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PubMed1990

Bacteroides gingivalis and Bacteroides intermedius recognize different sites on human fibrinogen.

Bacteroides (Porphyromonas) gingivalis and Bacteroides (Porphyromonas) intermedius have been implicated in the etiology of human periodontal diseases. These organisms are able to bind and degrade human fibrinogen, and these interactions may play a role in the pathogenesis of periodontal disease. In attempts to map the bacterial binding sites along the fibrinogen molecule, we have found that strains of B. gingivalis and B. intermedius, respectively, recognize spatially distant and distinct sites on the fibrinogen molecule. Isolated reduced and alkylated alpha-, beta-, and gamma-fibrinogen chains inhibited binding of 125I-fibrinogen to both Bacteroides species in a concentration-dependent manner. Plasmin fragments D and to some extent fragment E, however, produced a concentration-dependent inhibition of 125I-fibrinogen binding to B. intermedius strains but did not affect binding of 125I-fibrinogen to B. gingivalis strains. Radiolabeled fibrinogen chains and fragments were compared with 125I-fibrinogen with respect to specificity and reversibility of binding to bacteria. According to these criteria, gamma chain most closely resembled the native fibrinogen molecule in behavior toward B. gingivalis strains and fragments D most closely resembled fibrinogen in behavior toward B. intermedius strains. The ability of anti-human fibrinogen immunoglobulin G (IgG) to inhibit binding of 125I-fibrinogen to B. intermedius strains was greatly reduced by absorbing the IgG with fragments D. Absorbing the IgG with fragments D had no effect on the ability of the antibody to inhibit binding of 125I-fibrinogen to B. gingivalis strains. A purified staphylococcal fibrinogen-binding protein blocked binding of 125I-fibrinogen to B. intermedius strains but not to B. gingivalis strains.

Bacteroides↗

PubMed1985

Evaluation of xylan fermentation for the identification of Bacteroides ovatus and Bacteroides thetaiotaomicron.

The reliability of xylan fermentation in distinguishing Bacteroides ovatus from Bacteroides thetaiotaomicron was examined with strains previously identified by DNA homology studies or bacteriophage sensitivity testing or both. All of the 12 B. ovatus strains produced acid ranging from pH 5.0 to 5.6 in peptone-yeast-xylan broth; none of the 19 B. thetaiotaomicron strains produced acid, and the final pH was 6.0 or 6.1. Xylan fermentation and bacteriophage sensitivity-DNA homology appear to be equivalent in their ability to differentiate these two Bacteroides species.

Bacteriophage Typing↗

PubMed1987

A cross-reactivity study of whole genomic DNA probes for Haemophilus actinomycetemcomitans, Bacteroides intermedius, and Bacteroides gingivalis.

In this study, we evaluated the sensitivity and specificity of whole genomic DNA probes for the periodontal pathogens Haemophilus actinomycetemcomitans, Bacteroides intermedius, and Bacteroides gingivalis. By means of these probes, DNA hybridizations were performed against other organisms found in the oral cavity and organisms previously determined to be genetically similar. All three probes were sensitive to 10(3) cells for their respective organism. The H. actinomycetemcomitans probe cross-reacted with several haemophilus strains, Wolinella, and Campylobacter, indicating that H. actinomycetemcomitans-specific sequences would have to be identified and cloned for accurate detection of this organism in heterogeneous patient samples. Only very low levels of cross-reactivity were observed between the B. intermedius probe and representative black-pigmented Bacteroides. This low level of cross-reactivity did not interfere with the accurate identification of B. intermedius in sample evaluations. The B. gingivalis probe showed no cross-reactivity. Whole genomic probes will be used for the detection of B. intermedius and B. gingivalis in patient samples.

Bacteroides↗

PubMed1991

Identity of Bacteroides isolates and previously named Bacteroides spp in clinical specimens of animal origin.

