title: A six-well plate method: less laborious and effective method for cultivation of obligate anaerobic microorganisms.
abstract: 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.
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan. knak2007@gifu-u.ac.jp
authors.forename: Kohei
authors.initials: K
authors.lastname: Nakamura
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Tamaki
authors.forename: Myung Suk
authors.initials: MS
authors.lastname: Kang
authors.forename: Hanako
authors.initials: H
authors.lastname: Mochimaru
authors.forename: Sung-Taik
authors.initials: ST
authors.lastname: Lee
authors.forename: Kazunori
authors.initials: K
authors.lastname: Nakamura
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Kamagata
mesh_terms: D000693:Anaerobiosis; D001105:Archaea / Q000254:growth & development* / Q000302:isolation & purification*; D001419:Bacteria / Q000254:growth & development* / Q000302:isolation & purification*; D015169:Colony Count, Microbial; D008828:Microbiological Techniques / Q000379:methods*