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Updated: Feb 7, 2026
Time-Lapse Epifluorescence Microscopy Imaging of Pseudomonas aeruginosa and Staphylococcus aureus Heterogeneous Phenotypes
Published on: February 14, 2025
Drosophila melanogaster as a polymicrobial infection model for Pseudomonas aeruginosa and Staphylococcus aureus
Young-Joon Lee1, Hye-Jeong Jang1, In-Young Chung1
1Department of Pharmacy, College of Pharmacy and Institute of Pharmaceutical Sciences, CHA University, Seongnam, 13488, Republic of Korea.
Abstract:
Non-mammalian infection models have been developed over the last two decades, which is a historic milestone to understand the molecular basis of bacterial pathogenesis. They also provide small-scale research platforms for identification of virulence factors, screening for antibacterial hits, and evaluation of antibacterial efficacy. The fruit fly, Drosophila melanogaster is one of the model hosts for a variety of bacterial pathogens, in that the innate immunity pathways and tissue physiology are highly similar to those in mammals. We here present a relatively simple protocol to assess the key aspects of the polymicrobial interaction in vivo between the human opportunistic pathogens, Pseudomonas aeruginosa and Staphylococcus aureus, which is based on the systemic infection by needle pricking at the dorsal thorax of the flies. After infection, fly survival and bacteremia over time for both P. aeruginosa and S. aureus within the infected flies can be monitored as a measure of polymicrobial virulence potential. The infection takes ~24 h including bacterial cultivation. Fly survival and bacteremia are assessed using the infected flies that are monitored up to ~60 h post-infection. These methods can be used to identify presumable as well as unexpected phenotypes during polymicrobial interaction between P. aeruginosa and S. aureus mutants, regarding bacterial pathogenesis and host immunity.
Insights
This study introduces a fruit fly model to analyze interactions between Pseudomonas aeruginosa and Staphylococcus aureus. It allows monitoring of fly survival and bacterial load to understand polymicrobial virulence.
Area of Science:
- Microbiology
- Pathogenesis Research
- Infectious Disease Modeling
Background:
- Non-mammalian models are crucial for understanding bacterial pathogenesis.
- Fruit flies (Drosophila melanogaster) offer conserved immunity and physiology, making them suitable models.
- Assessing polymicrobial interactions is vital for understanding complex infections.
Purpose of the Study:
- To present a simple protocol for studying in vivo polymicrobial interactions.
- To evaluate the interaction between Pseudomonas aeruginosa and Staphylococcus aureus in a host model.
- To enable the identification of virulence factors and host immune responses in polymicrobial infections.
Main Methods:
- Systemic infection of Drosophila melanogaster via needle prick.
- Monitoring of fly survival post-infection (up to ~60 hours).
- Quantification of Pseudomonas aeruginosa and Staphylococcus aureus bacteremia over time.
Main Results:
- The protocol allows for the assessment of polymicrobial virulence potential.
- Fly survival and bacterial load provide measurable outcomes for pathogen interaction.
- The model facilitates the study of both expected and unexpected phenotypes.
Conclusions:
- This fruit fly model provides a robust platform for studying polymicrobial pathogenesis.
- The protocol enables detailed analysis of bacterial interactions and host immune responses.
- It aids in identifying virulence mechanisms and potential targets for antibacterial strategies.
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