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Journal of Endodontics Oct 1997The occurrence of Prevotella intermedia and Prevotella nigrescens in endodontic infections was studied using sodium dodecyl sulfate polyacrylamide gel electrophoresis...
The occurrence of Prevotella intermedia and Prevotella nigrescens in endodontic infections was studied using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of whole cell protein to distinguish between the species. Previous studies have shown an association between black-pigmented bacteria (BPB) and endodontic infections and that Prevotella intermedia (previously known as Bacteroides intermedius) was the most commonly isolated BPB. Recently, however, strains identified as P. intermedia were shown to in fact be composed of two separate species, P. intermedia and P. nigrescens. Fifty-six strains of BPB isolated from endodontic infections and previously identified as P. intermedia were used in this study. Following SDS-PAGE, P. nigrescens showed a unique 18.6 kDa band that was used to differentiate P. nigrescens from P. intermedia. Of the 56 strains of BPB, 41 (73.2%) were identified as P. nigrescens and 15 (26.8%) as P. intermedia. This study confirms that P. nigrescens, and not P. intermedia, is the BPB most often isolated from infections of endodontic origin.
Topics: Amino Acid Sequence; Bacterial Proteins; Bacteroidaceae Infections; Dental Pulp Cavity; Dental Pulp Diseases; Electrophoresis, Polyacrylamide Gel; Humans; Molecular Sequence Data; Molecular Weight; Prevotella; Prevotella intermedia; Species Specificity
PubMed: 9587274
DOI: 10.1016/S0099-2399(97)80173-2 -
Oral Microbiology and Immunology Apr 1996A total of 344 Prevotella intermedia and nigrescens group isolates from 59 subjects were identified by hybridization with nonradioactively labeled species-specific...
A total of 344 Prevotella intermedia and nigrescens group isolates from 59 subjects were identified by hybridization with nonradioactively labeled species-specific oligonucleotide probes. Identification of 20 P. intermedia and 46 P. nigrescens isolates was confirmed by analyzing the electrophoretic mobilities of malate and glutamate dehydrogenase enzymes. A total of 111 isolates (32%) were identified as P. intermedia and 233 isolates (68%) as P. nigrescens. Identification performed with oligonucleotide probes and with malate and glutamate dehydrogenase electrophoresis matched perfectly. The distribution of oral P. intermedia and P. nigrescens in various periodontal status groups was investigated in periodontally healthy or diseased individuals. To reveal intra- and interindividual genetic diversity and possible intrafamilial transmission, P. intermedia and P. nigrescens isolates from 16 to 59 subjects, representing 8 married couples, were ribotyped. The stability of colonization was examined in 12 of the 59 subjects, of whom 6 received periodontal treatment and 6 were untreated. All children and periodontally healthy adults and most subjects with initial periodontitis (13/21) harbored only P. nigrescens. Of the 20 subjects with advanced periodontitis, 7 harbored both P. intermedia and P. nigrescens, 7 only P. intermedia and 6 only P. nigrescens. One or two ribotypes of P. intermedia and/or P. nigrescens were found intraindividually. The spouses in 5 of the 8 married couples shared an identical ribotype of P. intermedia or P. nigrescens, whereas ribotypes from unrelated subjects were mostly unique. Colonization was stable, since the same ribotypes were found 1-6 months apart in both periodontally treated and untreated subjects. In conclusion, the study indicates that P. intermedia and P. nigrescens may occur simultaneously in the oral cavity, the colonization is stable and P. intermedia is associated with periodontal diseases. Ribotyping revealed considerable genetic heterogeneity in unrelated subjects, whereas isolates obtained from spouses could represent the same ribotype, which suggests transmission of these species.
Topics: Adult; Age Factors; Aged; Bacterial Typing Techniques; Case-Control Studies; Child, Preschool; Disease Progression; Disease Transmission, Infectious; Ecosystem; Female; Genetic Heterogeneity; Genetic Variation; Humans; Male; Metronidazole; Middle Aged; Mouth; Nucleic Acid Hybridization; Oligonucleotide Probes; Periodontitis; Prevotella; Prevotella intermedia; Species Specificity; Spouses
PubMed: 8941760
DOI: 10.1111/j.1399-302x.1996.tb00342.x -
Microbiology (Reading, England) May 2002This paper describes a quantitative fluorescent in situ hybridization (FISH) assay for the differential identification of Prevotella intermedia and Prevotella nigrescens... (Comparative Study)
Comparative Study
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.
