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Journal of Oral Science Jun 1998Large quantities of Prevotella nigrescens (intermedia) ATCC 25261 (P. nigrescens) cells adhere to hydroxyapatite (HA) treated with citrate, but do not adhere...
Large quantities of Prevotella nigrescens (intermedia) ATCC 25261 (P. nigrescens) cells adhere to hydroxyapatite (HA) treated with citrate, but do not adhere experimental pellicle prepared from human whole saliva. To determine the nature of the citrate responsible for promoting P. nigrescens cell adhesion, the duration and frequency of citrate treatment of HA and the inhibitory effect of other carboxylates were tested. The citrate rapidly adhered to HA beads in less than 15 min. With a lower concentration (0.4 mM) of citrate, four treatments of HA were required to promote the maximum adherence to P. nigrescens cells. Citrate-enhanced P. nigrescens cell adherence to HA beads was also inhibited in the presence of cis-aconitate, oxaloacetate and oxalsuccinate. It was also found that P. nigrescens cells heated to 65 degrees C or higher for 5 min could no longer become attached to citrate-treated HA. These data suggest that citrate is one of the essential factors responsible for P. nigrescens cell attachment to apatitic surfaces, and that P. nigrescens' adhesion to citrate is extremely heat-sensitive.
Topics: Aconitic Acid; Adhesiveness; Adult; Bacterial Adhesion; Carboxylic Acids; Citrates; Durapatite; Hot Temperature; Humans; Oxaloacetates; Prevotella intermedia; Saliva; Succinates; Surface Properties; Time Factors
PubMed: 9680763
DOI: 10.2334/josnusd.40.65 -
Oral Microbiology and Immunology Jun 1996Restriction endonuclease analysis, rRNA gene restriction analysis (ribotyping), multilocus enzyme electrophoresis and lipase production were investigated for their... (Comparative Study)
Comparative Study
Restriction endonuclease analysis, rRNA gene restriction analysis (ribotyping), multilocus enzyme electrophoresis and lipase production were investigated for their potential to differentiate isolates belonging to the closely-related species Prevotella intermedia and Prevotella nigrescens. Of 122 strains identified originally as P. intermedia, 52 were assigned to P. intermedia and 68 to P. nigrescens using multilocus enzyme electrophoresis. All 39 P. intermedia and 52 out of 53 P. nigrescens tested produced lipase. Restriction endonuclease analysis identified clonal variants, but did not facilitate the differentiation of strains into species. Taq I ribotyping of 99 strains revealed that all P. intermedia demonstrated a species-specific fragment of 0.40 kbp, which was always associated with a second fragment of 0.57 kbp, and all P. nigrescens tested shared a species-specific fragment of 2.21 kbp. Two strains atypical by multilocus enzyme electrophoresis had none of the above species-specific fragments. Thus, lipase production and restriction endonuclease analysis did not distinguish between P. intermedia and P. nigrescens, but Taq I ribotyping did and also allowed the characterization of individual strains.
Topics: Bacterial Typing Techniques; DNA Restriction Enzymes; DNA, Bacterial; Electrophoresis; Genetic Heterogeneity; Humans; Lipase; Prevotella; Prevotella intermedia; RNA, Ribosomal; Restriction Mapping; Species Specificity
PubMed: 8941766
DOI: 10.1111/j.1399-302x.1996.tb00348.x -
Journal of Periodontology Nov 1999Smoking is a recognized risk factor for the initiation and progression of periodontitis. However, the mechanism by which smoking induces its negative effects on the...
BACKGROUND
Smoking is a recognized risk factor for the initiation and progression of periodontitis. However, the mechanism by which smoking induces its negative effects on the periodontium is not clear. This study aimed to test the hypothesis that synergy may occur between cotinine and bacterial products isolated from 3 putative periodontopathogens.
METHODS
A chick embryo toxin assay was used to investigate bacterial toxins (cell-free extracellular toxins and cell-free cell lysates) from 5 species with and without cotinine. A total of 9 putative periodontopathogens (3 species) and 2 non-oral controls (2 species) were studied. The periodontal species were: Prevotella intermedia (n = 4), Prevotella nigrescens (n = 4), and Porphyromonas gingivalis (n = 1). The control species tested were: Staphylococcus aureus (n = 1) and Escherichia coli (n = 1).
