口腔バイオフィルム初期定着菌群による歯周病の個体差別予防モデルの構築

• Project/Area Number: 15J30007
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 国内
• Research Field: Morphological basic dentistry
• Research Institution: Health Sciences University of Hokkaido
• Principal Investigator: 眞島 いづみ 北海道医療大学, 歯学部, 特別研究員(PD)
• Project Period (FY): 2015-04-24 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: Veillonella tobetsuensis / 新菌種Veillonella infantium / Veillonella属細菌 / 唾液細菌叢 / 口腔バイオフィルム初期形成期 / 比較ゲノム解析 / ドラフトゲノム解析 / Autoinducers / Streptococcus属 / Veillonella属 / メタゲノム解析 / 全ゲノム解析 / Autoinducer-2 / Cyclo-Leu-Pro / Autoinducer-1 / Veillonellaｹﾞﾉﾑﾌﾟﾛｼﾞｪｸﾄ / ﾄﾞﾗﾌﾄｹﾞﾉﾑｼｰｸｴﾝｽ

Outline of Annual Research Achievements

平成29年度は、平成28度の唾液細菌叢解析に引き続き、健康な児童50名分の唾液サンプルを更に追加して、出身地域別にその細菌叢を解析した。結果として、地区間に有意な差は認められなかったが、口腔清掃状態別にそれを解析すると、先の解析結果同様、清掃状態が悪化するに従い、Veillonella属細菌の割合の増加が認められた。これらの解析から、う蝕や歯周病の直接的原因である口腔バイオフィルムの初期形成に重要な役割を果たすVeillonella属細菌が、口腔清掃状態が悪化するに従い、定着しやすくなることが明らかになった。
更に、本口腔細菌叢解析の過程で、既報のVeillonella属細菌には分類されないVeillonella属細菌未同定株を分離した。ゲノムシークエンスデータによるAverage Nucleotide Identity算出、複数のハウスキーピング遺伝子の塩基配列解析、生化学的性状検査等を行った結果、本未同定株を新菌種Veillonella infantiumとして確立し、国際機関に株の寄託を行った（JCM 31738T、 TSD-88T）後、口腔内におけるその出現頻度を明らかにした。
また、研究実施計画に基づき、上記新菌種を含む口腔Veillonella全7菌種標準株のドラフトゲノム解析後、KEGGデータベースを用いて比較ゲノム解析を行った。特に口腔バイオフィルムの初期形成に影響を与えることが示唆されているVeillonella tobetsuensis由来Autoinducer (AI)-1、AI-2及びCyclo-Leu-Proの代謝経路を含めた機能解析、当該産物産生責任遺伝子の同定作業を遂行した。
これら研究成果により、口腔バイオフィルム初期形成菌であるVeillonella属細菌を応用した口腔感染症の予防及び治療法の確立の可能性が強く示唆された。

Research Progress Status

29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

29年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] SUNY at Buffalo(米国)
• [Int'l Joint Research] Mahidol University(Thailand)
• [Int'l Joint Research] Universitas Indonesia(Indonesia)
• [Int'l Joint Research] Mahidol University(Thailand)
• [Int'l Joint Research] University at Buffalo(米国)
• [Int'l Joint Research] Mahidol University(タイ)
• [Journal Article] Draft genome sequences of four strains of recently established novel Veillonella species isolated from human oral cavities (2018)
  • Author(s): Mashima I, Liao YC, Sabharwal A, Haase EM, Nakazawa F, Scannapieco FA
  • Journal Title: Genome Announcements Volume: 6 Issue: 15
  • DOI: 10.1128/genomea.00259-18
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Veillonella infantium sp. nov., an anaerobic, Gram-stain-negative coccus isolated from tongue biofilms of a Thai child (2018)
  • Author(s): Mashima I, Liao YC, Miyakawa H, Theodorea CF, Thaweboon B, Thaweboon S, Scannapieco FA, Nakazawa F
  • Journal Title: International Journal of Systematic Evolutionary Microbiology Volume: 68 Issue: 4 Pages: 1101-1106
  • DOI: 10.1099/ijsem.0.002632
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Molecular Detection of Oral Veillonella Species in the Saliva of Children with Different Oral Hygiene Statuses (2017)
  • Author(s): Theodorea CF, Mashima I, Thaweboon B, Thaweboon S, Nakazawa F
