Innovative cultivation methods targeting environmentally key but uncultivated microbial types

Research Project

Project/Area Number	15K14696
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Applied microbiology
Research Institution	Hiroshima University
Principal Investigator	Aoi Yoshiteru 広島大学, 先端物質科学研究科, 准教授 (40386636)
Co-Investigator(Kenkyū-buntansha)	金田一智規広島大学, 工学研究科, 助教 (10379901)
Project Period (FY)	2015-04-01 – 2018-03-31
Project Status	Completed (Fiscal Year 2017)
Budget Amount *help	¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000) Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000) Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords	分離培養 / アナモックス / ポリリン酸蓄積細菌 / 難培養 / 未培養 / セルソーター / アナモックス細菌 / 分離培養装置 / 分離 / 継代培養 / 集積培養 / 連続培養
Outline of Final Research Achievements	Objective of the study is to develop innovative cultivation method and trying to cultivate uncultivated microbial types, which are key microbial species in the field of environmental science and engineering. In this study, two uncultivated microbial types were selected as a target for cultivation, those are Anaerobic ammonia oxidizing (ANAMMOX) bacteria, and Accumilibacter phosphatis, which has never been succeeded for isolation since their discovery. For ANAMMOX, fallowing methods were used in combination; 1) enrichment cultivation, 2) selective isolation of Anammox cells by using cell sorting system, 4) automatic isolation method (microbial trap) and 4) multi-stage and successive scaling up cultivation. Although, keeping the culture in alive were failed, some of the method brought success in cultivation of Anammox. For PAO, in addition to above methods, new methods were developed and applied for providing alternating condition of anaerobic / aerobic conditions.