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1Departamento de Microbiología, Universidad de Navarra, Aptdo 177, Pamplona, Spain
2Servicio de Microbiología, Clínica Universitaria de Navarra, Pamplona, Spain
Author for correspondence: Ignacio Moriyón. Tel: +34 48 425600. Fax: +34 48 425649. e-mail: imoriyon{at}unav.es
ABSTRACT
The relatedness of Brucella spp. and Ochrobactrum anthropi was studied by protein profiling, Western blot, immunoelectrophoresis and 16S rRNA analysis. Whole-cell and soluble proteins of brucellae and O. anthropi showed serological cross-reactivities quantitatively and qualitatively more intense than those existing with similar extracts of Agrobacterium spp. Numerical analysis of Western blot profiles of whole-cell extracts showed that O. anthropi LMG 3301 was closer to Brucella spp. than to O. anthropi LMG 3331Ta result not obtained by protein profiling. These differences were not observed by Western blot with soluble fractions, and immunoelectrophoretic analyses suggested that this was due to destruction of conformational epitopes in Western blot procedures with the subsequent simplification of antigenic profile. Analysis of the 16S rRNA sequences of strains previously used in the species definition confirmed that strain LMG 3301, and also LMG 3306, were closer to the brucellae, and that LMG 3331Twas in a separate cluster. The LMG 3301 and the LMG 3331Tclusters could also be separated by their different colistin sensitivity and by PCR with 16S rRNA Brucella primers, and both methods showed strains of both clusters among clinical isolates classified as O. anthropi by conventional tests. These results and those of previous DNA-DNA hybridization studies [Holmes, B., Popoff, M., Kiredjian, M. & Kersters, K. (1988). Int J Syst Bacteriol 38, 406–416] show that the LMG 3301 cluster and related clinical isolates should be given a new species status for which the name Ochrobactrum intermedium sp. nov. is proposed (type strain is LMG 3301T= NCTC 12171T= CNS 2-75T).
Key Words: Brucella • Ochrobactrum • antigenic relatedness • 16S rRNA
The GenBank accession number for the sequence of LMG 3301 is U70978.
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