  • Journal Title: International Journal of Current Microbiology and Applied Sciences Volume: 6 Issue: 7 Pages: 449-461
  • DOI: 10.20546/ijcmas.2017.607.054
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Exploring the salivary microbiome of children stratified by the oral hygiene index (2017)
  • Author(s): Mashima I, Theodorea CF, Thaweboon B, Thaweboon S, Scannapieco FA, Nakazawa F
  • Journal Title: PLOS ONE Volume: 68 Issue: 9 Pages: 1101-1106
  • DOI: 10.1371/journal.pone.0185274
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Prospects of Novel Species of Oral Veillonella in Human Saliva (2017)
  • Author(s): Theodorea CF, Mashima I, Nakazawa F
  • Journal Title: Advances in Biotechnology & Microbiology Volume: 5 Issue: 4 Pages: 555667-555667
  • DOI: 10.19080/aibm.2017.05.555667
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Application of Dental Implants Coated with Titanium Nitride; The Experimental Study with Porphyromonas gingivalis Infection (2017)
  • Author(s): Yamazaki K, Mashima I, Nakazawa F, Nakanishi Y, Ochi M.
  • Journal Title: International Journal of Current Microbiology and Applied Sciences Volume: 6 Issue: 1 Pages: 130-142
  • DOI: 10.20546/ijcmas.2017.601.017
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Role of an Autoinducer-2-like Molecule from Veillonella tobetsuensis in Streptococcus gordonii Biofilm Formation (2017)
  • Author(s): Mashima I, Nakazawa F.
  • Journal Title: Journal of Oral Biosciences Volume: 印刷中
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Identification of Veillonella Species in the Tongue Biofilm by Using a Novel One-step Polymerase Chain Reaction Method (2016)
  • Author(s): Mashima I, Theodorea CF, Thaweboon B, Thaweboon S, Nakazawa F.
  • Journal Title: PLoS ONE Volume: 11 Issue: 6 Pages: 1-16
  • DOI: 10.1371/journal.pone.0157516
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] The role of AI-2 like substance from Veillonella tobetsuensis (2016)
  • Author(s): Mashima I, Miyakawa H, Nakazawa F.
  • Journal Title: Bacterial Adherence & Biofilm Volume: 29
• [Journal Article] Monitoring the decontamination efficacy of the novel Poseidon-S disinfectant system in dental unit water lines (2016)
  • Author(s): Mari FUJITA, Izumi MASHIMA, Futoshi NAKAZAWA
  • Journal Title: Journal of Microbiology, Immunology and Infection Volume: xxx Issue: 3 Pages: 270-276
  • DOI: 10.1016/j.jmii.2015.05.006
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Draft Genome Sequence of Veillonella tobetsuensis ATCC BAA-2400T Isolated from Human Tongue Biofilm (2015)
  • Author(s): Izumi MASHIMA, Futoshi NAKAZAWA
  • Journal Title: Genome Announcements Volume: 3 Issue: 4 Pages: 1-2
  • DOI: 10.1128/genomea.00808-15
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The Distribution and Frequency of Veillonella spp. associated with Chronic Periodontal Diseases (2015)
  • Author(s): Mashima I, Fujita M, Nakatsuka Y, Kado T, Furuichi Y, Herastuti S, Nakazawa F.
