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At least 19 recordsLinked to original sources
PubMed1977

Chemical composition, serological reactivity and endotoxicity of lipopolysaccharides extracted in different ways from Bacteroides fragilis, Bacteroides melaninogenicus and Bacteroides oralis.

Lipopolysaccharides (LPS) extracted from strains of Bacteroides fragilis, Bacteroides melaninogenicus and Bacteroides oralis with phenol-water, trichloroacetic acid, EDTA or liquid phenol-chloroform-petroleum ether (PCP) and isolated by ultracentrifugation, varied considerably in their quantitative chemical composition. Negligible yields of LPS were obtained by PCP-extraction. All preparations were more or less serologically active. All methods (except PCP) extracted the same O-antigenic determinants from B. fragilis. Endotoxic activity, as measured by primary skin inflammations in rabbits, was low but was present in all preparations. Proteins (and/or lipoproteins) co-precipitated with LPS in the ultracentrifuge.

Bacteroides↗
PubMed1979

Ultrastructure of Bacteroides species: Bacteroides asaccharolyticus, Bacteroides fragilis, Bacteroides melaninogenicus subspecies melaninogenicus, and B. melaninogenicus subspecies intermedius.

Representative strains of two subspecies of Bacteroides melaninogenicus (subspecies melaninogenicus and subspecies intermedius) and Bacteroides asaccharolyticus as well as B. asaccharolyticus strain 536B isolated from a human perirectal abscess and Bacteroides fragilis ATCC 25285 were examined by glutaraldehyde-osmium fixation, ruthenium red fixation and staining, and thorium hydroxyde staining as well as by the physical preparative techniques of critical point drying--transmission electron microscopy (CPD--TEM) and scanning electron microscopy (SEM). All strains, with the exception of B. fragilis 25285, possessed an electron-dense material external to their outer membranes. Ruthenium red staining further revealed a layer, external to the surface of the outer membrane, that was distinct for each species examined. Thorium hydroxide, as well as CPD--TEM and SEM, showed the cells to be interconnected by thin fibers that not only connected adjacent cells but also traversed several microns to connect cell aggregates.

Bacteroides↗
PubMedDate not supplied

Participation of immunoglobulin and the alternative complement pathway in opsonization of Bacteroides fragilis and Bacteroides thetaiotaomicron.

Studies were conducted to determine the requirements for immunoglobulin and complement in opsonization of Bacteroides fragilis and Bacteroides thetaiotaomicron. The ability of human sera depleted of immunoglobulin or of components of complement to promote the phagocytosis and intracellular killing of the two strains of Bacteroides by human leukocytes was measured in vitro under anaerobic conditions. Neither hypogammaglobulinemic sera nor pooled normal human serum that was heated at 56 C for 30 min supported phagocytosis and killing of the two strains of Bacteroides. Neither sera depleted of terminal complement components by treatment with inulin or cobra venom factor nor human serum deficient in C8 supported phagocytosis of the tested strains. In addition, pooled normal human serum depleted of C3, factor B, or factor D did not support phagocytosis of either strain. Dose-dependent restoration of the opsonic activity of factor B-depleted serum was accomplished by purified human factor B but not by human C2. The results indicate that immunoglobulin and components of the alternative comed in this study.

Animals↗
PubMed1978

Participation of immunoglobulin and the alternative complement pathway in opsonization of Bacteroides fragilis and Bacteroides thetaiotaomicron.

Studies were conducted to determine the requirements for immunoglobulin and complement for opsonization of Bacteroides fragilis and Bacteroides thetaiotaomicron. The ability of human sera depleted of immunoglobulin or complement components to promote phagocytosis and intracellular killing of the strains of Bacteroides by human leukocytes was measured in vitro under anaerobic conditions. Neither hypogammaglobulinemic sera nor pooled normal human serum (PNHS) heated at 56 C for 30 min supported phagocytosis and killing of the strains of Bacteroides. Sera depleted of terminal complement components by treatment with inulin or cobra venom factor and C8-deficient human serum did not support phagocytosis of the test strains. PNHS depleted of C3, factor B, or factor D also did not support phagocytosis of either strain. Dose-dependent restoration of the opsonic activity of factor B-depleted serum was accomplished by purified human factor B but not by human C2. The results indicated that immunoglobulin and components of the alternative complement pathway participate in opsonization of the strains of Bacteroides tested in this study.

Agammaglobulinemia↗
PubMed1979

Comparative serology of two clinical isolates of Bacteroides fragilis and Bacteroides thetaiotaomicron.

Antiserum prepared in rabbits against Bacteroides fragilis showed numerous bands when reacted with B. fragilis antigen in Ouchterlony plates. This antiserum also reacted with Bacteroides thetaiotaomicron and showed one band of apparent identity with B. fragilis. The band of identity common for both organisms was lost if the antigen was heated at 80 degrees C for 30 min. Antisera prepared against B. thetaiotaomicron did not react with B. fragilis.

