interrogans Bataviae – - – - Selleckchem Defactinib – + + – + – L. kirschneri Grippotyphosa – - – - – + – - + + The displayed peaks are based on visual comparison of the algorithms analysis results of the software. All strains were screened twice using the QuickClassifier (QC)/Different average and SNN algorithms. The used symbols stand for: no peak found: – peak present: + peak set with high intensity: ++ Table 5 Differentiating peaks based on the statistical analysis of ClinProTools within the species L. borgpetersenii genomospecies peak mass (m/z) representing

the protein size in Dalton 3,759 5,765 5,779 6,388 7,519 7,547 L. borgpetersenii Ballum + – + – + – L. borgpetersenii Javanica + – + – + – L. borgpetersenii Sejroe + – + – + – L. borgpetersenii Saxkoebing – - + + – + L. borgpetersenii Tarassovi + + – + + – The displayed peaks

are based on visual comparison of the algorithm analysis results of the software. All strains were screened twice using the QuickClassifier (QC)/Different average and SNN algorithms. The used symbols stand for: MDV3100 cost no peak found: – peak present: + The additional statistical tool Principal component analysis (PCA) included in ClinProTools was applied to the analyzed datasets to visualize the homogeneity and heterogeneity of the protein spectra. PCA reduces the variables of a complex dataset on the basis of different statistical tests. The reduced datasets, the so-called PCs (principle components) can be displayed in a score plot illustration. Twenty individual protein spectra of the Silibinin L. interrogans strains and the L. kirschneri strain are displayed in three-dimensional PCA in Figure 2. Each dot stands for a displayed protein spectrum. The colors indicate the calculated cluster membership in which each dot represents one measured protein spectrum

profile for each sample. A clear separation of the serovars Pomona and Copenhageni is apparent. Conversely, L. kirschneri serovar Grippotyphosa did not cluster separately in PCA analysis, even if specific peaks could be detected for L. kirschneri in the peak statistics (see Table 4). For the genomospecies L. borgpetersenii the separation of the serovars Saxkoebing, Sejroe and Tarassovi was apparent when PCA was performed (Figure 3). Figure 2 Principle Component Analysis (PCA) of the analyzed strains of the genomospecies. L. interrogans and L. kirschneri using the software tool ClinProTools. Figure 3 Principle Component Analysis (PCA) of the analyzed strains of the genomospecies. L. borgpetersenii using the software tool ClinProTools. Strain confirmation and molecular sequencing Sequence analysis of the 28 leptospiral reference strains was performed on the basis of MLST analysis (Figure 4) and 16S rRNA gene sequencing (Figure 5). Confirmation of the field isolates relied on 16S rRNA gene sequencing. Species identity of all used strains was confirmed. Furthermore, the constructed phylogentic trees (Figures 4 and 5) revealed comparable clustering of the leptospiral strains.