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PubMed1990

Evidence that Bacteroides nodosus belongs in subgroup gamma of the class Proteobacteria, not in the genus Bacteroides: partial sequence analysis of a B. nodosus 16S rRNA gene.

The taxonomic status of the anaerobe Bacteroides nodosus has for some time been uncertain. To resolve this uncertainty, the distal portion of a 16S rRNA gene from this important ovine pathogen was cloned, mapped, and sequenced. A comparison of the sequence with the sequences of 16S rRNA molecules from other bacteria indicated that B. nodosus is more closely related to Escherichia coli and other members of the class Proteobacteria than to Bacteroides fragilis or the bacteroides-flavobacterium-cytophaga phylum. The evidence from the comparison of sequence signatures suggests that B. nodosus is not a member of the genus Bacteroides but that it belongs in subgroup gamma of the class Proteobacteria.

Animals↗

PubMed1990

Prevotella, a new genus to include Bacteroides melaninogenicus and related species formerly classified in the genus Bacteroides.

It was recently proposed that the genus Bacteroides should be restricted to Bacteroides fragilis (the type species) and closely related organisms (viz., B. caccae, B. distasonis, B. eggerthii, B. merdae, B. ovatus, B. stercoris, B. thetaiotaomicron, B. uniformis, and B. vulgatus). By contrast, the moderately saccharolytic, predominantly oral Bacteroides species, which include B. melaninogenicus, B. oralis, and related species, form a phenotypically and phylogenetically coherent group of species which differ so significantly from the emended description of the genus Bacteroides that they should not be classified in the same genus. Therefore, we formally propose that these species be reclassified in a new genus, Prevotella. The type species is Prevotella melaninogenica.

Bacteria↗

PubMed2005

Bacteroides plebeius sp. nov. and Bacteroides coprocola sp. nov., isolated from human faeces.

Nine strains of Gram-negative, anaerobic rod were isolated from human faeces. Based on phylogenetic analysis and specific phenotypic characteristics, these strains were included within the Bacteroides cluster and were divided into two clusters. Strains from the two clusters showed 16S rRNA gene sequence similarities of 90.4 and 92.7% to the nearest recognized species, Bacteroides vulgatus. The strains also formed two clusters exhibiting a 16S rRNA gene sequence divergence of approximately 6%. DNA-DNA hybridization studies confirmed that the two novel strain clusters were distinct from each other. Based on the phenotypic and phylogenetic findings, two novel species, Bacteroides plebeius sp. nov. and Bacteroides coprocola sp. nov., are proposed, each representing one of the two strain clusters. The DNA G+C content of the type strains were 43.9 mol% for B. plebeius (M12(T)=JCM 12973(T)=DSM 17135(T)) and 42.4 mol% for B. coprocola (M16(T)=JCM 12979(T)=DSM 17136(T)).

Bacterial Typing Techniques↗

PubMed1986

Bacteroides pectinophilus sp. nov. and Bacteroides galacturonicus sp. nov.: two pectinolytic bacteria from the human intestinal tract.

Studies on the physiological characteristics of two obligately anaerobic, rod-shaped bacteria from the human intestinal tract indicated that the organisms represented two previously undescribed species of Bacteroides, for which we propose the names Bacteroides pectinophilus (type strain, N3) and Bacteroides galacturonicus (type strain, N6). Both strains were pectinophilic; that is, they utilized as fermentable substrates for growth only pectin and a few related compounds. The two species differed significantly from each other in guanine plus cytosine content of the DNA, in substrate utilization patterns, and in other phenotypic characteristics. Both species deesterified pectin by means of an extracellular pectinesterase (EC 3.1.1.11) activity. Polygalacturonate (the main component of deesterified pectin) was depolymerized extracellularly with formation of unsaturated products by both species. The depolymerizing activity required Ca2+, functioned at a higher rate when polygalacturonate was the substrate as compared with pectin, and had an alkaline pH optimum. These data, as well as viscosity decrease studies and identification of products formed from polygalacturonate, indicated that the extracellular depolymerizing activity of either species was characteristic of an exopectate (exopolygalacturonate) lyase. The exopectate lyase activity had an unusual action pattern that resulted in terminal cleavage of unsaturated trigalacturonic acid units from polygalacturonate. An unsaturated trimer was the major product that accumulated in cell-free reaction mixtures, where it was not cleaved further. Growing cells of both Bacteroides species released the exopectate lyase into the external environment by processes that did not involve cell lysis to any significant extent.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteroides↗

PubMed1985

Bacteroides endodontalis and other black-pigmented Bacteroides species in odontogenic abscesses.

