Br.027) lineage of the B.Br.013 group phylogenetic tree in (A), and the letter corresponds to MLVA genotypes indicated in Table 2 and in Additional file 4. Subclade and MLVA genotypes are also shown for the two Crimean isolates, indicated by an arrow pointing in the direction of the Crimean peninsula (upper left). To understand the relationship of the Georgian lineage to selleck products other Eastern European lineages, we genotyped 132 geographically diverse group B.Br.013 isolates collected in Central and Eastern Europe across the B.Br.026 and B.Br.027 canSNP assays (selleckchem Figure 2A, see additional file 3). All resulting genotypes from this analysis were phylogenetically consistent with no observed homoplasy. With just two exceptions,
all of these isolates were assigned to the B.Br.026 lineage. The exceptions were two isolates from the Crimean region of Ukraine that were assigned to the Georgian lineage. Subsequent, additional canSNP analyses assigned AZD1152 cell line these two isolates to the basal B.Br.027/028 subclade within the Georgian lineage. These results indicate that the Georgian isolates, as well as the two isolates from Crimea, are phylogenetically distinct from the previously described F. tularensis subsp. holarctica
subpopulations. The subclades within the Georgian lineage did not display a differentiated phylogeographic pattern but, rather, were spatially dispersed in a mixed fashion throughout Eastern Georgia and the Crimean region of Ukraine (Figure 2B). The assignment of the Crimean isolates to the basal B.Br.027/028 subclade within the Georgian lineage (Figure 2A) confirms that this lineage is not geographically restricted to Georgia, and is Urocanase suggestive of a north to south dispersal pattern. That said, the overall geographic extent of the Georgian lineage is currently unknown due to the limited sampling in adjacent countries. Further discrimination using MLVA MLVA was used to examine genetic variation within each identified subclade of the Georgian lineage (Table 2; Additional file 4). Five unique MLVA genotypes were identified among the 25 Georgian
isolates (Table 2) that were distinct from the MLVA genotypes of strains found north of Georgia. Calculations of MLVA diversity (D = G/N) within each subclade (see methods for calculation) showed decreasing levels of diversity within higher resolution subclades (Figure 2A). The most basal Georgian subclade, B.Br.027/028 (D = 0.67) (Figure 2A), was comprised of a single Georgian isolate that was distinguishable from the two Crimean isolates in the same subclade due to a distinct MLVA genotype. There were three MLVA genotypes among the seven Georgian isolates within subclade B.Br.028/029 (D = 0.43). A single MLVA genotype was shared by all seven Georgian isolates in subclade B.Br.029/030 (D = 0.14), and the two other intermediate subclades (B.Br.030/031 and B.Br.031/032) contained only a single isolate each.