Primary teachers’ attitudes have been shown to strongly influence both their intention to teach science, and ultimately student engagement and learning outcomes, with previous research consistently showing preservice teachers hold negative attitudes towards science and/or science teaching. A recently developed instrument informed by a theoretical framework for attitudes towards science teaching (van Aalderen-Smeets et al. 2012) has shown promise in effectively identifying and evaluating primary teachers’ attitudes. This study explored 96 Australian preservice primary teachers’ attitudinal constructs toward teaching science in a single-semester science education course. Data sources included participants’ pre- and posttest responses to the Dimensions of Attitude towards Science (DAS) instrument, and interview responses from a smaller subset of participants (n = 5). Results indicated a significant improvement in participants’ confidence in teaching science, their perceptions of the relevance of science teaching, and their enjoyment of science teaching at the conclusion of the course. Participants were less anxious about teaching science and expressed improved intentions to include science activities in their classrooms. Self-efficacy was the most improved attitudinal construct, with findings indicating the science education course facilitated cognitive content mastery and cognitive pedagogical mastery (Palmer 2006). Evidence from this study supports the assertion that significant improvements in preservice primary teachers’ attitudes toward teaching science are possible in courses considered to be ‘typical’ science education courses in Australia, and future courses should focus on maximising opportunities for preservice teachers to improve their confidence via cognitive content mastery experiences and cognitive pedagogical mastery experiences.