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PubMed2003

Randomized controlled trial of the effect of bifidobacteria-fermented milk on ulcerative colitis.

BACKGROUND: Alterations of intestinal flora, such as reduction in the concentration of bifidobacteria and increase in that of Bacteroides species, are apparently associated with the severity of ulcerative colitis. OBJECTIVE: We conducted a randomised clinical trial of the use of a bifidobacteria-fermented milk (BFM) supplement as a dietary adjunct in the treatment of ulcerative colitis. METHODS: The subjects were randomly divided into two groups: a group with BFM supplementation (BFM group, 11 subjects) and a control group (control group, 10 subjects). The BFM group was given 100 mL/day of BFM for one year. Colonoscopies, general blood markers and examinations of intestinal flora including the analysis of fecal organic acids were performed at the commencement of the study and after one year. RESULTS: Exacerbation of symptoms was seen in 3 out of 11 subjects in the BFM group and in 9 out of 10 in the control group. Log rank statistic analysis of the cumulative exacerbation rates showed a significant reduction in exacerbations for the BFM group (p = 0.0184). The analysis of microflora and the organic acids in the feces showed a significant reduction in the relative proportion of B. vulgatus in Bacteroidaceae and butyrate concentration, respectively, after supplementation with BFM, in comparison with before. CONCLUSION: Supplementation with the BFM product was successful in maintaining remission and had possible preventive effects on the relapse of ulcerative colitis.

Adult↗
PubMed1993

[The nutritive effect of Bacillus cereus as a probiotic in the raising of piglets. 2. Effect and microbial count, composition and resistance determination of gastrointestinal and fecal microflora].

After a feeding trial of 42 days with 4 x 12 piglets 4 x 6 piglets were slaughtered and the influence of spores of the Bacillus cereus strain FH 1457 S added to the feed on the microorganism counts of Lactobacillus/Bifidobacterium, Eubacteria, Bacteroidaceae, E. coli, Enterococcus and passants (Bacillus cereus) in duodenum, jejunum, ileum, caecum and colon was investigated. Beside a negative control the feed was supplemented with 10(7), 10(8) and 10(9) CFU Bacillus cereus/kg. The feeding trial included two periods each of 21 days. In each period faeces was collected from all animals and the influence on the microorganism counts and also the frequency of resistance of the E. coli and Enterococcus germs against selected antibiotics and chemotherapeutics investigated. The addition of 10(8) CFU Bacillus cereus decreased E. coli counts in duodenum and jejunum, however increased them in ileum, caecum and colon. In the highest dosage the counts of Bacillus cereus in duodenum, caecum, colon and faeces were significantly higher. The addition of Bacillus cereus reduced the counts of E. coli and Enterococcus in faeces in the first period. In the second period the dosage 10(8) CFU showed significant increased counts of E. coli and the sums of microorganisms of the main and satellite flora. The additions had no influence on the frequency of resistance of E. coli and enterococci against the tested antibiotics and synthetic chemotherapeutics. From the addition of Bacillus cereus in this case a selection of factors influencing resistance is not to be expected.

Animal Feed↗
PubMed1993

Recent taxonomic changes for anaerobic gram-positive and selected gram-negative organisms.

The strains formerly classified as Streptococcus anginosus or "Streptococcus milleri" have now been recognized as three distinct species, Streptococcus anginosus, Streptococcus constellatus, and Streptococcus intermedius. Streptococcus morbillorum has been transferred into the genus Gemella. Four new species within the genus Peptostreptococcus were recently named. A recent addition to the genus Clostridium is Clostridium argentinense, which includes Clostridium botulinum type G. Two new species of Actinomyces have been introduced: Actinomyces georgiae and Actinomyces gerencseriae. Arachnia propionica was shown to be related to propionibacteria and has been renamed Propionibacterium propionicum. Eubacterium yurii was named to contain "test-tube brush" bacteria found in subgingival plaque. Lactobacillus uli and Lactobacillus rimae are obligately anaerobic lactobacilli found in periodontal pockets. Bilophila is a new genus of gram-negative bacilli. Wolinella recta and Wolinella curva are now included in the genus Campylobacter. The taxonomic position of Mobiluncus, currently assigned to the family Bacteroidaceae, remains uncertain.

Bacteroides↗
PubMed1993

Stimulation of B lymphocytes by lipopolysaccharides from anaerobic bacteria.

