Evaluation of the biorisk on the use of microorganisms for bioremediation in the environment

• Project/Area Number: 15310055
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Environmental technology/Environmental materials
• Research Institution: Osaka University
• Principal Investigator: FUJITA Masanori Osaka University, Graduate School of Engineering, Dept.of Environmental Engineering, Professor, 大学院・工学研究科, 教授 (70029289)
• Co-Investigator(Kenkyū-buntansha): IKE Michihiko Osaka University, Graduate School of Engineering, Dept.of Environmental Engineering, Associate Professor, 大学院・工学研究科, 助教授 (40222856) ; SODA Satoshi Osaka University, Graduate School of Engineering, Dept.of Global Architecture, Assistant Professor, 大学院・工学研究科, 助手 (30322176) ; SEI Kazunari Osaka University, Graduate School of Engineering, Dept.of Environmental Engineering, Assistant Professor, 大学院・工学研究科, 助手 (80324177)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥15,700,000 (Direct Cost: ¥15,700,000); Fiscal Year 2004: ¥7,000,000 (Direct Cost: ¥7,000,000); Fiscal Year 2003: ¥8,700,000 (Direct Cost: ¥8,700,000)
• Keywords: bioremediation / exogenously introduced microorganisms / genetically engineered microorganisms(GEMs) / horizontal gene transfer / pathogens / microbial community structure / nitrogen cycling function / biorisk assessment

Research Abstract

For the accurate assessment of biorisk on the use of microorganisms including genetically engineered microorganisms (GEMs) for the bioremediation of polluted soil/aquatic environment, scientific information was accumulated and systemized through experimental investigations and model simulations.
In the first year, biorisks on the horizontal transfer of the genes derived from exogenously introduced microorganisms to the indigenous harmful microorganisms such as pathogens were focused and investigated the factors which decide the frequency of the horizontal gene transfer as follows using model microcosm systems : 1.Survival of the GEMs introduced into natural environment, 2.Existence of the transmissible plasmids which have potential to transfer genetically engineered DNA sequences(GEDS), 3.Frequency and factors of plasmid transfer. The results were as follows : 1.GEMs introduced into soil and aquatic microcosms immediately decreased mainly because of the competition with indigenous micro organisms but after that survived at low level for relatively long periods, 2.There existed a certain level of microorganisms possessing tra gene, and therefore, the ability to transfer GEDS in natural environment, 3.GEDS could be transferred to wide range of indigenous microorganisms. These results implied that the GEDS can be transferred to indigenous microorganisms when GEMs possessing GEDS are released into the environment. The frame of the model to explain the behavior of the GEDS was also developed.
In the second year, 1.The transferability of the GEDS by conjugation, 2.The influence of the various environmental factors on the frequency of the transfer were evaluated in detail using soil microcosms, and 3.The behavior of the GEDS was analyzed by model. Furthermore, 4.The behavior of exogenously introduced microorganisms and the influence on indigenous microbial community were evaluated at the real bioremediation fields. The results were as follows : 1.The possibility of the conjugative gene transfer was considerably lower in the soil environments than under the optimal laboratory condition, 2.The frequency of the transfer was influenced by the levels of the organic carbons, metals, and water contents, 3.The behavior of GEDS by conjugative transfer could be expressed by the modified mass action model, and 4.The exogenously introduced microorganisms decreased rapidly and no adverse effects were observed during bioremediation of real crude oil and dioxin contaminated soils.

Research Products

• [Journal Article] Comparative evaluation of quantitative PCR methods for routine enumeration of specific bacterial DNA in aquatic samples. (2005)
  • Author(s): Inoue, D., Wada, K., Sei, K., Ike, M., Fujita, M.
  • Journal Title: World Journal of Microbiology and Biotechnology (未定)(印刷中)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Transferability of plasmids by conjugation in soil and influence of exogenous plasmid-harboring bacteria on the bacterial community : a model study for risk assessment on the use of genetically modified organisms. (2005)
  • Author(s): Inoue, D., Murashige, K., Sei, K., Ike, M., Fujita, M.
  • Journal Title: IBBS-13, 13^<th> International Biodeterioration and Biodegradation Symposium (印刷中)(未定)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Comparative evaluation of quantitative PCR methods for routine enumeration of specific bacterial DNA in aquatic samples. (2005)
  • Author(s): Inoue, D., Wada, K., Sei, K., Ike, M., Fujita, M.
  • Journal Title: World Journal of Microbiology and Biotechnology (In Press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Transferability of plasmids by conjugation in soil and influence of exogenous plasmid-harboring bacteria on the bacterial community : a model study for risk assessment on the use of genetically modified organisms. (2005)
  • Author(s): Inoue, D., Murashige, K., Sei, K., Ike, M., Fujita, M.
  • Journal Title: IBBS-13, 13^<th> International Biodeterioration and Biodegradation Symposium (Accepted)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Comparison evaluation of quantitative PCR methods for general routine enumeration of specific bacterial DNA in aquatic samples (2005)
  • Author(s): Inoue, D., Wada, K, Sei, K., Ike, M., Fujita, M.
  • Journal Title: World Journal of Microbiology and Biotechnology 未定
• [Journal Article] Monitoring behaviour of catabolic genes and change of microbial community structures in seawater microcosms during aromatic compounds degradation (2004)
  • Author(s): Sei, K., Inoue, D., Wada, K., Mori, K., Ike, M., Kohno, T., Fujita, M.
  • Journal Title: Water Research 38 Pages: 4405-4414
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Conjugation plasmid transfer from exogenously introduced genetically engineered microorganisms to environmental bacteria for fundamental investigation of bio-risk potential. (2004)
  • Author(s): Fujita, M., Ike, M., Sei, K., Inoue, D., Murashige, K.
  • Journal Title: Proceedings of 7^<th> Biennial Symposium of International Society of Environmental Biotechnology
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Monitoring behavior of catabolic genes and change of microbial community structures in seawater microcosms during aromatic compounds degradation. (2004)
  • Author(s): Sei, K., Inoue, D., Wada, K., Mori, K., Ike, M., Kohno, T., Fujita, M.
  • Journal Title: Water Research 38-20 Pages: 4405-4414
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Conjugative plasmid transfer from exogenously introduced genetically engineered microorganisms to environmental bacteria for fundamental investigation of bio-risk potential. (2004)
  • Author(s): Fujita, M., Ike, M., Sei, K., Inoue, D., Murashige, K.
  • Journal Title: Proceedings of 7^<th> Biennial Symposium of International Society of Environmental Biotechnology O-46
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Monitoring behavior of catabolic genes and change of microbial community structures in seawater microcosms during aromatic compounds degradiation. (2004)
  • Author(s): Sei, K., Inoue, D., Wada, K., Mori, K., Ike, M., Kohno, T., Fujita, M
  • Journal Title: Water Research Vol.38(No.20) Pages: 4405-4414
• [Journal Article] Conjugative plasmid transfer from exogenously introduced genetically engineered micro-organisms to environmental bacteria for fundamental investigation of bio-risk potential (2004)
  • Author(s): Fujita, M., Ike, M., Sei, K., Inoue, D., Murashige, K.
  • Journal Title: Proceedings of 7^<th> Biennial Symposium of International Society of Environmental Biotechnology