title: Brucella microti sp. nov., isolated from the common vole Microtus arvalis.
abstract: Two Gram-negative, non-motile, non-spore-forming, coccoid bacteria (strains CCM 4915(T) and CCM 4916), isolated from clinical specimens of the common vole Microtus arvalis during an epizootic in the Czech Republic in 2001, were subjected to a polyphasic taxonomic study. On the basis of 16S rRNA (rrs) and recA gene sequence similarities, both isolates were allocated to the genus Brucella. Affiliation to Brucella was confirmed by DNA-DNA hybridization studies. Both strains reacted equally with Brucella M-monospecific antiserum and were lysed by the bacteriophages Tb, Wb, F1 and F25. Biochemical profiling revealed a high degree of enzyme activity and metabolic capabilities not observed in other Brucella species. The omp2a and omp2b genes of isolates CCM 4915(T) and CCM 4916 were indistinguishable. Whereas omp2a was identical to omp2a of brucellae from certain pinniped marine mammals, omp2b clustered with omp2b of terrestrial brucellae. Analysis of the bp26 gene downstream region identified strains CCM 4915(T) and CCM 4916 as Brucella of terrestrial origin. Both strains harboured five to six copies of the insertion element IS711, displaying a unique banding pattern as determined by Southern blotting. In comparative multilocus VNTR (variable-number tandem-repeat) analysis (MLVA) with 296 different genotypes, the two isolates grouped together, but formed a separate cluster within the genus Brucella. Multilocus sequence typing (MLST) analysis using nine different loci also placed the two isolates separately from other brucellae. In the IS711-based AMOS PCR, a 1900 bp fragment was generated with the Brucella ovis-specific primers, revealing that the insertion element had integrated between a putative membrane protein and cboL, encoding a methyltransferase, an integration site not observed in other brucellae. Isolates CCM 4915(T) and CCM 4916 could be clearly distinguished from all known Brucella species and their biovars by means of both their phenotypic and molecular properties, and therefore represent a novel species within the genus Brucella, for which the name Brucella microti sp. nov. with the type strain CCM 4915(T) (=BCCN 07-01(T)=CAPM 6434(T)) is proposed.
authors.affiliation: Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, D-80937 Munich, Germany. Holger1Scholz@bundeswehr.org
authors.forename: Holger C
authors.initials: HC
authors.lastname: Scholz
authors.forename: Zdenek
authors.initials: Z
authors.lastname: Hubalek
authors.forename: Ivo
authors.initials: I
authors.lastname: Sedlácek
authors.forename: Gilles
authors.initials: G
authors.lastname: Vergnaud
authors.forename: Herbert
authors.initials: H
authors.lastname: Tomaso
authors.forename: Sascha
authors.initials: S
authors.lastname: Al Dahouk
authors.forename: Falk
authors.initials: F
authors.lastname: Melzer
authors.forename: Peter
authors.initials: P
authors.lastname: Kämpfer
authors.forename: Heinrich
authors.initials: H
authors.lastname: Neubauer
authors.forename: Axel
authors.initials: A
authors.lastname: Cloeckaert
authors.forename: Marianne
authors.initials: M
authors.lastname: Maquart
authors.forename: Michel S
authors.initials: MS
authors.lastname: Zygmunt
authors.forename: Adrian M
authors.initials: AM
authors.lastname: Whatmore
authors.forename: Enevold
authors.initials: E
authors.lastname: Falsen
authors.forename: Peter
authors.initials: P
authors.lastname: Bahn
authors.forename: Cornelia
authors.initials: C
authors.lastname: Göllner
authors.forename: Martin
authors.initials: M
authors.lastname: Pfeffer
authors.forename: Birgit
authors.initials: B
authors.lastname: Huber
authors.forename: Hans-Jürgen
authors.initials: HJ
authors.lastname: Busse
authors.forename: Karsten
authors.initials: K
authors.lastname: Nöckler
mesh_terms: D000818:Animals; D003411:Arvicolinae / Q000382:microbiology*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D002002:Brucella / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D002006:Brucellosis / Q000382:microbiology / Q000662:veterinary*; D004269:DNA, Bacterial / Q000032:analysis; D020459:Genes, rRNA; D005838:Genotype; D018598:Minisatellite Repeats / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010641:Phenotype; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D011938:Rec A Recombinases / Q000235:genetics; D012376:Rodent Diseases / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D011938:Rec A Recombinases