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At least 19 recordsLinked to original sources

PubMed2002

Dipeptide utilization by the periodontal pathogens Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Fusobacterium nucleatum.

Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Fusobacterium nucleatum, which can frequently be isolated from periodontal pockets, preferentially utilize proteins and peptides as growth substrates. In this study, we determined the size of peptide that is preferentially utilized as a source of energy and material for cell growth by P. gingivalis, P. intermedia, P. nigrescens and F. nucleatum using various sizes of poly amino acids consisting of two to approximately 100 molecules of aspartate or glutamate. Resting cells of P. gingivalis, P. intermedia and P. nigrescens utilized aspartylaspartate, while cells of P. gingivalis and F. nucleatum utilized glutamylglutamate. The addition of aspartylaspartate to the culture medium increased the growth of P. gingivalis, P. intermedia and P. nigrescens, while the addition of glutamylglutamate promoted the growth of P. gingivalis and F. nucleatum. These results clearly indicate that dipeptides such as aspartylaspartate and glutamylglutamate can be utilized as growth substrates for P. gingivalis, P. intermedia, P. nigrescens and F. nucleatum.

Aspartic Acid↗

PubMed2001

Effects of glucose on formation of cytotoxic end-products and proteolytic activity of Prevotella intermedia, Prevotella nigrescens and Porphyromonas gingivalis.

Black-pigmented bacteria which produce cytotoxic metabolic end-products and cell membrane-associated proteases have been reported to play an important role in the pathogenesis of periodontal diseases. These bacterial virulence factors can be modified by the environmental conditions including nutrients supplied variously into the oral cavity. Although glucose is one of the most essential nutrients for oral bacteria, the exogenous supply of glucose may be discontinuous and the glucose concentration in a periodontal pocket may be influenced by the depth of the periodontal pocket. Therefore, effects of glucose as an environmental factor on the virulence factors of Prevotella intermedia, Prevotella nigrescens and Porphyromonas in the presence o glucose.,bo t P. intermedia and P. nigrescens markedly decreased the production of cytotoxic end-products including succinate.,isobutyrate,isovalerate and ammonia, although their growth was increased. Furthermore, the proteolytic activities such as immunoglobulin- albumin- and casein-degrading activities of these bacteria were decreased in the presence of glucose. On the other hand, no effect of glucose on the metabolic activity of P gingivalis was observed. These results suggest that pathogenicity of P. intermedia P. nigrescens may be decreased by the presence of glucose.

Ammonia↗

PubMed1999

Beta-lactamase production in Prevotella intermedia, Prevotella nigrescens, and Prevotella pallens genotypes and in vitro susceptibilities to selected antimicrobial agents.

The present study investigated the beta-lactamase production of 73 Prevotella intermedia, 84 Prevotella nigrescens, and 14 Prevotella pallens isolates and their in vitro susceptibilities to six antimicrobial agents. The P. intermedia and P. nigrescens isolates were recovered from oral and extraoral samples obtained from subjects in two geographic locations from 1985 to 1995. The clonality of the beta-lactamase-positive and beta-lactamase-negative isolates and the clustering of the genotypes were studied by arbitrarily primed-PCR fingerprinting. beta-Lactamase production was detected in 29% of P. intermedia isolates, 29% of P. nigrescens isolates, and 57% of P. pallens isolates. No difference in the frequencies of beta-lactamase production by P. intermedia and P. nigrescens between isolates from oral and extraoral sites, between isolates obtained at different time periods, or between P. intermedia isolates from different geographic locations was observed. However, the P. nigrescens isolates from the United States were significantly more frequently (P = 0.015) beta-lactamase positive than those from Finland. No association between the genotypes and beta-lactamase production or between the genotypes and the sources of the isolates was found. The penicillin G MICs at which 90% of the isolates were inhibited were 8 microg/ml for P. intermedia, 8 microg/ml for P. nigrescens, and 16 microg/ml for P. pallens. For the beta-lactamase-negative isolates, the corresponding values were 0.031, 0.031, and 0.125 microg/ml, and for the beta-lactamase-positive isolates, the corresponding values were 16, 8, and 32 microg/ml. All isolates were susceptible to amoxicillin-clavulanate, cefoxitin, metronidazole, azithromycin, and trovafloxacin. The MICs of amoxicillin-clavulanate and cefoxitin were relatively higher for the beta-lactamase-positive population than for the beta-lactamase-negative population.

