Small inter and intra-scaffold gaps were selleck kinase inhibitor closed by PCR and Sanger sequencing. Seven larger gaps were closed using long range PCR and Illumina sequencing. Illumina reads were assembled

using Velvet [87], and the optimum assembly was determined using the N50 statistic. Annotation of the genome assembly was performed using the NCBI Prokaryotic Genomes Automatic Annotation Pipeline (PGAAP) and Blast2GO v.2.5.0 (E value cut-off = 1e-6 PX-478 nmr and minimum amino acid alignment length cut-off [hsp-length] = 33) [88] (annotations are shown in Additional file 2). This Whole Genome Shotgun project has been deposited at DDBJ/EMBL/GenBank under the accession AIDX00000000. The version described in this paper is the first version, AIDX01000000. Homologous gene clustering The MCL algorithm [89] as implemented in the MCLBLASTLINE pipeline (available at http://​micans.​org/​mcl) was used to delineate homologous protein sequences among 214 Streptococcus GSK3326595 chemical structure genomes including S. canis (see Additional file 3). Based

on sequence similarity, the pipeline uses Markov clustering (MCL) to assign genes to homologous clusters. Similarity was obtained from a reciprocal BLASTp within and between all genome pairs using an E value cut-off of 1e-5. The MCL algorithm was implemented using an inflation parameter of 1.8. Simulations have shown this value to be generally robust to false positives and negatives [90]. Virulence factors Amino acid sequences for all S. canis CDS were searched against the VFDB using BLASTp. We used an E value cut-off of 1e-5 and retained the single best hit. The search was refined by repeating the BLASTp search against a database that contained only Streptococcus virulence factors (88 genes). Population Oxymatrine genetics Including the strain genome sequenced here, a total of 83 S. canis isolates were obtained from bovine (n = 56), canine (n = 26),

and feline (n = 1) hosts (Table 1). Isolates of canine/feline origin included 25 canine isolates from patients of Cornell University’s College of Veterinary Medicine, Ithaca, NY, USA, one canine isolate from Belgium, and one isolate from a cat living on a dairy farm in upstate New York. The feline isolate was the likely source of a mastitis outbreak at the same farm. Canine isolates from NY originated from dermis (n = 1), ear swabs (n = 7), eye (n = 1), hock abscess (n = 1), lip (n = 1), pharyngeal swabs (n = 5), urine (n = 1), and vaginal swabs (n = 8), and were collected from December 2003 to May 2004. The canine isolate from Belgium originated from wound exudate [1] and the feline isolate originated from a nasal swab taken from a cat with chronic sinusitis [12].