Twenty-eight odontogenic abscesses were examined for the presence of black-pigmented Bacteroides spp. Of the 28 samples, 26 were found to contain one or more species of black-pigmented Bacteroides. Abscesses were divided into three categories according to the tissue of origin: endodontal, periodontal, and pericoronal. Four abscesses which developed after extraction were also examined. It was found that Bacteroides endodontalis, a newly described species of asaccharolytic black-pigmented Bacteroides, was isolated almost exclusively from periapical abscesses of endodontal origin. B. intermedius proved to be the most frequently isolated species in all of the samples. B. gingivalis was present in all of the periodontal abscesses studied, as well as in two endodontal abscesses. B. melaninogenicus was recovered once from a pericoronal abscess. Precautions for the isolation of B. endodontalis are discussed.

Adolescent↗

PubMed1986

Tn4351 transposes in Bacteroides spp. and mediates the integration of plasmid R751 into the Bacteroides chromosome.

The gene for resistance to erythromycin and clindamycin, which is carried on the conjugative Bacteroides plasmid, pBF4, has been shown previously to be part of an element (Tn4351) that transposes in Escherichia coli. We have now introduced Tn4351 into Bacteroides uniformis 0061 on the following two suicide vectors: (i) the broad-host-range IncP plasmid R751 (R751::Tn4351) and (ii) pSS-2, a chimeric plasmid which contains 33 kilobases of pBF4 (including Tn4351) cloned into the IncQ plasmid RSF1010 and which is mobilized by R751. When E. coli HB101, carrying either R751::Tn4351 or R751 and pSS-2, was mated with B. uniformis under aerobic conditions, Emr transconjugants were detected at a frequency of 10(-6) to 10(-5) (R751::Tn4351) or 10(-8) to 10(-6) (R751 and pSS-2). In matings involving pSS-2, all Emr transconjugants contained simple insertions of Tn4351 in the chromosome, whereas in matings involving R751::Tn4351, about half of the Emr transconjugants had R751 cointegrated with Tn4351 in the chromosome. Of the Emr transconjugants, 13% were auxotrophs. Bacteroides spp. which had R751 cointegrated with Tn4351 in the chromosome did not transfer R751 or Tn4351 to E. coli HB101 or to isogenic B. uniformis, nor did the intergrated R751 mobilize pE5-2, an E. coli-Bacteroides shuttle vector that contains a transfer origin that is recognized by R751.

Bacteroides↗

PubMed2002

Isolation and characterization of cLV25, a Bacteroides fragilis chromosomal transfer factor resembling multiple Bacteroides sp. mobilizable transposons.

Horizontal DNA transfer contributes significantly to the dissemination of antibiotic resistance genes in Bacteroides fragilis. To further our understanding of DNA transfer in B. fragilis, we isolated and characterized a new transfer factor, cLV25. cLV25 was isolated from B. fragilis LV25 by its capture on the nonmobilizable Escherichia coli-Bacteroides shuttle vector pGAT400DeltaBglII. Similar to other Bacteroides sp. transfer factors, cLV25 was mobilized in E. coli by the conjugative plasmid R751. Using Tn1000 mutagenesis and deletion analysis of cLV25, two mobilization genes, bmgA and bmgB, were identified, whose predicted proteins have similarity to DNA relaxases and mobilization proteins, respectively. In particular, BmgA and BmgB were homologous to MocA and MocB, respectively, the two mobilization proteins of the B. fragilis mobilizable transposon Tn4399. A cis-acting origin of transfer (oriT) was localized to a 353-bp region that included nearly all of the intergenic region between bmgB and orf22 and overlapped with the 3' end of orf22. This oriT contained a putative nic site sequence but showed no significant similarity to the oriT regions of other transfer factors, including Tn4399. Despite the lack of sequence similarity between the oriTs of cLV25 and Tn4399, a mutation in the cLV25 putative DNA relaxase, bmgA, was partially complemented by Tn4399. In addition to the functional cross-reaction with Tn4399, a second distinguishing feature of cLV25 is that predicted proteins have similarity to proteins encoded not only by Tn4399 but by several Bacteroides sp. transfer factors, including NBU1, NBU2, CTnDOT, Tn4555, and Tn5520.

