Application of a single‐colony coculture technique to the isolation of hitherto unculturable gut bacteria. (February 2015)

• Record Type: Journal Article
• Authors: Tanaka, Yoshiki; Benno, Yoshimi
• Is Part Of: Microbiology and immunology. Volume 59:Number 2(2015:Feb.)
• Page Start: 63
• Page End: 70
• Publication Date: 2015-02
• Subjects: Microbiology -- Periodicals; Immunology -- Periodicals; Allergy and Immunology -- Periodicals; Microbiology -- Periodicals; Microbiologie -- Périodiques; Immunologie -- Périodiques; 579
• DOI: 10.1111/1348-0421.12220
• Languages: English
• ISSNs: 0385-5600

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>ABSTRACT</title> <sec id="mim12220-sec-0001" sec-type="section"> <p>Molecular studies have led to postulation of a relationship between gut microbiota and certain diseases. However, because studies of hitherto uncultured species <italic>in vivo</italic> are essential for characterizing the biology and pathogenic properties of gut bacteria, techniques for culturing and isolating such bacteria must be developed. Here, a technique is described that partially overcomes the obstacles that prevent detection of interbacterial communication <italic>in vitro</italic> and are thus responsible for the failure to culture certain bacterial species. For this purpose, a ring with a membrane filter at the bottom was designed and a relatively simple nutrient medium was used instead of conventional media. Gut bacteria were cocultivated in soft agar separated by the membrane filter to simulate interbacterial communication <italic>in vitro</italic>. Use of this soft agar coculture technique led to the successful isolation of hitherto uncultured bacteria and the demonstration of multistage interbacterial communication among gut bacteria <italic>in vitro</italic>. Cultivation and isolation of single colonies of bacteria that require other bacteria for growth will enhance efforts to better understand the physiological and pathogenic roles of gut microbiota.</p> </sec> </abstract>