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Anaerobe Oct 2012We found that a 38-kDa protein was released from erythrocyte membranes lysed by hemolysin of Prevotella oris, although hypotonic hemolysis did not show such a...
We found that a 38-kDa protein was released from erythrocyte membranes lysed by hemolysin of Prevotella oris, although hypotonic hemolysis did not show such a phenomenon. The 38-kDa protein was identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) by N-terminal amino acid sequencing. This study discusses the relationship between GAPDH and hemolysis.
Topics: Erythrocyte Membrane; Erythrocytes; Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+); Hemolysin Proteins; Humans; Molecular Weight; Prevotella; Sequence Analysis, Protein
PubMed: 22967794
DOI: 10.1016/j.anaerobe.2012.08.008 -
Journal of Oral and Maxillofacial... Aug 2012Historically, the identification of microorganisms has been limited to species that could be cultured in the microbiology laboratory. The purpose of the present study...
PURPOSE
Historically, the identification of microorganisms has been limited to species that could be cultured in the microbiology laboratory. The purpose of the present study was to apply molecular techniques to identify microorganisms in orofacial odontogenic infections (OIs).
MATERIALS AND METHODS
Specimens were obtained from subjects with clinical evidence of OI. To identify the microorganisms involved, 16S rRNA sequencing methods were used on clinical specimens. The name and number of the clones of each species identified and the combinations of species present were recorded for each subject. Descriptive statistics were computed for the study variables.
RESULTS
Specimens of pus or wound fluid were obtained from 9 subjects. A mean of 7.4 ± 3.7 (standard deviation) species per case were identified. The predominant species detected in the present study that have previously been associated with OIs were Fusobacterium spp, Parvimonas micra, Porphyromonas endodontalis, and Prevotella oris. The predominant species detected in our study that have not been previously associated with OIs were Dialister pneumosintes and Eubacterium brachy. Unculturable phylotypes accounted for 24% of the species identified in our study. All species detected were obligate or facultative anaerobes. Streptococci were not detected.
CONCLUSIONS
Molecular methods have enabled us to detect previously cultivated and not-yet-cultivated species in OIs; these methods could change our understanding of the pathogenic flora of orofacial OIs.
Topics: Bacteria; Bacterial Typing Techniques; Bacteroidaceae Infections; Cohort Studies; Coinfection; Eubacterium; Fusobacterium Infections; Gram-Negative Anaerobic Straight, Curved, and Helical Rods; Gram-Negative Bacterial Infections; Gram-Positive Bacterial Infections; Humans; Molecular Biology; Peptostreptococcus; Porphyromonas endodontalis; Prevotella; Prospective Studies; RNA, Bacterial; RNA, Ribosomal, 16S; Sequence Analysis, RNA; Tooth Diseases
PubMed: 22326175
DOI: 10.1016/j.joms.2011.09.009 -
PloS One 2011The complexity of the human microbiome makes it difficult to reveal organizational principles of the community and even more challenging to generate testable hypotheses....
The complexity of the human microbiome makes it difficult to reveal organizational principles of the community and even more challenging to generate testable hypotheses. It has been suggested that in the gut microbiome species such as Bacteroides thetaiotaomicron are keystone in maintaining the stability and functional adaptability of the microbial community. In this study, we investigate the interspecies associations in a complex microbial biofilm applying systems biology principles. Using correlation network analysis we identified bacterial modules that represent important microbial associations within the oral community. We used dental plaque as a model community because of its high diversity and the well known species-species interactions that are common in the oral biofilm. We analyzed samples from healthy individuals as well as from patients with periodontitis, a polymicrobial disease. Using results obtained by checkerboard hybridization on cultivable bacteria we identified modules that correlated well with microbial complexes previously described. Furthermore, we extended our analysis using the Human Oral Microbe Identification Microarray (HOMIM), which includes a large number of bacterial species, among them uncultivated organisms present in the mouth. Two distinct microbial communities appeared in healthy individuals while there was one major type in disease. Bacterial modules in all communities did not overlap, indicating that bacteria were able to effectively re-associate with new partners depending on the environmental conditions. We then identified hubs that could act as keystone species in the bacterial modules. Based on those results we then cultured a not-yet-cultivated microorganism, Tannerella sp. OT286 (clone BU063). After two rounds of enrichment by a selected helper (Prevotella oris OT311) we obtained colonies of Tannerella sp. OT286 growing on blood agar plates. This system-level approach would open the possibility of manipulating microbial communities in a targeted fashion as well as associating certain bacterial modules to clinical traits (e.g.: obesity, Crohn's disease, periodontal disease, etc).
