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Fluoroquinolone-resistant Achromobacter xylosoxidans clinical isolates from Serbia: high prevalence of the aac-(6′)-Ib-cr gene among resistant isolates

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Abstract

The aim of this study was to evaluate the contribution of plasmid-mediated genes and efflux to fluoroquinolone resistance in collection of Achromobacter spp. gathered during a 3-year period. Susceptibility to ciprofloxacin and levofloxacin was tested by disk diffusion and microdilution tests for a collection of 98 Achromobacter spp. clinical isolates. Identification of fluoroquinolone-resistant isolates was performed by sequencing and phylogenetic analyses of the nrdA gene. Genetic relatedness among resistant isolates was determined by pulsed-field gel electrophoresis (PFGE) analysis. The influence of an H+ conductor cyanide m-chlorophenyl hydrazone (CCCP) and a resistance-nodulation-division-type efflux pump inhibitor phenylalanine-arginine beta-naphthylamide (PAβN) on minimal inhibitory concentration (MIC) value was evaluated by broth microdilution. The presence of the plasmid-mediated qnrA, qnrB, qnrC, qnrS, and aac-(6′)-Ib-cr genes was investigated by PCR and sequencing. Achromobacter spp. isolates that were resistant or intermediately resistant to fluoroquinolones in disk diffusion tests (44/98) were subjected to microdilution. As a result, 20/98 isolates were confirmed to be resistant to ciprofloxacin while 10/98 was resistant to levofloxacin. CCCP decreased twofold MIC value for ciprofloxacin in six isolates and more than 16 times in one isolate, while MIC value for levofloxacin was decreased in all isolates (twofold to more than eightfold). Fluoroquinolone-resistant isolates were identified as A. xylosoxidans with the nrdA gene sequencing. PFGE revealed that resistant isolates belonged to seven different genotypes. Ten isolates belonging to four genotypes were positive for the aac-(6′)-Ib-cr gene. Although resistance to fluoroquinolones was not widespread among analyzed isolates, detected contribution of efflux pumps and the presence of the aac-(6′)-Ib-cr gene present a platform for emergence of more resistant strains.

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Funding

This study was funded by the Ministry of Education, Science and Technological Development of the Republic of Serbia (grant no. 173019).

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Authors and Affiliations

  1. Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, P.O. Box 23, Belgrade, 11010, Serbia

    Branislav Lilić, Brankica Filipić, Milka Malešević, Katarina Novović, Milan Kojić & Branko Jovčić

  2. Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, Belgrade, 11221, Serbia

    Brankica Filipić

  3. Department of Clinical Microbiology, Mother and Child Health Care Institute of Serbia “Dr. Vukan Čupić”, Radoja Dakića 8, Belgrade, 11070, Serbia

    Zorica Vasiljević

  4. Faculty of Biology, University of Belgrade, Studentski trg 16, Belgrade, 11000, Serbia

    Branko Jovčić

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  1. Branislav Lilić
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  2. Brankica Filipić
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  3. Milka Malešević
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  4. Katarina Novović
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  5. Zorica Vasiljević
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  6. Milan Kojić
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  7. Branko Jovčić
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Correspondence to Branko Jovčić.

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Ethics Committee of the Mother and Child Health Care Institute “Dr. Vukan Cupic” (approval no. 8/6)

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Lilić, B., Filipić, B., Malešević, M. et al. Fluoroquinolone-resistant Achromobacter xylosoxidans clinical isolates from Serbia: high prevalence of the aac-(6′)-Ib-cr gene among resistant isolates. Folia Microbiol 64, 153–159 (2019). https://doi.org/10.1007/s12223-018-0639-7

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  • DOI: https://doi.org/10.1007/s12223-018-0639-7

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