Lipopolysaccharides (LPSs) from anaerobic gram-negative bacteria, including those of low endotoxic activity that are isolated from Bacteroides, Prevotella, and Porphyromonas are potent inducers of DNA replication and polyclonal immunoglobulin production in murine B lymphocytes. The activation is dose-dependent and T cell-independent. Replication of DNA and production of immunoglobulins were also stimulated by lipid A and by the LPS heteropolysaccharide that were isolated by mild acid hydrolysis of the LPSs of Bacteroides fragilis and Fusobacterium nucleatum. Combinations of LPS, lipid A, and acid-degraded polysaccharide amplified the blastogenic response. Antibodies that react with the polysaccharide part of LPSs isolated from members of the Bacteroidaceae are present in healthy human serum.

Animals↗
PubMedDate not supplied

Selective inhibition of enterobacteriaceae in fecal flora of children by oral trimethoprim-sulfamethoxazole.

Fecal flora of nine pediatric inpatients who were receiving trimethoprim-sulfamethoxazole were studied by quantitative aerobic and anaerobic culture methods. During the course of trimethoprim-sulfamethoxazole, the number of Enterobacteriaceae decreased from 10(8-9) to 10(4-5) bacteria per g feces. The isolation rate and the number of Veillonella were also reduced. Other prominent aerobic and anaerobic flora, including Streptococcus, Bacteroidaceae and Bifidobacterium, were not affected. No overgrowth of Pseudomonas, Staphylococcus, Candida or Clostridium was noted. These results support the usefulness of oral trimethoprim-sulfamethoxazole for the prophylaxis of bacterial infection in the compromised host.

Child↗
PubMed1995

Evidence for the placement of the gram-negative Catonella morbi (Moore and Moore) and Johnsonella ignava (Moore and Moore) within the Clostridium subphylum of the gram-positive bacteria on the basis of 16S rRNA sequences.

Comparative 16S rRNA analysis was used to determine the phylogenetic positions of Catonella morbi and Johnsonella ignava, which are members of two monospecific genera of gram-negative anaerobic bacilli isolated from human gingival crevices. Both of these genera were found to belong to cluster XIVa (M. D. Collins, P. A. Lawson, A. Willems, J. J. Cordoba, J. Fernandez-Garayzabal, P. Garcia, J. Cai, H. Hippe, and J. A. E. Farrow, Int. J. Syst. Bacteriol. 44:812-826, 1994) of the Clostridium subphylum of gram-positive bacteria. Within this cluster, which contains several Clostridium, Coprococcus, Eubacterium, and Ruminococcus species, C. morbi and J. ignava formed two distinct lines that were separate from all other taxa. Our findings support the separate generic status of the genera Catonella and Johnsonella and show that these genera do not belong to the family Bacteroidaceae but instead belong to the gram-positive Clostridium subphylum.

Base Sequence↗
PubMed1982

Detection of species-specific and cross-reactive cell-surface antigens of Bacteroides species by an indirect enzyme-linked immunosorbent assay.

EDTA-released outer-membrane antigen complexes were prepared from 172 laboratory and reference strains and 43 fresh clinical or faecal isolates representing 20 species or subspecies of Bacteroides, together with 13 species of other genera. These antigens were titrated against antisera to whole, live cells of 21 species or subspecies of Bacteroidaceae in an indirect enzyme-linked immunosorbent assay (ELISA). The presence of species-specific antigens was investigated and cross reactions between species were noted. Results showed that a high proportion of the species possess species-specific antigens with little significant cross reactivity. We believe that the ELISA system described here detects antigens that represent the whole cell surface of bacteroides organisms. We have exploited the sensitivity of the system and its quantitative potential to define approaches for those wishing to use serological approaches for either the identification of Bacteroides species or for the titration of serum antibodies in patients with a possible bacteroides infection.

Antigens, Bacterial↗
PubMed2003

Cloning and functional expression of dipeptidyl peptidase IV from the ruminal bacterium Prevotella albensis M384(T).

Ruminal bacteria of the genus Prevotella play a crucial role in peptide breakdown in the rumen, a component of protein catabolism that leads to the inefficient use of dietary protein by ruminant animals. This is the first report of the cloning of a peptidase gene from a ruminal bacterium. Part of the dipeptidyl peptidase type IV (DPP-IV) gene from Prevotella albensis M384(T) was cloned using degenerate primers designed from conserved regions found within other known DPP-IV sequences. Flanking regions were determined by genomic walking. The DPP-IV gene was expressed in Escherichia coli. The cloned enzyme required a free N terminus and catalysed the removal of X-Pro dipeptide from proline-containing oligopeptides, where proline was the second residue from the N terminus. It was inhibited by serine protease inhibitors and the substrate analogue for mammalian DPP-IV, diprotin A. The properties of the cloned enzyme were similar to those of the native form in P. albensis and, in general, DPP-IVs from other organisms. The enzyme contained a conserved motif which is associated with the S9 class of prolyl oligopeptidases. The DPP-IV gene appeared not to be part of a contiguous operon. Regions with similarity to other putative promoters of Prevotella spp. were also identified. Construction of a phylogenetic tree demonstrated that the DPP-IV of P. albensis clusters with other DPP-IVs found in bacteria of the Cytophaga-Flexibacter-Bacteroidaceae (CFB) phylum, which are more closely related to eukaryotic DPP-IVs than the DPP-IV-like enzyme (PepX) of the lactic acid bacteria.

