A Phage Therapy against Pseudomonas aeruginosa Infection in Zebrafish Embryos

Abstract

Source: Cafora, M., et al., A. Phage Therapy Application to Counteract Pseudomonas aeruginosa Infection in Cystic Fibrosis Zebrafish Embryos. J. Vis. Exp. (2020)

The video demonstrates the phage therapy technique targeting Pseudomonas aeruginosa infection in cystic fibrosis zebrafish embryos. Phage therapy effectively diminishes bacterial load and mitigates proinflammatory immune responses in CF zebrafish embryos infected with bacteria, leading to reduced CF embryo lethality.

Protocol

1. Microinjection of zebrafish embryos with bacteria and phage cocktail NOTE: To perform a systemic infection, the embryo must have blood circulation that usually starts after 26 hpf. After 26 hpf, dechorionate embryos with pronase solution prepared by dissolving pronase powder in E3 medium at a concentration of 2 mg/mL. Gently pipette the embryos to break the chorion. Discard the pronase/E3 medium and rinse the dish several times with fresh E3 to …

Materials

Bacto Agar	BD	214010
Dulbecco's phospate buffered saline PBS	Sigma-Aldrich	D8537
Ethyl 3-aminobenzoate methanesulfonate	Sigma-Aldrich	886-86-2	Common name tricaine
Femtojet Micromanipulator	Eppendorf	5247
Microinjection needles	Harvard apparatus
N-Phenylthiourea >=98%	Aldrich-P7629	103-85-5
Pronase	Sigma-Aldrich	9036-06-0
Z-MOLDS Microinjection	Word Precision Instruments