JoVE Logo

Sign In

Concept
Experiment

A Phage Therapy against Pseudomonas aeruginosa Infection in Zebrafish Embryos


Transcript


Take anesthetized dechorionated cystic fibrosis zebrafish embryos susceptible to Pseudomonas aeruginosa infection.

Inject fluorescent protein-expressing Pseudomonas aeruginosa into a major blood vessel, the duct of Cuvier.

Transfer injected embryos to a plate with media and a pigmentation-blocking agent to inhibit embryo pigmentation and visualize bacteria. Incubate.

Injected bacteria move through the circulation and invade organs. 

In response, the host immune system gets activated, releasing pro-inflammatory mediators and recruiting immune cells.

Following incubation, inject a cocktail of virulent phages against Pseudomonas aeruginosa into the duct of Cuvier and transfer the embryos to a plate containing media and the pigmentation-blocking agent.

Incubate. Phages infect the bacteria and use host machinery to produce new phage particles.

Bacteria lyse, releasing phages that infect neighboring bacteria, resulting in a reduced pro-inflammatory response.

Using a fluorescent microscope, observe the reduced bacterial fluorescence in the zebrafish embryos over time, indicating phage effectiveness against Pseudomonas aeruginosa.

USAGE STATISTICS
JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved