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Journal of Medical Microbiology Oct 1992Collection strains (21) and non-pigmented clinical isolates (96) provisionally identified as Prevotella spp. were classified numerically on the basis of pyrolysis mass... (Comparative Study)
Comparative Study
Collection strains (21) and non-pigmented clinical isolates (96) provisionally identified as Prevotella spp. were classified numerically on the basis of pyrolysis mass spectrometry (PMS) data and reaction patterns in conventional tests (CTRPs) for volatile and non-volatile fatty acids, pre-formed enzymes and biochemical activity. PMS and CTRP classifications were compared with a previous classification based on visual analysis of SDS-PAGE patterns. Although the order of clusters differed, cross-tabulation of cluster membership revealed strong correlations between classifications. Cluster membership in the PMS classification correlated particularly well with SDS-PAGE results. CTRP clusters corresponded largely to the recognised species of Prevotella, but PMS and SDS-PAGE divided two species into sub-groups: two in P. buccae and five in P. veroralis. The latter subgroups could be discriminated by small but consistent differences in CTRPs. An undesignated, well differentiated cluster of strains appeared closest to the main group of P. buccae strains in PMS and CTRPs. B. (P.) capillus could not be distinguished from P. buccae; these species are regarded as synonymous. Strains of P. zoogleoformans and B. (P.) pentosaceus were well separated from other strains in PMS. A complex comprising clusters of P. disiens, P. oralis, P. veroralis, P. loescheii and a further undesignated group similar to P. melaninogenica was well differentiated from P. buccae and P. oris in PMS; clusters corresponding to P. bivia, P. corporis, P. intermedia and P. denticola formed another complex.
Topics: Bacterial Typing Techniques; Bacteroides; Classification; Gas Chromatography-Mass Spectrometry; Periodontal Diseases; Statistics as Topic
PubMed: 1404327
DOI: 10.1099/00222615-37-4-273 -
International Journal of Systematic... Oct 1992During studies of human periodontal disease, a number of bacterial strains were encountered that, on the basis of results of standard biochemical tests, appeared to be...
During studies of human periodontal disease, a number of bacterial strains were encountered that, on the basis of results of standard biochemical tests, appeared to be Prevotella buccalis, Prevotella denticola, Prevotella melaninogenica, or Prevotella loescheii. However, use of the standard biochemical tests, cellular fatty acid analyses, and the polyacrylamide gel electrophoresis patterns of soluble proteins resulted in conflicting identifications of these strains. The results of tests for cellobiose fermentation, inulin fermentation, and pigment production were responsible for most of the discordant results. Cellular fatty acid analyses in which the Microbial Identification System was used did not differentiate these strains from validly described species, even though separate library entries were created for them. DNA reassociation determinations in which the S1 nuclease procedure was used showed that cellobiose fermentation and pigment production are variable among strains of P. melaninogenica and P. denticola and that fermentation of xylan is not a reliable characteristic for differentiating P. buccalis from Prevotella veroralis. In contrast to previous indications, most strains of P. veroralis do not ferment xylan. These species can be differentiated by DNA-DNA reassociation and by cellular fatty acid analysis, using the Microbial Identification System, but differentiation by currently described phenotypic characteristics is not reliable. Similarly, P. loescheii and the genetically distinct (but closely related) D1C-20 group cannot be distinguished reliably from each other or from P. veroralis, P. denticola, and P. melaninogenica on the basis of currently described phenotypic tests other than cellular fatty acid composition or, for some species, electrophoretic patterns of soluble whole-cell proteins.
Topics: Bacterial Typing Techniques; Bacteroides; Cellobiose; Culture Media; DNA, Bacterial; Fatty Acids; Fermentation; Humans; Phenotype; Xylans
PubMed: 1390106
DOI: 10.1099/00207713-42-4-536 -
Oral Microbiology and Immunology Aug 1992Soluble sonic extracts of Prevotella loescheii caused a dose-dependent inhibition of human peripheral blood lymphocyte proliferation by mitogen and of the proliferation...