RESULTS
The toxicity kill was significantly greater than expected by simple addition alone (P <0.05, Fisher's exact test) between cotinine (800 ng/ml) and 1) the cell-free extracellular toxins of P. nigrescens MH1 and 2) the cell-free cell lysates of P. intermedia MH2. Synergy occurred with cotinine plus the cell-free extracellular toxins in all but 3 periodontal isolates, and the cell-free cell lysates in all but 2 periodontal isolates. Cotinine significantly (P <0.05, Fisher's exact test) enhanced the effects of cell-free extracellular toxins and cell lysates from one control species (E. coli), but not the other (S. aureus).
CONCLUSIONS
These findings indicate that synergy in an in vitro assay can occur between cotinine and toxins from putative periodontopathogens. This may be one important mechanism by which smoking increases the severity of periodontitis.
Topics: Animals; Bacterial Toxins; Cell-Free System; Chick Embryo; Cotinine; Drug Synergism; Endotoxins; Periodontitis; Porphyromonas gingivalis; Prevotella; Prevotella intermedia; Toxicity Tests; Virulence
PubMed: 10588489
DOI: 10.1902/jop.1999.70.11.1269 -
Oral Microbiology and Immunology Jun 1995Degradation of immunoglobulins is thought to be an important factor in the causation of periodontal diseases by hindering local host defenses and by providing nutrients...
Degradation of immunoglobulins is thought to be an important factor in the causation of periodontal diseases by hindering local host defenses and by providing nutrients to the periodontal microflora. In this study, we characterized the proteolytic activity against human immunoglobulin G (IgG) of 20 strains of Prevotella intermedia and Prevotella nigrescens isolated from periodontal pockets and oral abscesses. IgG degradation was studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. All strains degraded IgG within 48 h after growth in trypticase-yeast extract medium (TY) supplemented with 0.3% IgG. Incorporating IgG in TY broth enhanced bacterial growth. Protease profiles (zymography), which revealed the presence of 1-4 IgG-degrading proteolytic bands in bacterial cell extracts, became more complex after growth in the presence of IgG. A 38-kDa protease capable of degrading IgG nonspecifically was present in almost all strains. The proteolytic activity was mainly located on the surface of the cell envelope. Two strains of P. intermedia and P. nigrescens ATCC 33563 were selected for further studies. Bacterial cell suspensions in phosphate-buffered saline completely degraded human IgG, IgA and IgM within 24 h. This activity depended on reducing conditions and was inhibited at temperatures above 50 degrees C. The pH optimum of immunoglobulin degradation was at pH 7. Strains cultured at 42 degrees C showed a markedly reduced capacity to degrade IgG. Inhibition studies revealed that breakdown of IgG was caused by a cysteine protease(s). The capacity of P. intermedia and P. nigrescens to degrade immunoglobulins may explain their association with polymicrobial oral diseases.
Topics: Animals; Antibodies, Bacterial; Bacterial Proteins; Cattle; Cysteine Endopeptidases; Dogs; Electrophoresis, Polyacrylamide Gel; Humans; Hydrogen-Ion Concentration; Hydrolysis; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Molecular Weight; Peptide Hydrolases; Prevotella; Prevotella intermedia; Protease Inhibitors; Serine Endopeptidases; Time Factors
PubMed: 7567062
DOI: 10.1111/j.1399-302x.1995.tb00134.x -
Australian Endodontic Journal : the... Dec 2000The purpose of the present study was to evaluate the prevalence of Prevotella intermedia and Prevotella nigrescens in symptomatic and asymptomatic endodontic infections...
The purpose of the present study was to evaluate the prevalence of Prevotella intermedia and Prevotella nigrescens in symptomatic and asymptomatic endodontic infections from a Brazilian population. DNA extracted from samples obtained from 28 cases of endodontic infection were examined by the 16S rDNA-directed Polymerase Chain Reaction (PCR) method. PCR detected P. intermedia in 7.1% of the cases (2 out of 28 teeth) and P. nigrescens in only one sample (3.6%). The low prevalence of P. intermedia and P. nigrescens as reported in the present study is probably justified for geographical reasons.
