A Six-well Plate Method: Less Laborious and Effective Method for Cultivation of Obligate Anaerobic Microorganisms

作者：Nakamura Kohei; Tamaki Hideyuki; Kang Myung Suk; Mochimaru Hanako; Lee Sung Taik; Nakamura Kazunori; Kamagata Yoichi

来源：Microbes and Environments, 2011, 26(4): 301-306.

DOI：10.1264/jsme2.ME11120

• methanogen
• sulfate-reducing bacterium (SRB)
• gellan gum
• uncultured anaerobes

We developed a simple, less laborious method to cultivate and isolate obligate anaerobic microorganisms using a six-well plate together with the AnaeroPack System, designated as the six-well plate method. The cultivation efficiency of this method, based on colony-forming units, colony formation time, and colony size, was evaluated with four authentic obligate anaerobes (two methanogenic archaea and two sulfate-reducing bacteria). The method was found to be comparable to or even better than the roll tube method, a technique that is commonly used at present for the cultivation of obligate anaerobes. Further experiments using 21 representative obligate anaerobes demonstrated that all examined anaerobes (11 methanogens, 5 sulfate- or thiosulfate-reducing bacteria, and 5 syntrophs) could form visible colonies on the six-well plate and that these colonies could be successfully subcultured in fresh liquid media. Using this method, an unidentified sulfate-reducing bacterium was successfully isolated from an environmental sample.

• 出版日期2011-12-1