title: Triclosan-induced aminoglycoside-tolerant Listeria monocytogenes isolates can appear as small-colony variants.
abstract: Exposure of the human food-borne pathogen Listeria monocytogenes to sublethal concentrations of triclosan can cause resistance to several aminoglycosides. Aminoglycoside-resistant isolates exhibit two colony morphologies: normal-size and pinpoint colonies. The purposes of the present study were to characterize the small colonies of L. monocytogenes and to determine if specific genetic changes could explain the triclosan-induced aminoglycoside resistance in both pinpoint and normal-size isolates. Isolates from the pinpoint colonies grew poorly under aerated conditions, but growth was restored by addition of antibiotics. Pinpoint isolates had decreased hemolytic activity under stagnant conditions and a changed spectrum of carbohydrate utilization compared to the wild type and isolates from normal-size colonies. Genome sequence comparison revealed that all seven pinpoint isolates had a mutation in a heme gene, and addition of heme caused the pinpoint isolates to revert to normal colony size. Triclosan-induced gentamicin-resistant isolates had mutations in several different genes, and it cannot be directly concluded how the different mutations caused gentamicin resistance. However, since many of the mutations affected proteins involved in respiration, it seems likely that the mutations affected the active transport of the antibiotic and thereby caused resistance by decreasing the amount of aminoglycoside that enters the bacterial cell. Our study emphasizes that triclosan likely has more targets than just fabI and that exposure to triclosan can cause resistance to antibiotics that enters the cell via active transport. Further studies are needed to elucidate if L. monocytogenes pinpoint isolates could have any clinical impact, e.g., in persistent infections.
authors.affiliation: National Food Institute, Technical University of Denmark, Kongens Lyngby, Denmark.
authors.forename: Vicky G
authors.initials: VG
authors.lastname: Kastbjerg
authors.affiliation: National Food Institute, Technical University of Denmark, Kongens Lyngby, Denmark.
authors.forename: Line
authors.initials: L
authors.lastname: Hein-Kristensen
authors.affiliation: Department of System Biology, Technical University of Denmark, Kongens Lyngby, Denmark gram@bio.dtu.dk.
authors.forename: Lone
authors.initials: L
authors.lastname: Gram
mesh_terms: D000617:Aminoglycosides / Q000494:pharmacology*; D000890:Anti-Infective Agents / Q000494:pharmacology*; D005839:Gentamicins / Q000494:pharmacology; D006801:Humans; D008089:Listeria monocytogenes / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D008088:Listeriosis / Q000188:drug therapy*; D008826:Microbial Sensitivity Tests; D014260:Triclosan / Q000494:pharmacology*
chemical_list: D000617:Aminoglycosides; D000890:Anti-Infective Agents; D005839:Gentamicins; D014260:Triclosan