1887

Abstract

Two Gram-negative, non-motile, non-spore-forming, coccoid bacteria (strains CCM 4915 and CCM 4916), isolated from clinical specimens of the common vole during an epizootic in the Czech Republic in 2001, were subjected to a polyphasic taxonomic study. On the basis of 16S rRNA () and gene sequence similarities, both isolates were allocated to the genus . Affiliation to was confirmed by DNA–DNA hybridization studies. Both strains reacted equally with 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 species. The and genes of isolates CCM 4915 and CCM 4916 were indistinguishable. Whereas was identical to of brucellae from certain pinniped marine mammals, clustered with of terrestrial brucellae. Analysis of the gene downstream region identified strains CCM 4915 and CCM 4916 as of terrestrial origin. Both strains harboured five to six copies of the insertion element IS, 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 . Multilocus sequence typing (MLST) analysis using nine different loci also placed the two isolates separately from other brucellae. In the IS-based AMOS PCR, a 1900 bp fragment was generated with the -specific primers, revealing that the insertion element had integrated between a putative membrane protein and , encoding a methyltransferase, an integration site not observed in other brucellae. Isolates CCM 4915 and CCM 4916 could be clearly distinguished from all known species and their biovars by means of both their phenotypic and molecular properties, and therefore represent a novel species within the genus , for which the name sp. nov. with the type strain CCM 4915 (=BCCN 07-01=CAPM 6434) is proposed.

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2008-02-01
2024-03-28
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