Soluble sonic extracts of Prevotella loescheii caused a dose-dependent inhibition of human peripheral blood lymphocyte proliferation by mitogen and of the proliferation of a leukemic cell line, BALL-1, when assessed by DNA synthesis (3H-thymidine incorporation). RNA (3H-uridine incorporation) and protein (3H-leucine incorporation) synthesis were similarly altered after exposure to the extract. There was no effect on cell viability as measured by either trypan blue exclusion or extracellular release of the cytoplasmic enzyme lactate dehydrogenase. Preliminary characterization indicates the suppressive factor(s) derived from P. loescheii to be a protein since it is heat-labile and trypsin-sensitive. The factor eluted in a peak on a high-pressure liquid chromatography gel filtration corresponding to a molecular weight of approximately 32,000. Since black-pigmented anaerobic rods have been implicated in the pathogenesis of periodontal disease, the data suggest that P. loescheii contributes to the disease process by suppressing lymphocyte function.
Topics: Bacterial Proteins; Bacteroides; Cell Division; DNA; Humans; Immunologic Factors; Lymphocyte Activation; Lymphocytes; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Sonication; Tumor Cells, Cultured; Virulence
PubMed: 1408357
DOI: 10.1111/j.1399-302x.1992.tb00030.x -
The International Journal of... Jun 19921. Porphyromonas gingivalis is believed an important pathogen of adult periodontitis. A gene library of P. gingivalis 381 was constructed in lambda phage vector L47.1....
1. Porphyromonas gingivalis is believed an important pathogen of adult periodontitis. A gene library of P. gingivalis 381 was constructed in lambda phage vector L47.1. The library was probed with serum obtained from patients of severe adult periodontitis. Two clones, lambda MDBG101 and lambda MDBG103 which were expressed, 200 and 160 kDa respectively, were selected and further studied. 2. The expressed antigens in these two clones were also reacted with rabbit antiserum against whole cells, capsular fraction and cell surface fraction of P. gingivalis. 3. Genes coding protein antigens in lambda MDBG101 and lambda MDBG103 were subcloned into high-copy-number plasmid vector pACYC184 and subclones obtained were designated as MD101 and MD103. Recombinant plasmids, pMD101 and pMD103, differed in their restriction endonuclease digestion. 4. Immunodiffusion analysis showed that cloned proteins from MD101 and MD103 reacted with antiserum against P. gingivalis but did not react with antiserum against Prevotella intermedia, Prevotella loescheii and Prevotella asaccharolyticus. 5. These data suggest that P. gingivalis species-specific antigens has been successfully cloned and expressed in Escherichia coli. Since these cloned specific antigens were recognized by adult periodontitis patient sera, the recombinant antigen will be useful material for the development of serodiagnosis of P. gingivalis infection in adults periodontitis.
Topics: Adult; Antigens, Bacterial; Cloning, Molecular; DNA Restriction Enzymes; Electrophoresis, Polyacrylamide Gel; Gene Library; Genetic Vectors; Humans; Immunodiffusion; Immunoenzyme Techniques; Periodontitis; Plasmids; Porphyromonas gingivalis; Species Specificity
PubMed: 1319357
DOI: 10.1016/0020-711x(92)90102-7 -
Infection and Immunity Apr 1992Prevotella loescheii PK1295 can grow on native hemoglobin as a source of heme. Supernatants of P. loescheii cultures hemolysed human erythrocytes and degraded native...
Prevotella loescheii PK1295 can grow on native hemoglobin as a source of heme. Supernatants of P. loescheii cultures hemolysed human erythrocytes and degraded native hemoglobin. These combined activities may provide heme (or iron) for the growth of P. loescheii and other dental plaque bacteria.
