Metagenomic Analysis of Silage

J Vis Exp. 2017 Jan 13:(119):54936. doi: 10.3791/54936.

Abstract

Metagenomics is defined as the direct analysis of deoxyribonucleic acid (DNA) purified from environmental samples and enables taxonomic identification of the microbial communities present within them. Two main metagenomic approaches exist; sequencing the 16S rRNA gene coding region, which exhibits sufficient variation between taxa for identification, and shotgun sequencing, in which genomes of the organisms that are present in the sample are analyzed and ascribed to "operational taxonomic units"; species, genera or families depending on the extent of sequencing coverage. In this study, shotgun sequencing was used to analyze the microbial community present in cattle silage and, coupled with a range of bioinformatics tools to quality check and filter the DNA sequence reads, perform taxonomic classification of the microbial populations present within the sampled silage, and achieve functional annotation of the sequences. These methods were employed to identify potentially harmful bacteria that existed within the silage, an indication of silage spoilage. If spoiled silage is not remediated, then upon ingestion it could be potentially fatal to the livestock.

Publication types

  • Video-Audio Media

MeSH terms

  • Animals
  • Bacteria / genetics
  • Cattle
  • Computational Biology
  • Metagenomics / methods*
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA / methods
  • Silage / microbiology*

Substances

  • RNA, Ribosomal, 16S