• Title/Summary/Keyword: Prevotella intermedia

Search Result 100, Processing Time 0.033 seconds

Cloning of a Potentially Strain-Specific DNA Probe of prevotella intermedia ATCC 25611 by Inverted Dot Blot Hybridization Screening Method

  • Kook, Joong-Ki;Han, Jin-Ju;Kim, Hwa-Sook;Seong, Jin-Hyo;Kim, Dong-Kie;Baek, Dong-Heon;Choe, Son-Jin
    • Journal of Microbiology and Biotechnology
    • /
    • v.13 no.2
    • /
    • pp.282-286
    • /
    • 2003
  • The purpose of this study was to isolate a specific DNA probe for the strain ATTC 25611 of the species Prevotella intermedia by using a new rapid screening mothod. The whole-genomic DNA of P. intermedia ATCC 25611 was isolated and purified. The HindIII-digested genomic DNAs from the strain were cloned by the random cloning method. To screen the strain-specific DNA probe, inverted dot blot hybridization tests were performed. In this assay, 20 ng of recombinant plasmids containing the HindIII-digested genomic DNA fragment were boiled and blotted onto a nylon membrane, and hybridized with digoxigenin-dUTP labeled genomic DNAs in a concentration of 100 ng/ml. Southern blot analysis was performed in order to confirm the results of the inverted dot blot hybridization tests. The data showed that a Pi34 probe (2.1 kbp; 1 out of 32 probes) was specific for P. intermedia strain ATCC 25611 and could be useful for the detection and identification of the strain, particularly in epidemiological studies of periodontal disease.

Antimicrobial Effect of Natural Plant Extracts against Periodontopathic Bacteria (치주염 원인균에 대한 천연 식물 추출물의 항균효과)

  • Lee, Seung-Hee;Kim, Min-Jeong
    • The Journal of the Korea Contents Association
    • /
    • v.19 no.1
    • /
    • pp.242-255
    • /
    • 2019
  • In this study, we examined the antimicroboal effect against Actinobacillus actinomycetemcomitans and Prevotella intermedia which were the bacteria causing the Periodontopathic by using 34 types of natural plant extracts. Therefore, this study measures growth inhibition activity and Minimum Inhibition Concentration (MIC) of a sample extract with the use of organic solvent extracts in order to analyze the antibacterial effect of natural plant extracts on periodontopathic bacteria. Each of the 34 types of natural plant extracts were extracted by using the ethanol, and subsequently, the size of growth inhibition zone(clear zone, ㎜) of respective extracts were measured through the disk diffusion method. As a result, it was found that the growth inhibitory activity was found for A. actinomycetemcomitans, which is the bacteria causing the Periodontitis, in 13 types of natural plant extracts such as Raphanus sativus, Akebia quinata, Paeonia lactiflora, Belamcanda chinensis, Inula britannics, Houttuynia cordata, Forsythia saxatilis, Gentiana macrophylla, Melia azedarach, Scutellaria baicalensis, Coptis chinensis, Phellodendron amurense, Kalopanax Pictus, etc. In the case of P. intermedia, the growth inhibitory activity was found in 13 types of natural plant extracts such as Raphanus sativus, Angelica acutiloba, Akebia quinata, Belamcanda chinensis, Inula britannics, Houttuynia cordata, Cinnamomum cassia, Aster tataricus, Melia azedarach, Scutellaria baicalensis, Coptis chinensis, Phellodendron amurense, Kalopanax Pictus etc. For A. actinomycetemcomitans, anti-bacterial effect was exhibited in Belamcanda chinensis, Cinnamomum cassia, Kalopanax Pictus, Phellodendron amurense, Coptis chinensis. The Coptis chinensis showed the most excellent growth inhibitory activity in all organic solvent fragment, while P. intermedia showed the growth inhibitory activity in Belamcanda chinensis, Cinnamomum cassia, Meliaazedarach, Phellodendron amurense, and Coptis chinensis.

