Ic cells. Purification by means of a 12 step sucrose gradient was performed prior to conditioning in vitro and in vivo.Introduction: Infections by two Gram-negative intracellular bacterial pathogens Piscirickettsia salmonis and Francisella noatunensis, are causing important issues in aquaculture world-wide. F. noatunensis sp hampers the improvement of fish farming determined by cod in and is deleterious to tilapia. P. salmonis infections have already been devastating for salmon aquaculture. As of nowadays no powerful therapies are readily available against the diseases. Each P. salmonis and F. noatunensis secrete membrane vesicles (MV). Bacterial MV has been reported as potential vaccine candidates for a range of host such as humans, mice and fish against infection PPARβ/δ Gene ID triggered by intracellular pathogenic bacteria as they induce each a humoral and cellular immunity.ISEV2019 ABSTRACT BOOKMethods: We have isolated MVs from both Francisella and Piscirickettsia by the ultracentrifugation Approach. The MVs have been characterized by their size distribution, by transmission electron microscopy (TEM) and proteomics. Their toxicity had been tested by injecting MVs into both our zebrafish vaccine and challenge model also as in cod, AMPA Receptor Inhibitor site tilapia and salmon. A vaccine trail was performed initially in our zebrafish model, and then in cod, tilapia and salmon. Outcomes: The MV size evaluation showed that the MVs size distribution ranged from 2050 nm in size with most ranging from 7000 nm. Both single and double membrane MV were located inside the population as investigated by TEM. Additional, immune-gold labelling revealed the presence of DNA in each populations. Proteomics evaluation revealed that the MV content material varied amongst bacterial strains. Immunization with MV gave protection against disease triggered by each P. salmonis and F. noatunensis in our zebrafish model, on the other hand, didn’t guard cod, tilapia nor salmon. Summary/Conclusion: The MVs from P. salmonis and F. noatunensis revealed a related size distribution and that the content includes different bacterial virulence factors as well as DNA that may be transferred to the host. As for their immunogenic properties this seems to differ between the vaccine and challenge model in comparison to the organic hosts. The usage of the MVs as vaccines in their organic hosts including strain-specificity and cross-immunity have to have further investigation. Funding: Analysis Council of Norway (RCN) and University of Oslo.OF14.Bacterial membrane vesicles enter polarised epithelial cells and provide their protein cargo to exosomes Lorinda Turnera, Nestor Solisb, Georg Rammc, Viola Oorschotc, Amanda De Paolia, Hassan Chaudhrya, Stuart Manneringd, Stuart Cordwellb, Maria Kaparakis-Liaskose and Richard Ferreroaa Hudson Institute of Healthcare Investigation, Melbourne, Australia; bThe University of Sydney, Sydney, Australia; cMonash University, Melbourne, Australia; dSt. Vincent’s Institute of Healthcare Analysis, Melbourne, Australia; 5Department of Physiology, Anatomy and Microbiology, La Trobe University, Melbourne, Australiaresistance and apical-basolateral polarity of normal epithelium. For this, colonic epithelial cells of your T84 line have been grown on Transwell filters to create transepithelial electrical resistance (TEER), a measure of epithelial monolayer integrity. The cells were then cocultured with Alexa Fluor-labelled OMVs from the gastric pathogen, Helicobacter pylori. Final results: We showed that H. pylori OMVs readily entered polarised epithelial cells, but had no impact on the TEER nor permeability.