Folia Microbiologica

, Volume 64, Issue 2, pp 153–159 | Cite as

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

  • Branislav Lilić
  • Brankica Filipić
  • Milka Malešević
  • Katarina Novović
  • Zorica Vasiljević
  • Milan Kojić
  • Branko JovčićEmail author
Original Article


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.



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

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

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

Supplementary material

12223_2018_639_MOESM1_ESM.doc (84 kb)
Supplementary Table 1 (DOC 84 kb)


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Copyright information

© Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2018

Authors and Affiliations

  1. 1.Institute of Molecular Genetics and Genetic EngineeringUniversity of BelgradeBelgradeSerbia
  2. 2.Faculty of PharmacyUniversity of BelgradeBelgradeSerbia
  3. 3.Department of Clinical MicrobiologyMother and Child Health Care Institute of Serbia “Dr. Vukan Čupić”BelgradeSerbia
  4. 4.Faculty of BiologyUniversity of BelgradeBelgradeSerbia

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