BACKGROUND: Today, considering the importance of bioterrorism, it may be time to assess the risk of bioterrorism as an important priority. Nurses, as the broadest group of therapy group, are very influential in this regard. General aim: To determine the impact of bioterrorism education through two methods of lectures and booklets on the knowledge and attitude of nursing staff. Practical Aim: Minimizing the damage caused by bioterrorism attacks by empowering nursing staff. MATERIALS AND METHODS: This two-group semiexperimental intervention study was performed as a before and after education intervention with the participation of 80 nursing staff of Shams Al-Shomus Nezaja Hospital affiliated to Health Administration of Islamic Republic of Iran Army Ground Forces. The study population included all hospital nursing staff in 2021. The inclusion criteria included having a willingness and informed consent to participate in the study and exclusion criteria included unwillingness to continue cooperation and failure to complete more than 10% of the questionnaire by the subject. This study was performed on 77 nurses in 2021. The nurses were divided into two groups: lecture and booklet. First, the pretest was completed by the nurses. After the educational intervention, both groups completed the posttest. Both groups then completed the questionnaire again after about 2 weeks. First, all the data were measured for normal distribution by the Kolmogorov–Smirnoff test. Statistical description was expressed for the variables with normal distribution as the ratio of geometric mean ± standard error and for abnormal variables as mean ± standard error. To compare the means of the studied quantitative variables, t-test and Wilcoxon matched-paired statistical tests were used. The significance level was considered <0.05. SPSS.16 and Graph Pad Prism 8.0.1 softwares were used to analyze the statistical data. RESULTS: Using both methods is effective in improving the level of knowledge and betterment attitude, but according to the paired t-test in comparison of the two groups, at intervals immediately after training and 2 weeks after training, a significant increase in the average knowledge score of the lecture group Statistically shows (P < 0.001). However, the analysis of attitude scores was performed between the two groups of lectures and booklets, which did not show statistically significant changes between them. CONCLUSIONS: One of the most important issues in the performance of medical staff in bioterrorism attacks is useful, effective, correct, and sufficient training. According to the results of the present study, bioterrorism training leads to increasing the level of knowledge and betterment the attitude of nurses in the field of bioterrorism. Therefore, the inclusion of training courses in medical centers, especially military medical centers, is necessary and sensitive.