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Published Online: 26 March 2013

In Vitro Development and Analysis of Escherichia coli and Shigella boydii Azithromycin–Resistant Mutants

Publication: Microbial Drug Resistance
Volume 19, Issue Number 2

Abstract

The aim of this study was to develop and analyze in vitro azithromycin (AZM)-resistant mutants of Escherichia coli and Shigella boydii. Three clinical isolates of E. coli and one S. boydii isolated from feces samples collected from children under 5 years of age with diarrhea in Lima, Peru were inoculated onto Mueller-Hinton plates containing increasing serial dilutions of AZM ranging from their specific minimal inhibitory concentration (2 or 4 mg/l) to 64 mg/l. From these plates, 16 AZM-resistant mutants were selected to determine the stability of the resistance and the presence of cross resistance with other antibiotics. The role of Phe-Arg-β-Naphthylamide (PAβN)-inhibitible efflux pumps as well as the presence of mutations in the rplV, rplD, and rrlH (23S rRNA) genes and alterations in the outer membrane profiles were determined in these 16 mutants. The rate of mutation ranged from < 2.70×10−10 to 2.17×10−7 for E. coli and from < 9.58×10−10 to 1.05×10−8 for S. boydii. E. coli mutants showed an increase in the AZM-MIC up to sixfold with one strain achieving a MIC >256 mg/l. In contrast, S. boydii only presented increases of up to twofold in MIC levels. All the strains obtained, but one showed stable AZM resistance. In the presence of PAβN, the AZM MICs decreased to parental levels in Shigella mutants, while no MIC returned to parental levels among the E. coli mutants. No cross resistance to other classes of antibiotics was found. These results show the relevance of PAβN-inhibitible efflux pumps in the basal levels and development of AZM resistance. Further studies to characterize the remaining unidentified mechanisms of AZM resistance are needed.

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Published In

cover image Microbial Drug Resistance
Microbial Drug Resistance
Volume 19Issue Number 2April 2013
Pages: 88 - 93
PubMed: 23176550

History

Published in print: April 2013
Published online: 26 March 2013
Published ahead of print: 23 November 2012

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Cláudia Gomes
Barcelona Center for International Health Research (CRESIB, Hospital Clínic-Universitat de Barcelona), Barcelona, Spain.
Maria J. Pons
Barcelona Center for International Health Research (CRESIB, Hospital Clínic-Universitat de Barcelona), Barcelona, Spain.
Ariel Magallon-Tejada
Barcelona Center for International Health Research (CRESIB, Hospital Clínic-Universitat de Barcelona), Barcelona, Spain.
David Durand
Institute of Tropical Medicine Alexander von Humboldt, University Cayetano Heredia, Peru.
Angela Lluque
Institute of Tropical Medicine Alexander von Humboldt, University Cayetano Heredia, Peru.
Susan Mosquito
Institute of Tropical Medicine Alexander von Humboldt, University Cayetano Heredia, Peru.
Maribel Riveros
Institute of Tropical Medicine Alexander von Humboldt, University Cayetano Heredia, Peru.
Erik Mercado
Institute of Tropical Medicine Alexander von Humboldt, University Cayetano Heredia, Peru.
Ana Prada
Institute of Tropical Medicine Alexander von Humboldt, University Cayetano Heredia, Peru.
Theresa J. Ochoa
Institute of Tropical Medicine Alexander von Humboldt, University Cayetano Heredia, Peru.
Center for Infectious Diseases, University of Texas School of Public Health, Houston, Texas.
Joaquim Ruiz
Barcelona Center for International Health Research (CRESIB, Hospital Clínic-Universitat de Barcelona), Barcelona, Spain.
CIBER Epidemiología y Salud Pública (CIBERESP), Spain.
Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain.

Notes

Address correspondence to:Joaquim Ruiz, MSc, PhDBarcelona Centre for International Health Research (CRESIB)Hospital Clínic-Universitat de BarcelonaC/.Rosselló 149-153Barcelona 08036Spain
E-mail: [email protected]

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The authors have no competing interests to disclose.

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