Abstract: To address the challenges in experimental teaching of isolation and identification of pathogenic bacteria from aquatic animals, such as complex content, high technical difficulty, high costs, and time-consuming operations that hinder students’ hands-on practice, this study develops a virtual simulation experiment system for artificial infection, isolation and identification of pathogenic bacteria in seawater and freshwater. The system covers key aspects including symptoms of aquatic animals infected by pathogens, pathological changes, isolation and identification of pathogenic bacteria, pathogenicity testing, artificial infection, and harmless disposal of experimental fish and bacterial strains. Through interactive experimental operations, students can fully understand and apply “Koch’s postulates” for the isolation and identification of aquatic animal pathogens. This approach effectively expands students' learning motivation and practical production skills, while fostering their scientific innovation spirit.