Mobile genetic elements associated with multi-drug resistance in the bacteria from aquaculture environments

• Project/Area Number: 15K07531
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Aquatic bioproduction science
• Research Institution: Dokkyo Medical University
• Principal Investigator: Lisa Nonaka 獨協医科大学, 医学部, 講師 (70363265)
• Co-Investigator(Kenkyū-buntansha): 矢野 大和 東北大学, 生命科学研究科, 講師 (20646773)
• Research Collaborator: Suzuki Satoru ; Maruyama Fumito
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2017: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2016: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2015: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
• Keywords: 薬剤耐性菌 / 多剤耐性 / 遺伝子伝達 / プラスミド / トランスポゾン / チロシンリコンビナーゼ / 養殖場 / ビブリオ属 / 接合 / pSEA1 / Tn6283 / 染色体 / 部位特異的挿入 / bcp遺伝子 / 薬剤耐性 / 染色体への組み込み / インテグラーゼ

Outline of Final Research Achievements

Here, we show a new pattern of interspecies antimicrobial resistance genes (ARGs) transfer involving a ‘non-conjugative’ integrative element. We conducted whole-genome sequencing of a transconjugant obtained by mating between Escherichia coli and V. ponticus. This revealed integration of a plasmid (pSEA1) into the chromosome, consisting of ARGs, and a 13.8-kb integrative element Tn6283. Molecular genetics analysis suggested a two-step gene transfer model. First, Tn6283 integrates into the recipient chromosome during suicidal plasmid transfer, followed by homologous recombination between the Tn6283 copy in the chromosome and that in the newly transferred pSEA1. Tn6283 is unusual among integrative elements in that it apparently does not encode transfer function and its excision barely generates unoccupied donor sites. Overall, this study reveals the presence of a previously unrecognized type of MGE in a marine organism, highlighting diversity in the mode of interspecies gene transfer.

Academic Significance and Societal Importance of the Research Achievements

細菌間で行われる遺伝子の伝達は薬剤耐性菌の出現・拡大を理解するための重要なファクターである。本研究ではこれまで現象のみが知られていた、伝達性プラスミドが受容菌へ伝達された後にその染色体へ潜り込むメカニズムを明らかにした。学術性意義としてはプラスミド上のトランスポゾンがプラスミド全長の染色体への組み込みをアシストしているというトランスポゾンの新規機能を明らかにした点である。一方、耐性遺伝子がこれまで考えられてきた以上に多様かつ複雑なメカニズムで細菌間を移動している可能性を示した点は薬剤耐性菌問題を考える上で重要な知見であり非常に社会的意義が高い。

Research Products

• [Int'l Joint Research] Chulalongkorn University/Quality Vet Product(Thailand)
• [Int'l Joint Research] National Taiwan Ocean University/National Cheng Kung University(Taiwan)
• [Int'l Joint Research] Chulalongkorn University/Quality Vet Product(タイ)
• [Int'l Joint Research] National Taiwan Ocean University/National Cheng Kung University(台湾)
• [Journal Article] Interplay of a non-conjugative integrative element and a conjugative plasmid in the spread of antibiotic resistance via suicidal plasmid transfer from an aquaculture Vibrio isolate. (2018)
  • Author(s): Nonaka, L., Yamamoto, T., Maruyama, F., Hirose, U., Onishi, Y., Kobayashi, T., Suzuki, S., Nomura, N., Masuda, M. and Yano, H.
  • Journal Title: PLos One Volume: 13 Issue: 6 Pages: e0198613-e0198613
  • DOI: 10.1371/journal.pone.0198613
  • NAID: 120006715750
  • Peer Reviewed / Open Access
• [Journal Article] The novel mef (C)-mph (G) macrolide resistance genes are conveyed in the environment on various vectors (2017)
  • Author(s): Sugimoto Yuta、Suzuki Satoru、Nonaka Lisa、Boonla Chanchai、Sukpanyatham Nop、Chou Hsin-Yiu、Wu Jer-Horng
  • Journal Title: Journal of Global Antimicrobial Resistance Volume: 10 Pages: 47-53
  • DOI: 10.1016/j.jgar.2017.03.015
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The novel mef(C)-mph(G) macrolide resistance genes are conveyed in the environment on various vectors (2017)
  • Author(s): Y. Sugimoto, S Suzuki, L Nonaka, C Boonla, N Sukpanyatham, H-Y Chou, J-H W
  • Journal Title: Journal of Global Antimicrobial Resistance Volume: 印刷中
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] 養殖場由来Vibrio属細菌が保有する伝達性多剤耐性プラスミドpSEA1のTn6283を介した受容菌染色体への組み込み機構 (2018)
  • Author(s): 野中里佐、山本達也、丸山史人、広瀬 侑、大西勇輝、小林 剛、鈴木 聡、野村暢彦、増田道明、矢野大和
  • Organizer: 日本微生物生態学会
• [Presentation] Novel transferable multi-drug resistance plasmid and macrolide resistance genes from Photobacterium damselae subsp. damselae isolated from seawater of an aquaculture site (2015)
  • Author(s): Lisa Nonaka
  • Organizer: 日本微生物生態学会第30回大会
  • Place of Presentation: 土浦市
  • Year and Date: 2015-10-17
  • Int'l Joint Research