Shotgun Metagenomic Sequencing Reveals Functional Genes and Microbiome Associated with Bovine Digital Dermatitis

PLoS One. 2015 Jul 20;10(7):e0133674. doi: 10.1371/journal.pone.0133674. eCollection 2015.

Abstract

Metagenomic methods amplifying 16S ribosomal RNA genes have been used to describe the microbial diversity of healthy skin and lesion stages of bovine digital dermatitis (DD) and to detect critical pathogens involved with disease pathogenesis. In this study, we characterized the microbiome and for the first time, the composition of functional genes of healthy skin (HS), active (ADD) and inactive (IDD) lesion stages using a whole-genome shotgun approach. Metagenomic sequences were annotated using MG-RAST pipeline. Six phyla were identified as the most abundant. Firmicutes and Actinobacteria were the predominant bacterial phyla in the microbiome of HS, while Spirochetes, Bacteroidetes and Proteobacteria were highly abundant in ADD and IDD. T. denticola-like, T. vincentii-like and T. phagedenis-like constituted the most abundant species in ADD and IDD. Recruitment plots comparing sequences from HS, ADD and IDD samples to the genomes of specific Treponema spp., supported the presence of T. denticola and T. vincentii in ADD and IDD. Comparison of the functional composition of HS to ADD and IDD identified a significant difference in genes associated with motility/chemotaxis and iron acquisition/metabolism. We also provide evidence that the microbiome of ADD and IDD compared to that of HS had significantly higher abundance of genes associated with resistance to copper and zinc, which are commonly used in footbaths to prevent and control DD. In conclusion, the results from this study provide new insights into the HS, ADD and IDD microbiomes, improve our understanding of the disease pathogenesis and generate unprecedented knowledge regarding the functional genetic composition of the digital dermatitis microbiome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biopsy
  • Cattle
  • Cattle Diseases / genetics*
  • Cattle Diseases / microbiology*
  • Chemotaxis
  • Copper / chemistry
  • Digital Dermatitis / genetics*
  • Digital Dermatitis / microbiology*
  • Digital Dermatitis / pathology
  • Drug Resistance, Bacterial
  • Female
  • Flagella / genetics
  • Metagenome
  • Metagenomics
  • Microbiota*
  • Phylogeny
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA
  • Treponema / genetics
  • Zinc / chemistry

Substances

  • RNA, Ribosomal, 16S
  • Copper
  • Zinc

Grants and funding

Funding was provided for research and publication by the USDA/NIFA Federal Formula Funds project NYC-478438. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.