The pathogenic strains represent the main causative serovars for Leptospirosis in humans and animals. The most common strains used in MAT panels belong to the three genomospecies L. interrogans, L. borgpetersenii and L. kirschneri (Table 1). All strains were cultured in Ellinghausen-McCullough-Johnson-Harris medium (Leptospira Medium Base EMJH BD, selleck compound DifcoTM and Leptospira Enrichment EMJH DifcoTM, NJ, USA) at 28°C. Cultures were controlled for growth and motility by darkfield microscopy and were periodically subcultured into fresh media. Bacteria used for MALDI-TOF MS measurements and protein reference spectra generation were cultured for seven days.

Table 1 Leptospira reference strains used for MALDI-TOF MS measurements and sequence analysis genomospecies serogroup serovar strain pathogenicity L. interrogans Australis Australis Ballico a, b pathogenic L. interrogans eFT508 concentration Australis Bratislava Jez Bratislava a, b pathogenic L. interrogans Autumnalis Autumnalis Akiyami Protein Tyrosine Kinase inhibitor A a, b pathogenic L. interrogans Bataviae Bataviae Swart a, b pathogenic L. interrogans Canicola Canicola Hond Utrecht IV a,

b pathogenic L. interrogans Hebdomadis Hebdomadis Hebdomadis a, b pathogenic L. interrogans Icterohaemorrhagiae Copenhageni M 20 a, b pathogenic L. interrogans Icterohaemorrhagiae Icterohaemorrhagiae Ictero I b pathogenic L. interrogans Pomona Pomona Pomona a,b pathogenic L. interrogans Pyrogenes Pyrogenes Salinem a,b pathogenic L. interrogans Sejroe Hardjo Hardjoprajitno a,b pathogenic L. kirschneri Grippotyphosa Grippotyphosa Moskva V a.b pathogenic L. borgpetersenii Sejroe Saxkoebing Mus 24

a, b pathogenic L. borgpetersenii Ballum Ballum Mus 127 a, b pathogenic L. borgpetersenii Sejroe Sejroe M 84 a, b pathogenic L. borgpetersenii Tarassovi Cytidine deaminase Tarassovi Perepelitsin a, b pathogenic L. borgpetersenii Javanica Javanica Veldrat Bataviae 46 b pathogenic L. alexanderi not defined Manhao 3 L60c pathogenic L. weilii not defined Celledoni Celledoni c pathogenic L. santarosai not defined Shermani LT 821 c pathogenic L. noguchii not defined Panama CZ 214 c pathogenic L. broomii not defined Not defined 5399 c intermediate L. fainei not defined Hurstbridge BUT 6 c intermediate L. inadai not defined Lyme 10 c intermediate L. biflexa Semaranga Patoc PatocI c non-pathogenic L. meyeri not defined Semaranga Veldrat S173 c non-pathogenic Turneriella parva not defined Parva H c non-pathogenic Leptonema illini not defined Illini 3055 c non-pathogenic a Acquired by purchase at the Federal Institute for Risk Assessment (BfR) Head of Unit. Diagnostics, Genetics and Pathogen Characterisation, Department Biological Safety Berlin, Germany. b Acquired by purchase at the WHO/FAO Collaborating Centre for Reference and Research on Leptospirosis, Biomedical Research, Royal Tropical Institute (KIT) Amsterdam, The Netherlands. c Acquired by purchase at the DSMZ – German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany.