title: Landscape of Resistance-Nodulation-Cell Division (RND)-Type Efflux Pumps in Enterobacter cloacae Complex.
abstract: In Gram-negative bacteria, the active efflux is an important mechanism of antimicrobial resistance, but little is known about theEnterobacter cloacaecomplex (ECC). It is mediated primarily by pumps belonging to the RND (resistance-nodulation-cell division) family, and only AcrB, part of the AcrAB-TolC tripartite system, was characterized in ECC. However, detailed genome sequence analysis of the strainE. cloacaesubsp.cloacaeATCC 13047 revealed to us that 10 other genes putatively coded for RND-type transporters. We then characterized the role of all of these candidates by construction of corresponding deletion mutants, which were tested for their antimicrobial susceptibility to 36 compounds, their virulence in the invertebrateGalleria mellonellamodel of infection, and their ability to form biofilm. Only the ΔacrBmutant displayed significantly different phenotypes compared to that of the wild-type strain: 4- to 32-fold decrease of MICs of several antibiotics, antiseptics, and dyes, increased production of biofilm, and attenuated virulence inG. mellonella In order to identify specific substrates of each pump, we individually expressed intransall operons containing an RND pump-encoding gene into the ΔacrBhypersusceptible strain. We showed that three other RND-type efflux systems (ECL_00053-00055, ECL_01758-01759, and ECL_02124-02125) were able to partially restore the wild-type phenotype and to superadd to and even enlarge the broad range of antimicrobial resistance. This is the first global study assessing the role of all RND efflux pumps chromosomally encoded by the ECC, which confirms the major role of AcrB in both pathogenicity and resistance and the potential involvement of other RND-type members in acquired resistance.
authors: Université de Caen Normandie, EA4655 U2RM (Équipe Antibio-Résistance), Caen, France CHU de Caen, Service de Microbiologie, Caen, France CNR de la Résistance aux Antibiotiques (Laboratoire Associé Entérocoques), Caen, France.
authors: François
authors: F
authors: Guérin
authors: Université de Caen Normandie, EA4655 U2RM (Équipe Antibio-Résistance), Caen, France.
authors: Claire
authors: C
authors: Lallement
authors: Université de Caen Normandie, EA4655 U2RM (Équipe Antibio-Résistance), Caen, France CHU de Caen, Service de Microbiologie, Caen, France.
authors: Christophe
authors: C
authors: Isnard
authors: Université de Caen Normandie, EA4655 U2RM (Équipe Antibio-Résistance), Caen, France.
authors: Anne
authors: A
authors: Dhalluin
authors: Université de Caen Normandie, EA4655 U2RM (Équipe Antibio-Résistance), Caen, France CHU de Caen, Service de Microbiologie, Caen, France CNR de la Résistance aux Antibiotiques (Laboratoire Associé Entérocoques), Caen, France cattoir-v@chu-caen.fr.
authors: Vincent
authors: V
authors: Cattoir
authors: Université de Caen Normandie, EA4655 U2RM (Équipe Antibio-Résistance), Caen, France.
authors: Jean-Christophe
authors: JC
authors: Giard
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000891:Anti-Infective Agents, Local / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects* / Q000254:growth & development; D002874:Chromosome Mapping; D004396:Coloring Agents / Q000494:pharmacology; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D016972:Enterobacter cloacae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D016680:Genome, Bacterial; D007814:Larva / Q000382:microbiology; D008826:Microbial Sensitivity Tests; D009036:Moths / Q000382:microbiology; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics* / Q000378:metabolism; D014774:Virulence
chemical_list: D000900:Anti-Bacterial Agents; D000891:Anti-Infective Agents, Local; D004396:Coloring Agents; D027425:ATP-Binding Cassette, Sub-Family C Proteins