A Phage Therapy against Pseudomonas aeruginosa Infection in Zebrafish Embryos

At 26 hours post-fertilization, load a microinjection needle with approximately 5 microliters of P. aeruginosa inoculum, and insert the needle dorsally to the starting point of the duct of Cuvier, at which the duct starts spreading over the yolk sac. Inject 1 to 3 nanoliters of the PAO1 inoculum into the embryo, making sure that the volume expands directly within the duct, and enters into the circulation.

After the injection, transfer the embryo into one of two new Petri dishes containing fresh E3 medium supplemented with phenylthiourea for a 30-minute or 3-hour incubation at 28 degrees Celsius. Next, load a microinjection needle with approximately 5 microliters of the phage cocktail, and fix the needle to the microinjector. Then, inject 1 to 3 nanoliters of phage cocktail into the duct of Cuvier of each embryo previously injected with bacteria, and place the embryos into one of two new Petri dishes containing fresh E3 medium supplemented with phenylthiourea at 28 degrees Celsius.