Ic cells. Purification by way of a 12 step sucrose gradient was performed before conditioning in vitro and in vivo.Introduction: Infections by two Gram-negative intracellular bacterial pathogens Piscirickettsia salmonis and Francisella noatunensis, are causing major difficulties in aquaculture world-wide. F. noatunensis sp hampers the development of fish farming depending on cod in and is deleterious to tilapia. P. salmonis infections have already been devastating for salmon aquaculture. As of currently no powerful treatments are obtainable against the diseases. Each P. salmonis and F. noatunensis secrete membrane vesicles (MV). Bacterial MV has been reported as possible vaccine candidates for any range of host which includes humans, mice and fish against infection triggered by intracellular pathogenic bacteria as they induce each a humoral and cellular immunity.ISEV2019 ABSTRACT BOOKMethods: We have IFITM1/CD225 Proteins Recombinant Proteins isolated MVs from each Francisella and Piscirickettsia by the ultracentrifugation Process. The MVs had been characterized by their size distribution, by transmission electron microscopy (TEM) and proteomics. Their toxicity were tested by injecting MVs into each our zebrafish vaccine and challenge model also as in cod, tilapia and salmon. A vaccine trail was performed initial in our zebrafish model, 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. Each single and double membrane MV had been found within the population as investigated by TEM. Additional, immune-gold labelling revealed the presence of DNA in each populations. Proteomics analysis revealed that the MV content material varied among bacterial strains. Immunization with MV gave protection against illness caused by both P. salmonis and F. noatunensis in our zebrafish model, however, did not defend cod, tilapia nor salmon. Summary/Conclusion: The MVs from P. salmonis and F. noatunensis revealed a related size distribution and that the content contains numerous bacterial virulence components too as DNA that can be transferred to the host. As for their immunogenic properties this appears to vary in between the vaccine and challenge model in comparison with the natural hosts. The usage of the MVs as vaccines in their natural hosts such as strain-specificity and cross-immunity need 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, Siglec-7 Proteins site Hassan Chaudhrya, Stuart Manneringd, Stuart Cordwellb, Maria Kaparakis-Liaskose and Richard Ferreroaa Hudson Institute of Health-related Study, Melbourne, Australia; bThe University of Sydney, Sydney, Australia; cMonash University, Melbourne, Australia; dSt. Vincent’s Institute of Health-related 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 the T84 line were grown on Transwell filters to create transepithelial electrical resistance (TEER), a measure of epithelial monolayer integrity. The cells had been then cocultured with Alexa Fluor-labelled OMVs in the gastric pathogen, Helicobacter pylori. Final results: We showed that H. pylori OMVs readily entered polarised epithelial cells, but had no effect around the TEER nor permeability.