Antigens, Bacterial↗
PubMed1979

Fluorescent antibody test kit for rapid detection and identification of members of the Bacteroides fragilis and Bacteroides melaninogenicus groups in clinical specimens.

The Fluoretec fluorescent antibody test kit (Pfizer Inc., New York, N.Y.), developed for the rapid detection of members of the Bacteroides fragilis and B. melaninogenicus groups, was evaluated by testing 58 stock cultures and 76 clinical specimens. The test reagents detected 100% of 40 B. fragilis and B. thetaiotaomicron stock culture strains, although only 22% of 18 B. vulgatus, B. distasonis, and B. ovatus strains showed positive fluorescence. The 76 clinical specimens were evaluated by examining fluorescent antibody-stained smears of 49 specimens of purulent material and smears of 27 blood cultures which were positive for gram-negative bacilli by Gram stain or subculture. The fluoretec reagent detected members of the B. fragilis group in 28 (97%) of the 29 specimens of purulent material and all (100%) of the 16 blood cultures in which these anaerobes were demonstrated by culture. Overall, the Fluoretec reagent detected members of B. fragilis group in 44 (98%) of the 45 clinical specimens which were shown by culture to contain these anaerobes. Two of the 76 clinical specimens gave positive fluorescence for members of the B. fragilis group but failed to yield these organisms by culture. Members of the B. melaninogenicus group were detected by culture in 15 specimens and in each case their presence was demonstrated by the Fluoretec reagent. No members of the B. melaninogenicus group were isolated from five clinical specimens that gave positive fluorescence with the B. melaninogenicus reagent.

Anaerobiosis↗
PubMed1977

Superoxide dismutase in Bacteroides fragilis and related Bacteroides species.

Superoxide dismutase (SOD) activity was demonstrated in cell-free extracts of Bacteroides fragilis, Bacteroides vulgatus, Bacteroides distasonis, Bacteroides ovatus, and Bacteroides thetaiotaomicron. The strains were grown under anaerobic conditions in Trypticase soy broth, and the specific activity of SOD in the extracts was, in most strains, higher than in cell-free extracts of Escherichia coli B grown under anaerobic conditions. Isoelectric focusing of the extracts in polyacrylamide gel demonstrated distinct forms of SOD in the different species.

Anaerobiosis↗
PubMed1979

Characterization and mode of action of a bacteriocin produced by a Bacteroides fragilis strain.

A Bacteroides fragilis strain produces a low-molecular-weight (13,500 to 18,700), proteinaceous bacteriocin during the stationary growth phase. The extracellular bacteriocin is not inducible by ultraviolet light or mitomycin C and is stable between pH 7.5 and 8.2. The majority of the bacteriocin is thermolabile, but a small proportion (3%) of the bacteriocin is stable after autoclaving at 121 degrees C for 15 min. Killing of sensitive bacteroides cells follows single-hit kinetics, and the interaction of a single molecule of bacteriocin with a target cell occurs in two stages. The killing of susceptible cells is affected by temperature and the growth state of the susceptible cells. The bacteriocin is unusual in that the primary event in its mode of action is the inhibition of RNA synthesis. The bacteriocin inhibits RNA synthesis immediately but has no effect on DNA synthesis or intracellular ATP levels. Protein synthesis is inhibited after a delay of 20 min, presumably as a result of the initial inhibition of RNA synthesis.

Adenosine Triphosphate↗
PubMedDate not supplied

Surface antigens as virulence factors in infection with Bacteroides fragilis.

Organisms of the genus Bacteroides represent the major group of obligate anaerobes involved in human infections. Bacteroides usually cause either bacteremia or localized abscesses. Of the numerous species of Bacteroides, Bacteroides fragilis is the single most frequent clinical isolate. B. fragilis and Bacteroides melaninogenicus have chemically incomplete lipopolysaccharides as compared with the lipopolysaccharides (endotoxins) of aerobic bacteria, and the lipopolysaccharides of Bacteroides lack the biologic potency characteristic of endotoxin. This inactivity may account for the very infrequent occurrence of disseminated intravascular coagulation or purpura that can accompany sepsis due to these organisms. Furthermore, strains of B. fragilis have an immunologically common capsular polysaccharide. In an animal model of intraabdominal sepsis, the encapsulated strains caused abscesses when given without other organisms, but abscess formation from unencapsulated strains of Bacteroides generally required the administration of a synergistic aerobe. The abscesses caused by encapsulated strains were shown to be directly attributable to the capsular polysaccharide, which is an important virulence factor of this organism. Patients or experimental animals infected with B. fragilis develop antibodies to the capsular polysaccharide, and these antibodies can be detected in a radioactive antigen-binding assay.

