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PubMed1999

Characterization of the Batl (Bacteroides aerotolerance) operon in Bacteroides fragilis: isolation of a B. fragilis mutant with reduced aerotolerance and impaired growth in in vivo model systems.

YT135.2.8, a Tn4400' insertion mutant of Bacteroides fragilis strain TM4000, grows poorly when used to infect Monika or Chinese hamster ovary (CHO) cell monolayers and is outcompeted by wild-type strains in mixed infections. YT135.2.8 also shows defects in the rat granuloma pouch model system in monoculture and is completely outcompeted by the wild-type strain in a mixed infection. In addition, this mutant shows defects in a new model system consisting of CHO suspension cell columns. All of these defects may be explained by the finding that YT135.2.8 shows decreased tolerance to exposure to atmospheric oxygen (less aerotolerant). The monolayer growth defect (MGD) of YT135.2.8 can be influenced significantly by the presence of sulphur-containing reducing agents (cysteine, dithiothreitol, thiodiglycol) or the non-sulphur reducing agent Tris-(2-carboxylethyl)phosphine (TCEP). The defects in YT135.2.8 can be complemented by a 6.6 kb fragment of the B. fragilis chromosome. DNA sequencing of this fragment and of the regions flanking the Tn4400' insertion in the B. fragilis chromosome revealed the presence of five open reading frames, corresponding to genes bat (Bacteroides aerotolerance) A, B, C, D, E, which form the Batl operon; Tn4400' inserted within batD. All of the hypothetical proteins possess one or more membrane-spanning domains. BatA and BatB show high similarity to each other but, like BatD, they show no match to sequences of known function in the databases. BatC and BatE contain 2-4 repeated sequences similar to the tetratricopeptide repeats (TPRs) seen in many eukaryotic proteins. The function of TPR sequences in protein interactions in other systems leads to the suggestion that the Bat proteins form a complex. The Batl complex may be involved in the generation or export of reducing power equivalents to the periplasm of the B. fragilis cell.

Amino Acid Sequence↗

PubMed1991

Wolinella recta, Wolinella curva, Bacteroides ureolyticus, and Bacteroides gracilis are microaerophiles, not anaerobes.

Although the nonfermentative, asaccharolytic, putative anaerobes Wolinella curva, Wolinella recta, Bacteroides ureolyticus, and Bacteroides gracilis are phylogenetically related to the true campylobacters, the type strains of these species exhibited O2-dependent microaerophilic growth in brucella broth and on brucella agar. The optimum O2 levels for growth of these strains ranged from 4 to 14% in brucella broth and from 2 to 8% on brucella agar, when H2 was provided as the electron donor. No growth occurred under 21% O2, and scant or no growth occurred under anaerobic conditions unless fumarate or nitrate was provided as a terminal electron acceptor. Aspartate, asparagine, and malate also served as apparent electron acceptors. The organisms were catalase negative and, except for B. gracilis, oxidase positive. Catalase added to brucella broth enhanced growth. O2 uptake by all species was inhibited by cyanide and 2-heptyl-4-hydroxyquinoline N-oxide. We concluded that these organisms are not anaerobes but instead are microaerophiles, like their campylobacter relatives.

Anaerobiosis↗

PubMed1995

Chemotaxonomic analyses of Bacteroides gracilis and Bacteroides ureolyticus and reclassification of B. gracilis as Campylobacter gracilis comb. nov.

The cellular fatty acids, respiratory quinones, and proteins of the generically misnamed taxa Bacteroides gracilis and Bacteroides ureolyticus were analyzed and compared with the corresponding chemotaxonomic features of their closest relatives, the campylobacters. Our results and previously published data for genotypic and phenotypic characteristics were used in a polyphasic approach to reconsider the classification of these organisms. We transfer B. gracilis to the genus Campylobacter as Campylobacter gracilis comb. nov. B. ureolyticus can be considered a campylobacter on genotypic grounds; in contrast, the proteolytic metabolism and fatty acid components of this taxon exclude it from the genus Campylobacter. We prefer to consider this taxon a species incertae sedis pending the isolation and characterization of additional B. ureolyticus-like bacteria.

Bacterial Proteins↗

PubMed1991

Bacteroides ureolyticus (NTU) medium for the selective recovery of Bacteroides gracilis.

