Conventional culture methods with commercially available media unveil the presence of novel culturable bacteria

Gut Microbes. 2019;10(1):77-91. doi: 10.1080/19490976.2018.1491265. Epub 2018 Aug 17.

Abstract

Recent metagenomic analysis has revealed that our gut microbiota plays an important role in not only the maintenance of our health but also various diseases such as obesity, diabetes, inflammatory bowel disease, and allergy. However, most intestinal bacteria are considered 'unculturable' bacteria, and their functions remain unknown. Although culture-independent genomic approaches have enabled us to gain insight into their potential roles, culture-based approaches are still required to understand their characteristic features and phenotypes. To date, various culturing methods have been attempted to obtain these 'unculturable' bacteria, but most such methods require advanced techniques. Here, we have tried to isolate possible unculturable bacteria from a healthy Japanese individual by using commercially available media. A 16S rRNA (ribosomal RNA) gene metagenomic analysis revealed that each culture medium showed bacterial growth depending on its selective features and a possibility of the presence of novel bacterial species. Whole genome sequencing of these candidate strains suggested the isolation of 8 novel bacterial species classified in the Actinobacteria and Firmicutes phyla. Our approach indicates that a number of intestinal bacteria hitherto considered unculturable are potentially culturable and can be cultured on commercially available media. We have obtained novel gut bacteria from a healthy Japanese individual using a combination of comprehensive genomics and conventional culturing methods. We would expect that the discovery of such novel bacteria could illuminate pivotal roles for the gut microbiota in association with human health.

Keywords: 16S rRNA gene metagenomic sequencing; Human gut microbiota; commercially available media; novel bacteria; unculturable bacteria; whole genome sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / classification
  • Bacteria / genetics
  • Bacteria / growth & development*
  • Bacteria / isolation & purification*
  • Bacteriological Techniques*
  • Culture Media
  • Feces / microbiology
  • Gastrointestinal Microbiome / genetics
  • Gastrointestinal Microbiome / physiology*
  • Genome, Bacterial / genetics
  • Humans
  • Metagenomics
  • Phylogeny
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA

Substances

  • Culture Media
  • RNA, Ribosomal, 16S

Grants and funding

Japan Agency for Medical Research and Development [JP16fk0108119]; Japan Agency for Medical Research and Development [JP17fk0108121]; Japan Agency for Medical Research and Development [JP17fk0108219]; Japan Agency for Medical Research and Development [JP16fk0108305].