The positive isolation rates of spirochete from Apodemus agrarius was 17.65% (3 strains

isolated from 17 Apodemus agrarius) for the site in Jingping, and 6.25% (1 strain isolated from 16 Apodemus agrarius) for the site in Liping (Table 2). Results of serogroup buy Ku-0059436 identification of leptospiral isolates MAT was performed using a battery of anti-serum against the Chinese reference strains belonging to 15 serovars in 15 serogroups. All the four strains agglutinated with anti-serum against reference strain 56601 belonging to serovars Lai of serogroup Icterohaemorrhagiae with titres ≥100, and no positive results of MAT were observed with anti- serum against to strains belong to the other serogroups (Table 3), according to the determine standard Luminespib mouse that samples with titres ≥100 were recognized as positive. Table 3 Results of MAT identification for leptospires isolated from Apodemus agrarius in Guizhou Province Anti-serum against the Chinese reference strains belonging

to 15 serovars in 15 serogroups MAT results (titres) of isolated strains Anti-Serum No. Strain Serovar Serogroup JP13 JP15 JP19 LP62 56601 Lai Lai Icterohaemorrhagiae + (1:800) + (1:800) + (1:800) + (1:400) 56602 M10 Javanica Javanica – - – - 56603 Lin Canicola Canicola – - – - 56604 Pishu Ballum Ballum – - – - 56605 4 Pyrogenes Pyrogenes – - – - 56606 Lin 4 Autumnalis Autumnalis – - – - 56607 Sep-65 Australis Australis – - – - 56608 Luo Pomona Pomona – - – - 56609 Lin 6 Linhai Grippotyphosa – - – - 56610 P7 Hebdomadis Hebdomadis – - – - 56612 L37 Paidjian Bataviae – - – - 56613 65-52 Tarassovi Tararrovi – - – - 56615 L 105 Cingshui Manhao – - – - 56635 L 138 Sejroe Wolffi – - – - 56655 Nan 10 Mini Mini – - – - +: Positive; -: Negative. MLST pattern of leptospiral isolates Seven MLST loci based primers were used to amplify the chromosome DNA of leptospiral isolates, and all of the seven loci were successfully amplified from the four isolates. The Isotretinoin MLST pattern showed that the four isolates produced a same size of PCR segment

at the same locus (Figure 1). Figure 1 PCR products from the seven selected MLST loci of four leptospiral strains isolated from Jinping and Liping County, Guizhou province. PCR products were electrophoresised through a 1.2% agarose gel. M: 100 bp DNA Ladder; 1, Leptospira isolate JP13; 2, Leptospira isolate JP15; 3, Leptospira isolate JP19; 4, Leptospira isolate JP62. ST of leptospiral isolates Seven loci (pntA, sucA, fadD, tpiA, pfkB, mreA, and glmU) of the chromosome DNA of the four leptospiral isolates were successfully sequenced. The sequences were analysed following the standard MLST protocol which can be accessed at http://​leptospira.​mlst.​net, an allele number was assigned to all the allele of different leptospiral strains and the allelic profile (string of seven integers) was defined as sequence type 1 (ST1: 1-1-1-1-1-1-1) (Figure 2).