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Small Colony Variants of Pseudomonas aeruginosa Display Heterogeneity in Inhibiting Aspergillus fumigatus Biofilm

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Abstract

Pseudomonas aeruginosa and Aspergillus fumigatus are major microbes in cystic fibrosis (CF). We reported non-mucoid P. aeruginosa isolates more inhibitory to A. fumigatus than mucoid ones. Another CF P. aeruginosa phenotype, small colony variants (SCVs), is an unknown factor in intermicrobial competition with A. fumigatus. Clinical SCV isolates and reference CF non-mucoid isolate (Pa10, producing normal-sized colonies) were compared. Live cells of P. aeruginosa or filtrates from P. aeruginosa planktonic or biofilm cultures were co-incubated with A. fumigatus growing under conditions allowing biofilm formation or with preformed biofilm. Metabolic activity of A. fumigatus biofilm was then measured. When necessary, assays were done after adjustment for growth differences by adding fresh medium to the planktonic culture filtrate. Pyoverdine determinations were performed spectrophotometrically on the planktonic culture filtrates. In all experimental conditions (live cells and planktonic or biofilm culture filtrates of P. aeruginosa versus A. fumigatus biofilm formation or preformed biofilm), three SCV isolates were less inhibitory than Pa10, two equal or more inhibitory. Adjusting planktonic culture filtrates for growth differences showed SCV inhibition differences variably related to growth or deficient inhibitor production. Studies suggested the principal P. aeruginosa inhibitor to be pyoverdine. SCV isolates appear heterogeneous in their capacity to inhibit A. fumigatus biofilm. SCV isolates can be important in the CF microbiome, because they are capable of intermicrobial inhibition.

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Acknowledgements

This work was presented in part to the 4th Meeting of the ECMM/ISHAM Working group, Fungal respiratory infections in Cystic Fibrosis, in Osuna (Seville), Spain, June 5–6, 2017. These studies were partially supported by a grant from the Child Health Research Institute, Stanford Transdisciplinary Initiatives Program, CIMR no. 3777, and by a gift from John Flatley, CIMR no. 3770. The funding sources had no involvement in study design, data handling or interpretation, writing or decision to publish.

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  1. Rajesh Anand

    Present address: Department of Biotechnology, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur, C.G, 495009, India

Authors and Affiliations

  1. California Institute for Medical Research, 2260 Clove Dr., San Jose, CA, 95128, USA

    Rajesh Anand, Gabriele Sass, Karl V. Clemons, Marife Martinez & David A. Stevens

  2. Division of Infectious Disease and Geographic Medicine, Department of Medicine, Stanford School of Medicine, Stanford University, Stanford, CA, 94305, USA

    Rajesh Anand, Gabriele Sass, Karl V. Clemons & David A. Stevens

  3. Division of Pulmonology, Department of Pediatrics, Stanford School of Medicine, Stanford University, Stanford, CA, 94305, USA

    Richard B. Moss

  4. Department of Pathology, Stanford School of Medicine, Stanford University, Stanford, CA, 94305, USA

    Niaz Banaei

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Correspondence to Rajesh Anand.

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CF isolates from patient respiratory cultures were obtained after written informed consent for biobanking of the patients’ specimens and subsequent use, approved by the Stanford Institutional Review Board.

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Anand, R., Moss, R.B., Sass, G. et al. Small Colony Variants of Pseudomonas aeruginosa Display Heterogeneity in Inhibiting Aspergillus fumigatus Biofilm. Mycopathologia 183, 263–272 (2018). https://doi.org/10.1007/s11046-017-0186-9

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  • DOI: https://doi.org/10.1007/s11046-017-0186-9

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