Bacteroides fragilis↗

PubMed1986

Effects of anti-Bacteroides-antibodies on Escherichia coli and different Bacteroides species in vitro and vivo.

Bacteroides species are known to cause synergistic pathogenicity in mixed infections with Enterobacteriaceae. In vitro studies showed that anaerobes may compete with aerobes for opsonisation thus interfering with the phagocytosis and killing of the latter by leukocytes. The present study investigated the effect of anti-Bacteroides-antibodies on in vitro phagocytosis of E. coli and on experimental mixed infections. It was shown that in vitro, B. fragilis suppressed the phagocytosis of E. coli by leukocytes only moderately. The addition of specific antibodies to the phagocytosis mixture could enhance this effect. However, antibodies in the absence of B. fragilis were also active suggesting a nonspecific mechanism such as blockade of Fc receptors. Prophylactic treatment with the immunoglobulin preparations failed to protect mice from experimental infections with E. coli and Bacteroides species.

Animals↗

PubMed1980

Detection of Bacteroides fragilis and Bacteroides melaninogenicus by direct immunofluorescence.

A new diagnostic kit, which contains a polyvalent antiserum for either Bacteroides fragilis or Bacteroides melaninogenicus, was tested for reliability and specificity on 146 clinical samples of different origin. A correlation between the culture and immunofluorescence was observed for B. fragilis in 87.39% of cases and for B. melaninogenicus in 81.48% of cases. When pure cultures were tested, aerobically as well as anaerobically, false-positive reactions were observed with staphylococci and Clostridium ramosum spores. The well-defined morphology of these bacteria and spores allows for the elimination of any diagnostic error. The method is rapid, and the margin for error is limited. The test gives a semiquantitative idea of the number of bacteroides organisms present in the clinical specimens even in the presence of a mixed flora.

Antibody Specificity↗

PubMed1993

Chromosomal DNA probes for the identification of Bacteroides tectum and Bacteroides fragilis from the oral cavity of cats.

A dot-blot hybridisation assay using high molecular weight DNA as whole chromosomal probes was used to differentiate Bacteroides tectum from Bacteroides fragilis. 32P-labelled probes were compared with digoxigenin (DIG)-labelled probes. The whole chromosomal probes were specific--differentiating B. tectum from B. fragilis and both from a variety of other species (including other members of the genera Bacteroides, Fusobacterium, Eubacterium, and Prevotella) found in normal and abnormal mouths of cats and horses. However, even at very high stringencies, B. tectum homology groups I, II and III were not distinguishable from one another using either 32P-labelled or DIG-labelled probes. Thus, DIG-labelled whole chromosome probes directed against cellular DNA released directly onto nitrocellulose membranes is considered a useful method for diagnostic veterinary laboratories wishing to identify B. tectum and distinguish it from B. fragilis and other oral anaerobic flora of cats.

Animals↗

PubMed1991

Susceptibilities of 394 Bacteroides fragilis, non-B. fragilis group Bacteroides species, and Fusobacterium species to newer antimicrobial agents.

