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Journal of Clinical Microbiology, September 2004, p. 4214-4222, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.4214-4222.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Phenotypic and Molecular Characteristics of Streptococcus agalactiae Isolates Recovered from Milk of Dairy Cows in Brazil

Rafael S. Duarte,1 Otávio P. Miranda,1 Bruna C. Bellei,1 Maria Aparecida V. P. Brito,2 and Lúcia M. Teixeira1*

Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro,1 Empresa Brasileira de Agropecuária (Embrapa), Juiz de Fora, Minas Gerais, Brazil2

Received 23 December 2003/ Returned for modification 15 February 2004/ Accepted 31 May 2004

Information on the characteristics of Streptococcus agalactiae obtained from bovine sources in Brazil is still very limited. The aim of this study was to assess the phenotypic and genotypic diversity among S. agalactiae isolates from milk of dairy cows presenting clinical or subclinical mastitis in the southeast region of Brazil. Phenotypic characterization was based on physiological and serological tests. Antimicrobial susceptibility tests were carried out by the disk method. Genetic diversity was evaluated by using random amplified polymorphic DNA-PCR (RAPD-PCR) (by using the primer 1254) and pulsed-field gel electrophoresis (PFGE) (by using SmaI as the restriction enzyme) and by PCRs for detection of genes associated with resistance to erythromycin and tetracycline as well as PCRs for detection of genes coding for cell surface-associated proteins. According to the results of physiologic tests, 45 (52.9%) isolates showed beta-hemolysis and 44 (51.7%) were susceptible to bacitracin. Fourteen different biotypes were detected. The two most frequent biotypes comprised strains that were non-beta-hemolytic; fermented galactose, lactose, and salicin; produced protease; and were negative for DNase production. Serotype III was predominant (66 isolates [77.6%]), followed by serotypes II, Ia, Ib, and VI. Resistance to tetracycline and erythromycin was found in 38 (44.7%) and 9 (10.5%) isolates, respectively, with tet(O) (31.7%) and erm(B) (100%) being the most frequently occurring resistance genes. Three genes coding for surface proteins, bca, lmb, and scpB, were detected in 55 (64.7%), 7 (8.2%), and 43 (50.5%) isolates, respectively. In most cases, isolates from animals in the same herd presented closely related genetic profiles (determined by either RAPD-PCR or PFGE), which were distinct from those of isolates from different herds.

* Corresponding author. Mailing address: Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, CCS, Bloco I, Cidade Universitária, Rio de Janeiro, RJ 21941-590, Brazil. Phone: 55 21 2260 4193. Fax: 55 21 2560 8344. E-mail: lmt2{at}micro.ufrj.br.

Journal of Clinical Microbiology, September 2004, p. 4214-4222, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.4214-4222.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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