Bacteroides gracilis is a gram-negative anaerobic bacillus which requires formate and fumarate for growth; it has been implicated in periodontal disease and serious infections of the head and neck. In this study, Bacteroides ureolyticus (NTU) medium was tested for its ability to allow the growth of B. gracilis and other formate-fumarate requiring gram-negative anaerobes and to enable the recovery of these organisms from clinical specimens. All reference strains grew on NTU medium with the exception of Wolinella recta and formate-fumarate requiring organisms were isolated from 18 of 20 samples of subgingival dental plaque from patients with chronic periodontitis. B. gracilis was the commonest species isolated (14 of the 29 isolates); B. ureolyticus was not found.

Bacteroides↗

PubMed1989

The occurrence of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, Bacteroides intermedius and spirochaetes in the subgingival microflora in relation to the early onset of periodontitis in a group of adolescents.

Subgingival samples were obtained from mesial and distal surfaces of first molars in 100 subjects aged 14 years at baseline. Sites were monitored at 6-monthly intervals for clinical loss of attachment. Six subjects showed greater than 1 mm loss of attachment at one site 18-24 months later. Further samples were obtained from these sites together with a comparison sample from the contralateral site and equivalent sites in a matched control subject. Actinobacillus actinomycetemcomitans was more frequently isolated from sites following loss of attachment, but it was also found in one comparison site and one control subject. Bacteroides gingivalis was found in only one subject at a site which subsequently experienced loss of attachment. A higher median proportion of spirochaetes was observed in sites after diagnosis of loss of attachment. The median proportion of Bacteroides intermedius was higher at both the baseline and 12-month examinations in subjects who subsequently lost attachment. Further work is required to substantiate the differences which preceded diagnosis of loss of attachment.

Actinobacillus↗

PubMed1990

Black-pigmented, asaccharolytic Bacteroides species resembling Porphyromonas gingivalis (Bacteroides gingivalis) from beagle dogs.

Black-pigmented, asaccharolytic Bacteroides strains, which positively reacted with anti-Bacteroides gingivalis (now reclassified as Porphyromonas gingivalis) serum, were isolated from beagle dogs, and their characteristics were studied and compared with those of P. gingivalis. The strains from dogs were different from P. gingivalis in their catalase activity, nutritional requirements and oxygen tolerance, and double-immunodiffusion tests showed serological dissimilarity between the strains from dogs and P. gingivalis. However, the strains from dogs had guanine-plus-cytosine contents of 49.0 to 49.3 mol %, which were very similar to P. gingivalis, and they showed strong DNA-DNA hybridization with P. gingivalis. These results indicated that beagle dogs had Porphyromonas species analogous to P. gingivalis in their oral cavities.

Animals↗

PubMed1990

Ingestion of Bacteroides buccae, Bacteroides oris, Porphyromonas gingivalis, and Fusobacterium nucleatum by human polymorphonuclear leukocytes in vitro.

The phagocytic ingestion of clinical isolates and reference strains of Bacteroides buccae, Bacteroides oris, Porphyromonas gingivalis and Fusobacterium nucleatum by polymorphonuclear leukocytes (PMNs) was studied. Special attention was focused on the hydrophobicity of the strains. B. buccae strains, less or equally hydrophobic than PMNs, were poorly ingested without opsonization. Hydrophobic, but not hydrophilic, strains of B. oris and both hydrophilic and hydrophobic P. gingivalis and F. nucleatum strains were readily ingested without opsonization. Hydrophobicity thus contributes to the adherence of bacteria to PMNs in some, but not all, species tested. Normal human serum enhanced the ingestion of B. buccae, but failed to do so after heat-inactivation. Heat-inactivation of the immune serum to B. buccae strain ES57 did not reduce opsonic activity suggesting that specific antibodies enhanced the ingestion of B. buccae.

Bacterial Adhesion↗

PubMed1980

Antimicrobial effect of fluorides on Bacteroides melaninogenicus subspecies and Bacteroides asaccharolyticus.

This study was undertaken to determine whether the suspected periodontopathogens Bacteroides melaninogenicus ss melaninogenicus and intermedius and Bacteroides asaccharolyticus are killed by clinically used topical fluorides. The viability of each organism was examined after exposure to three fluoride compounds, stannous (SnF2), acidulated phosphate (APF), and neutral sodium (NaF), for 1 and 8 h. Stannous fluoride was found to be effective at lowest fluoride concentrations with APF next in effectiveness. NaF required the highest concentrations of fluoride to kill test organisms. Longer exposure times were found to be more effective at lower concentrations. Data obtained in this study with gram-negative organisms support and extend previous information regarding the antibacterial effect of fluorides against gram-positive periodontopathogens.