The susceptibilities of 374 selected beta-lactamase-producing gram-negative anaerobes (including 22 cefoxitin-resistant strains and 36 strains refractory to the enhancing effect of beta-lactamase inhibitors) and 20 beta-lactamase-negative strains were tested by agar dilution against selected new agents. The organisms included 217 Bacteroides fragilis group strains, 137 non-B. fragilis group Bacteroides spp., and 40 fusobacteria. All strains were susceptible to piperacillin-tazobactam, imipenem, and meropenem. For the B. fragilis group, 96% were susceptible to ampicillin-sulbactam, 95% were susceptible to amoxicillin-clavulanate and cefoperazone-sulbactam, 94% were susceptible to tosufloxacin, 91% were susceptible to cefoxitin, 88% were susceptible to trospectomycin, and 73% were susceptible to cefotetan. For the beta-lactamase-positive non-B. fragilis group Bacteroides spp., greater than or equal to 94% were susceptible to cefoxitin, amoxicillin-clavulanate, ampicillin-sulbactam, cefoperazone-sulbactam, and trospectomycin, 90% were susceptible to cefotetan, and 85% were susceptible to tosufloxacin (the most resistant strains were B. bivius and B. disiens). For the beta-lactamase-positive fusobacteria, greater than or equal to 97% were susceptible to amoxicillin-clavulanate, ampicillin-sulbactam, cefoperazone-sulbactam, trospectomycin, and cefoxitin, 90% were susceptible to cefotetan, and 89% were susceptible to tosufloxacin. All agents showed excellent activity against beta-lactamase-negative strains (for trospectomycin, 95% were susceptible; for all other drugs, 100% were susceptible). Overall, both carbapenems and piperacillin-tazobactam were most active. Amoxicillin-clavulanate, ampicillin-sulbactam, and cefoperazone-sulbactam lacked activity against some cefoxitin-resistant B. fragilis group strains but had excellent activity against other organisms. Tosufloxacin, a new quinolone, had very good activity against B. fragilis group strains (94% susceptible), good activity against other beta-lactamase-positive strains (less than or equal 85% susceptible), and excellent activity against beta-lactamase-negative strains (100% susceptible; MIC for 90% of strains, 0.5 microgram/ml). Trospectomycin was active against >90% of all strains except for B. fragilis group strains (88% susceptible; MIC for 90% of strains, 32 microgram/ml). Clinical studies are required to delineate the role of newer agents in the therapy of anaerobic infections.

Anti-Bacterial Agents↗

PubMed2004

Pharmacodynamics of pulse dosing versus standard dosing: in vitro metronidazole activity against Bacteroides fragilis and Bacteroides thetaiotaomicron.

Pulse dosing is a novel approach to dosing that produces escalating antibiotic levels early in the dosing interval followed by a prolonged dose-free period. Antibiotic is frontloaded by means of four sequential bolus injections, after which antibiotic levels are allowed to diminish until the next dose. This study compares standard thrice-daily dosing and pulse dosing of metronidazole against Bacteroides spp. in an in vitro model. Two American Type Culture Collection Bacteroides fragilis isolates (metronidazole MIC for each organism = 1 mg/liter) were exposed to metronidazole for 48 or 96 h. Human pharmacokinetics were simulated for an oral 500-mg dose given every 8 h (maximum concentration of drug [C(max)] = 12 mg/liter; half-life = 8 h; area under the curve [AUC] = 294 mg . h/liter) and for pulse dosing. Pulses, each producing an increase in metronidazole concentration of 9 mg/liter, were administered at times 0, 2, 4, and 6 h of each 24-h cycle, with a targeted half-life of 8 h (AUC = 347 mg . h/liter). A metronidazole-resistant B. fragilis strain (metronidazole MIC = 32 mg/liter) was exposed to both dosing regimens and, additionally, to a regimen of 1,500 mg administered once daily (C(max) = 36 mg/liter; AUC = 364 mg . h/liter). Furthermore, regimens against one B. fragilis isolate and one B. thetaiotaomicron isolate corresponding to one-fourth and one-eighth of the thrice-daily and pulse dosing regimens, mimicking peak metronidazole concentrations achieved in abscesses, were simulated in 48-h experiments (metronidazole MIC = 1 mg/liter). Time-kill curves were generated for each experiment and analyzed for bactericidal activity, defined as a bacterial burden reduction >/= 3 log(10) CFU/ml. The results of paired (Wilcoxon matched-pair signed-rank test) and nonpaired (Mann-Whitney test) statistical analyses conducted on time to 3 log(10) kill data and area under the kill curve data from each of the thrice-daily dosing experiments versus each of the pulse dosing experiments were considered not significant for a given isolate-dosing regimen combination. The thrice-daily dosing, pulse dosing, and once-daily dosing regimens all exhibited bactericidal activity. Metronidazole administered in standard or pulse dosing fashion was highly active against both susceptible and resistant strains of Bacteroides spp.

