• Restorative Dentistry and Endodontics
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• v.27 no.1
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• pp.54-65
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• 2002

Black-pigmented bacteria have been implicated in the endodontic infections. This group of microorganisms includes Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. The organisms display a wide variety of virulence factors that may be pertinent to acute endodontic infections. The aim of this study was to identify P. endodontalis, P. gingivalis, P. intermedia, and P. nigrescens by using special potency disk test, filter paper spot test, 165 rRNA gene-directed PCR, and API 32A. Microbial samples were collected from root canals of 33 intact teeth with necrotic pulp and/or apical periodontitis. Conventional laboratory methods were used for identification of the strains of black pigmented bacteria. Eighteen of 33 samples were positive for the growth of black-pigmented bacteria Five colonies were cultured from each pure cultured colonies from Brucella agar plate. Seventy seven colonies were positive for the growth of black-pigmented bacteria. Thirty three of 77(42.6%) were identifed as P. nigrescens, 10 of 77(12.9%)were P. gingivalis, 6 of 77(7.8%) were P. endodontalis, 10 of 77(12.9%) were P. intermedia. On the contrary the reference strains of P. nigrescens, experimental strains of P nigrescens was sensitive to kanamycin in special potency disk test. 165 rRNA gene PCR and API test after rapid presumptative identification methods, such as special potency disk test and filter paper spot test, would be accurate detection methods for black-pigemented bacteria.

• Journal of Periodontal and Implant Science
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• v.36 no.1
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• pp.155-165
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• 2006

The results of this study confirm that the availability of hemin influences the expression of selected membrane proteins of Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is hemin regulated was identified and purified in P. gingivalis. A strong hemin-binding function was found by LDS-PAGE and TMBZ staining when P. gingivalis cells were grown under hemin-limited conditions. A 50 kDa cell envelope associated protein, whose expression is hemin regulated, is considered to be a putative hemin binding protein from P. intermedia and P. nigrescens, respectively. N-terminal amino acid sequence analysis of CNBr-digested 24 kDa hemin binding protein from P. gingivalis revealed that this protein belongs to a new, so far undescribed hemin-binding class of proteins. N-terminal amino acid sequence of a 50 kDa putative hemin binding protein from P. intermedia was identical with Enolase from Streptococcus intermedia. Work is in progress to further characterize the molecular structure of these proteins.

• International Journal of Oral Biology
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• v.35 no.1
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• pp.13-19
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• 2010

The DNA probes Pn17 and Pn34 were evaluated for their ability to specifically detect clinical strains of P. intermedia and P. nigrescens from a Korean population by dot blot hybridization. These probes were sequenced by extension termination and their specificity was determined by Southern blot analysis. The results revealed that the Pn17 sequence (2,517 bp) partially encodes an RNA polymerase beta subunit (rpoB) and that Pn34 (1,918 bp) partially encodes both rpoB (1-169 nts) and the RNA polymerase beta subunit (rpoB'; 695-1918 nts). These probes hybridized with both HindIII- and PstI-digested genomic DNAs from the strains of P. intermedia and P. nigrescens used in this study. Interestingly, each of the hybrid bands generated from the HindIII-digested genomic DNAs of the two bacterial species could be used to distinguish between them via restriction fragment length polymorphism. These results thus indicate that Pn17 and Pn34 can simultaneously detect P. intermedia and P. nigrescens.

• Microbiology and Biotechnology Letters
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• v.31 no.2
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• pp.205-209
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• 2003

Recently, we introduced a new method for rapid screening of bacterial species- or subspecies-specific DNA probes, named “inverted dot blot hybridization screening method”. We then applied this method to develop species- or strain- specific DNA probes for Prevotella intermedia and Prevotella nigrescens. In those studies, among 96 candidate DNA probes which were screened by the new method, 5 probes were confirmed as being putatively strain-specific : 3 probes for P. nigrescens 9336 (ATCC 33563), one for each p. intermedia ATCC 25611 and one for P. nigrescens G8-9K-3 (ATCC 49046). In the present study, we evaluated by Southern blot analysis a DNA probe Pi29-L, one of the 96 candidate probes described above, whether it is specific for the strain ATCC 25611 off. intermedia. Our data show that the probe Pi29-L is potentially P. intermedia ATCC 25611-specific, which can be useful for the detection and identification of the strain, particularly in maintenance of the strain.