Topics: Algorithms; Bacteroides; Bayes Theorem; Biofilms; DNA; Dental Plaque; Hot Temperature; Humans; Metagenome; Models, Biological; Models, Genetic; Models, Theoretical; Nucleic Acid Hybridization; Oligonucleotide Array Sequence Analysis; Periodontitis; Principal Component Analysis; Systems Biology
PubMed: 22163302
DOI: 10.1371/journal.pone.0028438 -
Wounds : a Compendium of Clinical... Oct 2011A 34-year-old Haitian man presented with a 1-year history of a gradually en- larging, ulcerating nodule on the right posterior ankle that...
A 34-year-old Haitian man presented with a 1-year history of a gradually en- larging, ulcerating nodule on the right posterior ankle that bled after trauma. The patient denied any history of prior trauma at the site of the lesion and foreign travel.There were no HIV risk factors or personal or family history of skin cancer.The patient was otherwise healthy with no additional complaints. Physical examination revealed a 1.4 cm firm, hyperpigmented nodule with a yellow crateriform center on the right posterior ankle (Figure 1).A punch biopsy was performed (Figures 2, 3). Histopathologic examination revealed a superficial and mid-dermal, multi- nodular proliferation of bland epithelioid cells with occasional foci of ductal differentiation embedded in a fibrotic stroma. Focal epidermal connection and mucinous metaplasia were noted. Immunohistochemical stains revealed diffuse positive staining of the lesional cells with cytokeratins (high and low) and variable positivity with epithelial membrane antigen (EMA), as well as negative S100P. Fungal, viral, and mycobacterial tissue cultures were negative and RPR was non-reactive. Bacterial tissue culture was positive for Prevotella oris and Staphylococcus aureus.
PubMed: 25881109
DOI: No ID Found -
Journal of Endodontics Jul 2011The characterization of microbial communities infecting the endodontic system in each clinical condition may help on the establishment of a correct prognosis and...
INTRODUCTION
The characterization of microbial communities infecting the endodontic system in each clinical condition may help on the establishment of a correct prognosis and distinct strategies of treatment. The purpose of this study was to determine the bacterial diversity in primary endodontic infections by 16S ribosomal-RNA (rRNA) sequence analysis.
METHODS
Samples from root canals of untreated asymptomatic teeth (n = 12) exhibiting periapical lesions were obtained, 16S rRNA bacterial genomic libraries were constructed and sequenced, and bacterial diversity was estimated.
RESULTS
A total of 489 clones were analyzed (mean, 40.7 ± 8.0 clones per sample). Seventy phylotypes were identified of which six were novel phylotypes belonging to the family Ruminococcaceae. The mean number of taxa per canal was 10.0, ranging from 3 to 21 per sample; 65.7% of the cloned sequences represented phylotypes for which no cultivated isolates have been reported. The most prevalent taxa were Atopobium rimae (50.0%), Dialister invisus, Prevotella oris, Pseudoramibacter alactolyticus, and Tannerella forsythia (33.3%).
CONCLUSIONS
Although several key species predominate in endodontic samples of asymptomatic cases with periapical lesions, the primary endodontic infection is characterized by a wide bacterial diversity, which is mostly represented by members of the phylum Firmicutes belonging to the class Clostridia followed by the phylum Bacteroidetes.
Topics: Adolescent; Adult; Bacteria; Bacteroidetes; Clostridium; Dental Pulp Cavity; Genomic Library; Humans; Microbial Consortia; Middle Aged; Periapical Diseases; RNA, Bacterial; RNA, Ribosomal, 16S; Ruminococcus; Sequence Analysis, RNA; Young Adult
PubMed: 21689545
DOI: 10.1016/j.joen.2011.04.007 -
Journal of Clinical Periodontology Apr 2011Because the absorption of stimulants of Toll-like receptor (TLR)2 and TLR4 from the gastrointestinal tract into the circulation has been proposed to promote the... (Comparative Study)
Comparative Study
AIM
Because the absorption of stimulants of Toll-like receptor (TLR)2 and TLR4 from the gastrointestinal tract into the circulation has been proposed to promote the development of atherosclerosis and insulin resistance, we aimed to quantify the abundance of stimulants of TLR2 and TLR4 in human saliva.