Acidulated Phosphate Fluoride↗

PubMed1986

The occurrence of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius in destructive periodontal disease in adults.

A total of 235 subgingival sites, including 104 progressive deep lesions from 61 untreated patients, 26 progressive deep lesions from 10 treated patients, 33 nonprogressive deep sites from 20 untreated patients, and 72 nonprogressive sites from 55 treated patients were examined for Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius. The periodontal disease progression was mainly determined on the basis of radiographic changes in the crestal alveolar bone level. A. actinomycetemcomitans isolation was carried out using the selective TSBV medium and B. gingivalis and B. intermedius isolations were performed using a nonselective blood agar medium. 1 or more of the 3 bacteria studied appeared in 99.2% of progressive periodontal lesions but only in 40.0% of nonprogressive sites. Culture-positive progressive periodontal sites in comparison with culture-positive nonprogressive sites showed higher median recovery rates of A. actinomycetemcomitans (0.5% vs 0.3%), B. gingivalis (30.5% vs 0.3%) and B. intermedius (4.9% vs 0.5%). Of total progressive lesions, 12.3% yielded solely A. actinomycetemcomitans, 21.5% demonstrated solely B. gingivalis, and 20.8% revealed solely B. intermedius. The A. actinomycetemcomitans--B. intermedius combination was found in 24.6% of progressive lesions. A. actinomycetemcomitans appeared in significantly higher prevalence in treated-progressive lesions (80.8%) than in nontreated-progressive lesions (42.3%). 32 of the 42 culture-positive nonprogressive sites yielded B. intermedius as the sole test organism. The main conclusion is that A. actinomycetemcomitans, B. gingivalis and B. intermedius are closely related to disease-active periodontitis, and more closely than to periodontal pocket depth. This finding is important in understanding periodontal disease etiology and pathogenesis and may also aid in a clinical setting to differentiate progressing and nonprogressing periodontal sites.

Actinobacillus↗

PubMed1987

The capability of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius to indicate progressive periodontitis; a retrospective study.

This study evaluated the statistical association of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius with progressive periodontitis. 146 adults with a history of advanced periodontitis contributed 105 "nonprogressing" and 130 "progressing" periodontal sites. Periodontal disease activity was assessed by radiographic changes in crestal alveolar bone level. The subgingival proportion of the 3 test bacteria was determined by selective and nonselective culturing. The relationship between bacterial proportions and disease progression was evaluated using subgrouping and multiple-regression analyses. All 3 test bacteria had to be considered in order to distinguish nonprogressing and progressing periodontitis with a reasonably high sensitivity. A recovery rate below 0.01% for A. actinomycetemcomitans, 0.1% for B. gingivalis and 2.5% for B. intermedius defined a site with nonprogressing disease with 87% sensitivity and 84% specificity. By utilizing transformed values of the bacterial recovery rates and optimal test criteria determined by multiple regression analysis, it was possible to obtain sensitivities between 83% and 95% and specificities between 86% and 69%. These 3 bacterial species might serve as valuable components of a periodontitis activity test based on microbiological variables.

Actinobacillus↗

PubMed1988

Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans in human periodontal diseases.

Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans seem to be major pathogens in advancing periodontitis in man. First, these organisms are recovered in higher prevalence and proportions from progressive periodontitis lesions than from quiescent periodontal sites. Second, antibody levels against B. gingivalis and A. actinomycetemcomitans are markedly elevated in serum and gingival crevice fluid of periodontitis patients compared to normal controls. Third, B. gingivalis and B. intermedius elaborate potent proteases and A. actinomycetemcomitans various noxious substances which have the potential to perturb important host defenses and to disintegrate key constituents of the periodontal tissues. Monitoring these bacteria in advanced periodontal lesions may greatly assist the assessment of treatment efficacy and risk of further periodontal breakdown.

Actinobacillus↗

PubMed1977

Beta-Lactamase activity in strains of Bacteroides melaninogenicus and Bacteroides oralis.

beta-Lactamase from strains of Bacteroides melaninogenicus and Bacteroides oralis hydrolyzed penicillin more rapidly than ampicillin or carbenicillin. Cephalothin and a chromogenic cephalosporin (87/312) were also hydrolyzed by the enzyme. Activity was found only in beta-lactam-resistant strains, but there was considerable variation in activity among strains having the same minimal inhibitory concentrations of antibiotic. beta-Lactamase activity was cell bound and appeared to be tightly associated with the cell envelope since detergents were required to elute this activity.

