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Study on the mechanism of toxin production by Clostridium difficile

Research Project

Project/Area Number 06670286
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Bacteriology (including Mycology)
Research InstitutionKanazawa University

Principal Investigator

NAKAMURA Shinichi  Kanazawa Univ., Sch. Med., Prof., 医学部, 教授 (90019620)

Co-Investigator(Kenkyū-buntansha) YAMAKAWA Kiyotaka  Kanazawa Univ., Sch. Med., Asst. Prof., 医学部, 講師 (20110629)
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsC.difficile / defined medium / amino acid / vitamin / toxin A / toxin B / toxin / biotin
Research Abstract

The mechanism of toxin production by Clostridium difficile was investigated from the stand poit of nutritional requirements. By the single-amino acid omission and addition methods with a basal defined medium consisting of 18 amino acids, it was found that valine, cysteine, tryptophan, proline, methionine, isoleucine, leucine, glycine and threonine were essential for good toxin production.
Further, by qualitative analysis, a defined medium (6xMADM) consisting (g/L) of tryptophan 0.6, methionine 1.2, valine 1.8, isoleucine 1.8, proline 1.8, leucine 2.4, glycine 0.2, threonine 0.4 and cysteine 0.5 was found to be fairly effective for stimulating toxin production. In a comparison of 6xMADM and m-BHI,which is excellent comple medium for toxin production, 13 of 20 strains tested produced the same amounts of toxin in both media, suggesting that the 9 amino acids mentioned above were particularly important for toxin production. By single-vitamin omission method with 6xMADM,it was found that pyridoxine, pantothenate and biotin were essential for good growth and toxin production and that biotin deficiency might stimulate toxin production. The effect of biotin was analyzed in detail with a strain KZ 1647. When toxin production was examined in relation to biotin concentration, it was found that with decreasing concentration of biotin bacterial growth decreased, but toxin production was remarkably increased, particularly with 0.05 nM biotin.
The time course of toxin production in biotin-limited conditions was similar to that in biotin-enriched conditions. The biotin effect on toxin production was also observed in 18 out of 19 strains tested, suggesting that the effect is a general phenomenon amongst toxigenic C.difficile strains.
These findings suggest that biotin play an important role in the development of C.difficile colitis.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] K.Yamakawa: "Toxin production by Clostridium difficile in a defined medium with limited amino acids" Journal of Medical Microbiology. 41. 319-323 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Karasawa: "A defined growth medium for Clostridium difficile" Journal of General Microbiology. 141. 371-375 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Yamakawa: "Enhancement of Clostridium difficile toxin production in biotin-limited conditions" Journal of Medical Microbiology. 44. 111-114 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 山川 清孝: "ビオチンによるClostriclium difficile毒素産生の増強効果" 日本細菌学雑誌. 50. 278 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 唐澤 忠宏: "Clostridium difficileの増殖に必要なアミノ酸とビタミン" 日本細菌学雑誌. 50. 167 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 中村 信一: "消化管エコロジー" 小橋恭一編集, 108 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Yamakawa: "Toxin production by Clostridium difficile in a defined medium with limited amino acids" Journal of Medical Microbiology. 41. 319-323 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Karasawa: "A defined medium for Clostridium difficile" Journal of General Microbiology. 141. 371-375 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Yamakawa: "Enhancement of Clostridium difficile toxin production in biotin-limited conditions" Journal of Medical Microbiology. 44. 111-114 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Karasawa: "A defined growth medium for Clostridium difficile" Journal of General Microbiology. 141. 371-375 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 山川清孝: "ビオチンによるClostridium difficile毒素産生の増強効果" 日本細菌学雑誌. 50. 278 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 唐澤忠宏: "Clostridium difficileの増殖に必要なアミノ酸とビタミン" 日本細菌学雑誌. 50. 167 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K.Yamakawa: "Enhancement of Clostridium difficile toxin production in biotin-limited conditions" Journal of Medical Microbiology. 44. 111-114 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] K.Yamakawa: "Toxin production by Clostridium difficile in a defined medium with limited amino acids" Journal of General Microbiology. 41. 319-323 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Karasawa: "A defined growth medium for clostridium difficile" Journal of Medical Microbiology. 141. 371-375 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 山川清孝: "ビチオンによるClostridium difficile毒素産生の増強効果" 日本細菌学雑誌. 50. 278 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 唐澤忠宏: "Clostridium difficileの増殖に必要なアミノ酸とビタミン" 日本細菌学雑誌. 50. 167 (1995)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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