BackgroundIn recognition of the interconnected nature of complex challenges such as COVID-19, a collaborative, multisectoral, and transdisciplinary approach, referred to as One Health, has been employed to address sustainable development and strengthen global health security. Although significant investments have been made to build global health capacity, characterization of the One Health is absent from the literature.Methods and findingsWe collected and analyzed perspectives from students, graduates, workers, and employers in One Health through a multinational online survey across health disciplines and sectors. Respondents were recruited through professional networks. A total of 828 respondents from 66 countries participated, representing governmental and academic institutions and students, among others; 57% were female, and 56% had completed professional health degrees. Interpersonal communication, communication with non-scientific audiences, and the ability to work in transdisciplinary teams were valued in the workplace and were considered essential competencies to build an interdisciplinary health workforce. Employers indicated difficulty recruiting workers, while workers indicated limited availability of positions. Employers identified limited funding and ill-defined career pathways as prominent challenges for retaining One Health workers.ConclusionsSuccessful One Health workers use interpersonal skills and scientific knowledge to address complex health challenges. Aligning the definition of One Health will likely improve the matching of job seekers and employers. Encouraging the employment of the One Health approach for a diverse range of positions, even if they do not explicitly include “One Health” in the job title, and clarifying the expectations, roles and responsibilities within a transdisciplinary team will lead to building a stronger workforce. As One Health has evolved to address food insecurity, emerging diseases, and antimicrobial resistance, it holds promise for supporting an interdisciplinary global health workforce that can make substantial progress on Sustainable Development Goals and improve global health security for all.