Effects of Clarithromycin at Sub-Minimum Inhibitory Concentrations on Early ermB Gene Expression, Metabolic Activity and Growth of an erm(B)-Expressing Macrolide-Resistant Strain of Streptococcus pneumoniae

Riana Cockeran, H. C. Steel, N. Wolter, L. de Gouveia, A. von Gottberg, K. P. Klugman, A. T. Leanord, D. J. Inverarity, T. J. Mitchell, C. Feldman, R. Anderson

Division of Infection and Immunity, Glasgow Biomedical Research Centre, University of Glasgow, Glasgow, UK.
Division of Pulmonology, Department of Internal Medicine, Charlotte Maxeke Johannesburg Academic Hospital and University of the Witwatersrand, Johannesburg, South Africa.
Hubert Department of Global Health, Rollins School of Public Health, and Division of Infectious Diseases, School of Medicine, Emory University, Atlanta, USA.
Medical Research Council (MRC) Unit for Inflammation and Immunity, Department of Immunology, Faculty of Health Sciences, University of Pretoria, and Tshwane Academic Division of the National Health Laboratory Service, Pretoria, South Africa.
Microbiology Department, Southern General Hospital, Glasgow, UK.
Respiratory and Meningeal Pathogens Research Unit, National Institute for Communicable Diseases of the National Health Laboratory Service and Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.

DOI: 10.4236/ojrd.2012.21001                

Abstract

Aim: To investigate the effects of exposure of a macrolide-resistant [erm (B)-expressing] strain of Streptococcus pneumoniae (strain 2507) to clarithromycin (0.5 and 5 mg/L) added at the outset and 6 hours after initiation of culture on early gene expression, energy metabolism, and growth. Methods: Bacterial growth was determined by turbidometric and colony counting procedures, energy metabolism by measurement of ATP, while analysis of gene expression was performed using reverse transcription-PCR and sequencing. Results: Addition of clarithromycin, at either concentration, at the outset of culture, caused transient suppression of growth of 10 - 12 hours duration, while delayed addition of antibiotic (during the logarithmic phase) resulted in an abrupt halt in growth followed by recovery. These inhibitory effects of clarithromycin on bacterial growth were associated with up-regulation of expression of erm(B), decreased ATP and protein synthesis, and were unaffected by inclusion of either catalase (500 and 1000 kunits/L), or competence-stimulating peptide (CSP-1, 0.5 mg/L). The inhibitory effects could, however, be overcome by pre-exposure of the bacteria to the antibiotic. Moreover, clarithromycin appeared to potentiate the antimicrobial actions of ceftriaxone, at sub-MIC concentrations, for strain 2507. Conclusions: Unlike several other common bacterial pathogens, the full expression of erm(B)-mediated macrolide resistance by the pneumococcus has a slow onset, which is associated with transient susceptibility to macrolides and inhibition of growth.

Keywords

Clarithromycin; Macrolide-Resistance; Pneumococcus

Conflicts of Interest

The authors declare no conflicts of interest.

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