Abstract
Filamentous fungi and bacteria form mixed-species biofilms in nature and diverse clinical contexts (Frey-Klett et al., 2011; Peleg et al., 2010). The interactions between fungi and bacteria, often mediated by secreted metabolites, have important ramifications for the biology of the interacting partners (Frey-Klett et al., 2011). This is particularly true for the bacterium Pseudomonas aeruginosa (P. aeruginosa) and the fungus Aspergillus fumigatus (A. fumigatus) which often reside in the same niche such as lungs of cystic fibrosis (CF) patients. Some studies have reported that co-infection with P. aeruginosa and A. fumigatus could lead to a decrease in lung function relative to their respective single species infection (Amin et al., 2010; Peleg et al., 2010). Metabolite extraction and analysis allow for the characterization of specific microbial metabolites in the polymicrobial biofilm. This protocol describes how to prepare the Pseudomonas-Aspergillus co-culture biofilm on solid medium in preparation for metabolite extraction.
Keywords: Aspergillus fumigatus, Pseudomonas aeruginosa, Biofilm, Cross Kingdom communication
Materials and Reagents
Equipment
Procedure
Representative data
Figure 5. Representative image of the co-culture biofilm of Pseudomonas aeruginosa PA14 wild-type (center colony) and Aspergillus fumigatus AF293 wild-type (wrinkled biofilm surrounding bacterial colony) after 6 days of incubation at 25 °C
Notes
Recipes
Acknowledgments
This work was supported by startup and ISEN funding from Northwestern University (to Y. W.), NIH Grant R01 GM 067725 (to N. L. K.), and National Science Foundation Grant Emerging Frontiers in Research and Innovation 1136903 (to N. P. K.). This protocol was adapted from Zheng et al. (2015).
References
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