Amino Acid Sequence↗
PubMed1987

Effect of the two-year milk-feeding on the gastrointestinal microflora of the cynomolgus monkey (Macaca fascicularis).

The effect of the two-year milk-feeding on the gastrointestinal (GI) microflora of a cynomolgus monkey was determined. Bifidobacterium spp. in the animal fed with cow's milk alone were distributed in higher number in the stomach, duodenum, cecum, and rectum. The increasingly higher numbers of Lactobacillus spp. and Candida spp. were counted in all regions of the GI tract of the animal fed with the formula diet. Bacteroidaceae, Enterobacteriaceae, and Streptococcus spp. were detected from the upper to lower intestines in the monkey given only milk, whereas in the monkey given the formula, those bacterial species were localized in only the lower intestines.

Animals↗
PubMed1989

Outer membrane proteins as major antigens of Fusobacterium nucleatum.

The immunochemical reactions of rabbit polyclonal antibodies directed to different preparations of Fusobacterium nucleatum i.e, whole cells, peptidoglycan associated proteins, a peptidoglycan-protein complex and a purified 40 kiloDalton (kDa) protein, were investigated on outer membrane preparations of Fusobacterium species and a restricted number of Leptotrichia buccalis after their separation on sodium dodecyl sulphate polyacrylamide gels and electrotransfer to nitrocellulose. All F. nucleatum strains had identical reaction patterns with the immune sera tested. Surface exposed parts of a restricted number of proteins with apparent molecular weights at 70 kDa (a doublet band), 60 kDa, 55 kDa and 40 kDa seemed to be major immunogens. Antigenic related proteins either of identical or slightly deviating electrophoretic mobilities to the 40-kDa protein were observed with the other members of Bacteroidaceae tested. The characteristic 70-kDa protein doublet seemed to be restricted to F. nucleatum although single protein bands of near identical molecular weights belonging to the other species tested also reacted. The data also indicate that the 60-kDa and 55-kDa polypeptides might be present in other species of Fusobacterium.

Animals↗
PubMed1980

Utilization of ammonia nitrogen by intestinal bacteria isolated from pigs.

In a medium containing ammonia, proteose peptone, and cysteine as nitrogen sources, 17 of 24 Bacteroidaceae strains, 3 of Selenomonas strains, 1 of 7 curved rods, 3 of 7 Spirochaetaceae strains, 8 of 20 Eubacterium strains, 8 of 13 Peptococcaceae strains, 3 of 4 Clostridium strains, 19 of 20 Enterobacteriaceae strains, and 1 of 8 Streptococcus strains utilized ammonia nitrogen preferentially to proteose peptone nitrogen. To determine the ability of intestinal microbes to synthesize amino acids from ammonia, ammonia utilization by Bacteroides ruminicola strain 9 was studied in defined media containing ammonia and other nitrogen sources. In another medium containing ammonia, proteose peptone, and cysteine as nitrogen sources, ammonia was preferentially utilized even when the proteose peptone nitrogen content was eight times greater than that of ammonia nitrogen. In a medium containing ammonia, an amino acid, and cysteine, the lowest uptake of ammonia nitrogen was observed when the medium contained aspartic acid, glutamic acid, threonine, or alanine; but ammonia was utilized more effectively than any of the amino acids. Incorporation of (15)N from [(15)N]ammonia into bacterial amino acids was studied. (15)N was incorporated into every amino acid of B. ruminicola strain 9, and the highest uptake was observed in aspartic acid and alanine.

Amino Acids↗
PubMed1997

Glutamate dehydrogenase activity profiles for type strains of ruminal Prevotella spp.