Suppression of nitric oxide and interleukin-6 production by methanol extract of Sophorae Flos in macrophage cells (괴화 추출물이 대식세포에서의 nitric oxide와 interleukin-6의 생성에 미치는 영향)

  • Lee, Ji-Eun;Lee, Ju-Youn;Choi, Jeom-Il;Kim, Chong-Kwan;Kim, Sung-Jo
    • Journal of Periodontal and Implant Science
    • /
    • v.35 no.1
    • /
    • pp.9-19
    • /
    • 2005
  • Both nitric oxide (NO) and interleukin-6 (IL-6) have been thought to have a role in the pathogenesis of inflammatory periodontal disease as it does in other inflammatory diseases, and the inhibitors of NO and IL-6 production have been considered as potential anti-inflammatory agents. In this study, we evaluated methanol extract of Sophorae Flos for inhibition of NO and IL-6 production in Prevotella intermedia LPS-induced mouse macrophages RAW264.7 cells. Dried Sopharae Flos was sliced, and extracted with 100% methanol. LPS from p. intermedia ATCC 25611 was prepared by the standard hot phenol-water method. NO production was assayed by measuring the accumulation of nitrite in culture supematants and IL-6 was measured using mouse IL-6 ELISA kit. Western blot analysis of iNOS and analysis of reverse transcription (RT)-PCR products were carried out. The methanol extract of Sophorae Flos concentration-dependently reduced the production of NO and the expression of iNOS protein and mRNA in RAw264.7 cells treated with P. intermedia LPS. Sophorae Flos also suppressed IL-6 production and the expression of IL-6 mRNA in RAw264.7 cells stimulated by P. intermedia LPS. The inhibition of NO and IL-6 production by Sophorae Flos may be useful in the therapy of inflammatory diseases such as periodontitis. This hypothesis, however, remains to be tested.

DETECTION OF BLACK-PIGMENTED BACTERIA IN INFECTED ROOT CANALS

  • Kwon, Eun-Kyoung;Lee, Su-Jong;Im, Mi-Kyung
    • Proceedings of the KACD Conference
    • /
    • 2001.11a
    • /
    • pp.582.2-582
    • /
    • 2001
  • Black-pigmented bacteria anaerobes 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.(omitted)

  • PDF

Antimicrobial Effect of Mouthwash against Streptococcus mutans by Visual Staining Method (시각적 염색 방법을 이용한 마우스워시의 구강균에 대한 항균효과 확인)

  • Park, Taehun;Cho, Jeong Hun;Sung, Youngeun;Cho, Jun-Cheol;Shin, Kyeho
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.40 no.2
    • /
    • pp.187-193
    • /
    • 2014
  • Dental caries are one of the most common oral diseases and Streptococcus mutans (S. mutans) plays an important role in the initiation and progression of dental caries. Oral malodor is primarily the result of microbial metabolism such as Porphyromonas gingivalis (P. gingivalis) that produce volatile sulfur compounds (VSCs), causing oral malodor. Prevotella intermedia (P. intermedia) is known as typical periodontopathic bacteria, and periodontal disease is chronic inflammatory disease that leads to damage of gingival connective tissue and alveolar bone, eventually loss of teeth. In this study, we investigated antimicrobial effect of mouthwash containing cetylpyridinium chloride (CPC), sodium fluoride (NaF), green tea water extract and pine needles water extract against cariogenic and periodontopathic bacteria sucn as S. mutans, P. gingivalis and P. intermedia. As a result, the reduction ratios of S. mutans and P. gingivalis were 4.00 Log and 4.68 Log reduction for 30 s, and P. intermedia were 2.40 Log reduction for 30 s and 2.70 Log reduction for 60 s. Dentocult SM Strip mutans (SM Strip) provides easy detection of visual data showing a significant inhibition on S. mutans. In conclusion, we expected that mouthwash containing CPC, NaF, green tea water extract and pine needles water extract could help preventing the dental disease like dental caries and oral malodor.

TRANSMISSION OF PREVOTELLA INTERMEDIA BY GENOMIC DAN FINGERPRINTING (P.intermedia의 유전자 이종성과 가족내 전이에 관한 연구)

