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

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.

[1]  T Berger,et al.  Injectional anthrax - new presentation of an old disease. , 2014, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[2]  P. Gajer,et al.  Phylogeography of Bacillus anthracis in the Country of Georgia Shows Evidence of Population Structuring and Is Dissimilar to Other Regional Genotypes , 2014, PloS one.

[3]  S. Derzelle,et al.  Application of High-Throughput Sequencing: Discovery of Informative SNPs to Subtype Bacillus anthracis , 2014 .

[4]  Markus H. Antwerpen,et al.  Injectional Anthrax in Heroin Users, Europe, 2000–2012 , 2014, Emerging infectious diseases.

[5]  G. Vergnaud,et al.  High-throughput sequencing of Bacillus anthracis in France: investigating genome diversity and population structure using whole-genome SNP discovery , 2014, BMC Genomics.

[6]  Koichiro Tamura,et al.  MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. , 2013, Molecular biology and evolution.

[7]  S. Derzelle,et al.  Genetic diversity of Bacillus anthracis in Europe: genotyping methods in forensic and epidemiologic investigations. , 2013, Biosecurity and bioterrorism : biodefense strategy, practice, and science.

[8]  R. Grunow,et al.  Anthrax among heroin users in Europe possibly caused by same Bacillus anthracis strain since 2000. , 2013, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[9]  J. Kalinowski,et al.  Draft Genome Sequence of Bacillus anthracis UR-1, Isolated from a German Heroin User , 2012, Journal of bacteriology.

[10]  N. Bergman,et al.  Molecular Epidemiologic Investigation of an Anthrax Outbreak among Heroin Users, Europe , 2012, Emerging infectious diseases.

[11]  Dawn N. Birdsell,et al.  Melt Analysis of Mismatch Amplification Mutation Assays (Melt-MAMA): A Functional Study of a Cost-Effective SNP Genotyping Assay in Bacterial Models , 2012, PloS one.

[12]  Hengliang Wang,et al.  Curing of Plasmid pXO1 from Bacillus anthracis Using Plasmid Incompatibility , 2012, PloS one.

[13]  A. Hoffmaster,et al.  Molecular Epidemiology of Anthrax Cases Associated with Recreational Use of Animal Hides and Yarn in the United States , 2011, PloS one.

[14]  P. Le Flèche,et al.  Characterization of Genetic Diversity of Bacillus anthracis in France by Using High-Resolution Melting Assays and Multilocus Variable-Number Tandem-Repeat Analysis , 2011, Journal of Clinical Microbiology.

[15]  Mihai Pop,et al.  Bacillus anthracis comparative genome analysis in support of the Amerithrax investigation , 2011, Proceedings of the National Academy of Sciences.

[16]  S. Banno,et al.  Genome-Wide Single Nucleotide Polymorphism Typing Method for Identification of Bacillus anthracis Species and Strains among B. cereus Group Species , 2010, Journal of Clinical Microbiology.

[17]  P. Keim,et al.  The genome and variation of Bacillus anthracis. , 2009, Molecular aspects of medicine.

[18]  M. Hugh-jones,et al.  The ecology of Bacillus anthracis. , 2009, Molecular aspects of medicine.

[19]  J. Foster,et al.  Pre-Columbian Origins for North American Anthrax , 2009, PloS one.

[20]  J. Ravel,et al.  Bacillus anthracis in China and its relationship to worldwide lineages , 2009, BMC Microbiology.

[21]  Paul Keim,et al.  Single Nucleotide Polymorphism Typing of Bacillus anthracis from Sverdlovsk Tissue , 2008, Emerging infectious diseases.

[22]  Lynn Y. Huynh,et al.  Global Genetic Population Structure of Bacillus anthracis , 2007, PloS one.

[23]  W. Ryan Easterday,et al.  Strain-Specific Single-Nucleotide Polymorphism Assays for the Bacillus anthracis Ames Strain , 2006, Journal of Clinical Microbiology.

[24]  D. Pauze,et al.  Clinical presentation of inhalational anthrax following bioterrorism exposure: report of 2 surviving patients. , 2001, JAMA.

[25]  M. Jensenius,et al.  Injectional anthrax in a heroin skin-popper , 2000, The Lancet.

[26]  M. Hugh-jones,et al.  1996–97 global anthrax report , 1999, Journal of applied microbiology.