• Journal of the Korean Society of Environmental Restoration Technology
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• v.11 no.6
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• pp.112-119
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• 2008

In order to elucidate growth characteristics, physiological responses and leaf color changes of Physocarpus opulifolius 'Diabolo' and Ophiopogon planiscapus 'Nigrescens'. These experiments were investigated under various shading levels. Growth of P.opulifolius 'Diabolo' was better at 30% shading level but physiological activities were double at strong light condition. Dark purple color was observed at 85% shading level and color was dim down when shading level decreased. Color was deep purple(RHS 202A) at 85% shading and green(RHS 139A) at 0%, These results imply that ornamental value was increased when shading level increased for P. opulifolius 'Diabolo'. Growth and highest physiological activity of O. planiscapus 'Nigrescens' were observed at 0% shading level and dark red color(RHS 202A) was also observed at 0% shading level. These results imply that ornamental value was increased when shading level decreased for O. planiscapus 'Nigrescens'.

• Journal of Korean society of Dental Hygiene
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• v.21 no.5
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• pp.555-565
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• 2021

Objectives: This study was conducted to analyze peri-implantitis bacteria and identify their associations with health status and health activities. Methods: Gingival sulcus fluid at the implant's periodontal pockets sampled from the participants were analyzed by multiplex real time PCR. Results: Participants had strains in the order of 100% F. nucleatum, 98.0% E. corrodens, and 96.0% P. micra, and the correlation between C. rectus and E. nodatum was high (p<0.01). Diabetic group (P. gingivalis, P. nigrescens) hypertension (P. nigrescens), group with four or more periodontal pockets (P. gingivalis, T. dentica, P. intermedia, E. nodatum, and C. rectum), smoking (P. micra, E. corrodens), drinking (T. dentola), and scaling groups (C. rectus) were found to have more strains (p<0.05). Conclusions: Representative pathogenic microorganisms detected in periodontal pockets of implants were similar to dental periodontal pockets; however there were differences in the amount and distribution of microorganisms, and they were affected by health status and health behavior.

• Journal of Oral Medicine and Pain
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• v.47 no.1
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• pp.10-26
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• 2022

Purpose: The purpose of this study was to investigate whether various saliva collection methods affect the observed salivary microbiome and whether microbiomes of stimulated and unstimulated saliva and plaque differ in richness and diversity. Methods: Seven sampling methods for unstimulated saliva, stimulated saliva, and plaque samples were applied to six orally and systemically healthy participants. Bacterial 16S ribosomal RNA genes of 10 major oral bacterial species, namely, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Fusobacterium nucleatum, Prevotella intermedia, Prevotella nigrescens, Streptococcus mitis, Streptococcus sobrinus, and Lactobacillus casei, were analyzed by real-time polymerase chain reaction. We comprehensively examined the dependence of the amount of bacterial ribosomal DNA (rDNA), bacterial-community composition, and relative abundance of each species on sample collection methods. Results: There were significant differences in the bacterial rDNA copy number depending on the collection method in three species: F. nucleatum, P. nigrescens, and S. mitis. The species with the highest richness was S. mitis, with the range from 89.31% to 100.00%, followed by F. nucleatum, P. nigrescens, T. denticola, T. forsythia, and P. intermedia, and the sum of the proportions of the remaining five species was less than 1%. The species with the lowest observed richness was P. gingivalis (<0.1%). The Shannon diversity index was the highest in unstimulated saliva collected with a funnel (4.449). The Shannon diversity index was higher in plaque samples (3.623) than in unstimulated (3.171) and stimulated (3.129) saliva and in mouthwash saliva samples (2.061). Conclusions: The oral microbial profile of saliva samples can be affected by sample collection methods, and saliva differs from plaque in the microbiome. An easy and rapid technique for saliva collection is desirable; however, observed microbial-community composition may more accurately reflect the actual microbiome when unstimulated saliva is assayed.

• Proceedings of the KACD Conference
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• 2001.11a
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• pp.582.1-582
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• 2001

IL-6 and IL-10 are known to be inflammatory cytokines that mediate host response to invading microorganisms or pathogenic antigen. But the roles of these cytokines in pulpal inflammation are not well established. The objective of this study was to investigate the concentrations and the roles of IL-6 and IL-10 in the pulpal inflammation associated with gram-negative bacteria, P. nigrescens. We exposed the pulps of rat mandibular incisors and inserted sterile cotton in control groups and inoculated P. nigrescens in experimental groups.(omitted)

• Journal of Periodontal and Implant Science
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• v.34 no.2
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• pp.255-268
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• 2004