  • Lee, Seoung-Min;Kim, Kack-Kyun;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
    • /
    • v.25 no.1
    • /
    • pp.89-98
    • /
    • 1995
  • P. intermedia are considered an important pathogen in adult periodontitis, rapidly progressing periodontitis, refractory periodontitis, pregnancy gingivitis, acute necrotizing ulcerative gingivitis, pubertal gingivitis. So far 2 DNA homology groups and 3 serotypes of P. intermedia have been reported but there is no data available as yet regarding genetic diversity for the species P. intermedia. The purpose of this study is to investigate, using bacterial DNA restriction endonuclease analysis, genetic diversity between individual strains of P. intermedia which are indistinguishable by serotyping and biotyping, occurrence of an intrafamilial transmission and genetic heterogeneity between P. intermedia strains isolated within a patient and within the same serotypes. The families who have had no systemic disease, no experience of periodontal treatment for the previous 1 year and no experience of antibiotics for the previous 6 months were selected and subgingival plaque was collected at 4 sites in each person and incubated in the anaerobic chamber. P. intermedia were identified by colony shape, gram stain, biochemical test, SK-I03(Sunstar Inc.) test and IIF using monoclonal antibody was perfomed for the determination of serotypes. P. intermedia strains were grown in BHI broth and whole genomic DNA was extracted and digested by restriction endonuclease. The resulting DNA fragments were separated by agarose gel electrophoresis, stained and photographed under UV. As the results of this study, intrafamilial vertial transmissions could be assessed in 2 families and horizintal transmissions in another 2 families. There were different DNA digest patterns within a patient, so P. intermedia showed that individuals could be colonized by multiple clonal types at anyone time. And different serotypes could be found within a patient and in the same serotype within a patient, obvius genetic heterogeneity could not be assessed. But in the same serotype in different famies, there were differences in the DNA digest patterns.

  • PDF

Multiplex Real-Time PCR for Simultaneous Detection of 6 Periodontopathic Bacteria (Multiplex Real-Time PCR을 이용하여 6종의 주요 잇몸질환 유발 미생물을 동시에 검출하는 기법)

  • Cho, Hong-Bum
    • Korean Journal of Microbiology
    • /
    • v.49 no.3
    • /
    • pp.292-296
    • /
    • 2013
  • This study utilized an analysis method for detecting six microorganisms, such as Actinobacillus actinomycetemcomitans, Campylobacter rectus, Porphyromonas gingivalis, Tannerella forsythus, Treponema denticola, and Prevotella intermedia, triggering periodontal disease, using multiplex real-time polymerase chain reaction (PCR). The analysis including internal control was made by dividing the six species into two groups using four fluorescence dyes, and it was verified that there was no interference or cross-reaction between the target species and different kinds of oral microbial species. Qualitative and quantitative analyses were conducted on each microorganism in various samples, such as saliva and the plaque, using the multiplex real-time PCR and comparative analysis between periodontitis patients and healthy people, revealing obvious differences between them.

EFFECTS OF HEAT-KILLED AND SONIC EXTRACTS OF MICROORGANISM ON CULTURED CELLS (세균액 및 세균단백질 추출물이 배양 세포에 미치는 영향)

  • Yu, Young-Dae;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
    • /
    • v.25 no.4
    • /
    • pp.606-618
    • /
    • 2000
  • Dental pulp infection is most commonly caused by extensive dental caries, and some bacterial species invade root canals; bacterial components and products are thought to be associated with the pathogenesis of periapical periodontitis. A principle driving force behind pulpal disease response appears to lie in the host immune system's to bacteria and their products. We examined the production of interleukin $1{\beta}$ (IL-$1{\beta}$) and tumor necrosis factor ${\alpha}$(TNF-${\alpha}$) from human peripheral mononuclear cells, lymphocytes and monocytes stimulated by heat-killed Acitnobacillus actinomycetemcomitans (ATCC 29523), Porphyromonas gingivalis (ATCC 33277) and Prevotella intermedia (ATCC 25611), and also by their sonicated bacterial extracts (SBE), respectively. The effects of three strains of heat-killed bacteria and their SBEs on the morphology of cultured blood cell lines HL-60 (KCLB 10240) and J774A.1 (KCLB 40067) were observed under the inverted microscope. Ultrastructural changes of J774A.1 exposed to heat-killed P. intermedia and its SBE were investigated using transmission electron microscopy. Production of IL-$1{\beta}$ was reduced in human peripheral mononuclear cells after stimulation by sonic bacterial extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia. Heat-killed and sonic extract of P. gingivalis inhibited the production of TNF-${\alpha}$ in peripheral mononuclear cells. Production of TNF-${\alpha}$ was inhibited in peripheral monocytes after stimulation by sonic extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia. HL-60 and J 774A.1 cells showed granular degeneration after treatment with heat-killed and sonic extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia Chromatin margination and shrinkage were observed in 774A.1 treated with heat-killed P. intermedia. Cell wall structure and organelles were destroyed and vacuoles were formed in cytoplasm in J774A.1 treated with P. intermedia sonic extract. These results suggest that A actinomycetemcomitans, P gingivalis and P intermedia may have an important role in the formation and progression of pulpal diseases via both modulation of production of IL-$1{\beta}$ and TNF-${\alpha}$ from blood mononuclear cells and cytopathic effects.

  • PDF