Introduction: In the biological outbursts, nurses are at the frontline of response. They need education about biological disasters. Objective: The present study aimed to compare the effectiveness of two methods including lecture-based training and multimedia software on military nurses’ knowledge about the triage of biological incident victims. Material and Methods: This study was quantitative, quasi-experimental, and interventional. Participants included 50 nurses of emergency wards or Members of the Crisis Committee in two military hospitals. They were randomly divided into two groups of multimedia software or lecture-based training about triage in biological disasters. Furthermore, Pre-test and post-test were performed. The researchermade questionnaire was valid and reliable. Data were analyzed by SPSS 25. The significance level of the tests was less than 5%. Results: The participants of both groups were homogeneous in terms of age, gender, education level, history of membership in disaster team, shift work, work experience, and participation in training courses. Mean knowledge scores of both groups did not show a significant difference before and after the intervention (P>0. 05). Each group had a significant difference in knowledge before and after the intervention (P=0. 001). Thus, the knowledge score had increased after the education in both groups. Discussion and Conclusion: Either of the methods (lecture-based and multimedia training) can be used due to their same effects. However, implementing virtual and distance learning methods in key items such as triage will be necessitated due to the rapid spread of the disease, the time-consuming nature of classical education, dependence on time, place, and coordination required in the formation of the class, and the lack of human resources as one of the challenges facing the hospital in such disasters. Evaluating the effectiveness and professional development of multimedia educational software, animation, and remote interactive training is recommended in teaching the content required in biological disasters.