The glutamate dehydrogenase (GDH) activities for the type strains of Prevotella ruminicola (strain 23), Prevotella brevis (strain GA33), and Prevotella bryantii (strain B(1)4) were assessed by a combination of enzyme assays and analysis of migration patterns of GDH proteins following nondenaturing polyacrylamide gel electrophoresis. Unlike results with most other prokaryotes, but similar to results with other members of the family Bacteroidaceae, NADPH-utilizing specific activity was greatest in all species following ammonia-limited growth. Similar also to previous findings with P. bryantii, the NAD(P)H-utilizing GDH activity of P. ruminicola can be attributed to a single protein. However, P. brevis produces an additional GDH protein(s) in response to growth with peptides. These results conclusively demonstrate that all type strains of the ruminal Prevotella sp. grouping possess GDH activity.

Ammonia↗
PubMed1998

Ammonia-hyperproducing bacteria from New Zealand ruminants.

Pasture-grazed dairy cows, deer, and sheep were tested for the presence of ammonia-hyperproducing (HAP) bacteria in roll tubes containing a medium in which tryptone and Casamino Acids were the sole nitrogen and energy sources. Colonies able to grow on this medium represented 5.2, 1.3, and 11.6% of the total bacterial counts of dairy cows, deer, and sheep, respectively. A total of 14 morphologically distinct colonies were purified and studied further. Restriction fragment length polymorphisms of 16S rRNA genes indicated that all isolates differed from the previously described HAP bacteria, Clostridium aminophilum, Clostridium sticklandii, and Peptostreptococcus anaerobius. Carbon source utilization experiments showed that five isolates (C2, D1, D4, D5, and S1) were unable to use any, or very few, of the carbon sources tested. Biochemical tests and phylogenetic analyses of 16S ribosomal DNA sequences indicated that all isolates were monensin sensitive; that D1 and S1 belonged to the genus Peptostreptococcus, that D4 and D5 belonged to the family Bacteroidaceae, where D4 was similar to Fusobacterium necrophorum; and that C2 was most similar to an unidentified species from the genus Eubacterium. Growth on liquid medium containing tryptone and Casamino Acids as the sole nitrogen and energy source showed that D1, D4, and S1 grew rapidly (specific growth rates of 0.40, 0.35, and 0.29 h-1, respectively), while C2 and D5 were slow growers (0.25 and 0.10 h-1, respectively). Ammonia production rates were highest in D1 and D4, which produced 945.5 and 748.3 nmol/min per mg of protein, respectively. Tests of individual nitrogen sources indicated that D1 and D4 grew best on tryptone, S1 grew equally well on Casamino Acids or tryptone, and C2 and D5 grew poorly on all nitrogen sources. The intact proteins casein and gelatin did not support significant growth of any of the isolates. These isolates extend the diversity of known HAP rumen bacteria and indicate the presence of significant HAP bacterial populations in pasture-grazed New Zealand ruminants.

Ammonia↗
PubMed1992

Sucrose fermentation by Fusobacterium mortiferum ATCC 25557: transport, catabolism, and products.

Studies of sucrose utilization by Fusobacterium mortiferum ATCC 25557 have provided the first definitive evidence for phosphoenolpyruvate-dependent sugar:phosphotransferase activity in the family Bacteroidaceae. The phosphoenolpyruvate-dependent sucrose:phosphotransferase system and the two enzymes required for the dissimilation of sucrose 6-phosphate are induced specifically by growth of F. mortiferum on the disaccharide. Monomeric sucrose 6-phosphate hydrolase (M(r), 52,000) and a dimeric ATP-dependent fructokinase (subunit M(r), 32,000) have been purified to electrophoretic homogeneity. The physicochemical and catalytic properties of these enzymes have been examined, and the N-terminal amino acid sequences for both proteins are reported. The characteristics of sucrose 6-phosphate hydrolase and fructokinase from F. mortiferum are compared with the same enzymes from both gram-positive and gram-negative species. Butyric, acetic, and D-lactic acids are the end products of sucrose fermentation by F. mortiferum. A pathway is proposed for the translocation, phosphorylation, and metabolism of sucrose by this anaerobic pathogen.

Acetates↗
PubMed1982

Controlled evaluation of hypertonic sucrose medium for detection of bacteremia and fungemia in supplemented peptone broth.

Because the value of hypertonic media in detection of bacteremia and fungemia is controversial, we evaluated supplemented peptone broth (SPB) with 0.03% sodium polyanetholsulfonate with and without 10% sucrose in 5,439 paired blood cultures from adult patients. The aerobic atmosphere, 1:10 ratio of blood to broth, and methods for processing blood cultures were identical. Only cultures with adequate blood samples (greater than or equal to 4 ml) were compared statistically. More clinically important bacteria were recovered from SPB with sucrose (P less than or equal to 0.001), including Staphylococcus epidermidis, Enterobacteriaceae, and Bacteroidaceae. However, only one of nine isolates of Neisseria gonorrhoeae grew in SPB with sucrose. Staphylococci (P less than 0.001), Enterobacteriaceae (P less than 0.01), Pseudomonas aeruginosa (P less than 0.01), and yeasts (P less than 0.05) were detected 1 or more days earlier in SPB with sucrose. The effect of sucrose on blood cultures appears to be medium dependent, based on comparisons of our results with those of published reports.

