Establishment of Three Francisella Infections in Zebrafish Embryos at Different Temperatures

Treatment of Francisella infections via PLGA- and lipid-based nanoparticle delivery of antibiotics in a zebrafish model

“…The control groups were injected with corresponding volumes of PBS. A detailed description of the procedure was reported earlier (Brudal et al 2014).…”

“…Optical density at 600 nm was measured with Genesys 20 Spectrophotometer (ThermoFisher Scientific) and ad justed to the value of 1.0. This corresponds to approximately 4.5 × 10 3 colony-forming units being injected into each embryo, as estimated by Brudal et al (2014). The control zebrafish embryo group was injected with PBS.…”

“…orientalis strain routinely used in these experiments expresses a fluorophore (mCherry) and kanamycin resistance gene (Brudal et al 2014). Frozen stock was inoculated on Eugon chocolate agar plates (Brudal et al 2013), and once the colonies were formed, they were transferred into liquid Eugon broth supplemented with iron (Sigma-Aldrich).…”

“…Within hours after injecting the bacteria, they were actively taken up by the fish macrophages, which then assembled into granuloma-like structures. The infection led to a pro-inflammatory immune response dominated by the up-regulation of the proinflammatory cytokines TNF-α and IL-1β, followed by rapid mortality of the infected embryos within 3 to 5 d (Brudal et al 2014).…”

“…Moreover, pathogenesis of many bacterial species in zebrafish closely reflects that of diseased wild and farmed fish and even humans (Phelps & Neely 2005, Sullivan & Kim 2008, Meijer & Spaink 2011, Rowe et al 2014. Brudal et al (2014) established zebrafish embryo infection models of 3 Gram-negative intracellular Francisella spp. bacteria at different temperatures.…”