Topics: Cell Division; Heme; Hemoglobins; Hemolysis; Humans; In Vitro Techniques
PubMed: 1548099
DOI: 10.1128/iai.60.4.1721-1723.1992 -
Oral Microbiology and Immunology Feb 1992The occurrence of oral gram-negative anaerobes was examined in 30 edentulous infants (mean age 3 months, range 1-7 months). One pooled swab sample from mucosal surfaces...
The occurrence of oral gram-negative anaerobes was examined in 30 edentulous infants (mean age 3 months, range 1-7 months). One pooled swab sample from mucosal surfaces (cheeks, palate, tongue) and one saliva sample was taken from each infant. The samples were cultured aerobically and anaerobically using non-selective and selective media. Prevotella (Bacteroides) melaninogenica was the most frequently isolated anaerobe, found in 70% of the infants. The other common anaerobes were Fusobacterium nucleatum, Veillonella spp. and nonpigmented Prevotella (Bacteroides) spp., found in 60%, 57% and 57% of the infants, respectively. Of corroding bacilli, Bacteroides gracilis was detected in 23% of the infants, Wolinella spp. and microaerophilic Eikenella corrodens in one infant (3% each). Leptotrichia spp., microaerophilic Capnocytophaga spp., Prevotella (Bacteroides) loescheii and Prevotella (Bacteroides) intermedia were found in 17%, 13%, 13% and 7% of the infants, respectively. In addition to these 30 infants, 21 edentulous infants were investigated for the presence of Actinobacillus actinomycetemcomitans only. A. actinomycetemcomitans was not detected in any of the 51 edentulous infants. The number of different anaerobic bacterial species in the same mouth varied from 0 to 7. No anaerobic bacteria were detected in 3 of 30 children (10%). These data suggest that various anaerobic bacterial species readily colonize the edentulous mouth in infants.
Topics: Bacteroides; Female; Fusobacterium nucleatum; Gram-Negative Anaerobic Bacteria; Humans; Infant; Male; Mouth Mucosa; Mouth, Edentulous; Saliva; Veillonella
PubMed: 1528621
DOI: 10.1111/j.1399-302x.1992.tb00016.x -
Journal of Clinical Microbiology Sep 1991A rapid method for presumptive identification of black-pigmented gram-negative anaerobic rods was developed. Using filter paper spot tests for indole production,...
A rapid method for presumptive identification of black-pigmented gram-negative anaerobic rods was developed. Using filter paper spot tests for indole production, sialidase, alpha-glucosidase, beta-glucosidase, alpha-fucosidase, and trypsinlike enzyme activities, 100% of Porphyromonas gingivalis, Prevotella intermedia, and Bacteroides levii and 89% of Prevotella corporis isolates were correctly identified to the species level. Porphyromonas asaccharolytica and Porphyromonas endodontalis could not be differentiated from each other but could be distinguished from all other species tested. Similarly, Prevotella denticola, Prevotella loescheii, and Prevotella melaninogenica could not be differentiated from each other. The methods described are based on 4-methylumbelliferone derivatives of the various substrates and are simple to perform, rapid (less than 15 min), and applicable to difficult-to-cultivate anaerobic rods.
Topics: Bacteriological Techniques; Evaluation Studies as Topic; Glucosidases; Gram-Negative Anaerobic Bacteria; Hymecromone; Indoles; Neuraminidase; Pigmentation; alpha-L-Fucosidase
PubMed: 1774320
DOI: 10.1128/jcm.29.9.1955-1958.1991 -
Journal of Dental Research Oct 1988Bacteroides intermedius includes two distinct groups of organisms that are phenotypically indistinguishable by conventional methods. These two groups are represented by...