Anti-Bacterial Agents↗

PubMed1989

Modulation of colonization by black-pigmented Bacteroides species in squirrel monkeys by immunization with Bacteroides gingivalis.

Periodontal diseases are inflammatory responses thought to be triggered by specific microorganisms colonizing in the gingival crevice. Theoretically, periodontal diseases could be prevented if the etiologic organisms were not allowed to colonize the subgingival area. The humoral immune response is one mechanism which may modulate bacterial colonization in the gingival crevice. To test the effect of systemic humoral immunity on subgingival colonization by bacteria, squirrel monkeys (Saimiri sciureus) were immunized with Bacteroides gingivalis, a black-pigmented Bacteroides sp. and putative periodontal pathogen. Immunized and sham-immunized monkeys were orally inoculated with 10(10) viable B. gingivalis during ligation of five teeth in one quadrant with bacterium-soaked suture material and distribution over the entire dentogingival margin. Immunization resulted in an increased level of immunoglobulin G anti-B. gingivalis in serum and was associated with a strong trend toward a statistically significant reduction in colonization of the gingival crevice by black-pigmented bacteroides.

Animals↗

PubMed1987

Bacteriologic and clinical study of Bacteroides oris and Bacteroides buccae.

We characterized clinical isolates previously identified in our laboratory as Bacteroides ruminicola, the human strains of which are now classified as Bacteroides oris and Bacteroides buccae. A total of 72 isolates (55 B. buccae isolates and 17 B. oris isolates) recovered over a 10-year period were studied. They were differentiated from each other by special-potency antibiotic disks and the RapID-ANA system. The two organisms were associated with a variety of infections, the majority being pleuropulmonary (29.2%) and infections of the head and neck region (27.8%). The infections were always polymicrobial, usually with more than five organisms per specimen. A total of 44% of the B. oris strains and 27% of the B. buccae strains were resistant to penicillin G (breakpoint, 2 U/ml), and this correlated with the presence of beta-lactamase. Although B. oris and B. buccae are found with some frequency in human infections, they are present primarily as components of a mixed flora.

Acetylglucosaminidase↗

PubMed1990

Occurrence of Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans in severe periodontitis in relation to age and treatment history.

A total of 242 subjects including 138 untreated severe periodontitis patients and 104 patients with refractory periodontal disease, previously treated for severe periodontitis, were examined for the occurrence of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius. Pooled subgingival samples of representative periodontal lesions were used for anaerobic cultivation on blood agar and for the enumeration of A. actinomycetemcomitans on selective TSBV medium. 97% of the untreated patients were infected with one or more of the test micro-organisms. In this patient group, the occurrence of A. actinomycetemcomitans, B. gingivalis and B. intermedius was 54%, 48% and 63%, respectively. The prevalence of A. actinomycetemcomitans positive patients appeared to be age related and decreased with increasing age. Likewise, the number of patients solely infected with A. actinomycetemcomitans decreased with increasing age. The prevalence of B. gingivalis infected patients appeared to increase with increasing age. These phenomena were not observed in the refractory periodontitis patients. The occurrence of A. actinomycetemcomitans, B. gingivalis and B. intermedius in the refractory periodontitis group was 55%, 27% and 59%, respectively. A statistical significant difference in the prevalence of B. gingivalis was found between the untreated and the refractory periodontitis patients. In both patient groups, the relative proportion of A. actinomycetemcomitans was significantly higher in subjects with this bacterium as the sole indicator micro-organism than in patients who, besides being infected with A. actinomycetemcomitans, were also infected with black-pigmented Bacteroides species. Furthermore, in comparison with untreated patients, unsuccessfully treated patients solely infected with A. actinomycetemcomitans had on average a lower number but also a higher mean % of this bacterium.(ABSTRACT TRUNCATED AT 250 WORDS)

Actinobacillus↗

PubMed1989

Prevalence of Bacteroides forsythus and Bacteroides gingivalis in subgingival plaque of prosthodontically treated patients on short recall.

