Cuba is located among a group of countries with high dengue incidence. Following several epidemics in the last 10 years, the country designed, implemented, and evaluated a participatory strategy based on the Ecohealth approach. The aim was to promote inter-sector ecosystem management to decrease Aedes aegypti infestation and prevent dengue transmission in the municipality of Cotorro, in Havana city. The study adopted a participatory research methodology. The strategy ensured active participation by the community, diverse sectors, and government in the production of healthy ecosystems. Timely and integrated measures for prevention and control were developed, thereby decreasing the risk of vector proliferation and local dengue transmission. The approach allowed holistic problem analysis, priority setting, and administration of solutions. The strategy has been sustained two years after concluding the process.