The objective of this exploratory study was to identify the content of preservice teachers’ practicalknowledge and the influence of their acculturation experiences on their practical knowledge. Informed byteacher occupational socialisation theory, a mixed method design was used to examine 26 first-year preservicephysical education and health (PEH) teachers’ practical knowledge construction during their first PEH course.Data collection included a written assignment in which the preservice teachers, based on their learningexperiences during the PEH course, were asked to describe 10 significant didactical milestones (DMs) (practicalknowledge) that would guide their future teaching in PEH. The early stage of teacher socialisation, theiracculturation phase, was examined and analysed by using a questionnaire gathering information about theirbackground and previous experiences. Results showed that the content of these preservice PEH teachers’practical knowledge was mainly pedagogical and focused most often on instructional strategies. The preserviceteachers showed a relative lack of focus on knowledge concerning curriculum aims and contextual issues relatedto school and society. Furthermore, the results showed significant individual differences in the content of theirpractical knowledge. Consequently, we must acknowledge that acculturation and professional socialisation ofindividual preservice teachers will not be similar and that there is fundamentally multifaceted interactionbetween the structure of teacher education, different stages, and content. Teacher educators need to consider thecomplexity between individual learning experiences and the importance of how the content is organised in adifferentiated approach.