Amidohydrolases↗

PubMed1990

Characterization of beta-lactamases from non-Bacteroides fragilis group Bacteroides spp. belonging to seven species and their role in beta-lactam resistance.

Twelve beta-lactamase-positive non-Bacteroides fragilis group Bacteroides spp. belonging to seven different species were examined by MIC determination and enzyme characterization. MICs of most beta-lactams except cefoxitin, cefotetan, imipenem, and meropenem were relatively high or very high. All enzymes hydrolyzed cephaloridine (Vmax, 100%; Km, 12 to 70 microM), cephalothin (Vmax, 25 to 826%; Km, 8 to 143 microM), cefamandole (Vmax, 13 to 158%; Km, 17 to 170 microM), and cefuroxime (hydrolysis rate, 19 to 98%), and 11 of 12 hydrolyzed cefotaxime (Vmax, 26 to 145%; Km, 13 to 127 microM); no hydrolysis of cefoxitin or moxalactam was observed. Penicillins were hydrolyzed at lower rates, with Vmax values less than or equal to 20% of that obtained with cephaloridine. Addition of clavulanate, sulbactam, or tazobactam led to a 4- to 2,048-fold lowering of MICs of penicillins as well as cephalosporins. All enzymes were inhibited by clavulanate (50% inhibitory concentration [IC50], 0.01 to 1.8 microM), sulbactam (IC50, 0.02 to 1.9 microM), tazobactam (IC50, 0.001 to 0.9 microM), cefoxitin (IC50, 0.002 to 0.35 microM), and moxalactam (IC50, 0.03 to 6.6 microM). No enzymes were inhibited by 100 microM EDTA or p-chloromercuribenzoic acid; an enzyme of one strain of B. loescheii was inhibited by 100 microM cloxacillin (IC50, 2.35 microM). Ten enzymes had optimal activity at pH 5.0 to 6.0, and two had optimal activity at pH 8.0. Isoelectric focusing revealed pIs between 4.2 and 5.6. These enzymes seem to belong to a previously unclassified group of beta-lactamases, related (but not identical) to beta-lactamases of the B. fragilis group.

Anti-Bacterial Agents↗

PubMed1988

Effect of phenolic monomers on the growth and beta-glucosidase activity of Bacteroides ruminicola and on the carboxymethylcellulase, beta-glucosidase, and xylanase activities of Bacteroides succinogenes.

trans-p-Coumaric acid inhibited the growth of Bacteroides ruminicola on both cellobiose and glucose, while trans-ferulic acid and vanillin retarded growth. The phenolic monomers varied in their potential to inhibit the Bacteroides succinogenes beta-glucosidase, carboxymethylcellulase, and xylanase, with p-coumaric acid being the most inhibitory. The B. ruminicola beta-glucosidase was inhibited less than 10% by all three compounds.

Bacteroides↗

PubMed1980

Participation of normal human immunoglobulins M, G, and A in opsonophagocytosis and intracellular killing of Bacteroides fragilis and Bacteroides thetaiotaomicron by human polymorphonuclear leukocytes.

Restoration of the ability of hypogammaglobulinemic serum to support opsonophagocytosis and intracellular killing of Bacteroides fragilis and Bacteroides thetaiotaomicron by human polymorphonuclear leukocytes was achieved by supplementation with normal human immunoglobulin M, but not with normal human immunoglobulin G. Polymorphonuclear leukocyte bactericidal activity in the presence of immunoglobulin A-deficient human serum was equivalent to that observed in the presence of normal human serum.

Bacteroides Infections↗

PubMed1981

Serum antibodies to oral Bacteroides asaccharolyticus (Bacteroides gingivalis): relationship to age and periondontal disease.