Topics: Bacterial Infections; Bacterial Typing Techniques; DNA, Bacterial; Dental Plaque; Fluorescent Antibody Technique; Humans; In Situ Hybridization, Fluorescence; Periodontitis; Prevotella; Prevotella intermedia; RNA, Ribosomal, 16S; Sensitivity and Specificity
PubMed: 11988511
DOI: 10.1099/00221287-148-5-1379 -
Frontiers in Cellular and Infection... 2017High-throughput sequencing has helped to reveal the close relationship between and periodontal disease, but the roles of subspecies diversity and genomic variation...
High-throughput sequencing has helped to reveal the close relationship between and periodontal disease, but the roles of subspecies diversity and genomic variation within this genus in periodontal diseases still need to be investigated. We performed a comparative genome analysis of 48 and isolates that from the same cohort of subjects to identify the main drivers of their pathogenicity and adaptation to different environments. The comparisons were done between two species and between disease and health based on pooled sequences. The results showed that both and have highly dynamic genomes and can take up various exogenous factors through horizontal gene transfer. The major differences between disease-derived and health-derived samples of and were factors related to genome modification and recombination, indicating that the isolates from disease sites may be more capable of genomic reconstruction. We also identified genetic elements specific to each sample, and found that disease groups had more unique virulence factors related to capsule and lipopolysaccharide synthesis, secretion systems, proteinases, and toxins, suggesting that strains from disease sites may have more specific virulence, particularly for . The differentially represented pathways between samples from disease and health were related to energy metabolism, carbohydrate and lipid metabolism, and amino acid metabolism, consistent with data from the whole subgingival microbiome in periodontal disease and health. Disease-derived samples had gained or lost several metabolic genes compared to healthy-derived samples, which could be linked with the difference in virulence performance between diseased and healthy sample groups. Our findings suggest that and may serve as "crucial substances" in subgingival plaque, which may reflect changes in microbial and environmental dynamics in subgingival microbial ecosystems. This provides insight into the potential of and as new predictive biomarkers and targets for effective interventions in periodontal disease.
Topics: Adult; Bacteroidaceae Infections; Base Sequence; Cohort Studies; Female; Gene Transfer, Horizontal; Genetic Variation; Genome, Bacterial; Gingiva; Humans; Male; Middle Aged; Periodontitis; Prevotella intermedia; Prevotella nigrescens; Virulence Factors
PubMed: 28983469
DOI: 10.3389/fcimb.2017.00409 -
Anaerobe Dec 2018The accuracy of a phenotypic scheme to recognize periodontal Prevotella intermedia/nigrescens group clinical isolates on primary isolation culture plates was assessed...
The accuracy of a phenotypic scheme to recognize periodontal Prevotella intermedia/nigrescens group clinical isolates on primary isolation culture plates was assessed with matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS). A total of 84 fresh subgingival isolates from 23 chronic periodontitis patients were presumptively recognized on anaerobically-incubated enriched Brucella blood agar primary isolation plates as P. intermedia/nigrescens based on their dark-pigmented colony morphology, brick-red autofluorescence under long-wave ultraviolet light, and a negative fluorescence test for lactose production. The presumptive P. intermedia/nigrescens clinical isolates were subjected to MALDI-TOF MS analysis using Bruker MALDI Biotyper analytic software containing mass spectra for P. intermedia and Prevotella nigrescens in its reference library of bacterial protein profiles. Using a ≥1.7 log score agreement threshold, 60 (71.4%) of the presumptive P. intermedia/nigrescens clinical isolates were confirmed as either P. intermedia (25 isolates) or P. nigrescens (35 isolates). All isolates with a <1.7 log score were also identified as P. intermedia or P. nigrescens from the top choice designated on the MALDI Biotyper most likely species identification list. These MALDI-TOF MS findings document the ability of the phenotypic scheme to correctly recognize most periodontal P. intermedia/nigrescens group clinical isolates on primary isolation culture plates.
