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P1-001 白癬症起因菌の同定に対する Nucleotide Tag P1-002 MLVA Typing of Leptospira Interrogans and L

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P1-001 白癬症起因菌の同定に対する Nucleotide Tag P1-002 MLVA Typing of Leptospira Interrogans and L P1-001 白癬症起因菌の同定に対する Nucleotide tag P1-002 MLVA typing of Leptospira interrogans and L. 配列の有用性 borgpetersenii isolated from small mammals in East Asia ○山西 千晶1，佐藤 一朗1，佐野 文子3，中村 郁郎2，槇村 浩一1（1帝 ○小泉 信夫1，泉谷 秀昌1，Jung-Jung Mu2，岡野 祥3，中島 千絵4， 京大・医・大学院・宇宙環境・医・研究室，2千葉大・園芸・大学院， 鈴木 定彦4，武藤（水谷）麻紀1,5，大西 真1（1感染研・細菌第一， 3琉球大・農学部） 2Taiwan CDC，3沖縄衛環研・感染症，4北大人獣センター・バイオリソー ス，5関東学院大・理工・生命） Utilization of the RNA polymerase I partial sequence ○Nobuo Koizumi1, Hidemasa Izumiya1, Jung-Jung Mu2, Shou (Ntag) as a molecular marker of dermatophytes 3 4 4 1,5 ○ 1 1 3 2 Okano , Chie Nakajima , Yasuhiko Suzuki , Maki Muto Mizutani , Chiaki Yamanishi , Kazuo Sato , Ayako Sano , Ikuo Nakamura , 1 1 2 3 1 1 Makoto Ohnishi ( Dept. Bacteriolo. I, NIID, Taiwan CDC, Dept. Koichi Makimura ( Lab. Space Environ. Med., Grad. Sch. Med, 4 2 3 Biol. Sci., Okinawa Pref. Inst. Health Environ., Div. Bioresources, Teikyo Univ., Horticul., Grad. Sch., Chiba Univ., Dept. Agriculture, 5 Ryukyus Univ.) Hokkaido Univ. CZC, Dept. Biosci., Kanto Gakuin Univ.) 近年，RNA ポリメラーゼ I 複合体を構成する最大サブユニット Leptospira spp. are the causative agents of a worldwide zoonosis, （POLA1）の C 末端領域に存在するアミノ酸配列（Protein tag, leptospirosis, maintained by various mammals. We investigated the Ptag）が，真核生物の系統関係を反映し，また種特異性を示す事 molecular characteristics of L. interrogans and L. borgpetersenii which が報告された。Ntag は Ptag をコードする DNA 塩基配列を指し， were isolated from small feral and wild animals in Japan, Philippines, 同様に高い種特異性を示す。白癬の起因菌は系統的に極めて近 Taiwan, and Vietnam using multiple-locus variable-number tandem 縁関係にあり，既存の同定マーカー（Internal transcribed spacer repeat analysis (MLVA). 1 等）で分類された DNA 塩基配列も酷似している。そのため分 MLVA using 11 and four loci were performed on 110 L. interrogans 子系統解析の実施および種の同定指標として用いる場合，他に and 52 L. borgpetersenii, respectively. In L. interrogans, serogroups 数種類の DNA 塩基配列を解析する必要があり，時間的・経済 Autumnalis and Hebdomadis appeared more genetically diverse than 的な問題があった。Ntag 配列は種特異性が高いので，この問題 serogroups Bataviae, Grippotyphosa, Icterohaemorrhagiae, Pomona, を解決する可能性がある。そこで今回我々は，Ntag 配列を用い or Pyrogenes. The former serogroup strains with the exception of た白癬菌の種間変異と種内変異について，分子系統解析を行い， one Hebdomadis strain were isolated from Apodemus speciosus while 既存の分子系統と比較することによって，Ntag 配列の白癬症起 all the latter serogroup strains with the exception of Grippotyphosa 因菌に対する系統分類と菌種同定の有用性を検証した。 were isolated from Rattus norvegicus. L. borgpetersenii was isolated （会員外共同研究者：中村郁郎 千葉大学大学院園芸学研究科） from at least 11 animal species while L. interrogans was isolated from five species, which may suggest a wider host range for L. borgpetersenii. Broad host preference in a single genotype was also observed, which colonized not only different species of the same genera but multiple animal genera. This study demonstrates that there may be variability in the range of genetic diversity among different Leptospira serogroups, which may be attributed to maintenance host animals and environmental factors. P1-003 Mycobacterium avium の系統分岐に伴う大規 P1-004 フサオマキザル口腔から分離された新菌種 模なゲノム構造の変化 Streptococcus oricebi の解析 ○岩本 朋忠1，矢野 大和2，西内 