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

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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 NeuroTrack ^TM (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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Strategies for combating bacterial biofilms: A focus on anti-biofilm agents and their mechanisms of action

Ranita Roy, Monalisa Tiwari, Gianfranco Donelli … (2017)

ABSTRACT Biofilm refers to the complex, sessile communities of microbes found either attached to a surface or buried firmly in an extracellular matrix as aggregates. The biofilm matrix surrounding bacteria makes them tolerant to harsh conditions and resistant to antibacterial treatments. Moreover, the biofilms are responsible for causing a broad range of chronic diseases and due to the emergence of antibiotic resistance in bacteria it has really become difficult to treat them with efficacy. Furthermore, the antibiotics available till date are ineffective for treating these biofilm related infections due to their higher values of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), which may result in in-vivo toxicity. Hence, it is critically important to design or screen anti-biofilm molecules that can effectively minimize and eradicate biofilm related infections. In the present article, we have highlighted the mechanism of biofilm formation with reference to different models and various methods used for biofilm detection. A major focus has been put on various anti-biofilm molecules discovered or tested till date which may include herbal active compounds, chelating agents, peptide antibiotics, lantibiotics and synthetic chemical compounds along with their structures, mechanism of action and their respective MICs, MBCs, minimum biofilm inhibitory concentrations (MBICs) as well as the half maximal inhibitory concentration (IC50) values available in the literature so far. Different mode of action of anti biofilm molecules addressed here are inhibition via interference in the quorum sensing pathways, adhesion mechanism, disruption of extracellular DNA, protein, lipopolysaccharides, exopolysaccharides and secondary messengers involved in various signaling pathways. From this study, we conclude that the molecules considered here might be used to treat biofilm-associated infections after significant structural modifications, thereby investigating its effective delivery in the host. It should also be ensured that minimum effective concentration of these molecules must be capable of eradicating biofilm infections with maximum potency without posing any adverse side effects on the host.

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Pseudomonas aeruginosa pyocyanin and 1-hydroxyphenazine inhibit fungal growth.

N Høiby, G. Taylor, Stephen J Kerr … (1999)

To examine strains of Pseudomonas aeruginosa for specific antifungal factors. Two clinical strains of P aeruginosa with strong in vitro inhibition (by cross streak assay) of Candida albicans and Aspergillus fumigatus were examined. Both strains were isolated from sputum--one from a patient with cystic fibrosis and one from a patient with bronchiectasis. Bacterial extracts were fractionated by high performance liquid chromatography and examined by ultraviolet absorbance and mass spectroscopy. Antifungal activity against C albicans and A fumigatus was determined in a well plate assay. Pyocyanin was the major antifungal agent of P aeruginosa; 1-hydroxy-phenazine also possessed activity. Pyocyanin MICs for C albicans and A fumigatus were > 64 micrograms/ml. These phenazines were active against nine other yeast species pathogenic for man. Preliminary experiments also suggested possible inhibition of yeast mycelial transformation in C albicans by pyocyanin. There may be a role for pyocyanin and 1-hydroxyphenazine in the prevention of pulmonary candidiasis in patients colonised by P aeruginosa.

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Pseudomonas aeruginosa manipulates redox and iron homeostasis of its microbiota partner Aspergillus fumigatus via phenazines

Benoit Briard, Perrine Bomme, Beatrix E. Lechner … (2015)

The opportunistic fungal pathogen Aspergillus fumigatus is increasingly found as a coinfecting agent along with Pseudomonas aeruginosa in cystic fibrosis patients. Amongst the numerous molecules secreted by P. aeruginosa during its growth, phenazines constitute a major class. P. aeruginosa usually secreted four phenazines, pyocyanin (PYO), phenazine-1-carboxamide (PCN), 1-hydroxyphenazine (1-HP) and phenazine-1-carboxylic acid (PCA). These phenazines inhibited the growth of A. fumigatus but the underlying mechanisms and the impact of these four phenazines on A. fumigatus biology were not known. In the present study, we analyzed the functions of the four phenazines and their mode of action on A. fumigatus. All four phenazines showed A. fumigatus growth inhibitory effects by inducing production of reactive oxygen species (ROS), specifically O2 ·−, and reactive nitrogen species (RNS), ONOO−. A. fumigatus Sod2p was the major factor involved in resistance against the ROS and RNS induced by phenazines. Sub-inhibitory concentrations of PYO, PCA and PCN promote A. fumigatus growth by an independent iron-uptake acquisition. Of the four phenazines 1-HP had a redox-independent function; being able to chelate metal ions 1-HP induced A. fumigatus iron starvation. Our data show the fine-interactions existing between A. fumigatus and P. aeruginosa, which can lead to stimulatory or antagonistic effects.

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Journal

Journal ID (nlm-ta): Virulence

Journal ID (iso-abbrev): Virulence

Title: Virulence

Publisher: Taylor & Francis

ISSN (Print): 2150-5594

ISSN (Electronic): 2150-5608

Publication date (Electronic, pub): 5 October 2020

Publication date (Electronic, collection): 2020

Publication date PMC-release: 5 October 2020

Volume: 11

Issue: 1

Pages: 1329-1336

Affiliations

[a ]Department of Infectious Diseases, Infection Control and Employee Health, The University of Texas M.D. Anderson Cancer Center; , Houston, TX, USA

[b ]California Institute for Medical Research; , San Jose, CA, USA

[c ]INRS-Centre Armand-Frappier Santé Biotechnologie; , Laval, Quebec, Canada

[d ]Division of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine; , Stanford, CA, USA

Author notes

CONTACT Sebastian Wurster stwurster@ 123456mdanderson.org ; Dimitrios P. Kontoyiannis dkontoyi@ 123456mdanderson.org

[*]

These authors contributed equally to this work.

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Dimitrios P. Kontoyiannis https://orcid.org/0000-0002-8051-2940

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Publisher ID: 1827885

DOI: 10.1080/21505594.2020.1827885

PMC ID: 7549912

PubMed ID: 33017225

SO-VID: 7f96ac38-ef4a-4722-94e4-b07e4f44d28c

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This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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

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Microbial Genomics

Most cited references 22

Strategies for combating bacterial biofilms: A focus on anti-biofilm agents and their mechanisms of action

Pseudomonas aeruginosa pyocyanin and 1-hydroxyphenazine inhibit fungal growth.

Pseudomonas aeruginosa manipulates redox and iron homeostasis of its microbiota partner Aspergillus fumigatus via phenazines

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