The term One Health encompasses the medicine of humans, animals, the environment and the interrelations between them. There has been substantial progress in recognising the importance of collective agreements and thinking in healthcare and preventative medicine, particularly in the wake of the COVID-19 pandemic, with amplified acknowledgement of the role zoonoses increasingly play in our global health system. The 2021 Royal Society of Medicine’s One Health: The Missing Link conference brought together subject experts from across medical specialties to discuss cultures of coworking, key challenges and the future vision of One Health both nationally and on a global scale. This perspective article aims to summarise the discussions arising from the event, and highlights particular challenges in medical education, public and global health policy, antimicrobial resistance and stewardship and environmental medicine. This article will serve to consolidate the outcomes of this event and generate dialogue between clinical and non-clinical stakeholders within the One Health space. One Health Impact Statement This article summarises the discussions and contributions of the 2021 Royal Society of Medicine One Health symposium and linked events. Major discussion points are described including antimicrobial resistance, training of human and animal clinicians, the link to environmental medicine, and positioning one health efforts within the wider third sector. We outline current challenges in unifying a One Health approach as well as offering perspectives for collaboration and interdisciplinary work.