METHODS
A recently developed bioassay based upon measurement of NF-κB activation in TLR-deficient human embryonic kidney (HEK)-293 cells transfected with human TLR2 or TLR4 and calibrated with synthetic bacterial lipopeptide (Pam(3) CSK(4) ) or Escherichia coli lipopolysaccharide (LPS), was used to establish the normal range of TLR stimulants in saliva of 20 healthy subjects and 20 subjects with periodontal disease.
RESULTS
Median soluble stimulants of TLR2 and TLR4 were significantly higher in saliva of periodontitis patients compared with saliva of healthy subjects; 3450 versus 77 ng/ml Pam(3) CSK(4) equivalents (p<0.0001) and 138 versus 7 ng/ml LPS equivalents, respectively (p<0.0001). Salivary TLR stimulant levels remained relatively stable in healthy subjects over several days. Six strains of oral Gram-negative bacteria, including Tannerella forsythensis, Lysobacter enzymogenes, Prevotella intermedia, Prevotella oris and Porphyromonas gingivalis, from a panel of nine examined did not stimulate TLR4-dependent signalling.
CONCLUSIONS
Elevated salivary TLR stimulants may represent a novel mechanism by which periodontitis increases the risk of developing cardiovascular disease and insulin resistance.
Topics: Adult; Bacteroides; Case-Control Studies; Cells, Cultured; Chronic Periodontitis; Culture Media, Conditioned; Escherichia coli; Female; HEK293 Cells; Humans; Lipopeptides; Lipopolysaccharides; Lymphocyte Antigen 96; Lysobacter; Male; Porphyromonas gingivalis; Prevotella; Prevotella intermedia; Saliva; Salivary Proteins and Peptides; Toll-Like Receptor 2; Toll-Like Receptor 4; Toll-Like Receptor 5
PubMed: 21284689
DOI: 10.1111/j.1600-051X.2011.01702.x -
Anaerobe Oct 2010Head-and-neck infections often involve anaerobes such as Prevotella species. Aim of the present study was to assess the evolution and the factors associated with...
Head-and-neck infections often involve anaerobes such as Prevotella species. Aim of the present study was to assess the evolution and the factors associated with resistance in Prevotella species to penicillin, clindamycin, metronidazole, tetracycline and β-lactams/β-lactamase inhibitors (BL/BLIs). In total, 192 Prevotella strains, isolated from patients with oral and head-and-neck infections, were evaluated. Common isolates were Prevotella intermedia and Prevotella melaninogenica within the pigmented species as well as Prevotella oris and Prevotella oralis group within the non-pigmented species. Overall resistance was 43.2% for penicillin, 10.9% for clindamycin, 0% for metronidazole. Nonsusceptibility to tetracycline was 29.1% without significant differences in resistance rates between pigmented and other species. Penicillin resistant strains were β-lactamase positive. From 2003-2004 to 2007-2009, penicillin resistance rates increased about four-fold (from 15.4% to 60.6%). Clindamycin resistance did not show evolution, whereas tetracycline nonsusceptibility decreased from 43.3% in 2003-2004 to 20.7% in 2007-2009. Except for one (0.5%) P. oralis strain with intermediate susceptibility to BL/BLIs, the other strains were susceptible to the agents. In conclusion, in Prevotella strains from patients with head-and-neck infections, the resistance rate to penicillin increased, that to clindamycin remained stable and the nonsusceptibility rate to tetracycline decreased during the period. Activity against >99% of Prevotella strains was observed with metronidazole and BL/BLIs. The penicillin resistance and tetracycline nonsusceptibility were associated with the year of study, national antibiotic consumption and possibly with previous treatment (for tetracycline). The evolution of penicillin resistance in Prevotella strains was highly dynamic.
Topics: Anti-Bacterial Agents; Bacteroidaceae Infections; Bulgaria; Clindamycin; Drug Resistance, Multiple, Bacterial; Female; Humans; Male; Microbial Sensitivity Tests; Penicillins; Prevotella; Tetracycline
PubMed: 20670687
DOI: 10.1016/j.anaerobe.2010.07.004 -
International Journal of Systematic and... Dec 2007Three strains of anaerobic Gram-negative bacilli isolated from human oral sites were subjected to a comprehensive range of phenotypic and genotypic tests and were found...