Anti-Bacterial Agents↗

PubMed1999

Possible potentiation of toxins from Prevotella intermedia, Prevotella nigrescens, and Porphyromonas gingivalis by cotinine.

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.

Animals↗

PubMed2000

Genetic heterogeneity in Prevotella intermedia, Prevotella nigrescens, Prevotella corporis and related species isolated from oral and nonoral sites.

Prevotella intermedia (43 isolates), Prevotella nigrescens (55) and Prevotella corporis (8) from oral and nonoral sites were distinguished by species-specific DNA fragments, after hybridization of DNA fragments with ribosomal RNA (ribotyping). Eight strains previously identified as P. intermedia did not have these specific fragments. P. nigrescens, P. intermedia and P. corporis formed separate clusters in dendrograms constructed using clustering with an unweighted pair group method with arithmetic averages of similarity values derived from ribotype patterns, with 10 subclusters in P. intermedia isolates and 26 in P. nigrescens. Nine groups of P. intermedia isolates and 6 of P. nigrescens shared identical patterns. Specific ribotypes or species were not associated with particular diseases when all isolates were analyzed. However, results from organisms isolated by one laboratory using consistent clinical reporting indicated that P. intermedia was associated with more severe forms of periodontitis and P. nigrescens with mild to moderate disease.

Bacteroidaceae Infections↗

PubMed1999

Identification of a fimbriae-associated haemagglutinin from Prevotella intermedia.

Prevotella intermedia, a putative periodontopathic microorganism, possesses various types of fimbriae (surface appendages). Some of these surface structures mediate the adherence of the organism to several mammalian erythrocytes, resulting in the agglutination of the erythrocytes. Prevotella intermedia fimbriae were solubilized and separated by preparative SDS gel electrophoresis followed by preparative isoelectric focusing to determine which fimbrial component(s) were responsible for the haemagglutinating activity exhibited by these bacteria. Heat treatment of isolated fractions which exhibited haemagglutinating activity revealed the presence of two types of haemagglutinating activity which were either heat sensitive or resistant. Analysis of isolated fractions, which exhibited haemagglutinating activity that were heat labile, by Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of a 3.8 kD peptide. The purified peptide had a pI of 4.8-5.0. Examination of fractions containing this peptide by electron microscopy showed the presence of fimbriae. Immunogold labelling showed that mouse antibodies raised against the 3.8 kD peptide bound specifically and extensively to P. intermedia fimbriae. It appears that this peptide is a fimbriae-associated haemagglutinin and may represent a major constituent of the fimbriae. Further, fractions exhibiting haemagglutinating activity that were heat resistant, which were recovered at a pH of 3.5 in preparative isoelectric focusing of fimbrial samples, did not possess any detectable major protein bands as shown by analytical gel electrophoresis. However, silver stained gels for the detection of lipopolysaccharide (LPS) revealed the presence of LPS-like components in these fractions. In addition, LPS isolated from whole cells showed a similar electrophoretic pattern and exhibited the haemagglutinating activity that was heat resistant. The results of this study strongly suggest that P. intermedia possesses the machinery to agglutinate erythrocytes, which may be a contributing factor in its colonization in vivo.

Animals↗

PubMed1998

The binding and utilization of hemoglobin by Prevotella intermedia.

Prevotella intermedia, a putative periodontopathic microorganism, requires iron for growth. Hemoglobin can be a major source of iron for bacterial growth in vivo since it is present in the crevicular fluid collected from periodontitis sites. Experiments studying the growth of P. intermedia in iron-depleted Todd-Hewitt broth supplemented with human hemoglobin showed that the bacteria were able to utilize human hemoglobin as a source of iron. The uptake of iron from hemoglobin by P. intermedia appears to be initiated by the binding of hemoglobin to the bacteria as shown by direct binding studies using 125I-labeled human hemoglobin. Scatchard analysis of saturation binding data revealed that 125I-labeled human hemoglobin had a dissociation constant (Kd) of 2.53 x 10(-8) M for the receptor on P. intermedia. Binding of labeled hemoglobin to P. intermedia was competitively inhibited by unlabeled human hemoglobin showing that the binding was specific. The ability of bovine hemoglobin, but not hemin or non-hemoglobin heme-containing compounds, to inhibit binding competitively suggested that the globin moiety of the hemoglobin molecule is recognized by the hemoglobin binding receptors.