Animals↗
PubMedDate not supplied

The chemotactic effect of Bacteroides fragilis lipopolysaccharide.

The atypical Bacteroides fragilis lipopolysaccharide is chemotactic for polymorphonuclear leukocytes. The chemotactic activity is mediated by complement activated through the alternative pathway. This mediator has been isolated. It is a polypeptide with a molecular weight of 16,000 and is most likely the complement component C5a.

Animals↗
PubMed1978

Pathway of succinate and propionate formation in Bacteroides fragilis.

Cell suspensions of Bacteroides fragilis were allowed to ferment glucose and lactate labeled with (14)C in different positions. The fermentation products, propionate and acetate, were isolated, and the distribution of radioactivity was determined. An analysis of key enzymes of possible pathways was also made. The results of the labeling experiments showed that: (i) B. fragilis ferments glucose via the Embden-Meyerhof pathway; and (ii) there was a randomization of carbons 1, 2, and 6 of glucose during conversion to propionate, which is in accordance with propionate formation via fumarate and succinate. The enzymes 6-phosphofrucktokinase (pyrophosphate-dependent), fructose-1,6-diphosphate aldolase, phosphoenolpyruvate carboxykinase, malate dehydrogenase, fumarate reductase, and methylmalonyl-coenzyme A mutase could be demonstrated in cell extracts. Their presence supported the labeling results and suggested that propionate is formed from succinate via succinyl-, methylmalonyl-, and propionyl-coenzyme A. From the results it also is clear that CO(2) is necessary for growth because it is needed for the formation of C4 acids. There was also a randomization of carbons 1, 2, and 6 of glucose during conversion to acetate, which indicated that pyruvate kinase played a minor role in pyruvate formation from phosphoenolpyruvate. Phosphoenolpyruvate carboxykinase, oxaloacetate decarboxylase, and malic enzyme (nicotinamide adenine dinucleotide phosphate-dependent) were present in cell extracts of B. fragilis, and the results of the labeling experiments agreed with pyruvate synthesis via oxaloacetate and malate if these acids are in equilibrium with fumarate. The conversion of [2-(14)C]- and [3-(14)C]lactate to acetate was not associated with a randomization of radioactivity.

Acetates↗
PubMed1977

Carbohydrate repression of catalase synthesis in Bacteroides fragilis.

Catalase formation by Bacteroides fragilis was immediately stopped upon addition of glucose to a culture growing in peptone medium. Each of eight other carbohydrates fermented by the organism also repressed catalase formation. Without added carbohydrate, the strains produced relatively large amounts of catalase (25 to 50 U/mg of protein).

Bacteroides fragilis↗
PubMed1979

Factors contributing to resistance to beta-lactam antibiotics in Bacteroides fragilis.

Fifteen strains of Bacteroides fragilis, five highly resistant, five moderately resistant, and five susceptible to benzylpenicillin and cephaloridine, were tested for beta-lactamase production. In the highly resistant and the moderately resistant groups of strains, a correlation between formation of beta-lactamase and minimal inhibitory concentrations was demonstrated. In the presence of the beta-lactamase inhibitors clavulanic acid and CP-45,899 at concentrations of 1 mug/ml, the susceptibility to cephaloridine increased fourfold or more in the beta-lactamase-producing strains. Crypticity measurements (beta-lactamase activity broken/intact cells) with cephaloridine as substrate could indicate a diffusion barrier in the cell wall.

Anti-Bacterial Agents↗
PubMed1978

Formation of glycosidases in batch and continuous culture of Bacteroides fragilis.

Nine strains of bacteroides fragilis were cultivated in stirred fermentors and tested for their ability to produce glycosidases. B. fragilis subsp. vulgatus B70 was used for optimizing the production of glycosidases. The highest bacterial yield was obtained in proteose peptone-yeast extract medium. The optimum pH for maximal bacterial yield was 7.0, and the optimum temperature for growth was 37 degrees C. The formation of glycosidases was optimal between pH 6.5 and 7.5, and the optimum temperature for synthesis of glycosidases was between 33 and 37 degrees C. Culture under controlled conditions in fermentors gave more reproducible production of glycosidases than static cultures in bottles. The strain was also grown in continuous culture at a dilution rate of 0.1 liter/h at pH 7.0 and 37 degrees C with a yield of 2.0 mg of dry weight per ml in the complex medium. The formation of glycosidases remained constant during the entire continuous process.

Bacteriological Techniques↗
PubMed1978

New medium for selection and presumptive identification of the Bacteroides fragilis group.

