Detection of optrA in the African continent (Tunisia) within a mosaic Enterococcus faecalis plasmid from urban wastewaters

J Antimicrob Chemother. 2017 Dec 1;72(12):3245-3251. doi: 10.1093/jac/dkx321.

Abstract

Objectives: Oxazolidinone resistance is a serious limitation in the treatment of MDR Enterococcus infections. Plasmid-mediated oxazolidinone resistance has been strongly linked to animals where the use of phenicols might co-select resistance to both antibiotic families. Our goal was to assess the diversity of genes conferring phenicol/oxazolidinone resistance among diverse enterococci and to characterize the optrA genetic environment.

Methods: Chloramphenicol-resistant isolates (>16 mg/L, n = 245) from different sources (hospitals/healthy humans/wastewaters/animals) in Portugal, Angola and Tunisia (1996-2016) were selected. Phenicol (eight cat variants, fexA, fexB) or phenicol + oxazolidinone [cfr, cfr(B), optrA] resistance genes were searched for by PCR. Susceptibility (disc diffusion/microdilution), filter mating, stability of antibiotic resistance (500 bacterial generations), plasmid typing (S1-PFGE/hybridization), MLST and WGS (Illumina-HiSeq) were performed for optrA-positive isolates.

Results: Resistance to phenicols (n = 181, 74%) and phenicols + oxazolidinones (n = 2, 1%) was associated with the presence of cat(A-8) (40%, predominant in hospitals and swine), cat(A-7) (29%, predominant in poultry and healthy humans), cat(A-9) (2%), fexB (2%) and fexA + optrA (1%). fexA and optrA genes were co-located in a transferable plasmid (pAF379, 72 918 bp) of two ST86 MDR Tunisian Enterococcus faecalis (wastewaters) carrying several putative virulence genes. MICs of chloramphenicol, linezolid and tedizolid were stably maintained at 64, 4 and 1 mg/L, respectively. The chimeric pAF379 comprised relics of genetic elements from different Gram-positive bacteria and origins (human/porcine).

Conclusions: To the best of our knowledge, we report the first detection of optrA in an African country (Tunisia) within a transferable mosaic plasmid of different origins. Its identification in isolates from environmental sources is worrisome and alerts for the need of a concerted global surveillance on the occurrence and spread of optrA.

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Chloramphenicol / pharmacology
  • Cities
  • DNA, Bacterial / genetics
  • Disk Diffusion Antimicrobial Tests
  • Drug Resistance, Bacterial*
  • Enterococcus faecalis / drug effects
  • Enterococcus faecalis / genetics*
  • Enterococcus faecalis / isolation & purification*
  • Gene Order*
  • Genes, Bacterial*
  • Multilocus Sequence Typing
  • Oxazolidinones / pharmacology
  • Plasmids / isolation & purification*
  • Polymerase Chain Reaction
  • Tunisia
  • Wastewater / microbiology*
  • Whole Genome Sequencing

Substances

  • Anti-Bacterial Agents
  • DNA, Bacterial
  • Oxazolidinones
  • Waste Water
  • Chloramphenicol