Existing research highlights conflicting results attempting to capture gender preferences with respect to unimodal versus multimodal designs in online learning. As online learning continues to expand, more research examining the role of gender in multimedia design holds considerable potential. However, the presence of multimedia in online learning contexts presents a more complex problem when taking into consideration the concept of learner interest. The current study sought to investigate gender differences in situational interest (SI) with respect to three different but related designs of an online, multimedia learning environment for continuing, professional education in emergency medical services. The content in these designs was identical, but applied different combinations of the redundancy and modality multimedia principles. A two-way analysis of variance was used to examine the types of SI by gender and type of multimedia design. While no significant difference or interaction effect was identified between gender and triggered SI, an interaction effect was observed for females and the different multimedia designs. Specifically, females reported a higher maintained SI when animation, narration and text were effectively combined. The findings of this study provide insight into best practices for instructional and media designers developing multimedia learning environments as well as future research implications.