This study explored the self-reported behaviors and perceptions of Australian paramedics in relation to their environmental hygiene practices. A national online survey was conducted with Paramedics Australasia members (N = 417). Participants reported working in ambulances often contaminated with body fluids. Widespread noncompliance with routine and deep cleaning of ambulances, and misunderstandings about environmental hygiene practices were apparent. Improvements to environmental hygiene practices of Australian paramedics are recommended to avoid pathogen transmission and ensure patient safety.