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Background   Brucella  spp. are highly similar, having identical 16S RNA. However, they have important phenotypic differences such as differential susceptibility to antibiotics binding the ribosome. Neither the differential susceptibility nor its basis has been rigorously studied. Differences found among other conserved ribosomal loci could further define the relationships among the classical  Brucella  spp.    Results  Minimum inhibitory concentration (MIC) values of  Brucella  reference strains and three marine isolates to antibiotics binding the ribosome ranged from 0.032 to >256 μg/ml for the macrolides erythromycin, clarithromycin, and azithromycin and 2 to >256 μg/ml for the lincosamide, clindamycin. Though sequence polymorphisms were identified among ribosome associated loci 23S  rrn ,  rpl V,  tuf -1 and  tuf -2 but not  rplD , they did not correlate with antibiotic resistance phenotypes. When spontaneous erythromycin resistant (ery R ) mutants were examined, mutation of the peptidyl transferase center (A2058G Ec) correlated with increased resistance to both erythromycin and clindamycin.  Brucella  efflux was examined as an alternative antibiotic resistance mechanism by use of the inhibitor L-phenylalanine-L-arginine β-naphthylamide (PAβN). Erythromycin MIC values of reference and all ery R  strains, except the  B. suis  ery R  mutants, were lowered variably by PAβN. A phylogenetic tree based on concatenated ribosomal associated loci supported separate evolutionary paths for  B. abortus, B. melitensis , and  B. suis/B. canis , clustering marine  Brucella  and  B. neotomae  with  B. melitensis . Though  Brucella ovis  was clustered with  B. abortus , the bootstrap value was low.    Conclusion  Polymorphisms among ribosomal loci from the reference  Brucella  do not correlate with their highly differential susceptibility to erythromycin. Efflux plays an important role in  Brucella  sensitivity to erythromycin. Polymorphisms identified among ribosome associated loci construct a robust phylogenetic tree supporting classical  Brucella  spp. designations.

bmc microbiology

Intrinsic and selected resistance to antibiotics binding the ribosome: analyses of Brucella 23S rrn, L4, L22, EF-Tu1, EF-Tu2, efflux and phylogenetic implications.