With the time frame 1912–1949, and according to the recording in the government gazettes and Chinese press, this essay systematically discusses the powerful and effective measures and achievements in cholera prevention, epidemic prevention, and infectious disease control of the Macao-Portuguese government. Meanwhile, it analyzes and investigates the cooperation and interaction in cholera prevention between the Macao and Hong Kong governments and emphasizes the critical significance of regional developments of Macao’s cholera prevention and control, that is, with the internal control to lower incidence and external control to minimize importation and diffusion, the aim to control cholera was achieved thus allayed the public panic among Macao and perimeter zone, creating advantages to stabilize economic development, social order and peaceful living environment in Macao as well as other Cantonese regions. As one of the seaport cities that adopted the European medical system and model and implemented free cholera vaccination for all, the fighting process and measures of Macao-Portuguese government adopted to control the cholera epidemic show that cholera, a dreadful infectious disease, can be effectively managed as long as scientific and appropriate methods are taken. Sorting out the types and characteristics of infectious diseases in Macao during this period and the mechanism of combining medical and preventive treatment at that time, and analyzing the effectiveness of the administration of disease prevention and control by the Macao-Portuguese government and the limitations of governance under the conflict between Chinese and Portuguese will have important academic significance for understanding the efficiency and deficiency of Macao-Portuguese governance towards disease control, the construction, development and change of the urban public health emergency management system and the urban governance. The more important significance will bring to the urban public health emergency management system and the urban governance in modern China.