  • Journal Title: International Journal of Current Microbiology and Applied Sciences Volume: 4 Pages: 150-160
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The interaction between Streptococcus spp. and Veillonella tobetsuensis in oral biofilm formation at early stage (2015)
  • Author(s): Izumi MASHIMA, Futoshi NAKAZAWA
  • Journal Title: Journal of Bacteriology Volume: 197 Issue: 13 Pages: 2104-2111
  • DOI: 10.1128/jb.02512-14
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Identification of oral Veillonella species by using One-step PCR method isolated from saliva of Japanese children (2018)
  • Author(s): Theodorea CF, Djais AA, Mashima I, Otomo M, Saitoh M, Nakazawa F
  • Organizer: 北海道医療大学歯学会第36回学術大会
• [Presentation] Bioindicator for oral hygiene status (2018)
  • Author(s): Theodorea CF, Mashima I, Nakazawa F
  • Organizer: International Symposium on Development of Human Resources in Practical Oral health and Treatment - Basic, Clinical and Dental Public Health innovation for Better Oral Health -
  • Int'l Joint Research
• [Presentation] Effects of oral Veillonella on Streptococcus biofilm formation (2018)
  • Author(s): Ramadhani R, Mashima I, Theodorea CF, Nakazawa F
  • Organizer: International Symposium on Development of Human Resources in Practical Oral health and Treatment - Basic, Clinical and Dental Public Health innovation for Better Oral Health -
  • Int'l Joint Research
• [Presentation] Exploring the Microbial Community in Saliva from Children (2017)
  • Author(s): Mashima I, Theodorea CF, Nakazawa F
  • Organizer: The 95th General Session & Exhibition of the International Association for Dental Research (IADR)
  • Place of Presentation: Moscone Convention center, San Francisco, CA, USA
  • Year and Date: 2017-03-22
  • Int'l Joint Research
• [Presentation] Exploring the Microbiota in Saliva from Children (2017)
  • Author(s): Mashima I, Theodorea CF, Nakazawa F
  • Organizer: 第90回日本細菌学会総会
  • Place of Presentation: 仙台国際センター展示棟、仙台市
  • Year and Date: 2017-03-19
• [Presentation] The Phylogenetic Diversity of Unclassified Veillonella Isolates (2017)
  • Author(s): Theodorea CF, Msahima I, Nakazawa F
  • Organizer: 第59回歯科基礎医学会
• [Presentation] Oral Veillonella Profiles in Saliva of the Children with Different Oral Hygiene Statuses and Their Phylogenetic Diversity (2017)
  • Author(s): Theodorea CF, Msahima I, Nakazawa F
  • Organizer: 第84回日本細菌学会北海道支部学術大会
• [Presentation] RpoB and DnaK Sequences of Unidentified Veillonella Isolates (2017)
  • Author(s): Theodorea CF, Msahima I, Nakazawa F
  • Organizer: International Association of Dental Research-South-East Asia Division & 28th SEAADE Annual Scientific Meeting
  • Int'l Joint Research
• [Presentation] Veillonella childrensis Sp. Nov., an Anaerobic, Gram Negative Coccus Isolated from Tongue Biofilm of Thai Children (2017)
  • Author(s): Msahima I, Theodorea CF, Nakazawa F
  • Organizer: ASM Microbe 2017
  • Int'l Joint Research
• [Presentation] The effects of chemical treatments used following air-polishing on experimentally contaminated titanium surface (2017)
  • Author(s): Ichioka Y, Kado T, Mashima I, Nakazawa F, Endo K, Furuichi Y
  • Organizer: The 5th Lindhe International Symposium
  • Int'l Joint Research
• [Presentation] The Role of Oral Veillonella in Biofilm Formation - Its Significance and Prospects - (2017)
  • Author(s): Nakazawa F, Mashima I
  • Organizer: 2nd Regional Oral Biology Scientific Meeting/PBOI
  • Int'l Joint Research / Invited
• [Presentation] 口腔Veillonella全6菌種同定One Step PCR法の開発 (2016)
  • Author(s): 眞島いづみ、シトラ・F・セオドレア、中澤太
  • Organizer: 第83回日本細菌学会北海道支部学術総会
  • Place of Presentation: 北海道大学、札幌市
  • Year and Date: 2016-09-17
• [Presentation] The strategy for oral biofilm -The best use of oral Veillonella- (2016)
  • Author(s): Mashima I, Theodorea CF, Miyakawa H, Nakazawa F
  • Organizer: 第58回歯科基礎医学会学術大会(メインシンポジウムB)
  • Place of Presentation: 札幌コンベンションセンター、札幌市
  • Year and Date: 2016-08-25
  • Invited
• [Presentation] タイ児童舌苔から分離されたVeillonella属新菌種の塩基配列解析 (2016)
  • Author(s): 眞島いづみ、シトラ・F・セオドレア、中澤太
  • Organizer: 第58回歯科基礎医学会学術大会
  • Place of Presentation: 札幌コンベンションセンター、札幌市
  • Year and Date: 2016-08-24
• [Presentation] Application of Titanium Nitride for Dental Implants: A Rabbit Study (2016)
  • Author(s): Mashima I, Yamazaki K, Nakazawa F, Nakanishi Y, Ochi M.