During the years 1984 through 1987 2,574 isolates of obligately anaerobic bacteria were isolated from samples submitted for analysis. The most common anaerobic isolates were members of the genus Bacteroides, representing 44.6% of the isolates. Of these, the most commonly isolated identifiable microorganisms were bile-resistant and nonpigmented, belonging to the B fragilis group of Bacteroides. Importantly, obvious predilections for any one species or group of Bacteroides were not apparent for animal or site (condition), except that the proportion of isolates belonging to the nonpigmented, bile-resistant group obtained from the respiratory tract was significantly (P less than 0.005) higher than that not belonging to this group. On the other hand, the proportion of isolates of the non-pigmented, bile-resistant group obtained from abscesses was significantly lower than that not belonging to this group.

Animals↗

PubMed1984

Synergism between penicillin, clindamycin, or metronidazole and gentamicin against species of the Bacteroides melaninogenicus and Bacteroides fragilis groups.

Clinical isolates of the Bacteroides melaninogenicus and Bacteroides fragilis groups were tested for in vitro and in vivo susceptibility to penicillin, clindamycin, and metronidazole, used singly or in combination with gentamicin. The in vitro tests consisted of determinations of minimal inhibitory concentrations (MICs) carried out with or without constant amounts of gentamicin. When used alone, gentamicin had negligible effects on the bacteria but significantly reduced the MICs of penicillin, clindamycin, and metronidazole against 11, 10, and 3, of the 15 strains of the B. melaninogenicus group, respectively. The 15 strains of the B. fragilis group were all beta-lactamase producers and were highly resistant to penicillin or the combination of penicillin and gentamicin. However, gentamicin reduced the MICs of clindamycin and metronidazole against 1 and 7 strains of this group, respectively. The in vivo tests were carried out in mice and consisted of measurements of the effects of the antimicrobial agents on the sizes and bacterial content of abscesses induced by subcutaneous injection of bacterial suspensions. The results of the in vivo tests were generally consistent with those obtained in vitro with strains of the B. melaninogenicus group. Synergism between gentamicin and penicillin, clindamycin, or metronidazole was shown in 13, 10, and 3 strains of this group, respectively. In vivo synergism was not clearly demonstrated with the strains of the B. fragilis group, possibly because clindamycin and metronidazole used alone were highly efficacious. We suggest that the synergistic effect of gentamicin is due to its increased transport into the bacterial cell in the presence of penicillin and, possibly, other antimicrobial agents. The newly recognized in vitro and in vivo synergism between penicillin and other antimicrobial agents and an aminoglycoside in B. melaninogenicus may have clinical implications that deserve to be investigated.

Abscess↗

PubMed1992

Analysis of rRNA restriction fragment length polymorphisms from Bacteroides spp. and Bacteroides fragilis isolates associated with diarrhea in humans and animals.

The Escherichia coli rRNA operon rrnB was used as a 32P-labeled hybridization probe in Southern blots of genomic DNAs from representative strains of the saccharolytic, gram-negative, obligate anaerobes of the genus Bacteroides. Control experiments with the B. fragilis type strain ATCC 25285 established that nearly identical rRNA fragment patterns were produced when either the E. coli rrnB gene probe or homologous rRNA isolated from B. fragilis was used as the probe. In addition, it was shown that a specific 16S or 23S rrnB gene probe also could be used to produce fragment patterns suitable for analysis. Thirty-one strains from 8 of the 10 recognized Bacteroides species were then examined. The resulting autoradiographs revealed specific fragment patterns for all but one (B. ovatus) of the species tested. Restriction fragment length polymorphisms were observed for many of the strains tested, but these differences did not hinder species classification. The five B. ovatus strains examined did not form a distinct group, and their rRNA fragment patterns displayed a marked heterogeneity. The same approach was applied to a unique set of enterotoxin-producing B. fragilis strains isolated from animals and humans with diarrhea. The results demonstrated that these strains were in fact B. fragilis and that they produce rRNA fragment patterns closely related to those of the type strain ATCC 25285. This set of strains did not appear to form a separate subgroup or genotype within the B. fragilis species, and there were no distinguishable restriction fragment length polymorphisms that could be used to specifically separate enterotoxin-producing strains from nonenterotoxigenic strains.

