Animal Biosafety Level 3 laboratory (Absl-3) is the basic supporting platform of the national biosafety system for new and recurrent infectious diseases, it provides an important hardware basis for the study of biological characteristics, prevention and treatment of highly pathogenic microorganisms. ABSL -3 laboratory also provides an important support for the combination of production, teaching and research. The normal operation of ABSL -3 laboratory and the smooth development of scientific research work can not be separated from the careful planning and effective implementation of emergency planning work. In addition, the reasonable design and layout of the laboratory is also a key factor to avoid and reduce accidents. Based on the practice of construction and management of animal biosafety level III Laboratory in Yangzhou University, starting with the preparation of emergency plan, this paper introduces the points for attention in the layout of laboratory construction, daily operation and emergency treatment, it is expected to provide useful reference for biosafety laboratories operating various pathogenic microorganisms in the future.