37 [20] FFIVC131 CDC2412-93 O139   1 1 0 1995 Human USA 1 1 1 1 1 1 2.43 [20] FFIVC133   O139   1 1 0 2003 unknown unknown 1 1 1 1 1 1 2.49 [20] 080025/FR Vib31 O141   1 1 1 1993 Human Spain singleton 8 7 3 2 9 2.24 [18] FFIVC050   non O1/O139   0 0 0   Mussels Norway singleton 8 9 9 11 5 2.28 [20] FFIVC084   non O1/O139   0 0 0 2003 Mussels Norway singleton 4 2 4 3 3 2.45 [20] FFIVC114   non O1/O139   0 0 0 2004 Water Norway 4 6 1 6 6 6 2.29 [20] FFIVC115   non O1/O139  

0 0 0 2004 Water Norway 4 6 1 6 6 6 2.39 [20] FFIVC137   non O1/O139   0 0 0   Human Norway singleton 7 5 8 10 4 2.41 [20] 2/110/2006   non O1/O139   0 0 0 1998 Water selleck Poland 5 10 4 2 12 4 2.25 [18] 3/110/2006   non O1/O139   0 0 0 1998 Water Poland 5 10 4 2 12 4 2.42 [18] 4/110/2006   non O1/O139   0 0 0 2004 Water Poland singleton

11 0 13 0 11 2.38 [18] 14/110/2006   non O1/O139   0 0 0 1998 Water Poland singleton 5 3 10 4 7 2.37 [18] 17/110/2006   non O1/O139   0 0 0 1998 Water Poland 6 3 6 7 5 10 2.47 [18] 22/110/2006   non O1/O139   0 0 0 2004 Water Poland 6 3 6 7 5 10 2.26 [18] 070256/J V. mimicus ATCC 33655 –   1 0 0     10 14 10 12 1 14 1.71 [18] a“0” means no PCR product was obtained. bMSP value: highest logarithmic value of the four generated MS-spectra score value compared to Biotyper reference library. cReference(s), in which the isolate EPZ5676 manufacturer is described previously. Confirmation of strain identification Identification of the isolates at species level was confirmed by MALDI-TOF MS using Biotyper 2.0 (Bruker Daltonics GmbH, Bremen, Germany) [11]. Serogroup and serotype were confirmed using the Vibrio cholerae E Agglutinating Sera kit containing specific antisera O1 polyvalent agglutination serum, Inaba agglutination serum, and Ogawa agglutination serum (Remel Europe Ltd. Darford, Kent, United Kingdom) according to the manufacturer’s guidelines. Genotyping of isolates with multilocus sequence typing (MLST) analysis MLST analysis was performed

according to Teh et al. [21]. Internal gene fragments of dnaE, lap, recA, gyrB, and cat were PCR amplified and sequenced. The gmd gene was not included in the analysis due to low discriminatory power [21]. Each sequence variant of a locus was assigned a distinct allele number. In the case that no PCR product could be obtained for a specific allele, the Selleck BI-2536 number zero was assigned. The allele profiles next were entered into BioNumerics version 6.6 software (Applied-Maths, Belgium) as character values, and the genetic relationship between isolates was constructed using the categorical coefficient and the Minimum Spanning Tree algorithm. Isolates that differed at two or fewer loci were considered genetically closely related, while single locus variants (SLV) were defined as having at least three alleles that were different from all other tested isolates.