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Open AccessArticle

Achromobacter Species Isolated from Cystic Fibrosis Patients Reveal Distinctly Different Biofilm Morphotypes

1
Department of Clinical Medicine, Aarhus University, DK-8200 Aarhus, Denmark
2
Department of Clinical Microbiology, Aarhus University Hospital, DK-8200 Aarhus, Denmark
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Costerton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, DK-2200 Copenhagen, Denmark
4
Department of Clinical Microbiology, Rigshospitalet, DK-2100 Copenhagen, Denmark
5
Interdisciplinary Nanoscience Center, Aarhus University, DK-8000 Aarhus, Denmark
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Department of Bioscience, Aarhus University, DK-8000 Aarhus, Denmark
*
Author to whom correspondence should be addressed.
Academic Editor: Roger Pickup
Microorganisms 2016, 4(3), 33; https://doi.org/10.3390/microorganisms4030033
Received: 15 July 2016 / Revised: 19 August 2016 / Accepted: 1 September 2016 / Published: 14 September 2016
(This article belongs to the Special Issue The Biofilm Matrix)
Achromobacter species have attracted attention as emerging pathogens in cystic fibrosis. The clinical significance of Achromobacter infection is not yet fully elucidated; however, their intrinsic resistance to antimicrobials and ability to form biofilms renders them capable of establishing long-term chronic infections. Still, many aspects of Achromobacter biofilm formation remain uncharacterized. In this study, we characterized biofilm formation in clinical isolates of Achromobacter and investigated the effect of challenging the biofilm with antimicrobials and/or enzymes targeting the extracellular matrix. In vitro biofilm growth and subsequent visualization by confocal microscopy revealed distinctly different biofilm morphotypes: a surface-attached biofilm morphotype of small aggregates and an unattached biofilm morphotype of large suspended aggregates. Aggregates consistent with our in vitro findings were visualized in sputum samples from cystic fibrosis patients using an Achromobacter specific peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) probe, confirming the presence of Achromobacter biofilms in the CF lung. High antibiotic tolerance was associated with the biofilm phenotype, and biocidal antibiotic concentrations were up to 1000 fold higher than for planktonic cultures. Treatment with DNase or subtilisin partially dispersed the biofilm and reduced the tolerance to specific antimicrobials, paving the way for further research into using dispersal mechanisms to improve treatment strategies. View Full-Text
Keywords: Achromobacter; cystic fibrosis; biofilm; biofilm morphology; dispersal; confocal microscopy; PNA-FISH; antimicrobial susceptibility testing Achromobacter; cystic fibrosis; biofilm; biofilm morphology; dispersal; confocal microscopy; PNA-FISH; antimicrobial susceptibility testing
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MDPI and ACS Style

Nielsen, S.M.; Nørskov-Lauritsen, N.; Bjarnsholt, T.; Meyer, R.L. Achromobacter Species Isolated from Cystic Fibrosis Patients Reveal Distinctly Different Biofilm Morphotypes. Microorganisms 2016, 4, 33.

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