Journal
LETTERS IN APPLIED MICROBIOLOGY
Volume 56, Issue 1, Pages 63-70Publisher
WILEY
DOI: 10.1111/lam.12019
Keywords
culture media; gellan gum; isolation; rumen bacteria
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Funding
- National Agriculture and Food Research Organization (NARO), Japan
- University of Tsukuba, Japan
- Japan Society for Promotion of Science (JSPS) [24580402]
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The aim of this study was to develop novel anaerobic media using gellan gum for the isolation of previously uncultured rumen bacteria. Four anaerobic media, a basal liquid medium (BM) with agar (A-BM), a modified BM (MBM) with agar (A-MBM), an MBM with phytagel (P-MBM) and an MBM with gelrite (G-MBM) were used for the isolation of rumen bacteria and evaluated for the growth of previously uncultured rumen bacteria. Of the 214 isolates composed of 144 OTUs, 103 isolates (83 OTUs) were previously uncultured rumen bacteria. Most of the previously uncultured strains were obtained from A-MBM, G-MBM and P-MBM, but the predominant cultural members, isolated from each medium, differed. A-MBM and G-MBM showed significantly higher numbers of different OTUs derived from isolates than A-BM (P<0.05). The Shannon index indicated that the isolates of A-MBM showed the highest diversity (H'=3.89) compared with those of G-MBM, P-MBM and A-BM (H'=3.59, 3.23 and 3.39, respectively). Although previously uncultured rumen bacteria were isolated from all media used, the ratio of previously uncultured bacteria to total isolates was increased in A-MBM, P-MBM and G-MBM.
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