Abstract

This article describes Bacillus anthracis strains isolated in Kazakhstan since the 1950s until year 2016 from sixty-one independent events associated with anthrax in humans and animals. One hundred and fifty-four strains were first genotyped by Multiple Locus VNTR (variable number of tandem repeats) Analysis (MLVA) using 31 VNTR loci. Thirty-five MLVA31 genotypes were resolved, 28 belong to the A1/TEA group, five to A3/Sterne-Ames group, one to A4/Vollum and one to the B clade. This is the first report of the presence of the B-clade in Kazakhstan. The MLVA31 results and epidemiological data were combined to select a subset of seventy-nine representative strains for draft whole genome sequencing (WGS). Strains from Kazakhstan significantly enrich the known phylogeny of the Ames group polytomy, including the description of a new branch closest to the Texas, United States A.Br.Ames sublineage stricto sensu. Three among the seven currently defined branches in the TEA polytomy are present in Kazakhstan, “Tsiankovskii”, “Heroin”, and “Sanitary Technical Institute (STI)”. In particular, strains from the STI lineage are largely predominant in Kazakhstan and introduce numerous deep branching STI sublineages, demonstrating a high geographic correspondence between “STI” and Kazakhstan, Central Asia. This observation is a strong indication that the TEA polytomy emerged after the last political unification of Asian steppes in the fourteenth century of the Common Era. The phylogenetic analysis of the Kazakhstan data and of currently available WGS data of worldwide origin strengthens our understanding of B. anthracis geographic expansions in the past seven centuries.

Highlights

  • Bacillus anthracis, a soil-borne bacterial species causing anthrax in animals and humans, combines two virulence plasmids, pXO1 and pXO2 and a Bacillus cereus sensu lato chromosomal background (Okinaka and Keim, 2016; Pena-Gonzalez et al, 2018)

  • Twenty-two and three strains were isolated from soil and manure from the place of keeping or forced slaughter of sick animals, respectively; two strains were isolated from cooked cattle meat; two strains were isolated from a shop or sausage factory, and one strain was isolated from a door handle (Supplementary Table 1)

  • Most loci are only moderately polymorphic in this collection, and a more cost-efficient assay appropriate for Kazakhstan could be devised using ten loci as previously proposed (Thierry et al, 2014; Shevtsov et al, 2019)

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Summary

Introduction

A soil-borne bacterial species causing anthrax in animals and humans, combines two virulence plasmids, pXO1 and pXO2 and a Bacillus cereus sensu lato chromosomal background (Okinaka and Keim, 2016; Pena-Gonzalez et al, 2018). It is hoped that “microbial forensics” capacities to trace back the source of an attack, might act as a deterrent vis-à-vis potential users (Schmedes et al, 2016). This goal has had a major impact regarding our knowledge of the genetic diversity of B. anthracis, and our capacity to investigate its phylogeography

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