A Phage Therapy against Pseudomonas aeruginosa Infection in Zebrafish Embryos

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.

At 26 hours post-fertilization, load a microinjection needl