New Tn916-related elements causing erm(B)-mediated erythromycin resistance in tetracycline-susceptible pneumococci
Ileana Cochetti1, Emily Tili, Manuela Vecchi
1Institute of Microbiology and Biomedical Sciences, Polytechnic University of Marche Medical School, 60020 Ancona, Italy.
Objectives:
To analyse the as yet unexplored genetic elements encoding erm(B)-mediated erythromycin resistance in tetracycline-susceptible pneumococci.
Methods:
Sixteen Streptococcus pneumoniae clinical isolates sharing erm(B)-mediated erythromycin resistance and susceptibility to tetracycline were used. Gene detection was performed by PCR using both established and specially designed primers. S. pneumoniae R6, Streptococcus pyogenes 12RF and Enterococcus faecalis JH2-2 were used as recipients in mating experiments.
Results:
Of the 16 test strains, 14 bore an unexpressed tet(M) gene which in 13 strains had a genetic linkage with erm(B). Three isolates yielded a 3.2 kb and 10 an 11.9 kb erm(B)/tet(M) amplicon. The former three showed genetic organizations similar to that of the composite element Tn3872, where the erm(B)-carrying Tn917 transposon is inserted into a Tn916-like element. Of the latter 10 isolates, 9 showed genetic organizations substantially overlapping with that of Tn6002, a newly sequenced erm(B)-containing Tn916-related transposon. The tenth isolate carried a novel composite element (designated Tn6003) resulting from the insertion into a Tn6002-like transposon of a fragment [designated macrolide-aminoglycoside-streptothricin (MAS) element] containing a second erm(B) (lacking the stop codon) and a variant of the aadE-sat4-aphA-3 cluster. The two tet(M)-negative isolates had different Tn3872-related elements, one containing a complete and one a deleted MAS fragment. Conjugative transfer was obtained from donors carrying Tn6002-related elements, not from donors carrying Tn3872-related elements.
Conclusions:
In tetracycline-susceptible pneumococci with erm(B)-mediated erythromycin resistance, the erm(B) gene is carried on a variety of Tn916-related genetic elements either lacking tet(M) or, more often, carrying an unexpressed tet(M) gene.
Insights
Erythromycin resistance in tetracycline-susceptible pneumococci is often linked to the erm(B) gene. This gene is frequently found on Tn916-related elements, sometimes alongside an unexpressed tetracycline resistance gene, tet(M).
Area of Science:
- Microbiology
- Genetics
- Molecular Biology
Background:
- Erythromycin resistance in Streptococcus pneumoniae is a growing public health concern.
- The erm(B) gene is a common mechanism for conferring erythromycin resistance.
- Understanding the genetic elements carrying erm(B) is crucial for tracking resistance dissemination.
Purpose of the Study:
- To investigate the genetic elements encoding erm(B)-mediated erythromycin resistance in tetracycline-susceptible pneumococci.
- To characterize the genetic organization and mobile genetic elements associated with erm(B) in these isolates.
Main Methods:
- Analysis of 16 clinical isolates of Streptococcus pneumoniae with erm(B)-mediated erythromycin resistance and tetracycline susceptibility.
- Polymerase Chain Reaction (PCR) using established and novel primers for gene detection.
- Conjugation experiments using S. pneumoniae R6, Streptococcus pyogenes, and Enterococcus faecalis as recipients.
Main Results:
- 14 out of 16 isolates harbored an unexpressed tet(M) gene, often genetically linked to erm(B).
- Genetic organizations similar to Tn3872 and Tn6002 were identified, with one isolate containing a novel composite element Tn6003.
- Conjugative transfer was observed from elements related to Tn6002, but not from those related to Tn3872.
Conclusions:
- In tetracycline-susceptible pneumococci, erm(B) is frequently located on Tn916-related genetic elements.
- These elements may either lack tet(M) or carry an unexpressed tet(M) gene.
- The diversity of these genetic elements influences their mobility and potential for spread.
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