Achromobacter Species Isolated from Cystic Fibrosis Patients Reveal Distinctly Different Biofilm Morphotypes.
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.
Department of Clinical Medicine, Aarhus University, DK-8200 Aarhus, Denmark. smni@clin.au.dk.|Department of Clinical Microbiology, Aarhus University Hospital, DK-8200 Aarhus, Denmark. smni@clin.au.dk.
Signe M
SM
Nielsen
Department of Clinical Microbiology, Aarhus University Hospital, DK-8200 Aarhus, Denmark. nielnoer@rm.dk.
Niels
N
Nørskov-Lauritsen
Costerton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, DK-2200 Copenhagen, Denmark. tbjarnsholt@sund.ku.dk.|Department of Clinical Microbiology, Rigshospitalet, DK-2100 Copenhagen, Denmark. tbjarnsholt@sund.ku.dk.
Thomas
T
Bjarnsholt
Interdisciplinary Nanoscience Center, Aarhus University, DK-8000 Aarhus, Denmark. rikke.meyer@inano.au.dk.|Department of Bioscience, Aarhus University, DK-8000 Aarhus, Denmark. rikke.meyer@inano.au.dk.
Rikke L
RL
Meyer
Achromobacter; PNA-FISH; antimicrobial susceptibility testing; biofilm; biofilm morphology; confocal microscopy; cystic fibrosis; dispersal
J Cyst Fibros. 2013 Mar;12(2):170-6
22944724
J Cyst Fibros. 2016 Jul;15(4):479-85
26412052
Mol Microbiol. 2006 Feb;59(4):1114-28
16430688
J Clin Microbiol. 2006 Sep;44(9):3415-7
16954289
Proc Natl Acad Sci U S A. 2016 Jul 19;113(29):8266-71
27382184
Respiration. 2000;67(3):239-47
10867591
Science. 1999 May 21;284(5418):1318-22
10334980
J Antimicrob Chemother. 1990 Dec;26(6):857-60
2081727
Int J Antimicrob Agents. 2010 Jan;35(1):68-71
19889520
Trends Microbiol. 2013 Sep;21(9):466-74
23827084
J Cyst Fibros. 2011 Dec;10(6):466-9
21835703
Nat Protoc. 2010 Jul;5(7):1236-54
20595953
Infect Immun. 2015 Jul;83(7):2686-93
25895968
J Cyst Fibros. 2013 Dec;12(6):638-43
23727271
Int J Food Microbiol. 2014 Sep 18;187:26-32
25043896
PLoS One. 2015 Aug 27;10(8):e0136790
26313451
PLoS One. 2011;6(11):e27943
22132176
PLoS One. 2013 Jul 22;8(7):e68484
23894309
Antimicrob Agents Chemother. 2015 Nov 02;60(1):650-2
26525791
J Clin Microbiol. 2012 Aug;50(8):2688-94
22675125
PLoS Genet. 2014 Oct 16;10(10):e1004651
25330091
Science. 2002 Feb 22;295(5559):1487
11859186
Curr Opin Biotechnol. 2015 Jun;33:73-80
25528382
Int J Biol Macromol. 1999 Oct;26(1):3-16
10520951
Antimicrob Agents Chemother. 2014 Nov;58(11):6851-60
25182651
J Vis Exp. 2011 Jan 30;(47):null
21307833
Nat Methods. 2012 Jul;9(7):671-5
22930834
Methods Mol Biol. 2014;1211:261-71
25218392
J Intern Med. 2007 Jan;261(1):5-16
17222164
Antimicrob Agents Chemother. 2013 May;57(5):2352-61
23478967
Curr Top Microbiol Immunol. 2008;322:85-105
18453273
Infect Immun. 2012 Aug;80(8):2601-7
22585963
Ann Am Thorac Soc. 2014 Sep;11(7):1120-9
25102221
Crit Rev Microbiol. 2015;41(3):341-52
24303798
Appl Environ Microbiol. 1994 Jul;60(7):2355-9
8074516
APMIS. 2008 Sep;116(9):837-41
19024606
J Cyst Fibros. 2013 May;12(3):298-301
23141756
Curr Opin Microbiol. 2008 Dec;11(6):560-6
18935979
J Cyst Fibros. 2005 Aug;4 Suppl 2:49-54
16023416
Curr Opin Microbiol. 2012 Feb;15(1):71-7
22137884
Semin Respir Crit Care Med. 2007 Jun;28(3):312-21
17562501
Curr Protoc Microbiol. 2011 May;Chapter 1:Unit 1B.2
21538301
J R Soc Med. 1995;88 Suppl 25:24-9
7776324
J Bacteriol. 2001 Sep;183(18):5395-401
11514525
J Cyst Fibros. 2006 Dec;5(4):245-51
16777495
N Engl J Med. 1994 Sep 8;331(10):637-42
7503821
J Cyst Fibros. 2010 Jan;9(1):51-8
19939747
Environ Microbiol Rep. 2013 Dec;5(6):778-86
24249286
Proc Natl Acad Sci U S A. 1990 Dec;87(23):9188-92
2251263
PLoS One. 2012;7(7):e41075
22815921
J Bacteriol. 2008 Apr;190(8):2767-76
18156255
Acta Paediatr. 2006 Sep;95(9):1070-4
16938752
J Bacteriol. 2007 Jan;189(1):28-37
17041046
BMC Microbiol. 2013 Apr 12;13:82
23586331