Topics: Adolescent; Adult; Bacteroidaceae Infections; Brazil; DNA, Bacterial; Dental Pulp Necrosis; Humans; Middle Aged; Polymerase Chain Reaction; Prevalence; Prevotella; Prevotella intermedia
PubMed: 11359249
DOI: 10.1111/j.1747-4477.2000.tb00293.x -
Oral Microbiology and Immunology Aug 2005he aim of this study was to investigate the presence of four black-pigmented bacteria, Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia and...
he aim of this study was to investigate the presence of four black-pigmented bacteria, Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia and Prevotella nigrescens, in endodontic infections by culture and polymerase chain reaction (PCR) analyses. Microbial samples were obtained from 50 teeth with untreated necrotic pulps (primary infection) and from 50 teeth with failing endodontic treatment (secondary infection). Microbiological strict anaerobic techniques were used for serial dilution, plating, incubation, and identification. For PCR detection, the samples were analyzed using species-specific primers of 16S rDNA and the downstream intergenic spacer region. Culture and PCR detected the test species in 13/100 and 50/100 of the study teeth, respectively. The organisms were cultured from 11/50 (22%) of primarily infected root canal samples and from 2/50 (4%) of secondary root canal samples. PCR detection identified the target species in 32/50 (64%) and 18/50 (36%) of primary and secondary infections, respectively. P. gingivalis was rarely isolated by culture methods (1%), but was the most frequently identified test species by PCR (38%). Similarly, P. endodontalis was not recovered by culture from any tooth studied, but was detected by PCR in 25% of the sampled teeth. PCR-based identification also showed higher detection rates of P. intermedia (33%) and P. nigrescens (22%) than culture (13%). In conclusion, P. gingivalis, P. endodontalis, P. intermedia, and P. nigrescens were identified more frequently in teeth with necrotic pulp than in teeth with failing endodontic treatment. Also, a higher frequency of black-pigmented species was detected by PCR than by culture.
Topics: Bacterial Typing Techniques; Bacteroidaceae Infections; Colony Count, Microbial; DNA, Bacterial; Dental Pulp Necrosis; Dental Restoration Failure; Female; Humans; Male; Polymerase Chain Reaction; Porphyromonas; Porphyromonas endodontalis; Porphyromonas gingivalis; Prevotella; Prevotella intermedia; Prevotella nigrescens
PubMed: 15943764
DOI: 10.1111/j.1399-302X.2005.00214.x -
FEMS Microbiology Letters Jul 1994Prevotella intermedia and Prevotella nigrescens are not easily distinguished, making it difficult to assess their roles in disease. This study examined the specificity...
Prevotella intermedia and Prevotella nigrescens are not easily distinguished, making it difficult to assess their roles in disease. This study examined the specificity of three monoclonal antibodies (mAbs) for these species. Differentiation between P. intermedia (13 isolates) and P. nigrescens (24 isolates) was by the electrophoretic mobility of their malate and glutamate dehydrogenase enzymes or by DNA homology grouping. All P. intermedia reacted strongly with mAb 40BI3.2.2 whereas P. nigrescens strains did not. Monoclonal antibodies 37BI6.1 and 39BI1.1.2 recognised all strains of both species but most P. nigrescens reacted weakly with mAb 39BI1.1.2. Monoclonal antibody 40BI3.2.2 therefore recognises an antigen specific for P. intermedia but not P. nigrescens and provides an easy and reliable means of distinguishing between these species. Three vaginal isolates identified biochemically as P. intermedia had enzymes with mobilities corresponding to neither P. intermedia nor P. nigrescens. These isolates were not recognised by mAbs 39BI1.1.2 or 40BI3.2.2 and may represent an undescribed taxon within this group of organisms.
Topics: Antibodies, Monoclonal; Antibody Specificity; Bacteroides; DNA; Electrophoresis; Nucleic Acid Hybridization; Species Specificity
PubMed: 8056301
DOI: 10.1111/j.1574-6968.1994.tb07014.x -
Oral Microbiology and Immunology Dec 2005This study investigated the mechanism of protein attachment to the surface of the putative periodontal pathogens Prevotella intermedia and Prevotella nigrescens in...
This study investigated the mechanism of protein attachment to the surface of the putative periodontal pathogens Prevotella intermedia and Prevotella nigrescens in artificial gingival crevicular fluid, and ways to increase protein attachment to the bacterial cells. The effects of cations on protein attachment, bacterial adhesion, and hemagglutination were examined, and cation-binding components on both bacterial species were identified. The presence of cations, especially zinc, copper and cerium, increased attachment of human serum proteins to both bacterial species. In contrast, the presence of hydrophobic inhibitors or sugars had little effect. Protein attachment was reduced by heat treatment of the bacterial cells. Pretreatment of bacteria with human serum proteins inhibited adhesion of both species to buccal epithelial cells and hemagglutination. These effects were enhanced by the presence of zinc and copper during pretreatment. Using a chelating column, specific zinc- and copper-binding proteins were identified on the surfaces of both bacterial species.