Animals↗

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↗

PubMed1995

Antibiotic susceptibility profiles of Bacteroides fragilis and Bacteroides thetaiotaomicron in Japan from 1990 to 1992.

The antimicrobial susceptibility of clinical isolates of Bacteroides fragilis and Bacteroides thetaiotaomicron collected from December 1990 through November 1992 was determined by the agar dilution technique. Metronidazole, imipenem, and cefoperazone/sulbactam remained highly active against both organisms. Rates of resistance to those agents were 0, 2%, and 0.9% in B. fragilis and 0, 0.9%, and 3% in B. thetaiotaomicron, respectively. Cefoxitin and other cephamycins were active against B. fragilis; rates of resistance to these agents did not tend to increase. With the inclusion of these data, the variation of rates of resistance to several agents was summarized for each year from 1987 to 1992. Rates of resistance to imipenem decreased in 1991 and 1992 among isolates of B. fragilis (2.3% in 1991, 1.8% in 1992) and B. thetaiotaomicron (2.4% in 1991, 0 in 1992). Rates of resistance to cefoxitin in B. thetaiotaomicron varied from 10% to 38% during these 6 years, though the distributional peak of MIC values did not change. The rate of resistance to ofloxacin in B. fragilis increased from 42% in 1989 to 81% in 1992. The rate of resistance to ampicillin in B. thetaiotaomicron was 68% in 1992--approximately 30% lower than before. Mostly, however, the rates of resistance to the antimicrobial agents examined did not change significantly.

Anti-Bacterial Agents↗

PubMedDate not supplied

Activation of Hageman factor by lipopolysaccharides of Bacteroides fragilis, Bacteroides vulgatus, and Fusobacterium mortiferum.

Preincubation of lipopolysaccharides (LPS) of Bacteroides fragilis, Bacteroides vulgatus, and Fusobacterium mortiferum with purified human Hageman factor (HF) followed by addition of purified human prekallikrein resulted in the activation of HF, as measured by the generation of kallikrein using the tripeptide substrate N-alpha-benzoyl-L-proline-L-phenylalanine-L-arginine-4-nitroanilide. Addition of purified HF, which had been preincubated with the LPS, to HF-deficient plasma resulted in the acceleration of the recalcification time. Addition of the LPS to HF-deficient plasma reconstituted with various concentrations of HF also resulted in the acceleration of the recalcification time; the magnitude of the acceleration correlated with the amount of HF used to reconstitute the deficient plasma. These results demonstrate that LPS of selected gram-negative anaerobes activate HF and thereby initiate the intrinsic pathway of coagulation.

Bacteroides↗

PubMed1994

Cloning and expression of the Bacteroides fragilis YCH46 neuraminidase gene in Escherichia coli and Bacteroides uniformis.

A neuraminidase-encoding gene nanH of Bacteroides fragilis strain YCH46 was cloned into the cosmid vector pHC79. The nanH gene was subcloned from the cosmid and was located within a 2.2-kb XhoI-KpnI fragment. Southern hybridization experiments demonstrated that the gene was present as a single copy on the bacterial chromosome. Neuraminidase activity expressed in the initial Escherichia coli clone was approximately 3600-fold lower than that expressed in B. fragilis YCH46. However, when nanH was transferred from E. coli to B. uniformis by mobilization of a shuttle plasmid, the transconjugant expressed 1100-fold higher activity than the E. coli donor did. These results suggest that modes of nanH expression in E. coli and Bacteroides are heterologous.

Bacteroides↗

PubMed1984

Alternative pathways for biosynthesis of leucine and other amino acids in Bacteroides ruminicola and Bacteroides fragilis.

