Abstract
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.
Highlights
Polymicrobial infections have increasingly become a focus of interest in infectious diseases research as inter-kingdom interplay of pathogens can mutually affect their virulence, susceptibility to antimicrobial therapy, and interactions with host immune surveil lance [1]
We compared mycelial growth inhibi tion by WT Pseudomonas aeruginosa (PA) filtrates depending on iron availability, as iron is a source of competition between Aspergillus fumigatus (AF) and PA in their in vivo environment [2]
Efficient techniques for longitudinal visualization and analysis of morphological endpoints could expand our understanding of the complex inter-kingdom relation ships of fungal and bacterial pathogens competing for the same ecological niches
Summary
Polymicrobial infections have increasingly become a focus of interest in infectious diseases research as inter-kingdom interplay of pathogens can mutually affect their virulence, susceptibility to antimicrobial therapy, and interactions with host immune surveil lance [1]. PA metabolites with iron-binding capacity, such as pyoverdine or pyocyanin, inhibit growth, bio film formation, and metabolism of AF [2,3,4]. Whereas most of these studies used conventional measurements such as growth inhibition or changes in metabolism [2], there is a need for robust and efficient experimental systems that assess morphogenesis, a key feature of fungal virulence [5], in the context of bacterial interactions. Currently available modalities to study mycelial expansion, hyphal morphology, and bio film formation, such as fluorescence or electron micro scopy [6,7,8] and spectroscopy techniques [9], have low throughput and are time-intensive. Most currently employed assays for inter-kingdom interac tion studies [2,3,4,6,7,8,9] present endpoint analyses that are not suitable for efficient longitudinal monitoring
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