Topics: Bacterial Adhesion; Bacterial Proteins; Blood Proteins; Carrier Proteins; Cerium; Copper; Electrophoresis, Polyacrylamide Gel; Epithelial Cells; Gingival Crevicular Fluid; Hemagglutination; Hot Temperature; Humans; Prevotella intermedia; Prevotella nigrescens; Protein Binding; Zinc
PubMed: 16238592
DOI: 10.1111/j.1399-302X.2005.00234.x -
Journal of Periodontal Research Aug 1996Black-pigmented anaerobes have been implicated as major pathogens in the aetiology of adult periodontitis but these organisms are also found in healthy sites. This study...
Black-pigmented anaerobes have been implicated as major pathogens in the aetiology of adult periodontitis but these organisms are also found in healthy sites. This study aimed to examine the relationship between genotypes of black-pigmented anaerobes and disease status of periodontal sites using restriction endonuclease analysis (REA) and ribotyping. The main black-pigmented species recovered from sites were Porphyromonas gingivalis, Prevotella intermedia and Prevotella nigrescens. Each of the 58 subjects investigated harboured distinct genotypes of these three species. Most subjects appeared to be colonized by a single genotype of P. gingivalis and Pr. intermedia, whereas multiple types of Pr. nigrescens colonized many individuals. Plasmids were only found in a few Pr. nigrescens strains. No association was found between the disease status of sites and any specific or group of genotypes of either species or presence of a plasmid. Since the same genotypes of P. gingivalis, Pr. intermedia and Pr. nigrescens were found at both diseased and non-diseased sites in a subject, adult periodontitis is not explained by the presence of specially virulent clones of these organisms. Their role in periodontitis, therefore, is likely to be opportunistic.
Topics: Adult; Bacterial Typing Techniques; Blotting, Southern; Clone Cells; DNA Restriction Enzymes; DNA, Bacterial; Dental Plaque; Electrophoresis, Polyacrylamide Gel; Female; Humans; Male; Mouth; Periodontal Pocket; Periodontitis; Plasmids; Porphyromonas gingivalis; Prevotella; Prevotella intermedia; Prohibitins; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Species Specificity; Virulence
PubMed: 8884636
DOI: 10.1111/j.1600-0765.1996.tb00511.x -
Microbiology (Reading, England) Jul 2003The haem pigment of Porphyromonas gingivalis is composed of micro -oxo bishaem, [Fe(III)PPIX](2)O, but the nature of that generated by Prevotella species has not been...
The haem pigment of Porphyromonas gingivalis is composed of micro -oxo bishaem, [Fe(III)PPIX](2)O, but the nature of that generated by Prevotella species has not been established. Mössbauer, Raman and UV-visible spectrophotometry were used to characterize the haem pigment of Prevotella intermedia and Prevotella nigrescens. Mössbauer and Raman spectroscopy revealed the major haem species to be monomeric iron protoporphyrin IX, Fe(III)PPIX.OH (haematin). The terminal growth pH of both species on blood agar was between 5.8 and 6.0, which favours the formation and maintenance of monomeric Fe(III)PPIX.OH. Incubation of Pr. nigrescens and Pr. intermedia with oxyhaemoglobin at pH 6.5 resulted in formation of aquomethaemoglobin which was degraded to generate Fe(III)PPIX.OH which in turn became cell-associated, whilst incubation at pH 7.5 resulted in formation of [Fe(III)PPIX](2)O. It is concluded that both Prevotella species degrade oxyhaemoglobin to form [Fe(III)PPIX](2)O as an intermediate, which is converted to Fe(III)PPIX.OH through a depression in pH. The low pH encourages cell-surface deposition of insoluble Fe(III)PPIX.OH which would act as a barrier against oxygen and reactive oxygen species, and also protect against H(2)O(2) through its inherent catalase activity.
Topics: Animals; Heme; Horses; Humans; Hydrogen Peroxide; Hydrogen-Ion Concentration; In Vitro Techniques; Oxyhemoglobins; Pigments, Biological; Prevotella; Prevotella intermedia; Protoporphyrins; Spectrophotometry; Spectroscopy, Mossbauer; Spectrum Analysis, Raman
PubMed: 12855722
DOI: 10.1099/mic.0.26258-0