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Live imaging and quantitative analysis of Aspergillus fumigatus growth and morphology during inter-microbial interaction with Pseudomonas aeruginosa.

Authors
  • Wurster, Sebastian1
  • Sass, Gabriele2
  • Albert, Nathaniel D1
  • Nazik, Hasan2
  • Déziel, Eric3
  • Stevens, David A2, 4
  • Kontoyiannis, Dimitrios P1
  • 1 Department of Infectious Diseases, Infection Control and Employee Health, The University of Texas M.D. Anderson Cancer Center , Houston, TX, USA.
  • 2 California Institute for Medical Research , San Jose, CA, USA.
  • 3 INRS-Centre Armand-Frappier Santé Biotechnologie , Laval, Quebec, Canada. , (Canada)
  • 4 Division of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine , Stanford, CA, USA.
Type
Published Article
Journal
Virulence
Publisher
Landes Bioscience
Publication Date
Dec 01, 2020
Volume
11
Issue
1
Pages
1329–1336
Identifiers
DOI: 10.1080/21505594.2020.1827885
PMID: 33017225
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Pseudomonas aeruginosa (PA) and Aspergillus fumigatus (AF) chronically colonize the airways of patients with cystic fibrosis or chronic immunosuppression and mutually affect each other's pathogenesis. Here, we evaluated IncuCyte time-lapse imaging and NeuroTrackTM (NT) analysis (Wurster et al., 2019, mBio) as a toolbox to study mycelial expansion and morphogenesis of AF during interaction with PA. Co-incubation of AF with supernatant filtrates of wild-type (WT) PA strains strongly inhibited hyphal growth and branching. Consonant with prior metabolic studies, pyoverdine-deficient PA mutants had significantly attenuated inhibitory capacity. Accordingly, purified PA products pyoverdine and pyocyanin suppressed mycelial expansion of AF in a concentration-dependent way. Using fluorescence-guided tracking of GFP-AF293 mycelia during co-culture with live WT PA cells, we found significant inoculum-dependent mycelial growth inhibition and robust precision of the NT algorithm. Collectively, our experiments position IncuCyte NT as an efficient platform for longitudinal analysis of fungal growth and morphogenesis during bacterial co-infection.

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