Employees in the equine industry are at occupational risk of zoonoses such as Hendra virus and equine chlamydiosis through exposure to infected materials. This study aimed to gain a deeper understanding of the views and experiences of employees, and the key drivers of infection control and personal biosecurity (PB) practices in the Thoroughbred breeding industry. Methods: An exploratory qualitative study was conducted in 2018 in New South Wales, Australia using interviews (9) and small group discussions (7). The 29 participants included veterinarians, veterinary nurses, foaling staff, stud managers and laboratory personnel working in a range of equine medicine settings. Interviews and focus groups were recorded, transcribed and analysed manually by at least two members of the research team. An iterative approach was used to derive themes. Results: Five main themes emerged: (i) greater awareness of current and emerging infectious risks promotes use of Personal Protective Equipment (PPE); (ii) currently available PPE is not comfortable, practical or well-suited to equine reproductive work in Australia's hot climate; (iii) creating supportive environments for PB reduces risk of exposure to infectious materials; (iv) strong leadership is required to implement sustainable change in workplace culture and practices; and (v) policy and economic factors play an important role in adopting biosecurity (BS) and PB measures in the workplace. Personnel working in the Australian Thoroughbred breeding industry face unique zoonotic risks in a challenging physical environment. A qualitative approach provided rich insights into social and physical factors motivating BS and PB in this occupational group. There is an opportunity for collaboration between Public Health services and industry partners to develop and implement strategies most likely to be effective in ensuring consistent uptake of PB measures in the workplace.