Assessing Pseudomonas virulence with nonmammalian host: zebrafish.

In the last years, the zebrafish (Danio rerio) has become an important vertebrate animal model to study host-pathogen interactions, especially in its embryonic stage. The presence of a fully developed innate immune system in the first days of embryogenesis, the facility of obtaining and manipulating large numbers of embryos, the optical transparency of the embryos that allow the direct visualization of bacterial infections, a wide range of genetic tools, and extensive mutant resources and collections of transgenic reporter lines are important advantages of the zebrafish-embryo model. Pseudomonas aeruginosa is able to lethally infect zebrafish embryos when the amount of cells injected exceeds the phagocytic capacity of the embryo. Different studies have proved the suitability of zebrafish embryos as a model to analyze P. aeruginosa infection. Here we describe the detailed protocols to establish a P. aeruginosa infection in zebrafish embryos and to image the interaction of the bacterium with this host with fluorescent microscopy.