由紀子3，有川 健太郎1，中島 千絵4， ○齋藤 真規，桑原 紀子，續橋 治，小林 良喜，落合 智子（日大・松 鈴木 定彦4，丸山 史人5（1神戸市環保研・感染症部，2東大・新領域・ 戸歯・微生物免疫） メディカルゲノム，3大阪市大・医・刀根山結核，4北大人獣センター・ バイオリソース，5京大・医・微生物感染） Streptococcus oricebi sp. nov., isolated from the oral cavity of tufted capuchin Changes in genomic architecture of Mycobacterium ○Masanori Saito, Noriko Kuwahara, Osamu Tsuzukibashi, Ryoki avium at the point of phylogenetic diversification Kobayashi, Tomoko Ochiai (Dept. Microbiol. Immunol., Sch. Dent. ○Tomotada Iwamoto1, Hirokazu Yano2, Yukiko Nishiuchi3, Kentaro Matsudo, Nihon Univ.) Arikawa1, Chie Nakajima4, Yasuhiko Suzuki4, Fumito Maruyama5 (1Dept. Infect. Dis., Kobe. Inst. Heal., 2Grad. Sch. Front. Sci., Univ. A Gram-positive, catalase-negative, coccus-shaped organism was of Tokyo, 3Toneyama Inst. Tuberculosis Res. Osaka City Univ. Med. isolated from the oral cavity of tufted capuchin (Cebus apella). Sch., 4Div. Bioresources, Hokkaido Univ. Res. Cent. Zoonosis Cont., A comparative 16S rRNA gene-sequencing analysis suggested 5Microbiol. Grad. Sch. Med., Kyoto Univ.) classification of the organism into the genus Streptococcus. Strain M8T was the most closely related to Streptococcus oralis (96.17%) followed Mycobacterium avium は，近年先進諸国を中心に患者数が急増している by Streptococcus massiliensis (95.90%) based on the 16S rRNA gene. MAC 症（難治性慢性肺疾患）の起因菌である。また，ブタや牛などの家 Strain M8T was the most closely related to S. massiliensis (86.58%) 畜にも疾病を引き起こす。本菌種は MLST や MLVA 等の遺伝子型別解析 based on the RNA polymerase β subunit-encoding gene (rpoB), the によって，遺伝子型が異なる複数の亜種や亜型から構成されていることが most closely related to Streptococcus tigurinus (81.26%) followed 知られている。本菌種は環境中の常在細菌であるといわれているが，宿主 by S. massiliensis (80.45%) based on the 60 kDa heat-shock protein 側でなく，本菌種側に病原性や宿主特異性の調節を可能にするような遺伝 gene (groEL), and the most closely related to S. massiliensis (79.6%) 的背景があるのかもしれない。そこで本研究では，M. avium の遺伝系統 based on a recombination and repair protein-encoding gene (recN), の多様性やその形成に寄与する環境要因についての情報を得るために，多 respectively. The phylogenetic trees of 16S rRNA, rpoB, groEL, 様な分離源由来の株（合計 69 株）のゲノム情報を用いて，比較ゲノム解 and recN gene sequences showed that strain M8T was most closely 析を行った。 related to S. massiliensis. Based on phenotypic characterization as コアゲノムを用いた系統樹解析により，各亜種ならびに亜系統群の進化的 well as the 16S rRNA gene and housekeeping gene (rpoB, groEL and 位置付けと分離由来との関連性を検討した。その結果，主にヒト・ブタ recN) sequence data, we here propose a novel taxon, Streptococcus から分離される M. avium subsp. hominissuis で，宿主および分離地域を反 oricebi sp. nov. (type strain M8T= JCM 30719T= DSM 100101T). This 映した 5 つの clade の存在が明らかになった。反芻動物にヨーネ病を引き work was supported by JSPS KAKENHI Grant Number 25870768. 起こす M. avium subsp. paratuberculosis では，Sheep 型と Bison/Cow 型は， それぞれ独立して分岐・派生したことが示された。さらにコアゲノム系統 樹から推定されるゲノム配列が急速に変化した系統分岐時においては，大 規模なゲノム構造の変化も伴い，それが継承されていることが判明した。 ゲノム構造変化の機構の詳細については現在解析中である。 これらの知見は，M. avium の疫学，臨床，治療薬の開発に有用な情報を もたらすものと思われる。 