Constraints to the introduction of enhanced biosecurity systems are rarely considered in sufficient detail when population medicine specialists initiate new control schemes. The main objective of our research was to investigate and compare the different attitudes constraining improvement in biosecurity for cattle and sheep farmers, practising veterinary surgeons and the auxiliary industries in Great Britain (GB). This study was carried out utilizing farmer focus groups, a questionnaire survey of veterinary practitioners and a telephone survey of auxiliary industry representatives. It appears that farmers and veterinarians have their own relatively clear definitions for biosecurity in relation to some major diseases threatening GB agriculture. Overall, farmers believe that other stakeholders, such as the government, should make a greater contribution towards biosecurity within GB. Conversely, veterinary practitioners saw their clients' ability or willingness to invest in biosecurity measures as a major constraint. Veterinary practitioners also felt that there was need for additional proof of efficacy and/or the potential economic benefits of proposed farm biosecurity practices better demonstrated. Auxiliary industries, in general, were not certain of their role in biosecurity although study participants highlighted zoonoses as part of the issue and offered that most of the constraints operated at farm level.