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Extracellular vesicles (EVs) are released from cells and carry protein and genetic cargo involved in cell communication. EVs isolated from bodyfluids, including plasma and serum, can also be used as markers of pathophysiological changes. Peptidylarginine deiminases (PADs) are phylogenetically conserved enzymes with physiological and pathophysiological roles and cause post-translational protein deimination. This can affect function of target proteins and deimination is also linked to EV release. Atlantic cod (Gadus morhua L.) reared at 4 °C and 9 °C respectively for 18 months, were here assessed for changes in serum derived EVs, including analysis of deiminated protein and micro-RNA cargo markers related to stress and growth. We found that cod reared at 9 °C showed significantly reduced numbers of EVs in serum, compared to cod reared at 4 °C. Some deiminated protein targets, including complement component C3, were found to be considerably higher in EVs of cod reared at 4 °C. Proteomic analysis revealed further differences in deiminated protein targets in EVs isolated from sera of the two temperature groups. Whole cod sera from the two temperature groups furthermore showed differences in deiminated protein targets, including C3, CRP and histone H3, which is a marker of neutrophil extracellular trap formation. MicroRNAs related to inflammation (miRNA-21) and stress (miRNA-155) were elevated in both total cod serum and serum-derived EVs from the 9 °C group, while the growth-related miRNA-206 was higher in the 4 °C group. Our findings highlight EVs as novel biomarkers to assess fish health in response to environmental rearing temperature.

aquaculture reports

Extracellular vesicles, deiminated protein cargo and microRNAs are novel serum biomarkers for environmental rearing temperature in Atlantic cod (Gadus morhua L.)