58 日本細菌学雑誌 71（ 1 ）， 2016 P1-005 北海道に生息するアライグマから分離されたレ P1-006 MIRU-VNTR Typing of Beijing MDR-TB プトスピラの遺伝学的解析 strains from Myanmar ○橋場 和野，高崎 真理枝，吉織 綾子，村田 亮，中野 良宣，菊池 直 ○Lai Lai San1，Nan Aye Thida Oo1，Wah Wah Aung2，Khin Saw 哉（酪農大・獣医・獣医細菌） Aye2，中島 千絵1,3，鈴木 定彦1,3（1北大・人獣セ・バイオリソース， 2ミャンマー保健省医学研究局，3北海道大学国際連携研究教育局） Genetic characterization of Leptospira isolated from ○Lai Lai San1, Nan Aye Thida Oo1, Wah Wah Aung2, Khin Saw Aye2, raccons (Procyon lotor) in Hokkaido, Japan 1,3 1,3 1 ○ Chie Nakajima , Yasuhiko Suzuki ( Divi. Biores., CZC, Hokkaido Wano Hashiba, Marie Takasaki, Ayako Yoshiki, Ryo Murata, 2 3 Yoshinori Nakano, Naoya Kikuchi (Dept. Pathobiol., Sch. Vet. Med., Univ., Dept. Med. Res., Min. Health, Myanmar, Hokkaido Univ. GI- Rakuno Gakuen Univ.) CoRE GZC) 【目的】レプトスピラは保有体であるネズミなどのげっ歯類など Introduction: Myanmar is one of the countries with the highest が尿中に排菌することで川や土壌などの環境を汚染し，ヒトや burden of tuberculosis (TB) as well as multidrug-resistant 家畜，野生動物などへの感染源となる。我々は北海道に生息す tuberculosis (MDR-TB). The previous studies on genotping in るアライグマからレプトスピラを分離し，本菌が広く浸潤して Myanmar pointed out that the Beijing strain has a significant いることを明らかにした。さらにそれらの分離株が Leptospira association with MDR-TB. The active transmission of MDR borgpetersenii 血清型Javanica と L.interrogans 血清型Autumnalis Mycobacterium tuberculosis (MTB) strains is the emerging problem, に近縁な血清型であることを明らかにした。本研究ではそれら hence, it is necessary to identify characteristics that can explain why の分離株についてさらに遺伝学的解析を実施した。【方法】アラ they keep spreading widely. イグマから分離されたレプトスピラ株 12 株を用いた。各分離株 Materials and Method: A total of 171 Beijing MDR-MTB isolates の遺伝子種を決定するために 16S rRNA・flaB・gyrB 遺伝子の塩 collected from Myanmar was analyzed by variable number of tandem 基配列の解析を行い，さらに菌株間の異同を推定するため PFGE repeat(s) of mycobacterial interspersed repetitive units (MIRU- を実施した。【成績と結論】血清型 Javanica と同定された 10 株 VNTR) typing. Fifteen loci of MIRU-VNTR (QUB26, Mtub21, は 16S rRNA・flaB 遺伝子の解析により L. borgpetersenii に近縁な MIRU31, MIRU10, QUB11b, MIRU26, MIRU4, MIRU39, MIRU40, 遺伝子種であり，遺伝学的にも血清型 Javanica と証明された。し Mtub4, QUB4156, Mtub30, MIRU39, ETR A, MIRU16) were used かし血清型 Autumnalis と推定された 2 株は L.interrogans に近縁 and the results were analyzed by bionumeric software. な遺伝子種であることが確認できたが，gyrB 遺伝子の解析では Results: There were 32 clusters in 171 isolates with the biggest 血清型 Canicola に近縁であり血清学的解析と異なる結果となっ cluster consisted of 5 isolates. VNTR set used in this study was た。PFGE の結果，10 株は L. borgpetersenii 血清型 Javanica の参 useful for the discrimination of MDR-TB Beijing genotype and 照株と類似した切断パターンを示し，2 株は L.interrogans 血清型 effective for the determination of transmission pattern of MDR-TB. Autumnalis の参照株と類似した切断パターンを示した。さらに Beijing MDR-MTB was highly diverse and it can be inferred that the 同じ血清型の切断パターンを比較すると，わずかに相違が認め outbreak of MDR-TB is less likely to occur. られたことから，北海道に生息するアライグマの保有するレプ Conclusion: MDR-MTB may be emerging in each patient トスピラは多様性を示していることが示唆された。 individually. So that we can stop spread of MDR-MTB with proper drug usage. P1-007 Evaluation of MIRU-VNTR typing of MDR- P1-008 Anaplasma phagocytophilum specific P44 MTB Beijing genotype isolates in Nepal epitopes identified by peptide array analysis ○Yogendra Shah1，Jeewan Thapa1，Ajay Poudel2，Bhagwan ○蘇 泓如1，呉 東興1，伊藤 圭祐2，河原崎 泰昌2，大橋 典男1（1静岡 Maharjan3，鈴木 定彦1,4（1北大・人獣セ・バイオリソース，2ネパール・ 県大・食品栄養科学・微生物，2静岡県大・食品栄養科学・生分工） 人民歯科大学，3ネパール・結核予防会・ドイツ－ネパール結核プロ 1 1 2 2 ジェクト，4北大・国際連携研究教育局） ○Hongru Su , Dongxing Wu , Keisuke Ito , Yasuaki Kawarasaki , Norio Ohashi1 (1Lab. Microbiol., Sch. Food Nutr. Sci., Univ. Shizuoka, ○Yogendra Shah1, Jeewan Thapa1, Ajay Poudel2, Bhagwan 2Biomol. Engneer. Lab., Sch. Food Nutr. Sci., Univ. Shizuoka) Maharjan3, Yasuhiko Suzuki1,4 (1Divi. Biores., CZC, Hokkaido Univ., 2Peoples Dental Col., Nepal, 3GENETUP, Nepal, 4Hokkaido Univ.
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