Background: In this study, we explore how a health education course may play a role in pre-service teachers' perceptions in teaching and integrating health education activities to nurture K-8 students' health literacy. Methods: We used mixed methods to examine the effect of a health education course in a teacher education program. Of 55 pre-service teachers, 41 voluntarily participated in the study. Quantitative data were obtained through an online questionnaire administered to participants at the beginning and end of the course. We conducted 6 focus groups at the end of the course. Results: The inferential analysis from a series of analysis of variance with repeated measures revealed significant differences in health knowledge (F = 113.39, p < 0.01, n[superscript 2] = 0.74), preparedness (F = 104.74, p < 0.01, n[superscript 2] = 0.73), attitudes (F = 15.02, p < 0.01, n[superscript 2] = 0.28), and beliefs (F = 8.87, p < 0.01, n[superscript 2] = 0.19) between time points. Qualitative data led to the conclusion that where one health education course is insufficient, such a course is the first step into future curriculum development and implementation. Conclusions: One health education course might be beneficial for general education teachers to increase their knowledge and preparation to teaching school health. On-going training is needed for program success.