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Global health trends, marked by increasing epidemics and pandemics, call for enhanced human resource preparedness to prevent and respond to emerging and future health problems. Indeed, according to the World Organization for Animal Health (WOAH), at least 75% of the pathogens responsible for emerging and re-emerging human infectious diseases originate from animals. These health problems involve environmental, economic and social factors. Because of their multidimensional nature, addressing these health problems requires the adoption of a One Health approach. To achieve this, training of human resources is a priority.This descriptive cross-sectional study was conducted from September 12 to 22, 2021 in 6 regions of Senegal, namely, Dakar, Thiès, Diourbel, Kaolack, Saint-Louis, and Ziguinchor. Questionnaires were administered online and in person to 217 professionals in human, environmental, animal health and agriculture. Data were statistically processed, and bivariate analyses enabled a better correlation between training needs for professionals according to their occupational group.The survey sample was 64% male and 36% female. More than half of the professionals surveyed (53.5%) had less than 10 years of experience. Human health workers were the most represented (46%), followed by animal health workers (34%) and environmental workers (16%). Agriculture, fisheries, and food security were weakly represented. Few had received training in the One Health approach and entry-level One Health skills predominated. The One Health competencies in which professionals want to build capacity differ by sector. Globally, public health and epidemiology, health risk management, basic of One Health concepts, animal health and biotechnology are the priorities.The development of training programs could then enable these expressed needs to be addressed.