  • Organizer: The 11th Scientific of the Asian Academy of Osseointegration (AAO 2016)
  • Place of Presentation: Mahidol University, Bangkok, Thailand
  • Year and Date: 2016-06-02
  • Int'l Joint Research
• [Presentation] Veillonella tobetsuensis由来AI-2の口腔ﾊﾞｲｵﾌｨﾙﾑ形成初期における影響 (2016)
  • Author(s): 眞島いづみ、中澤太
  • Organizer: 第89回日本細菌学会総会
  • Place of Presentation: 大阪国際交流センター、大阪
  • Year and Date: 2016-03-23
• [Presentation] The Role of Autoinducers from Veillonella in the Formation of Oral Biofilms (2015)
  • Author(s): Izumi MASHIMA, Futoshi NAKAZAWA
  • Organizer: the 7th ASM Conference on Biofilms
  • Place of Presentation: Hyatt Regency Chicago, Chicago, IL, USA
  • Year and Date: 2015-10-24
  • Int'l Joint Research
• [Presentation] The Roles of Oral Veillonella in Biofilm Formation (2015)
  • Author(s): Izumi MASHIMA, Hiroshi MIYAKAWA, Futoshi NAKAZAWA
  • Organizer: International Seminar
  • Place of Presentation: Department of Oral Biology, State University at Buffalo, NY, USA
  • Year and Date: 2015-10-23
  • Int'l Joint Research / Invited
• [Presentation] 口腔ﾊﾞｲｵﾌｨﾙﾑ初期形成期におけるcyclic dipeptideの影響 (2015)
  • Author(s): 眞島いづみ、宮川博史、中澤太
  • Organizer: 第57回歯科基礎医学会学術大会
  • Place of Presentation: 朱鷺メッセ、新潟
  • Year and Date: 2015-09-12
• [Presentation] Molecular Biological Control Method for Oral Biofilm at Early Stage (2015)
  • Author(s): Izumi MAHSIMA, Hiroshi MIYAKAWA, Futoshi NAKAZAWA
  • Organizer: 第57回歯科基礎医学会学術大会ｻﾃﾗｲﾄｼﾝﾎﾟｼﾞｳﾑ
  • Place of Presentation: 朱鷺メッセ、新潟
  • Year and Date: 2015-09-11
  • Invited
• [Presentation] Veillonella tobetsuensis由来Autoinducer-2様物質の役割 (2015)
  • Author(s): 眞島いづみ、宮川博史、中澤太
  • Organizer: 第29回日本ﾊﾞｲｵﾌｨﾙﾑ学会学術集会
  • Place of Presentation: ホテル竹島、蒲郡
  • Year and Date: 2015-07-10
• [Presentation] The Development of Molecular Biological Control Method for Dental Plaque (2015)
  • Author(s): Izumi MASHIMA
  • Organizer: Master Class of Antimicrobial Defenses chaired by Jules Hoffmann, Bruce Beutler in The 65th Lindau Nobel Laureates Meeting
  • Place of Presentation: Hotel Bayer. Hof, Lindau, Germany
  • Year and Date: 2015-06-29
  • Int'l Joint Research