title: Antifungal susceptibility of Candida biofilms: unique efficacy of amphotericin B lipid formulations and echinocandins.
abstract: Biofilms, likely the predominant mode of device-related microbial infection, exhibit resistance to antimicrobial agents. Evidence suggests that Candida biofilms have dramatically reduced susceptibility to antifungal drugs. We examined antifungal susceptibilities of Candida albicans and Candida parapsilosis biofilms grown on a bioprosthetic model. In addition to conventional agents, we determined if new antifungal agents (triazoles, amphotericin B lipid formulations, and echinocandins) have activities against Candida biofilms. We also explored effects of preincubation of C. albicans cells with subinhibitory concentrations (sub-MICs) of drugs to see if they could modify subsequent biofilm formation. Finally, we used confocal scanning laser microscopy (CSLM) to image planktonic- and biofilm-exposed blastospores to examine drug effects on cell structure. Candida biofilms were formed on silicone elastomer and quantified by tetrazolium and dry weight (DW) assays. Susceptibility testing of fluconazole, nystatin, chlorhexidine, terbenafine, amphotericin B (AMB), and the triazoles voriconazole (VRC) and ravuconazole revealed resistance in all Candida isolates examined when grown as biofilms, compared to planktonic forms. In contrast, lipid formulations of AMB (liposomal AMB and AMB lipid complex [ABLC]) and echinocandins (caspofungin [Casp] and micafungin) showed activity against Candida biofilms. Preincubation of C. albicans cells with sub-MIC levels of antifungals decreased the ability of cells to subsequently form biofilm (measured by DW; P < 0.0005). CSLM analysis of planktonic and biofilm-associated blastospores showed treatment with VRC, Casp, and ABLC resulted in morphological alterations, which differed with each agent. In conclusion, our data show that Candida biofilms show unique susceptibilities to echinocandins and AMB lipid formulations.
authors.affiliation: Division of Infectious Diseases, Department of Medicine, University Hospitals of Cleveland and Case Western Reserve University, Cleveland, Ohio 44106, USA.
authors.forename: D M
authors.initials: DM
authors.lastname: Kuhn
authors.forename: T
authors.initials: T
authors.lastname: George
authors.forename: J
authors.initials: J
authors.lastname: Chandra
authors.forename: P K
authors.initials: PK
authors.lastname: Mukherjee
authors.forename: M A
authors.initials: MA
authors.lastname: Ghannoum
mesh_terms: D000666:Amphotericin B / Q000008:administration & dosage / Q000494:pharmacology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D000935:Antifungal Agents / Q000008:administration & dosage / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects*; D002175:Candida / Q000187:drug effects* / Q000648:ultrastructure; D003470:Culture Media; D004352:Drug Resistance, Microbial; D054714:Echinocandins; D005656:Fungal Proteins; D008081:Liposomes; D008826:Microbial Sensitivity Tests; D018613:Microscopy, Confocal; D010455:Peptides; D010456:Peptides, Cyclic; D011090:Polyenes / Q000494:pharmacology; D016459:Prosthesis-Related Infections / Q000382:microbiology; D014230:Triazoles / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D003470:Culture Media; D054714:Echinocandins; D005656:Fungal Proteins; D008081:Liposomes; D010455:Peptides; D010456:Peptides, Cyclic; D011090:Polyenes; D014230:Triazoles; D000666:Amphotericin B; C007572:echinocandin B