The ST 13 was formed with 10 Group-Ia low-virulence strains and one strain (Lm74905) belonging to the comparative set (in white). The analysis of this strain revealed that it exhibited the PrfAK220T mutation and the same truncated InlA characterizing the genotypic Group-Ia. Likewise, the Lm85820 strain which grouped in the ST31 (in white) exhibited the same mutation in InlA than the low-virulence strains of this ST, but no mutation in PfrA. Remarkably, although all strains of the ST31 had InlA mutations, only half of these
strains also had the PrfAΔ174-237 mutation. In this analysis, the A23 strain selleck corresponds to a singleton (ST196) with only one mismatch with Group-IIIa and two with Group-Ia. It is related to Group-Ib through ST11. Figure 3 Minimum spanning tree based on allelic profiles by using BioNumerics version 4.6. (Applied-Maths, Sint-Martens-Latem, Belgium). AZD3965 solubility dmso The comparative set included 656 L. monocytogenes strains from the French Reference BVD-523 nmr Centre for Listeria and the WHO Collaborative Centre for Foodborne Listeriosis. The experimental set included 92 L. monocytogenes strains defined as virulent (“virulent to mice”) or low-virulence (phenotypic Groups “I to VI”) using a virulence test combining a PF assay in HT-29 cells and sub-cutaneous inoculation of mice. Each circle corresponds
to a sequence type (ST). ST numbers are given inside the circles. The lines between STs show inferred phylogenetic relationships and are represented by bold, continuous, dotted and pale dotted lines according to the number of allelic mismatches between profiles (1, 2, 3 and 4 or more, respectively); the discontinuous links are only indicative, as alternative links of equal weight may exist. Phenotypic Groups (I to VI) of low-virulence and virulent L. monocytogenes Phosphoprotein phosphatase strains are marked in color. The comparative set of L. monocytogenes strains are in white. Specific STs for Groups-Ia, -Ib and -IIIa and A23 strains are in an area shaded grey. Overall, half of the low-virulence strains (22 out of 43), belonging
to the genotyping Groups-Ia, -Ib and -IIIa, are likely to have descended from a single virulent 1/2a ancestral bacterium. In contrast, the other strains were distributed into five clonal complexes and 10 STs and may be regarded as virulence variants of L. monocytogenes strains. Contribution of the optical mapping To investigate the genomic relationship between the A23 strain and the closely related low-virulence strains belonging to Group-IIIa strains, two strains (BO43 and 416) were compared with the A23 strain using optical mapping and the in silico reference EGDe map (Figure 4). The EGDe optical map was approximately 20% different from the maps of the Group-IIIa and A23 strains, whereas the A23 strain showed 99% similarities with Group-IIIa.