Background: The microbiology component of Australian undergraduate nursing programmes varies considerably. Any actual or potential impact of this variation on infection control practice, as a nursing graduate, is relatively unknown. Aims: The aim of this study was to explore infection control professionals’ perceptions of the importance of microbiology and infection control training in undergraduate nursing curricula and the perceived retention of that knowledge and its transferability to practice. Method: Semi-structured interviews were conducted with eight infection control professionals from a range of hospital settings in Australia. Findings: Four main themes emerged: Theory versus practice, importance of role modelling, disjunction between university curricula and ‘the real world,’ and learning in context. Conclusion: As the underpinning element of infection control practice, the role of microbiology education and training in nursing education will benefit from review. Further discussions about the nature and timing of theoretical microbiology content and assessment of undergraduate students’ microbiology knowledge to ensure retention and appropriate application of that knowledge in practice are urgently needed.