Two Gram-negative, non-motile, non-spore-forming coccoid bacteria (strains F8/08-60 T and F8/ 08-61) isolated from clinical specimens obtained from baboons (Papio spp.) that had delivered stillborn offspring were subjected to a polyphasic taxonomic study. On the basis of 16S rRNA gene sequence similarities, both strains, which possessed identical sequences, were assigned to the genus Brucella. This placement was confirmed by extended multilocus sequence analysis (MLSA), where both strains possessed identical sequences, and whole-genome sequencing of a representative isolate. All of the above analyses suggested that the two strains represent a novel lineage within the genus Brucella. The strains also possessed a unique profile when subjected to the phenotyping approach classically used to separate species of the genus Brucella, reacting Abbreviations: MLSA, multilocus sequence analysis; MLVA, multilocus variable number of tandem repeat analysis; RTD, routine test dilution.The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA, omp2a and omp2b gene sequences of strains F8/08-60 T and F8/08-61 are HG932316 and HG932317 (16S rRNA gene), KJ493822 and KJ493823 (omp2a) and KJ510540 and KJ510541 (omp2b), respectively.A supplementary table and a supplementary figure are available with the online Supplementary Material.

Two slow-growing, Gram-negative, non-motile, non-spore-forming, coccoid bacteria (strains F60 T and F965), isolated in Austria from mandibular lymph nodes of two red foxes (Vulpes vulpes), were subjected to a polyphasic taxonomic analysis. In a recent study, both isolates were assigned to the genus Brucella but could not be attributed to any of the existing species. Hence, we have analysed both strains in further detail to determine their exact taxonomic position and genetic relatedness to other members of the genus Brucella. The genome sizes of F60 T and F965 were 3 236 779 and 3 237 765 bp, respectively. Each genome consisted of two chromosomes, with a DNA G+C content of 57.2 %. A genome-to-genome distance of .80 %, an average nucleotide identity (ANI) of 97 % and an average amino acid identity (AAI) of 98 % compared with the type species Brucella melitensis confirmed affiliation to the genus. Remarkably, 5 % of the entire genetic information of both strains was of non-Brucella origin, including as-yet uncharacterized bacteriophages and insertion sequences as well as ABC transporters and other genes of metabolic function from various soil-living bacteria. Core-genome-based phylogenetic reconstructions placed the novel species well separated from all hitherto-described species of the genus Brucella, forming a long-branched sister clade to the classical species of Brucella. In summary, based on phenotypic and molecular data, we conclude that strains F60 T and F965 are members of a novel species of the genus Brucella, for which the name Brucella vulpis sp. nov. is proposed, with the type strain F60 T (5BCCN 09-2 T 5DSM 101715 T ).Abbreviations: AAI, average amino acid identity; ANI, average nucleotide identity.The GenBank/EMBL/DDBJ accession numbers for the genome sequences (chr1 and chr2) of strains F60 T and F965 are LN997863-LN997864 and LN998033-LN998034, respectively, and can be retrieved from the European Nucleotide Archive under

An extended multilocus sequence analysis (MLSA) scheme applicable to the Brucella, an expanding genus that includes zoonotic pathogens that severely impact animal and human health across large parts of the globe, was developed. The scheme, which extends a previously described nine locus scheme by examining sequences at 21 independent genetic loci in order to increase discriminatory power, was applied to a globally and temporally diverse collection of over 500 isolates representing all 12 known Brucella species providing an expanded and detailed understanding of the population genetic structure of the group. Over 100 sequence types (STs) were identified and analysis of data provided insights into both the global evolutionary history of the genus, suggesting that early emerging Brucella abortus lineages might be confined to Africa while some later lineages have spread worldwide, and further evidence of the existence of lineages with restricted host or geographical ranges. The relationship between biovar, long used as a crude epidemiological marker, and genotype was also examined and showed decreasing congruence in the order Brucella suis > B. abortus > Brucella melitensis. Both the previously described nine locus scheme and the extended 21 locus scheme have been made available at http://pubmlst.org/brucella/ to allow the community to interrogate existing data and compare with newly generated data.

ABSTRACTBacterial isolates from frogs were phenotypically identified asOchrobactrum anthropi, but 16S rRNA sequencing showed up to 100% identity withBrucella inopinata. Further analysis ofrecA,omp2a,omp2b,bcsp31, and IS711and multilocus sequence analysis (MLSA) verified a close relationship withBrucella, suggesting the isolates may actually represent novel members of this growing genus of zoonotic pathogens.

LETTERSgenotyping M. bovis strains isolated from farm animals to help elucidate the source of infection and transmission of M. bovis in Taiwan.