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Antimicrobial Agents and Chemotherapy, October 2003, p. 3187-3194, Vol. 47, No. 10
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.10.3187-3194.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Genetic Characterization of Optochin-Susceptible Viridans Group Streptococci

Antonio J. Martín-Galiano,1 Luz Balsalobre,1 Asunción Fenoll,2 and Adela G. de la Campa1*

Unidad de Genética Bacteriana (Consejo Superior de Investigaciones Científicas),1 Servicio de Bacteriología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain2

Received 31 January 2003/ Returned for modification 7 April 2003/ Accepted 20 June 2003

Two clinical isolates of viridans group streptococci (VS) with different degrees of susceptibility to optochin (OPT), i.e., fully OPT-susceptible (Opts) VS strain 1162/99 (for which the MIC was equal to that for Streptococcus pneumoniae, 0.75 µg/ml) and intermediate Opts VS strain 1174/97 (MIC, 6 µg/ml) were studied. Besides being OPT susceptible, they showed characteristics typical of VS, such as bile insolubility; lack of reaction with pneumococcal capsular antibodies; and lack of hybridization with rRNA (AccuProbe)-, lytA-, and pnl-specific pneumococcal probes. However, these VS Opts strains and VS type strains hybridized with ant, a gene not present in S. pneumoniae. A detailed characterization of the genes encoding the 16S rRNA and SodA classified isolates 1162/99 and 1174/97 as Streptococcus mitis. Analysis of the atpCAB region, which encodes the c, a, and b subunits of the F0F1 H+-ATPase, the target of optochin, revealed high degrees of similarity between S. mitis 1162/99 and S. pneumoniae in atpC, atpA, and the N terminus of atpB. Moreover, amino acid identity between S. mitis 1174/97 and S. pneumoniae was found in {alpha} helix 5 of the a subunit. The organization of the chromosomal region containing the atp operon of the two Opts VS and VS type strains was spr1284-atpC, with spr1284 being located 296 to 556 bp from atpC, whereas in S. pneumoniae this distance was longer than 68 kb. In addition, the gene order in S. pneumoniae was IS1239-74 bp-atpC. The results suggest that the full OPT susceptibility of S. mitis 1162/99 is due to the acquisition of atpC, atpA, and part of atpB from S. pneumoniae and that the intermediate OPT susceptibility of S. mitis 1174/97 correlates with the amino acid composition of its a subunit.

* Corresponding author. Mailing address: Unidad de Genética Bacteriana (Consejo Superior de Investigaciones Científicas), Centro Nacional de Microbiología, Instituto de Salud Carlos III, 28220 Majadahonda, Madrid, Spain. Phone: (341) 509-7904. Fax: (341) 509-7919. E-mail: agcampa{at}isciii.es.

Antimicrobial Agents and Chemotherapy, October 2003, p. 3187-3194, Vol. 47, No. 10
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.10.3187-3194.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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