Bacteroides ruminicola is one of several species of anaerobes that are able to reductively carboxylate isovalerate (or isovaleryl-coenzyme A) to synthesize alpha-ketoisocaproate and thus leucine. When isovalerate was not supplied to growing B. ruminicola cultures, carbon from [U-14C]glucose was used for the synthesis of leucine and other cellular amino acids. When unlabeled isovalerate was available, however, utilization of [U-14C]glucose or [2-14C]acetate for leucine synthesis was markedly and specifically reduced. Enzyme assays indicated that the key enzyme of the common isopropylmalate (IPM) pathway for leucine biosynthesis, IPM synthase, was present in B. ruminicola cell extracts. The specific activity of IPM synthase was reduced when leucine was added to the growth medium but was increased by the addition of isoleucine plus valine, whereas the addition of isovalerate had little or no effect. The activity of B. ruminicola IPM synthase was strongly inhibited by leucine, the end product of the pathway. It seems unlikely that the moderate inhibition of the enzyme by isovalerate adequately explains the regulation of carbon flow by isovalerate in growing cultures. Bacteroides fragilis apparently also uses either the isovalerate carboxylation or the IPM pathway for leucine biosynthesis. Furthermore, both of these organisms synthesize isoleucine and phenylalanine, using carbon from 2-methylbutyrate and phenylacetate, respectively, in preference to synthesis of these amino acids de novo from glucose. Thus, it appears that these organisms have the ability to regulate alternative pathways for the biosynthesis of certain amino acids and that pathways involving reductive carboxylations are likely to be favored in their natural habitats.

2-Isopropylmalate Synthase↗

PubMed1980

Production of phenylacetic acid by strains of Bacteroides asaccharolyticus and Bacteroides gingivalis (sp. nov.).

Strains of Bacteroides asaccharolyticus and Bacteroides melaninogenicus subspecies isolated from human and animal sources were examined for the production of phenylacetic acid. B. asaccharolyticus strains isolated from sites in humans and monkeys always produced phenylacetic acid. B. asaccharolyticus strains isolated from human nonoral sites consistently failed to produce this product. This metabolic difference correlates with the genetic dichotomy recently found to exist between oral and nonoral B. asaccharolyticus strains.

Animals↗

PubMed1981

Plasmid analyses in clinical isolates of Bacteroides fragilis and other Bacteroides species.

Plasmid analyses were performed on Bacteroides strains isolated from clinical specimens. Of 32 Bacteroides strains, 8 were found to contain plasmids. Seven of these eight strains were B. fragilis, and the other one was B. distasonis. Three of these eight strains harbored only a 3.0-megadalton plasmid. Two strains had only a 2.0-megadalton plasmid, and one had 2.0-, 3.0-megadalton plasmid. Of the remaining two strains, one had 2.0-, 3.0-, and 5.0-megadalton plasmids, and the other had 3.0- and 5.0-megadalton plasmids. Beta-Lactamase was produced by 93% of the clinical isolates. Seven of the eight plasmid-carrying strains were cadmium resistant, five were zinc resistant, four were mercury resistant, and two expressed a brick-red fluorescence under ultraviolet light. None of these traits could be associated with a plasmid after performing either curing experiments or genetic transfer experiments by cell-to-cell contact.

Anti-Bacterial Agents↗

PubMed1985

Further characterization of Bacteroides endodontalis, an asaccharolytic black-pigmented Bacteroides species from the oral S cavity.

In this study, the isolation, characterization, and identification of Bacteroides endodontalis is described. It was found that this asaccharolytic black-pigmented Bacteroides species is associated with infected dental root canals and oral submucous abscesses. B. endodontalis could be differentiated from B. gingivalis by a negative direct hemagglutination test and the absence of trypsin and N-acetyl-beta-glucosamidase. B. endodontalis could be differentiated from B. asaccharolyticus by the absence of alpha-fucosidase, its inability to grow in an atmosphere of 95% N2-5% H2, and a growth requirement for menadione. Immune serum raised against B. endodontalis strain HG 370T agglutinated only B. endodontalis cells. Precautions for the isolation of B. endodontalis are discussed.

Acetylglucosaminidase↗

PubMed1988

Refractory periodontitis: mixed infection with Bacteroides gingivalis and other unusual Bacteroides species. A case report.

