Characterization of the isolate
The isolate was serologically characterized using polyclonal antibodies according to previous recommendation (Dikken and Kmety, 1978). A panel of 34 specific antisera representing 28 serogroups was used (Andamana, Australis, Autumnalis, Ballum, Bataviae, Canicola, Celledoni, Codice, Cynopteri, Djasiman, Grippotyphosa, Hebdomadis, Holland, Icterohaemorrhagiae, Javanica, Louisiana, Lyme, Manhao, Mini, Panama, Pomona, Pyrogenes, Ranarum, Sarmin, Sejroe, Semaranga, Shermani and Tarassovi), and serogroup identification was confirmed by the identification of the highest titration against specific representative serovars included in the panel.
For molecular characterization, DNA from the isolated strain was extracted using Illustra Bacterium Genomic Prep Mini Spin kit following the manufacturer’s instructions (GE Healthcare, Sao Paulo, SP, Brazil). The complete genome was sequenced through an Ion Torrent Personal Genome Machine (PGM) (Life Technologies, Saint Aubin, France) sequencing platform, using a 316 chip (Life Technologies) and single-end libraries to generate readings of 400 bpon the BAM format, which were then converted to the FASTQ format with the bamTofastq tool from the BEDTools toolset. For the identification of Leptospira species, the generated contigs were aligned using BLAST against the Multilocus Sequence Typing (MLST) sequence sets proposed elsewhere (Boonsilp et al., 2013). The allelic profile and sequence type obtained (ST) was compared to other publicly available sequences from the PubMLST (http://pubmlst.org/leptospira).