Three strains of anaerobic Gram-negative bacilli isolated from human oral sites were subjected to a comprehensive range of phenotypic and genotypic tests and were found to comprise a homogeneous group. 16S rRNA gene sequence analysis revealed the strains to constitute a novel group within the genus Prevotella, most closely related to Prevotella oris and Prevotella salivae. A novel species, Prevotella maculosa sp. nov., is proposed to accommodate these strains. Prevotella maculosa is saccharolytic and produces acetic and succinic acids as end products of fermentation. The G+C content of the DNA of the type strain is 48 mol%. The type strain of Prevotella maculosa is W1609(T) (=DSM 19339(T) =CCUG 54766(T)).
Topics: Acetic Acid; Anaerobiosis; Bacterial Typing Techniques; Base Composition; Carbohydrate Metabolism; DNA, Bacterial; DNA, Ribosomal; Genes, rRNA; Humans; Molecular Sequence Data; Mouth; Phylogeny; Prevotella; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid; Succinic Acid
PubMed: 18048753
DOI: 10.1099/ijs.0.65281-0 -
FEMS Microbiology Letters Nov 2007Prevotella oris is a nonpigmented, Gram-negative, anaerobic bacterium that has been associated with several serious oral and systemic infections. Prevotella oris has...
Prevotella oris is a nonpigmented, Gram-negative, anaerobic bacterium that has been associated with several serious oral and systemic infections. Prevotella oris has been identified in clinical specimens by bacterial culture and biochemical tests, which are generally unreliable. The aim of this study was to develop a PCR assay for the direct detection of P. oris in clinical specimens. PCR primers specific for P. oris were identified by alignment of bacterial 16S rRNA genes from closely related species and selection of PCR primers specific for P. oris at their 3' ends. Amplification of a 1110-bp product indicated PCR positivity for P. oris. The primers were shown to be specific for P. oris DNA, because no PCR products were obtained when DNA from other oral bacteria, including closely related Prevotella species, were used as test species, and this was confirmed by digestion of PCR products with RsaI and MnlI. Prevotella oris DNA was detected in 17 (36.2%) of 47 pus samples from subjects with dentoalveolar abscesses and in all three pus samples from subjects with spreading odontogenic infections. This PCR assay provides a sensitive, specific and reliable method for identifying P. oris in clinical specimens.
Topics: Bacteroidaceae Infections; DNA Primers; Humans; Periapical Abscess; Polymerase Chain Reaction; Prevotella; RNA, Bacterial; RNA, Ribosomal, 16S; Sensitivity and Specificity; Suppuration
PubMed: 17937671
DOI: 10.1111/j.1574-6968.2007.00926.x -
Journal of Clinical Microbiology Sep 2007Multiple-displacement amplification (MDA) has been used to uniformly amplify bacterial genomes present in small samples, providing abundant targets for molecular...
Multiple-displacement amplification (MDA) has been used to uniformly amplify bacterial genomes present in small samples, providing abundant targets for molecular analysis. The purpose of this investigation was to combine MDA and checkerboard DNA-DNA hybridization to examine the microbiota of endodontic infections. Sixty-six samples were collected from teeth with endodontic infections. Nonamplified and amplified samples were analyzed by checkerboard DNA-DNA hybridization for levels and proportions of 77 bacterial taxa. Counts, percentages of DNA probe counts, and percentages of teeth colonized for each species in amplified and nonamplified samples were computed. Significance of differences for each species between amplified and nonamplified samples was sought with Wilcoxon signed-rank test and adjusted for multiple comparisons. The amount of DNA in the samples ranged from 6.80 (+/- 5.2) ng before to 6.26 (+/- 1.73) mug after MDA. Seventy of the 77 DNA probes hybridized with one or more of the nonamplified samples. All probes hybridized with at least one sample after amplification. Most commonly detected species at levels of >10(4) in both amplified and nonamplified samples were Prevotella tannerae and Acinetobacter baumannii at frequencies between 89 and 100% of samples. The mean number of species at counts of >10(4) in amplified samples was 51.2 +/- 2.2 and in nonamplified samples was 14.5 +/- 1.7. The endodontic microbiota was far more complex than previously shown, although microbial profiles at teeth with or without periradicular lesions did not differ significantly. Species commonly detected in endodontic samples included P. tannerae, Prevotella oris, and A. baumannii.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Bacteria; Bacterial Infections; Bacteriological Techniques; Biodiversity; Child; Colony Count, Microbial; Humans; Middle Aged; Nucleic Acid Amplification Techniques; Nucleic Acid Hybridization; Tooth Diseases
PubMed: 17634304
DOI: 10.1128/JCM.02618-06