Topics: Adult; Bacterial Typing Techniques; Bacteroidaceae Infections; Chronic Periodontitis; Female; Humans; Male; Phenotype; Prevotella intermedia; Prevotella nigrescens; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 29913204
DOI: 10.1016/j.anaerobe.2018.06.007 -
International Journal of Medical... 2021is an oral pathogen that is frequently observed in the subgingival plaque of periodontitis patients. Interleukin-1β (IL-1β) is known to be involved in the...
is an oral pathogen that is frequently observed in the subgingival plaque of periodontitis patients. Interleukin-1β (IL-1β) is known to be involved in the immunopathology of periodontal diseases and has been implicated in the destruction of bone. In this study, we investigated the mechanism of IL-1β production by in murine bone marrow-derived dendritic cells (BMDCs). Our results showed that a host receptor, Toll-like receptor 2 (TLR2), but not TLR4 is required for pro-IL-1β induction and nucleotide-binding oligomerization domain like receptor pyrin domain containing 3 (NLRP3) priming in BMDCs in response to and activation of the NLRP3 inflammasome is necessary for processing of pro-IL-1β into mature IL-1β. In addition, an inhibitor assay revealed that production of reactive oxygen species, P2XR activity, and release of cathepsin B are involved in IL-1β production in BMDCs in response to
Topics: Animals; Cathepsin B; Cells, Cultured; Dendritic Cells; Disease Models, Animal; Gram-Negative Bacterial Infections; Humans; Inflammasomes; Interleukin-1beta; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Periodontitis; Prevotella nigrescens; Primary Cell Culture; Reactive Oxygen Species; Receptors, Purinergic P2X7; Signal Transduction; Toll-Like Receptor 2
PubMed: 33390812
DOI: 10.7150/ijms.47197 -
Journal of Endodontics May 1999Isolates previously thought to be Prevotella intermedia have been shown to be a closely related species now known as Prevotella nigrescens. The purpose of this study was...
Isolates previously thought to be Prevotella intermedia have been shown to be a closely related species now known as Prevotella nigrescens. The purpose of this study was to determine the efficacy of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and polymerase chain reaction (PCR) to differentiate endodontic isolates of P. nigrescens from P. intermedia. Fifty-six strains of black-pigmented bacteria isolated from endodontic infections and conventionally identified as P. intermedia were used in this study. Using SDS-PAGE, novel polypeptide bands were used to differentiate P. nigrescens from P. intermedia. PCR was accomplished with specific primers for the 16S ribosomal RNA gene of both strains. Of 56 endodontic isolates, 41 (73%) strains were identified by SDS-PAGE as P. nigrescens and 15 (27%) strains as P. intermedia. Of the 41 strains of P. nigrescens identified by SDS-PAGE, PCR identified 37 strains as P. nigrescens. Restriction endonuclease digestion of amplified 16S ribosomal RNA genes indicated that the remaining four strains originally identified by SDS-PAGE as P. nigrescens were actually strains of Prevotella distinct from P. nigrescens and P. intermedia. Of 15 strains of P. intermedia identified by SDS-PAGE, PCR identified 14 strains as P. intermedia; but, one strain was identified as P. nigrescens. The results indicated that PCR was a more precise method than SDS-PAGE to differentiate P. intermedia from P. nigrescens. This study confirms that P. nigrescens is more commonly isolated in pure culture from endodontic infections than P. intermedia.
