Abstract: The teaching content of isolation and identification for aquatic animal pathogens is complex, technical difficulty, high experimental cost, experiment time consuming, and the real experiment cannot be carried out. According to the actual teaching needs of courses such as “Microbiology Experiment of Aquatic Animal Pathogen”, the virtual simulation experimental system of artificial infection, isolation and identification of seawater and freshwater pathogens is constructed. The system includes six technologies: symptom of infected aquatic animals, pathological changes, isolation and identification of pathogenic bacteria, pathogenic examination, artificial infection experiment, disposal of experimental fish and strains. Through interactive experimental operation, students can fully understand and use the “Koch’s postulates” for the pathogen isolation and identification of aquatic animals. It effectively expands the breadth and depth of experimental teaching of marine and aquatic courses, and improves students' learning enthusiasm, scientific innovation spirit and practical ability.