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Antimicrobial Agents and Chemotherapy, November 2000, p. 2979-2984, Vol. 44, No. 11
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Genetic Diversity of the tet(M) Gene in Tetracycline-Resistant Clonal Lineages of Streptococcus pneumoniae

Neil Doherty, Krzysztof Trzcinski,dagger Paul Pickerill,Dagger Piotr Zawadzki,§ and Christopher G. Dowson*

Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, United Kingdom

Received 5 May 2000/Returned for modification 8 July 2000/Accepted 31 July 2000

The aim of the present study was to examine the stability and evolution of tet(M)-mediated resistance to tetracyclines among members of different clonal lineages of Streptococcus pneumoniae. Thirty-two tetracycline-resistant isolates representing three national (Spanish serotype 14, Spanish serotype 15, and Polish serotype 23F) and one international (Spanish serotype 23F) multidrug-resistant epidemic clones were all found to be tet(M) positive and tet(O), tet(K), and tet(L) negative. These isolates all carried the integrase gene, int, which is associated with the Tn1545-Tn916 family of conjugative transposons. High-resolution restriction analysis of tet(M) products identified six alleles, tet(M)1 to tet(M)6: tet(M)1 to tet(M)3 and tet(M)5 in isolates of the Spanish serotype 14 clone, tet(M)4 in both the Spanish serotype 15 and 23F clones, and tet(M)6, the most divergent allele, in the Polish 23F clone. This indicates that tet(M) variation can occur at the inter- and intraclone levels in pneumococci. Two alleles of int were identified, with int1 being found in all isolates apart from members of the international Spanish 23F clone, which carried int2. Susceptibility to tetracycline, doxycycline, and minocycline was evaluated for all isolates with or without preincubation in the presence of subinhibitory concentrations of tetracyclines. Resistance to tetracyclines was found to be inducible in isolates of all clones; however, the strongest induction was observed in the Spanish serotype 15 and 23F clones carrying tet(M)4. Tetracycline was found to be the strongest inducer of resistance, and minocycline was found to be the weakest inducer of resistance.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, United Kingdom. Phone: 44 2476 523534.Fax: 44 2476 523568. E-mail: c.g.dowson{at}warwick.ac.uk.

dagger Present address: National Institute of Hygiene, Chocimska 24, 00-791 Warsaw, Poland.

Dagger Present address: Zeneca Agrochemicals, Jeallots Hill Research Station, Bracknell RG42 6ET, United Kingdom.

§ Present address: Procter & Gamble, 65824 Schwalbach/Ts, Germany.

Antimicrobial Agents and Chemotherapy, November 2000, p. 2979-2984, Vol. 44, No. 11
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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