We describe the genetic diversity of 1327 Brucella strains from human patients in Kazakhstan using multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA). All strains were assigned to the Brucella melitensis East Mediterranean group and clustered into 16 MLVA11 genotypes, nine of which are reported for the first time. MLVA11 genotype 116 predominates (86.8%) and is present all over Kazakhstan indicating existence and temporary preservation of a “founder effect” among B. melitensis strains circulating in Central Eurasia. The diversity pattern observed in humans is highly similar to the pattern previously reported in animals. The diversity observed by MLVA suggested that the epidemiological status of brucellosis in Kazakhstan is the result of the introduction of a few lineages, which have subsequently diversified at the most unstable tandem repeat loci. This investigation will allow to select the most relevant strains for testing these hypotheses via whole genome sequencing and to subsequently adjust the genotyping scheme to the Kazakhstan epidemiological situation.

Tularemia is a highly dangerous zoonotic infection due to the bacteria Francisella tularensis. Low genetic diversity promoted the use of polymorphic tandem repeats (MLVA) as first-line assay for genetic description. Whole genome sequencing (WGS) is becoming increasingly accessible, opening the perspective of a time when WGS might become the universal genotyping assay. The main goal of this study was to describe F. tularensis strains circulating in Kazakhstan based on WGS data and develop a MLVA assay compatible with in vitro and in silico analysis. In vitro MLVA genotyping and WGS were performed for the vaccine strain and for 38 strains isolated in Kazakhstan from natural water bodies, ticks, rodents, carnivores, and from one migratory bird, an Isabellina wheatear captured in a rodent burrow. The two genotyping approaches were congruent and allowed to attribute all strains to two F. tularensis holarctica lineages, B.4 and B.12. The seven tandem repeats polymorphic in the investigated strain collection could be typed in a single multiplex PCR assay. Identical MLVA genotypes were produced by in vitro and in silico analysis, demonstrating full compatibility between the two approaches. The strains from Kazakhstan were compared to all publicly available WGS data of worldwide origin by whole genome SNP (wgSNP) analysis. Genotypes differing at a single SNP position were collected within a time interval of more than fifty years, from locations separated from each other by more than one thousand kilometers, supporting a role for migratory birds in the worldwide spread of the bacteria.

Almost every year in Kazakhstan, cases of diseases of animals and people with anthrax are recorded. The incidence rate of people with anthrax in the period from 2000 to 2018 ranged from 0.01 to 0.24 per 100 thousand people. In the territory of the East Kazakhstan region, the incidence rate is higher than in the republic.Objective: a retrospective analysis of anthrax outbreaks in the East Kazakhstan region from 2000 to 2018.Methods: a retrospective analysis using statistical, cadastral data, archival funds, the results of participation in the investigation of outbreaks of infection.Results. Since 1938, cases of animal and human disease of anthrax have been recorded in East Kazakhstan. Infection of humans occurs during the slaughter of animals. In Zharma, Urjar, and Ayagozskiy areas most of all are anthrax foci of soil. In 67 % of cases, outbreaks of anthrax were recorded in these areas.Conclusion. The relative incidence rate of anthrax in people in East Kazakhstan is from 0.07 to 0.27, since 2001 it is higher than the republican indicator.In the East Kazakhstan region from 1997 to 2018, 37 people fell ill with anthrax. The form of the disease is skin, in 8.1 % secondary sepsis. In the remaining patients, the diseases were mild (70.3 %), ended in recovery. The source of human infection is mainly cattle. The isolated strains of B. anthracis have typical properties, they are included in cluster A1a, A3b (MLVA-8). The strains isolated in 2016 are similar to the strains isolated in the Almaty region (MLVA-25). Grouped with a number of European, Asian and African strains from France, Germany, Italy, Tajikistan, Pakistan, Korea and Namibia. The presence of a significant number of soil foci of anthrax in the territory of the East Kazakhstan region, not fully the implementation of veterinary and sanitary preventive measures leads to an exacerbation of the situation for anthrax.

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.

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