Continuing the series of workshops on the role of extracellular vesicles (EVs) started in Nanoinnovation 2022 (Extracellular vesicles: The new era of the intercellular communication) and continued with the editions of 2023 (The intercellular communications by extracellular vesicles: a specialized and diversified network in physiological and pathological conditions) and 2024 (The emerging role of extracellular vesicles in reproduction: from gametogenesis to interaction with immune system), we decided in this edition of Nanoinnovation 2025 to develop the topic of EVs in the more general and inclusive context of One Health. Indeed, it is surprising that although EVs play a critical role in maintaining the homeostasis of normal physiological systems, most of our knowledge of EV signalling comes from pathological systems, such as the understanding of the role of EVs gained from their analysis in biofluids obtained in the clinic. The world of extracellular vesicles is therefore not limited to disease, but their role in physiological processes is crucial for good health, which in turn is closely linked to the health of the environment, animals and plants. Therefore, in order to interfere with EV signalling for therapeutic benefit, a more fundamental understanding of how EV populations change in different bio districts - in size, number and physicochemical composition - is crucial to unravel their functional roles in translational models in vitro and in vivo, which can then be fed back to the clinic.