Bacteroides intermedius includes two distinct groups of organisms that are phenotypically indistinguishable by conventional methods. These two groups are represented by the type strain of the species ATCC 25611T (B. intermedius type I) and by ATCC 33563 (B. intermedius type II). Members of each group can be distinguished from each other by analysis of the cellular protein composition by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and by DNA-DNA homology studies, because they share less than 40% homology. The purpose of this study was to prepare specific DNA probes for the two groups of Bacteroides intermedius and to test them against field isolates. Whole-cell DNA probes were prepared from B. intermedius types I and II and tested against 253 field strains of Bacteroides which had been identified by conventional phenotypic tests as B. intermedius. Of these, 170 (67%) hybridized with the B. intermedius type I DNA probe, 28 (11%) with the type II, and 23 (9%) failed to react with the B. intermedius probes but did hybridize with either B. melaninogenicus, B. loescheii, or B. corporis whole-cell DNA probes. The 32 (13%) remaining isolates failed to hybridize with any of the five Bacteroides probes or with probes to B. asaccharolyticus, B. buccae, B. buccalis, B. denticola, B. gingivalis, B. oralis, or B. oris. These data demonstrate the usefulness of whole-cell DNA probes for the identification of phenotypically similar or identical field isolates.
Topics: Bacteroides; DNA Probes; Nucleic Acid Hybridization; Prevotella melaninogenica; Sequence Homology, Nucleic Acid; Species Specificity
PubMed: 2902111
DOI: 10.1177/00220345880670100401 -
Journal of Dental Research Nov 1987Cross-inhibition within the group of black-pigmented Bacteroides, including both oral and non-oral strains, was studied by means of a membrane filter technique. It was...
Cross-inhibition within the group of black-pigmented Bacteroides, including both oral and non-oral strains, was studied by means of a membrane filter technique. It was found that B. gingivalis possessed the most extended inhibitory capacity among all species tested. B. gingivalis showed inhibitory activity against B. intermedius, B. endodontalis, B. loescheii, and B. melaninogenicus. B. endodontalis was active against some B. intermedius strains. Among the saccharolytic species, some B. melaninogenicus strains were inhibitory for some B. endodontalis strains, some B. gingivalis strains, and some B. intermedius strains. These inhibitory activities observed in vitro may play a role in the colonization of the periodontal pocket.
Topics: Abscess; Antibiosis; Bacteroides; Bacteroides Infections; Colony Count, Microbial; Culture Media; Culture Media, Conditioned; Dental Plaque; Dental Pulp Cavity; Filtration; Gingiva; Humans; Mouth; Mouth Diseases; Periodontal Pocket; Pigments, Biological; Porphyromonas gingivalis; Prevotella intermedia; Prevotella melaninogenica; Tongue
PubMed: 10872403
DOI: 10.1177/00220345870660111201 -
Journal of General Microbiology Jun 1987We compared 22 Bacteroides species by DNA-DNA homology studies using the S1 endonuclease method. None of the currently defined species shared more than 30% DNA homology...
We compared 22 Bacteroides species by DNA-DNA homology studies using the S1 endonuclease method. None of the currently defined species shared more than 30% DNA homology with any other species examined with the exception of B. buccae and B. capillus (which along with B. pentosaceus are now considered a single species), which shared 86% of their DNA sequences. Two clusters showed weak genetic relationships, with DNA homology greater than 10%. The first cluster included B. coporis, B. disiens, B. bivius, B. intermedius and B. melaninogenicus. The second cluster included B. fragilis, B. eggerthii, B. ovatus, B. thetaiotaomicron and B. uniformis. Five of the oral species, B. asaccharolyticus, B. gingivalis, B. loescheii, B. intermedius and B. melanogenicus, were chosen for study as whole chromosomal probes in dot blot assays. These were tested against 243 clinical strains biochemically identified as Bacteroides species. The DNA probes correctly identified 94% of the clinical strains. DNA probe and biochemical identification was 100% for two of the five species. In contrast, only 86% of the strains biochemically identified as B. intermedius were identified by the DNA probe. The DNA probes gave a species identification to seven strains which could not be biochemically identified.
Topics: Bacteroides; Bacteroides fragilis; Chromosomes, Bacterial; DNA, Bacterial; Genetic Markers; Prevotella melaninogenica; Sequence Homology, Nucleic Acid
PubMed: 2889792
DOI: 10.1099/00221287-133-6-1423