Animals↗

PubMed2005

Effects of zinc and copper on adhesion and hemagglutination of Prevotella intermedia and Prevotella nigrescens.

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.

Bacterial Adhesion↗

PubMed2002

Direct quantitative differentiation between Prevotella intermedia and Prevotella nigrescens in clinical specimens.

This paper describes a quantitative fluorescent in situ hybridization (FISH) assay for the differential identification of Prevotella intermedia and Prevotella nigrescens in clinical samples, and compares its performance with less discriminatory culture and quantitative immunofluorescence (IF) assays. Fluorescence-labelled oligonucleotide probes directed to specific 16S rRNA sequences of P. intermedia, P. nigrescens, Prevotella pallens and Prevotella denticola were hybridized under stringent conditions with cultured reference strains or plaque samples from deep periodontal pockets. Probe specificity was defined with strains from multiple oral Prevotella species. The lower detection level of the assays was approximately 3x10(3) target cells per ml of plaque-sample suspension. P. intermedia, P. nigrescens, P. pallens and P. denticola were detected in plaques with prevalences of 69, 67, 0 and 28%, respectively. On average, 3.9 x 10(6) P. intermedia, 3.1 x 10(6) P. nigrescens and 5.6 x 10(5) P. denticola cells were counted per positive sample. All three species were found almost exclusively in dense mixed aggregates. Quantitative FISH data agreed satisfactorily with corresponding IF data (r=0.711). Both FISH and IF enumerations of the sum of P. intermedia and P. nigrescens markedly exceeded the c.f.u. counts of black-pigmented colonies in Porphyromonas gingivalis-free cultured subgingival plaques. The results demonstrate the validity of this new assay. Unlike established IF, culture, PCR or checkerboard DNA hybridization assays, this FISH assay differentiates quantitatively between P. intermedia and P. nigrescens, provides visual accuracy control, and offers insights into the spatial distribution of the target cells within a clinical sample.

Bacterial Infections↗

PubMed2005

Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia and Prevotella nigrescens in endodontic lesions detected by culture and by PCR.

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.

Bacterial Typing Techniques↗

PubMed2000

Glucose metabolism by Prevotella intermedia and Prevotella nigrescens.

Glucose metabolism by Prevotella intermedia and Prevotella nigrescens were investigated. Glucose increased the anaerobic growth of these bacteria and promoted the accumulation of intracellular polysaccharide. The polysaccharide was confirmed to be glycogen-like glucan by the absorption spectrum of iodinepolysaccharide complex and the sugar composition. The washed cells consumed glucose anaerobically and converted a part of glucose into the metabolic end-products acetate, formate and succinate. The rest of glucose was confirmed to be accumulated as intracellular polysaccharide. The cells grown in the presence of glucose produced acetate, formate and succinate without exogenous glucose along with the consumption of intracellular polysaccharide. The metabolism of glucose and intracellular polysaccharide required bicarbonate. Prevotella cells had hexokinase and a set of the usual enzymes of the Embden-Meyerhof-Parnas pathway except that phosphofructokinase was pyrophosphate-dependent. A series of enzymes, including phosphoenolpyruvate carboxylase, phosphoenolpyruvate carboxykinase, malate dehydrogenase, fumarase and fumarate reductase, was found for succinate formation. Another series of enzymes, pyruvate oxidoreductase, pyruvate formate-lyase, phosphotransacetylase and acetate kinase was found for acetate and formate formation. Glucose 1,6-bisphosphate-dependent phosphoglucomutase and fructose 1,6-bisphosphate-activated UDP-glucose pyrophosphorylase were detected for glycogen synthesis, while glycogen phosphorylase was for glycogen degradation. The capacity of intracellular polysaccharide formation in addition to glucose fermentation could be advantageous for survival in the supragingival area as well as in the subgingival area.

Energy Metabolism↗

PubMed1998

Antimicrobial susceptibilities of Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens spp. isolated in Spain.

The susceptibilities of 143 Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens isolates to 18 antimicrobial agents were tested. All P. gingivalis isolates were susceptible. In contrast, some Prevotella spp. (17%) were resistant to beta-lactams, erythromycin, clindamycin, or tetracycline and carried resistance genes, ermF or tetQ, or beta-lactamases.