An enzyme-linked immunosorbent assay microplate method was used for measuring levels of antibody specific for the oral serotype of Bacteroides asaccharolyticus (Bacteroides gingivalis) in serum samples obtained from umbilical cords, infants, children, periodontally normal adults, and edentulous adults. Serum from patients with various periodontal diseases, including adult periodontitis, localized juvenile periodontitis, generalized juvenile periodontitis, post-localized juvenile periodontitis, and acute necrotizing ulcerative gingivitis, were also studied. A positive correlation between increase in age and increase in both prevalence and level of specific antibody in the G, A, and M classes of immunoglobulins was observed. This indicates that antibodies reactive with oral B. asaccharolyticus found in up to 84% of normal adults are natural antibodies, presumably with a protective role. Among the patient groups, those with adult periodontitis were found to have levels of immunoglobulin G antibodies to oral B. asaccharolyticus that were five times higher than the antibody levels found in control subjects. The levels of IgG antibodies to this organism in the other patient groups were comparable to the levels found in the control group. However, 50% of the individuals in the generalized juvenile periodontitis group had high levels of immunoglobulin G antibodies to B. asaccharolyticus, suggesting heterogeneity with respect to immune response in these patients. These results indicate that antibodies to oral B. asaccharolyticus (B. gingivalis) occur at low levels in most normal children and adults and that the rise in titer of the specific antibodies of each major class of immunoglobulins parallels the ontogenic change in serum levels of that isotype. In contrast, there is a marked increase in titer of immunoglobulin G antibodies to oral B. asaccharolyticus in the group of patients with adult periodontitis and in patients with the generalized form of juvenile periodontitis.

Adult↗

PubMed1982

Effects of estradiol and progesterone on Bacteroides melaninogenicus and Bacteroides gingivalis.

Bacteroides melaninogenicus subsp. intermedius increases in the subgingival microflora during pregnancy. These studies evaluated direct interactions between hormonal steroids and oral Bacteroides species. Resting cell suspensions of pure cultures of plaque organisms were incubated anaerobically with [14C]estradiol and [14C]progesterone. Uptake of labeled compound per microgram of bacterial protein was determined by thin-layer chromatography and liquid scintillation counting. B. melaninogenicus subsp. intermedius and B. melaninogenicus subsp. melaninogenicus took up 2.6 x 10(-4) to 5.4 x 10(-4) mumol of estradiol or progesterone per microgram of cell protein. Minimal steroid uptake was observed with B. gingivalis and five other organisms. Uptake of steroids by B. melaninogenicus subsp. intermedius was temperature dependent and resulted in a labeled product as detected on thin-layer chromatography. Growth curves indicated that intermedius and melaninogenicus subspecies of B. melaninogenicus but not B. gingivalis could substitute progesterone or estradiol for vitamin K, an essential growth factor. Growth of B. melaninogenicus subsp. intermedius in steroids was concentration dependent. Addition of fumarate to resting cells of B. melaninogenicus subspecies as well as B. gingivalis increased steroid uptake by 70 to 500% and resulted in the gas-liquid chromatographic detection of succinate. Cultures given fumarate alone or steroids alone produced no succinate. Steroids appeared to directly interact with the fumarate reductase system and foster the growth of B. melaninogenicus subsp. intermedius. This interaction may be of ecological significance.

Bacteroides↗

PubMed1992

Lipopolysaccharides of Bacteroides intermedius (Prevotella intermedia) and Bacteroides (Porphyromonas) gingivalis induce interleukin-8 gene expression in human gingival fibroblast cultures.

Lipopolysaccharides (LPS) prepared from Bacteroides intermedius (Prevotella intermedia) and Bacteroides (Porphyromonas) gingivalis by hot phenol-water extraction induced interleukin-8 (IL-8) mRNA in normal human gingival fibroblast cultures, as demonstrated by Northern (RNA) blot analysis. IL-8 mRNA levels began to increase after a 2-h exposure, reached a maximum after 12 h, and then dropped to the unstimulated level at 48 h. IL-8 mRNA levels were also enhanced in a dose-dependent manner. By contrast, LPS specimens from various Salmonella species with S and R chemotypes and bacterial [corrected] and synthetic lipid A preparations did not increase IL-8 mRNA levels in fibroblasts. Although recombinant human IL-1 alpha induced IL-8 mRNA expression in fibroblast cultures, an antiserum to recombinant human IL-1 alpha did not decrease the IL-8 mRNA accumulation induced by B. intermedius LPS. Fibroblasts primed with natural human gamma interferon (IFN-gamma) expressed higher IL-8 mRNA levels upon stimulation with B. intermedius LPS, but not with Salmonella LPS, compared with nontreated cells. Natural human IFN-beta exhibited a similar priming effect on the fibroblasts, and antiserum to IFN-beta added to the cultures together with B. intermedius LPS decreased the IL-8 mRNA levels. Therefore, endogenous IFN-beta enhanced IL-8 mRNA production in response to B. intermedius LPS in fibroblasts.

Bacteroides↗