The purpose of the research is to compare the distribution of Black-pigmented Bacteroides between Chronic Periodontitis and Aggressive Periodontitis. P. gingivalis, P. intermedia and P. nigrescens were examined in order to evaluate their distribution in patients with Chronic Periodontitis(CP) and Aggressive Periodontitis(AP). PCR and dot-blots hybridization of 16s rRNA gene were used to compare bacterial distribution of two groups - CP group and AP group, which were divided into two subgroups. Subgingival plaque taken from the diseased sites(pocket $depth{\geq}6$ mm) and healthy sites(pocket $depth{\leq}3$ mm) were grouped into the experimental group and the control group. The result are as follows ; 1. The distribution of P. gingivalis was 98.33% for chronic Periodotitis(CP), 94.17% for Aggressive Periodontitis(AP), the distribution of P. intermedia was 77.50% for CP, 64.17% for AP, and the distribution of P. nigrescens was 35.00%, 29.17%. In all 3 types of bacteria, CP group showed higher distribution compared to AP group, but only P. intermedia showed statistically significant difference. 2. In the case of CP, every type of bacteria showed higher distribution in the experimental group with statistically significant difference. 3. In the case of AP, every type of bacteria also showed higher distribution in the experimental group, but P. gingivalis and p..intermedia showed the result with statistically significant difference, and the other did not 4. In 3 all bacteria type, N-AP showed higher distribution than N-CP without statistically significant difference These results suggest that the comparison of the distribution of Bacteroides between Chronic Periodontitis and Aggressive Periodontitis has no statistically significant difference, except P. intermedia.

• Journal of Periodontal and Implant Science
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• v.30 no.3
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• pp.661-676
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• 2000

Gingivitis and periodontitis are infectious diseases in that microorganisms are the primary extrinsic cause of the diseases. the occurrence of gingivitis has been associated clearly with the presence of microorganisms at the disease site, and the histologic nature of the tissue involved is indicative of an inflammatory response induced by microorganisms. additional evidence for the microbial etiology of periodontal disease is that numerous antimicrobial agents are effective in reducing plaque accumulation and periodontal diseases. the purpose of this article is to analyze the antimicrobial effects of Pulsatilla koreana. Well-dried Pulsatilla koreana purchased from herbs distributor was ground and extracted into methanol(MeOH), ethylacetate(EtoAc), chlorform($CHCl_3$) and Butyl alcohol(BuOH). we have then applied each solution to the bacteria samples(Bacteroides forsythus, Streptococcus mutans, Streptococcus sanguis, Porphylomonas gingivalis, Actinobacillus actinomycetemcomitans, Eikenella corrodens, Prevotella intermedia, Actinomyces viscosus, Prevotella nigrescens , Rothia dentocariosa, Fusobacterium nucleatum, Pseudomonas aeruginosa, Staphylococcus aureus) collected from several organizations. To conduct susceptibility test(Kirby-Bauer method), plate contained each periodontopathic bacteria is spread extracted into methanol(MeOH), ethylacetate(EtoAc), chlorform($CHCl_3$) and Butyl alcohol(BuOH) and to measure the minimum inhibition concentration(MIC) of the bacteria against the solutions to ultimately determine antimicrobial effects of the solutions, insert bacteria sample into $20{\mu\ell}/{m\ell}$, $10{\mu\ell}/{m\ell}$, $5{\mu\ell}/{m\ell}$, $2.5{\mu\ell}/{m\ell}$ of each solution and control group(not contained solution) 1. Solution extracted into methanol did not show clear zone against all bacteria samples. Only P.nigrescens, S. mutans and S. sanguis in solution extracted into ethylacetate, S. mutans and S. anguis in solutions extracted into chlorform and Butyl alcohol showed clear zone against all bacteria samples. Solution extracted into Butyl alcohol showed clear zone against 13 types of bacteria, excluding P. gingivalis. 2. In Solution extracted into methanol, the bacteria samples grew in the highest concentrated plate, showing minimal variation from control group. 3. In Solution extracted into Butyl alcohol, S. aureus, P. intermedia, E. corrodens, A. actinomycetemcomitans, B. forsythus, P. gingivalis et al. showed decreased growth in the highest concentrated plate. P. auruginosa, R. dentocariosa, A. viscosus, P. nigrescens, S. mutans et al. showed decreased growth at MIC $20{\mu\ell}/{m\ell}$ and S. sanguis showed decreased growth at MIC $10{\mu\ell}/{m\ell}$. 4. By analyzing the MIC level through considering the results from Kirby-Bauer method, Solution extracted into methanol did not reveal any antimicrobial effects and Solution extracted into Butyl alcohol showed the highest antimicrobial effects In conclusion, it can be used the extracts of Pulsatilla koreana as wide spectrum antimicrobial agent.