Adult↗

PubMed1996

Characterization of Prevotella intermedia and Prevotella nigrescens by enzyme production, restriction endonuclease and ribosomal RNA gene restriction analyses.

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.

Bacterial Typing Techniques↗

PubMed1998

Incidence of Prevotella intermedia and Prevotella nigrescens in periodontal health and disease.

The incidence of black-pigmented rods (BPRs), especially Prevotella intermedia and Prevotella nigrescens, in periodontal health and disease were examined. Furthermore, the degradative enzyme activities of P. intermedia were compared among the strains from periodontal health and disease. Microbiological specimens were collected from subgingival crevice or periodontal pocket by paper point. The BPRs were found in 71.1% of periodontally healthy subjects (n=45), and in 47.1% of healthy sites (n=34) and 87.8% of active sites (n=41) among periodontally diseased patients. Porphyromonas gingivalis was detected only in active sites of periodontally diseased patients (17.8% of 180 strains). P. intermedia was the predominant BPR in both healthy and active sites (37.3 and 41.7%, respectively) of the patients. However, P. nigrescens was the predominant BPR (70.5% of 173 strains) in periodontally healthy subjects. The enzyme activities of esterase, esterase-lipase, acid-phosphatase and alpha-fucosidase of P. intermedia strains isolated from active sites in patients were significantly higher (P<0.05) than those of healthy subjects. The results suggest that P. intermedia might increase the activity of degradative enzymes under a certain condition and support the progression of periodontitis.

Adult↗

PubMed1999

Hemolytic and hemagglutinating activities of Prevotella intermedia and Prevotella nigrescens.

A total of 91 isolates of Prevotella intermedia or Prevotella nigrescens from subgingival sites were identified by PCR using primers specific for sequences of 16S rRNA. The hemolytic and hemagglutinating activities of the P. intermedia isolates exhibited significantly higher levels compared to those of the P. nigrescens isolates by quantitative analysis. The hemagglutinin gene (phg) was found in 23 of 26 P. intermedia isolates (88.5%), whereas it was found in only two of 44 isolates (4.5%) of P. nigrescens. The high hemolytic and hemagglutinating activities of P. intermedia may be involved in the pathogenicity of P. intermedia in the progression of periodontal disease.

Animals↗

PubMed1996

Prevotella intermedia and Prevotella nigrescens serotypes, ribotypes and binding characteristics.

Type strains and 62 clinical isolates of Prevotella intermedia and Prevotella nigrescens were typed with the use of genomic DNA fingerprints and rRNA gene probes. The strains were further serotyped with monoclonal antibodies and characterized with SDS-PAGE, enzymatic activities, hemolysis and hemagglutination and coaggregation with Streptococcus and Actinomyces spp. P. intermedia and P. nigrescens were found to have distinct ribotype patterns which correspond to previously defined serotyupes I and II/III, respectively. No clear phenotypic difference related to hemolysis, hemagglutination and coaggregation with streptococcus and actinomyces species, or expression of aminopeptides and lipase was found between P. intermedia and P. migrescens.

Animals↗

PubMed1998

Pathogenicity of facultative and obligate anaerobic bacteria in monoculture and combined with either Prevotella intermedia or Prevotella nigrescens.

The pathogenicity of obligate and facultative anaerobic bacteria commonly found in endodontic infections was tested using a mouse model. The capacity of inducing abscesses was evaluated seven days after subcutaneous injection of the bacteria in pure culture and in combinations with either Prevotella intermedia or Prevotella nigrescens. Nine of the fifteen bacterial strains tested were pathogenic in pure culture. No statistically significant differences were detected between these strains in pure culture and in mixtures with either P. intermedia or P. nigrescens. Synergism between the bacterial strains was only apparent when associating Porphyromonas endodontalis with P. intermedia or P. nigrescens. Histopathological examination of tissue sections from induced abscesses revealed an acute inflammatory reaction, dominated by polymorphonuclear leukocytes. Sections from the control group using sterile medium showed no evidence of inflammatory reaction.

Abscess↗

PubMed1996

Clonality of Porphyromonas gingivalis, Prevotella intermedia and Prevotella nigrescens isolated from periodontally diseased and healthy sites.

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.

Adult↗