Whole Genome-Sequencing and Phylogenetic Analysis of a Historical Collection of Bacillus anthracis Strains from Danish Cattle

PLoS One. 2015 Aug 28;10(8):e0134699. doi: 10.1371/journal.pone.0134699. eCollection 2015.


Bacillus anthracis, the causative agent of anthrax, is known as one of the most genetically monomorphic species. Canonical single-nucleotide polymorphism (SNP) typing and whole-genome sequencing were used to investigate the molecular diversity of eleven B. anthracis strains isolated from cattle in Denmark between 1935 and 1988. Danish strains were assigned into five canSNP groups or lineages, i.e. A.Br.001/002 (n = 4), A.Br.Ames (n = 2), A.Br.008/011 (n = 2), A.Br.005/006 (n = 2) and A.Br.Aust94 (n = 1). The match with the A.Br.Ames lineage is of particular interest as the occurrence of such lineage in Europe is demonstrated for the first time, filling an historical gap within the phylogeography of the lineage. Comparative genome analyses of these strains with 41 isolates from other parts of the world revealed that the two Danish A.Br.008/011 strains were related to the heroin-associated strains responsible for outbreaks of injection anthrax in drug users in Europe. Eight novel diagnostic SNPs that specifically discriminate the different sub-groups of Danish strains were identified and developed into PCR-based genotyping assays.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anthrax / microbiology
  • Anthrax / veterinary*
  • Bacillus anthracis / classification*
  • Bacillus anthracis / genetics
  • Bacillus anthracis / isolation & purification*
  • Bacterial Typing Techniques / methods
  • Cattle
  • Cattle Diseases / microbiology*
  • DNA, Bacterial / analysis
  • Europe
  • Genome, Bacterial
  • Molecular Typing / methods
  • Phylogeny
  • Phylogeography
  • Polymorphism, Single Nucleotide
  • Sequence Analysis, DNA / methods*


  • DNA, Bacterial

Grant support

This work was supported by the French National Research Agency (ANR) and Direction Générale de l'Armement (DGA) (project number ANR11ASTR0007) (URL: http://www.agence-nationale-recherche.fr/suivibilan/ingenierie-procedes-securite/accompagnement-specifique-des-travaux-derecherches-et-dinnovation-defense/). GG is a PhD student co-supported by Anses and DGA grants (convention number A537) (URL: http://www.ixarm.com/Theses-DGA). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.