Host immune response and the predominant subgingival microflora were evaluated in a 47-year-old male exhibiting severe, recurrent periodontitis. The patient's neutrophils were chemotactically elevated but other functions were within normal limits. Significantly, Bacteroides gingivalis and Bacteroides zoogleoformans constituted 80% of the cultivable microflora and total cell count in subgingival plaque. The remainder of the cultivable microbiota was comprised of Fusobacterium nucleatum and Haemophillis aprophillis. The present study provides additional evidence for an association between B. gingivalis and severe, recurrent periodontal disease.

Bacteroides↗

PubMed1999

[Toxins of Bacteroides fragilis and Bacteroides thetaiotaomicron rods as stimulators of adhesion molecule expression on the surface of vascular endothelial cells].

Lipopolysaccharides from four Bacteroides fragilis strains: one nonenterotoxigenic (NTBF) and three enterotoxigenic (ETBF), and from three B. thetaiotaomicron strains were extracted by hot phenol-water method and purified. B. fragilis enterotoxin was prepared according to the procedure of van Tassell et al. (1992). The influence of the examined toxins on the expression of adhesion molecules: ICAM-1, VCAM-1 and E-selectin on HMEC-1 (human dermal microvascular endothelial cells) was assayed in ELISA test with monoclonal antibodies. Four concentrations of toxins were applied: 0.01, 0.1, 1.0 and 10.0 (micrograms/ml). Endothelial cells were activated for 24 hours (ICAM-1 and VCAM-1 expression) and for 4 hours (E-selectin expression). The coloured product of immunoenzymatic reaction was measured by reading the absorbance at wavelength 492 nm. Two controls were performed in each experiment: with resting HMEC-1 and E. coli O55:B5 LPS (Sigma, USA). Bacteroides fragilis and B. thetaiotaomicron lipopolysaccharides stimulated three adhesion molecules under investigation. Their activity was comparable, but weaker than the activity of E. coli O55:B5 LPS. ICAM-1 was the most stimulated molecule. B. fragilis enterotoxin induced two adhesion molecules: VCAM-1 and E-selectin demonstrating weaker stimulatory activity than E. coli LPS. Stimulation of adhesion molecules on vascular endothelial cells should be considered to be a biological activity of B. fragilis and B. thetaiotaomicron endotoxins and B. fragilis enterotoxin.

Bacteroides↗

PubMed1980

Susceptibility of Bacteroides fragilis, Bacteroides thetaiotaomicron, and Fusobacteria in Antibacterial agents.

The minimum inhibitory concentrations of 45 antimicrobial agents were established by the agar dilution method for 37 strains of Bacteroidaceae: Bacteroides fragilis (18 strains), B. thetaiotaomicron (8 strains), Fusobacterium nucleatum (6 strains), and Fusobacterium spp. (5 strains). The fusobacteria tested showed good susceptibility to beta-lactam-antibiotics, tetracyclines, lincomycins, and nitroimidazole compounds. Bacteria of the B. fragilis group were resistant to most of the penicillins and cephalosporins and could not be suppressed with certainty by nitroimidazoles and older tetracyclines. Doxycycline and minocycline, however, showed sufficient activity, and all our Bacteroides strains tested were sensitive to clindamycin. Especially the good in vitro susceptibility of all Bacteroidaceae strains investigated to cefoxitin may offer new therapeutic possibilities.

Anti-Bacterial Agents↗

PubMed1989

Electroporation induced transformation of Bacteroides ruminicola and Bacteroides uniformis by plasmid DNA.

Frequencies of greater than 10(5) transformants per microgram DNA were achieved in Bacteroides ruminicola F101 by electroporation of cells under anaerobic conditions, using the 19.5 kbp tetracycline resistance plasmid pRRI4. Similar procedures gave frequences of 10(6) erythromycin resistant transformants per microgram DNA with the shuttle plasmid pDP1 (19 kbp) in Bacteroides uniformis. Transformation of B. uniformis occurred at a far lower frequency (10(3) micrograms) when pDP1 DNA was derived from E. coli rather than B. uniformis.

Bacteroides↗