three kinds of isolates were susceptible to penicillins, the isolates other than black-pigmented Bacteroides were less susceptible to cephems, tetracyclines, and macrorides with several resistant strains. All strains were uniformly resistant to aminoglycosides. Some differences in susceptibilities were observed among species of Eubacterium and Peptostreptococcus, while penicillins were effective for both species. Black-pigmented Bacteroides showed good susceptibilities to all agents except for aminoglycosides. The susceptibility of Bacteroides gingivalis was superior to that of Bacteroides intermedius. There were many resistant strains in non-black-pigmented but not in black-pigmented Bacteroides isolates. Penicillins were the most effective for Eubacterium, Peptostreptococcus, and Bacteroides, indicating that penicillins are suitable for treatment of root canals with acute apical periodontitis.

Aminoglycosides↗

PubMed1981

Serum agglutinins to Eubacterium and Peptostreptococcus species in Crohn's and other diseases.

Sera from patients suffering from Crohn's and other diseases and from healthy subjects were tested for agglutinins to anaerobic, gram-positive coccoid rods belonging to species of Eubacterium and Peptostreptococcus. Four strains labelled Eubacterium contortum (two strains), Eubacterium rectale and Peptostreptococcus productus were agglutinated by a higher percentage of sera from patients with Crohn's disease than from healthy subjects and from patients with liver and intestinal diseases (including ulcerative colitis), ankylosing spondylitis, granulomatous diseases, diseases of immunity and malignancies. The agglutinins were of the IgG and IgM classes and strain-specific; the titres were low. The results obtained with sera from patients with Crohn's disease and healthy people were subjected to discriminant analysis to estimate the probability, based on the combined results with the four strains, that a patient suffers from Crohn's disease. When sera giving an a posteriori probability greater than or equal to 0.95 (a priori probability = 0.5) were considered positive, the test with four strains had a sensitivity of 54% and a specificity of nearly 100%. The results with sera submitted for diagnosis showed that positive reactions in patients with a diagnosis apparently incompatible with Crohn's disease were within acceptable limits.

Adolescent↗

PubMed2002

Eubacterium bacteremia and colon cancer.

Eubacterium bacteremia is rare. We report a senile patient who presented with 3 episodes of bacteremia, caused by Clostridium perfringens, Eubacterium limosum and Escherichia coli, respectively, which led to the diagnosis of adenocarcinoma of the colon. A differential diagnosis of colon cancer should be considered in patients with eubacterium bacteremia.

Aged↗

PubMed1990

Intestinal flora of patients with rheumatoid arthritis: induction of chronic arthritis in rats by cell wall fragments from isolated Eubacterium aerofaciens strains.

The composition of the obligate anaerobic intestinal flora of patients with rheumatoid arthritis (RA) differed from that of healthy subjects (HS). Total numbers of aerobes as well as anaerobic coccoid rods were found elevated when compared with HS. Eubacterium species were found in all stool samples of both groups; Bifidobacterium species were present in seven (RA) and eight (HS) out of 10 subjects. From the flora of two RA patients and two HS Eubacterium species were isolated and identified. Cell wall fragments from four E. aerofaciens strains (two from RA, two from HS) were tested for arthritis induction in rats. All four strains induced chronic arthritis which was histologically confirmed. We concluded that in the normal intestinal flora of RA patients Eubacterium species are present in high numbers (i.e. greater than 10(9)/g faeces); cell walls from isolated E. aerofaciens strains had arthropathic properties.

Animals↗

PubMed1994

Characterization of Eubacterium coprostanoligenes sp. nov., a cholesterol-reducing anaerobe.

A small, anaerobic, gram-positive coccobacillus that reduces cholesterol to coprostanol was isolated from a hog sewage lagoon. This isolate, strain HLT (T = type strain) does not require cholesterol for growth, but it requires lecithin and has phospholipase activity. Much acid is produced by the fermentation of amygdalin, lactose, and salicin. Arabinose, cellobiose, fructose, glucose, mannose, and melibiose are fermented weakly. Acetic, formic, and succinic acids are produced, as is hydrogen. The isolate does not reduce nitrate, produce indole, or hydrolyze starch and gelatin. Esculin is hydrolyzed. The properties of strain HLT are most similar to those of members of the genus Eubacterium. Because strain HL (= ATCC 51222) has unique morphological and physiological properties, we propose that it should be the type strain of a new species in the genus Eubacterium, Eubacterium coprostanoligenes.

Anaerobiosis↗

PubMed1997

Phenotypic and phylogenetic characterization of some Eubacterium-like isolates containing a novel type B wall murein from human feces: description of Holdemania filiformis gen. nov., sp. nov.

A group of Eubacterium-like strains (designated group S14), isolated from the feces of healthy people, was characterized by biochemical tests, fatty acid analysis, cell wall murein analysis, and 16S rDNA analysis. Our results indicate that group S14 is phylogenetically a member of the Clostridium subphylum of the gram-positive bacteria. Despite a phenotypic resemblance to the genus Eubacterium, group S14 was shown to be phylogenetically distantly related to the type species of the genus, Eubacterium limosum. Group S14 showed a specific phylogenetic association with Erysipelothrix rhusiopathiae. Group S14 resembled Erysipelothrix in possessing the uncommon type B cell wall murein. Structural analyses, however, revealed the presence of a previously unknown B1 delta (L-Ala)-D-Glu-Gly-L-Lys murein type. Based on a 16S rRNA sequence divergence of greater than 10% with E. rhusiopathiae and the presence of a unique murein type, a new genus, Holdemania, is proposed for group S14, with one species, Holdemania filiformis. Type strain of H. filiformis is ATCC 51649.

Cell Wall↗

PubMed1996

Reduction of oxalate content of foods by the oxalate degrading bacterium, Eubacterium lentum WYH-1.

BACKGROUND: Urinary oxalate may contribute far more than urinary calcium to the pathogenesis of urinary calculi. Urinary oxalate may be reduced by restricting the intake of foods high in oxalate. The oxalate content foods might be reduced by oxalate-degrading bacteria. The purpose of this experiment was to reduce the oxalate content of foods with an oxalate-degrading bacterium which was isolated from the feces of Japanese male. METHODS: An artificial intestinal juice was prepared by modifying Rogosa medium. An infusion of black tea was prepared from a commercial tea bag. The oxalate-degrading bacteria used were Eubacterium lentum WYH-1 which were have isolated. To 5 ml of the above oxalate-containing artificial intestinal juice and infusion of black tea, 0.5 ml of the bacterial culture was added and incubated anaerobically at 37 degrees C. Oxalic acid in the supernatant of the culture medium was assayed by high-performance liquid chromatography. RESULTS: In 24 hours, 1 x 10(6) cells/ml of Eubacterium lentum WYH-1 decomposed 100% of 1 mg/ml oxalate in the artificial intestinal juice. The oxalate in the black tea infusion (1 mg/mL) was also decomposed completely within 48 hours by 1 x 10(7) cells/mL of bacteria. CONCLUSION: Eubacterium lentum WYH-1 was able to efficiently decompose the oxalate in foods.

Chromatography, High Pressure Liquid↗