The prevalence of Bacteroides forsythus and Bacteroides gingivalis in subgingival plaque of patients on short recall was analyzed in relation to the probing depth of the test sites. The subjects had excellent oral hygiene and therefore were unlikely to suffer from active periodontal destruction. Sixty-four subgingival plaque samples, taken from gingival or periodontal pockets with probing depths ranging from 1 to 8 mm, were quantitatively assessed for the presence of the two species using species-specific monoclonal antibodies in conjunction with a very sensitive indirect immunofluorescence technique. Both organisms were encountered in probes from sites as shallow as 2 mm, but the percentage of positive samples clearly rose in relation to the probing depth of the test sites. Overall, B. forsythus was found to colonize lesions earlier, that is at smaller probing depths, than B. gingivalis. Interestingly, whenever a sample was found to be positive for B. gingivalis it was also positive for B. forsythus. The numbers of B. forsythus and B. gingivalis and the total bacterial cell number found in the pockets were significantly correlated to the probing depth. However, with advancing probing depth the increase of the total cell numbers of the two Bacteroides species was considerably more pronounced than the increase of the total subgingival plaque cell number. The recall interval neither affected the frequency of sites positive for B. forsythus or B. gingivalis nor influenced significantly the proportions of the two species in subgingival plaque.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1991

Introduction of the Bacteroides ruminicola xylanase gene into the Bacteroides thetaiotaomicron chromosome for production of xylanase activity.

The xylanase gene from the ruminal bacterium Bacteroides ruminicola 23 is highly expressed in colonic Bacteroides species when carried on plasmid pVAL-RX. In order to stabilize xylanase expression in the absence of antibiotic selection, the xylanase gene was introduced into the chromosome of Bacteroides thetaiotaomicron 5482 by using suicide vector pVAL-7. Xylanase activity in the resulting strain, B. thetaiotaomicron BTX, was about 30% of that observed in B. thetaiotaomicron 5482 containing the xylanase gene on pVAL-RX. The data obtained from continuous culture experiments using antibiotic-free medium showed that expression of xylanase activity in strain BTX was extremely stable, with no demonstrated loss of the inserted xylanase gene over 60 generations, with dilution rates from 0.42 to 0.03 h-1. In contrast, the plasmid-borne xylanase gene was almost completely lost by 60 generations in the absence of antibiotic selection. Incubation of strain BTX with oatspelt xylan resulted in the degradation of more than 40% of the xylan to soluble xylooligomers. The stability of xylanase expression in B. thetaiotaomicron BTX suggests that this microorganism might be suitable for introduction into the rumen and increased xylan degradation.

Animals↗

PubMed1988

Tetracycline-dependent appearance of plasmidlike forms in Bacteroides uniformis 0061 mediated by conjugal Bacteroides tetracycline resistance elements.

Some human colonic Bacteroides strains carry conjugal tetracycline resistance (Tcr) elements, which are thought to be chromosomal. We have found that some of these Tcr elements can mediate the appearance of plasmidlike forms in Bacteroides uniformis 0061. When B. uniformis 0061, containing a conjugal Tcr element designated Tcr ERL, was grown in medium containing tetracycline (1 microgram/ml), two circular DNA forms were found in the alkaline plasmid preparations: NBU1 (10.3 +/- 0.5 kilobases) and NBU2 (11.5 +/- 0.5 kilobases). Restriction analysis of NBU1 and NBU2 showed that they were not identical, although Southern blot analysis indicated that they did contain some region(s) of homology. Results of Southern blot analysis also demonstrated that both NBU1 and NBU2 were normally integrated in the chromosome of B. uniformis or in some undetected large plasmid. Although we were unable to determine the exact structure and location of the integrated forms of NBU1 and NBU2 in B. uniformis, they appear to be in close proximity to each other. Neither NBU1 or NBU2 could be detected as a plasmidlike form in cells exposed to UV light, thymidine starvation, mitomycin C, or autoclaved chlortetracycline (50 micrograms/ml). Four conjugal Tcr elements other than the Tcr ERL element were able to mediate the appearance of NBU1 alone, and two Tcr elements did not mediate the excision of either NBU1 or NBU2. Three strains from different Bacteroides species contained some DNA sequences which had homology to NBU1 and NBU2.

Bacteroides↗