Bacteria↗
PubMed1997

Fecal microflora in a patient with short-bowel syndrome and identification of dominant lactobacilli.

Fecal microflora and lactate concentrations in blood and feces obtained from a patient (a 5 year-old boy) with short-bowel syndrome (SBS) were compared during acidosis to results for the normal condition (no SBS symptoms). The taxonomical position of the lactobacilli found predominantly in the feces sample obtained 2 days before the fifth attack was also studied. The D-lactate level in serum obtained 1 day after the fourth attack was 10-fold higher than that for the normal condition, although there was not a great difference in L-lactate levels. D-Lactate (3.91 mM) and L-lactate (2.86 mM) were also detected in the feces samples collected 2 days before the fifth attack, while no lactate was detected in the feces sample for the normal condition. The counts of total fecal bacteria, especially anaerobic bacteria such as members of the family Bacteroidaceae, were found to be low. The counts of lactobacilli and the total population of lactobacilli relative to total fecal bacteria in the feces 2 days before the fifth attack (40.4%) were extremely high. In this case, a majority of the lactobacilli were D-lactate producers as determined by homolactic fermentation. These lactobacilli were identified as Lactobacillus delbrueckii subsp. lactis. The percentages of bifidobacteria relative to total fecal bacteria in feces samples obtained both 2 days before the fifth attack (50.9%) and for normal condition (61.9%) were also high, although these bacteria were L-lactate producers. In the feces samples for the normal condition, the D-lactate producers decreased to less than 10(9) per g, while the counts of L- or DL-lactate producers were 100-fold higher than the numbers in feces samples obtained 2 days before the fifth attack. These results suggested that an increase in the level of D-lactate producers, such as L. delbrueckii subsp. lactis, in the colon may be associated with the clinical expression of metabolic acidosis.

Acidosis, Lactic↗
PubMed2004

Muscularis inflammation and the loss of interstitial cells of Cajal in the endothelin ETB receptor null rat.

Endothelin receptor null rats [ETB(-/-)] are a model for long-segment Hirschsprung's disease. These animals have significant intestinal distension (megaileum) proximal to a constricted region of the gastrointestinal tract lacking enteric ganglia. Experiments were performed to determine the pathophysiological changes that occur in these animals and to examine the tunica muscularis as a unique, immunologically active compartment. We observed abnormal intestinal flora in ETB(-/-) rats, which included a marked increase in gram-negative aerobes (Enterobacteriaceae) and anaerobes (Bacteroidaceae) in the distended region of the small intestine. Histochemical observations showed that neutrophilic infiltration was rarely or not observed, but the number of ED2 positive macrophages was increased in the tunica muscularis. Expression of IL-1beta and IL-6 mRNA was also significantly increased, and the level of CD14 (LPS receptors) were increased significantly in the tunica muscularis. Spontaneous phasic contractions were irregular in the distended intestinal regions of ETB(-/-) rats, and this was associated with an increased number of macrophages and damage to interstitial cells of Cajal (ICC) as revealed by using Kit-like immunoreactivity and electron microscopy. These results suggest that ED2-positive resident macrophages may play an important role in the inflammation of tunica muscularis in ETB(-/-) rats. Increased numbers and activation of macrophages may result in damage to ICC networks leading to disordered intestinal rhythmicity in regions of the gut in which myenteric ganglia are intact.

Animals↗
PubMed1985

Comparison of media for isolation of mouse anaerobic faecal bacteria.

A study was made to determine suitable plate-in-bottle media for enumeration and isolation of the anaerobic faecal bacteria of mice. Comparison of non-selective media indicated that colony counts were higher on M-SM10, M10 and M-M98-5 than on EG, W & E medium and BHI, and those on M-SM10 were always the highest. Bacteroidaceae counts on M-SM10, M10, M-M98-5 were particularly high. On the other hand, in chloroform-treated faeces which contain only Clostridial spores, colony counts were higher on EG and W & E medium than on M-SM10, M10 and M-M98-5. These results indicate that suitable non-selective media vary according to the bacterial species to be enumerated and isolated.

Animals↗