1887

Abstract

A Gram-negative, non-motile, non-spore-forming coccoid bacterium (strain BO1) was isolated recently from a breast implant infection of a 71-year-old female patient with clinical signs of brucellosis. Affiliation of strain BO1 to the genus was confirmed by means of polyamine pattern, polar lipid profile, fatty acid profile, quinone system, DNA–DNA hybridization studies and by insertion sequence (IS)-specific PCR. Strain BO1 harboured four to five copies of the -specific insertion element IS, displaying a unique banding pattern, and exhibited a unique 16S rRNA gene sequence and also grouped separately in multilocus sequence typing analysis. Strain BO1 reacted with M-monospecific antiserum. Incomplete lysis was detected with bacteriophages Tb (Tbilisi), F1 and F25. Biochemical profiling revealed a high degree of enzymic activity and metabolic capabilities. In multilocus VNTR (variable-number tandem-repeat) analysis, strain BO1 showed a very distinctive profile and clustered with the other ‘exotic’ strains, including strains isolated from marine mammals, and , biovar 5 and . Comparative and gene sequence analysis revealed the most divergent sequences identified to date for a strain. The gene sequence of strain BO1 differed in seven nucleotides from the consensus sequence. Using the species-specific multiplex PCR assay, strain BO1 displayed a unique banding pattern not observed in other species. From the phenotypic and molecular analysis it became evident that strain BO1 was clearly different from all other species, and therefore represents a novel species within the genus . Because of its unexpected isolation, the name with the type strain BO1 (=BCCN 09-01=CPAM 6436) is proposed.

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2010-04-01
2019-10-20
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Lysis of strain BO1 on agar by bacteriophage Tb at routine test dilution (RTD) and various other concentrations (10 to 10 × RTD). K=phage negative control.

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Two-dimensional thin-layer chromatogram of total polar lipids of strain BO1 . PME, phosphatidylmonomethylethanolamine; PE, phosphatidylethanolamine; PC, phosphatidylcholine; DPG, diphosphatidylglycerol; PG, phosphatidylglycerol; AL1, unknown aminolipid; PL, unknown phospholipid; APL2, unknown aminophospholipid; L3–7, unknown polar lipids.

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IS element fingerprinting by Southern blotting with RI digested chromosomal DNA of 16M (lane 1) (bv. 1), Ether (lane 2) (bv. 3), 1330 (lane 3) (bv. 1), Thomsen (lane 4) (bv. 2), 513 (lane 5) (bv. 5), 63/290 (lane 6), (lane 7), CCM 4915 (lane 8), B2/94 (lane 9), B1/94 (lane 10), and sp. nov., strain BO1 (lane 11), M=molecular mass marker.

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Condensed dendrogram of clustered MLVA-16 genotypes obtained with more than 470 isolates corresponding to 324 different genotypes. Missing data in strain BO1 was considered as 0. The markers were analysed in three panels, 1, 2A and 2B, given different weight, as described by Kattar (2008). Bar, percentage of sequence divergence.

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