A BSTRACT Purpose: Many types of research have been published on the history of biological warfare, the agents used, and the medical implications. However, no studies measure how people are aware of the magnitude of these health problems and international threats. The present study aimed to produce and make background about biological warfare information for health college students to be used as a basis for future studies or research and prepare the hospitals’ bases for similar disasters. Methods: This observational, cross-sectional, descriptive study was conducted among undergraduate students (N = 626) enrolled in health-related colleges at Jazan University, Saudi Arabia. A preliminary survey of 30 participants was then undertaken to improve the questionnaire’s understanding and validity. The questionnaire encompassed three primary sections, including (1) sociodemographic characteristics, (2) knowledge, and (3) awareness. Sociodemographic characteristics consisted of age, gender, college type, academic level, and specialty. All data were gathered using an online self-reported questionnaire using Google Forms and participants were recruited using a random sampling strategy. Results: The total participants were 626 students; 514 were females, whereas 112 were males. Knowledge and attitude indices were 3.8650 ± 0.48 and 4.06 ± 0.51 (maximum is 5). The indices showed variable statistical differences among sociodemographic factors. With adjusted and crude odds ratios of 0.53 and 0.54, attitude score is the sole significant ( P = 0.05) predictor of knowledge as analyzed using logistic regression. Conclusion: The results of the present study are the first of their kind in the region and can be used to shape public awareness among specialists and decision-makers, especially in light of the recent pandemic.