A medium, Bacteroides fragilis bile-esculin (BBE) agar, was designed for the selection and, presumptive identification of the B. fragilis group. BBE agar contains bile, esculin, ferric ammonium citrate, hemin, and gentamicin in a Trypticase soy agar base. Growth in the presence of 20% bile and esculin hydrolysis, detected by blackening of the medium, provide presumptive evidence for the identification of the B. fragilis group. In addition to stimulating the growth of many strains of the B. fragilis group, hemin provides the option of testing isolates for catalase production. Gentamicin and bile prevent the growth of most organisms other than the esculin-positive bacteroides that can tolerate bile. Of 160 clinical isolates of the B. fragilis group tested on BBE agar, 159 grew well on the medium and 157 blackened it. Other anaerobes, Enterobacteriaceae, and enterococci either failed to grow on BBE agar or did not produce the characteristic morphology and blackening associated with isolates of the B. fragilis group. In a clinical laboratory trial, 687 specimens from patients were inoculated onto BBE agar plates. The B. fragilis group was recovered from 81 (11.8%) of these specimens in 24 to 48 h. Use of BBE agar in the clinical laboratory enables earlier recovery and identification of this important pathogen.

Anaerobiosis↗
PubMed1976

The polysaccharide capsule of Bacteroides fragilis subspecies fragilis: immunochemical and morphologic definition.

A large-molecular-weight capsular polysaccharide was isolated from strains of Bacteroides fragilis subspecies fragilis. By means of electron microscopy and staining with ruthenium red, the thick polysaccharide capsule was also visualized. With use of a radioactive antigen-binding assay, antibody to this capsular polysaccharide was demonstrated in antisera prepared in rabbits to each of eight strains of B. fragilis fragilis. Antibody of similar specificity was not found in antisera prepared to Bacteroides melaninogenicus or to strains of Bacteroides fragilis subspecies vulgatus and Bacteroides fragilis subspecies distasonis; such antibody was found in antisera to only one of two strains of Bacteroides fragilis subspecies thetaiotaomicron. The radioactive antigen-binding assay is a sensitive test for the detection of antibody to capsular polysaccharide. This polysaccharide antigen may form the basis of a serogrouping system for B. fragilis.

Antigen-Antibody Complex↗
PubMed1975

NAD- and NADP-dependent 7alpha-hydroxysteroid dehydrogenases from bacteroides fragilis.

Twenty strains of Bacteroides fragilis were screened for hydroxysteroid oxidoreductase activity in cell-free preparations. Eighteen strains were shown to contain NAD-dependent 7alpha-hydroxysteroid dehydrogenase. Sixteen of the strains containing the NAD-dependent enzyme also contained NADP-depedent 7alpha-hydroxysteroid dehydrogenase, but invariably in lesser amounts. A strain particulary rich in both 7alpha-hydroxysteroid dehydrogenase activities was selected for further study. Measurement of activity as a function of pH revealed a fairly sharp optimal activity range of 9.5--10.0 for the NAD-dependent enzyme and a broad flat optimal range of 7.0--9.0 for the NADP-dependent enzyme. Michaelis constants for trihydroxy-bile acids for the NAD-dependent enzyme were in the range of 0.32--0.34 mM, whereas dihydroxy-bile acids gave a Km of 0.1 mM. Thin-layer chromatography studies on the oxidation product of 3alpha, 7alpha-dihydroxy-5beta-cholanoic acid (chenodeoxycholic acid) by the dehydrogenase revealed a band corresponding to that of synthetic 3alpha-hydroxy, 7-keto-5beta-cholanoic acid. Similarly the oxidation product of chenodeoxycholic acid by both 7alpha-hydroxysteroid dehydrogenase and commercially available 3alpha-hy-droxysteroid dehydrogenase revealed a band corresponding to that of synthetic 3,7-diketo-5beta-cholanoic acid. Neither of these two oxidation products could be distinguished from those by the Escherichia coli dehydrogenase oxidation previously reported. Disc-gel electrophoresis of a cell-free lyophilized preparation indicated one active band for NAD-dependent activity of mobility similar to that for the NADP-dependent E. coli enzyme. The NADP-dependent dehydrogenase was unstable and rapidly lost activity after polyacylamide disc-gel electrophoresis, ultracentrifugation, freezing on refrigeration at 4 degrees C. No 3 alpha- or 12alpha-oriented oxidoreductase activity was demonstrated in any of the strains examined.

Bacteroides↗
PubMed1978

Antibody response in thirteen patients with Bacteroides fragilis infections.

13 patients with Bacteroides fragilis infections were studied for antibody response against the infecting strain using indirect immunofluorescence (IFL), passive hemagglutination (HA) and tube agglutination (TA). With indirect IFL significant titre changes in IgG were found in 12/13 cases, in IgA in 8/13 and in IgM in 4/13 cases. With passive HA significant titre changes were found in 11/13 cases and with TA in 6/11 cases. The rise in antibody titre was fast and persisted for a long time in most cases. Of the methods used the indirect IFL was the most convenient for clinical use.

Adult↗