Topics: Bacterial Typing Techniques; DNA, Bacterial; Dental Pulp Diseases; Electrophoresis, Polyacrylamide Gel; Humans; Polymerase Chain Reaction; Prevotella; Prevotella intermedia
PubMed: 10530254
DOI: 10.1016/S0099-2399(06)81164-7 -
Oral Microbiology and Immunology Jun 1997The pathogenicity of 14 isolates identified as Prevotella intermedia or Prevotella nigrescens by serogrouping using monoclonal antibodies was compared in a tissue cage... (Comparative Study)
Comparative Study
The pathogenicity of 14 isolates identified as Prevotella intermedia or Prevotella nigrescens by serogrouping using monoclonal antibodies was compared in a tissue cage model in rabbits. Seven strains from periodontal abscesses, 5 strains from deep periodontal pockets and 2 strains from gingivitis were tested in the animal model comprising 6 Teflon tissue cages implanted on the back each of 34 rabbits. A total of 10(5)-10(8) cells of P. intermedia or P. nigrescens strains were inoculated alone or together with either Actinobacillus actinomycetemcomitans or Streptococcus mitis. Five strains of Porphyromonas gingivalis were used as a reference. The infectivity was recorded as pus formation and log viable count in aspirated material for 3, 7 and 14 days. None of the Prevotella strains inoculated in monoculture survived more than 3 days, and they had no capacity to produce abscess. P. intermedia or P. nigrescens strains in combination with A. actinomycetemcomitans produced abscesses in 33-100% and with S. mitis in 42-100%. No difference in abscess formation or log viable count in samples after 14 days was recorded between serogroup I (P. intermedia) and serogroup II and III (P. nigrescens). The infectivity of P. intermedia or P. nigresceas strains did not differ whether they were isolated from periodontal abscess, periodontal pocket or gingivitis. P. intermedia and P. nigrescens strains produced abscesses in combination with a facultative anaerobic strain and appears to have a similar pathogenicity in the wound chamber model in rabbits.
Topics: Aggregatibacter actinomycetemcomitans; Animals; Diffusion Chambers, Culture; Ecosystem; Humans; Periodontal Abscess; Porphyromonas gingivalis; Prevotella; Prevotella intermedia; Rabbits; Streptococcus; Superinfection; Virulence; Wound Infection
PubMed: 9467400
DOI: 10.1111/j.1399-302x.1997.tb00371.x -
Oral Microbiology and Immunology Jun 2000Glucose metabolism by Prevotella intermedia and Prevotella nigrescens were investigated. Glucose increased the anaerobic growth of these bacteria and promoted the...
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.
Topics: Energy Metabolism; Fermentation; Glucose; Glycolysis; Polysaccharides, Bacterial; Prevotella; Prevotella intermedia
PubMed: 11154402
DOI: 10.1034/j.1399-302x.2000.150307.x -
Journal of Periodontal Research Dec 2011Oral microorganisms may be involved in the development of cardiovascular diseases, and Porphyromonas gingivalis is one of the periodontal microorganisms that has been...
BACKGROUND AND OBJECTIVE
Oral microorganisms may be involved in the development of cardiovascular diseases, and Porphyromonas gingivalis is one of the periodontal microorganisms that has been found in carotid atheroma. The aim of this work was to study subgingival microorganisms and early carotid lesions in subjects with and without periodontitis.
MATERIAL AND METHODS
Eighty-eight subjects with periodontitis and 40 subjects without periodontitis underwent dental examinations in 2003. The presence of the periodontal microorganisms Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Tannerella forsythia was analyzed from subgingival plaque using PCR amplification. The common carotid artery was scanned using ultrasound and the calculated intima-media area (cIMA) was measured. The association between periodontitis, the cIMA value and the presence of periodontal microorganisms, together with several confounders, was studied in a multiple logistic regression model.
RESULTS
Smoking [odds ratio (OR) = 5.64; p = 0.001), level of education (OR = 5.02; p < 0.05) and the presence of P. gingivalis (OR = 6.50; p < 0.05) were associated with periodontitis. Explanatory factors for the increased cIMA were periodontitis (OR = 4.22; p < 0.05), hypertension (OR = 4.81; p < 0.05), high body mass index (OR = 5.78; p < 0.01), male gender (OR = 3.30; p < 0.05) and poor socioeconomic status (OR = 4.34; p < 0.05). P. nigrescens (OR 4.08; p < 0.05) and P. gingivalis (OR 7.63; p < 0.01) also appeared as explanatory variables associated with increased cIMA values.
CONCLUSION
This cross-sectional study showed that P. nigrescens and P. gingivalis were significantly associated with increased cIMA values.
Topics: Atherosclerosis; Carotid Artery Diseases; Carotid Artery, Common; Carotid Intima-Media Thickness; Carotid Stenosis; Case-Control Studies; Chi-Square Distribution; Cross-Sectional Studies; Female; Humans; Logistic Models; Male; Middle Aged; Periodontitis; Porphyromonas gingivalis; Prevotella nigrescens; Risk Factors
PubMed: 21793826
DOI: 10.1111/j.1600-0765.2011.01398.x