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Journal of Clinical Microbiology, September 2007, p. 2923-2928, Vol. 45, No. 9
0095-1137/07/$08.00+0     doi:10.1128/JCM.00822-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Molecular Epidemiological and Antibiotic Susceptibility Characterization of Brucella Isolates from Humans in Sicily, Italy

Dipartimento di Sanità Alimentare ed Animale, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy,¹ Azienda Ospedaliera Vittorio Emanuele, Dipartimento di Microbiologia, Via Plebiscito 628, 951254 Catania, Italy,² Azienda Ospedaliera Gravina, Dipartimento di Microbiologia, Via Portosalvo 1, 95041 Caltagirone, Italy,³ Azienda Unità Sanitaria Locale No. 3, Dipartimento di Prevenzione, Via Santa Maria La Grande 5, 95124 Catania, Italy,⁴ Istituto Zooprofilattico Sperimentale della Sicilia, Via S. Andrea 96, 98051 Barcellona Pozzo di Gotto, Italy⁵

Received 18 April 2007/ Returned for modification 18 June 2007/ Accepted 6 July 2007

Brucellosis is a serious problem in Sicily. Brucella melitensis was identified as the species most frequently isolated in humans in Italy. No data, however, are available about the molecular epidemiological characterization of Brucella isolates from humans. We have conducted this study to molecularly characterize clinical isolates of Brucella spp. and to evaluate their antimicrobial susceptibilities. Twenty Brucella isolates were studied. Differential growth characteristics and DNA polymorphisms such as the restriction patterns of the PCR-amplified omp2a and omp2b genes, rpoB nucleotide sequencing, and multiple-locus variable-number tandem repeat analysis of 16 loci (MLVA-16) were used to characterize the strains. In vitro antibiotic susceptibility was determined by the E-test method on two different agar media, and the results were compared. All isolates were identified as B. melitensis biovar 3. rpoB nucleotide sequence analysis allowed the identification of two different genotypes of B. melitensis biovar 3. On the other hand, the MLVA-16 typing assay recognized 17 distinct genotypes. All isolates were sensitive to all tested antibiotics (rifampin, doxycycline, ciprofloxacin, ceftriaxone, and trimethoprim-sulfamethoxazole), and the Mueller-Hinton agar plate is recommended for antibiotic susceptibility testing by the E-test method. Our findings identify B. melitensis biovar 3 as the etiological agent isolated in Sicily and encourage the use of both molecular methods, and in particular of the MLVA-16 assay, in epidemiological trace-back analysis. This study represents the first epidemiological data from molecular typing of Brucella strains circulating in Italy and, in particular, in eastern Sicily.

Published ahead of print on 18 July 2007.