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Updated: Sep 17, 2025
Modeling Persistent Pseudomonas aeruginosa Infection in Wounded Zebrafish Larvae
Published on: June 13, 2025
Modeling Persistent Pseudomonas aeruginosa Infection in Wounded Zebrafish Larvae
Nilly Flore1, Blanc-Potard Anne-Béatrice2
1Laboratory of Pathogens and Host Immunity (LPHI), CNRS, Inserm, Université de Montpellier; flore.nilly@umontpellier.fr.
Abstract:
Pseudomonas aeruginosa is a major human pathogen, particularly in chronic wound infections and chronic pulmonary infections (especially in patients with cystic fibrosis (CF)). Chronic bacterial infections are refractory to antibiotic treatments and there is an urgent need for in vivo chronic infection models amenable to drug screening to develop efficient therapies. Here, we describe a protocol of infection by a P. aeruginosa clinical isolate from a CF patient, expressing constitutively the Green Fluorescent Protein (GFP), for generating a persistent wound infection in zebrafish larvae. Tail fin-injured embryos are immersed in a bacterial suspension for 1.5 h, washed, and monitored for bacterial load for 3 days. The bacterial burden was quantified daily by counting fluorescent colony-forming units (CFU) from lysed infected larvae, which allowed a persistent infection. Moreover, persistent P. aeruginosa bacteria were refractory to antibiotic treatment. This novel in vivo model of persistent P. aeruginosa infection offers opportunities to evaluate the efficacy of innovative treatments against chronic infections.
Insights
A new zebrafish model mimics persistent Pseudomonas aeruginosa wound infections, crucial for testing new therapies against antibiotic-resistant bacteria in chronic conditions like cystic fibrosis.
Area of Science:
- Microbiology
- Infectious Diseases
- Zebrafish Models
Background:
- Pseudomonas aeruginosa is a significant pathogen causing difficult-to-treat chronic wound and pulmonary infections, especially in cystic fibrosis (CF) patients.
- Current antibiotic treatments are often ineffective against chronic bacterial infections, highlighting the need for novel therapeutic strategies.
- Developing reliable in vivo models is essential for screening potential drugs against persistent infections.
Purpose of the Study:
- To establish a novel zebrafish larvae model for persistent Pseudomonas aeruginosa wound infections.
- To utilize a clinical isolate expressing Green Fluorescent Protein (GFP) for tracking bacterial burden.
- To assess the potential of this model for evaluating antibiotic efficacy against chronic infections.
Main Methods:
- Zebrafish embryos were infected with a GFP-expressing P. aeruginosa clinical isolate via tail fin injury and immersion.
- Larvae were washed and monitored for bacterial load over 3 days post-infection.
- Bacterial burden was quantified daily by counting fluorescent colony-forming units (CFU) from lysed larvae.
Main Results:
- The protocol successfully generated a persistent P. aeruginosa wound infection in zebrafish larvae.
- Quantification of GFP-labeled bacteria confirmed a sustained bacterial burden over the observation period.
- The persistent P. aeruginosa infection in this model demonstrated resistance to antibiotic treatment.
Conclusions:
- This study presents a validated in vivo zebrafish model for persistent P. aeruginosa wound infections.
- The model effectively simulates chronic infection characteristics, including antibiotic refractoriness.
- This platform provides a valuable tool for screening and developing new treatments for chronic P. aeruginosa infections.
