Following the first ever equine influenza outbreak in Australia in 2007, a study was conducted involving 200 horse owners and managers to determine their perceptions about effectiveness of biosecurity measures and the factors associated with these perceptions. Face-to-face interviews were conducted with horse owners/managers to obtain information about their perceptions of the effectiveness of biosecurity practices, their sources of information about infection control during the outbreak and their horse industry involvement. Two outcome variables were created from horse owners’ responses to a 17-item question on the perceived effectiveness of various recommended equine influenza biosecurity measures: (a) a binary outcome variable (Low/High biosecurity effectiveness) and (b) a continuous outcome variable (the proportion of the 17 measures considered ‘very effective’). These outcomes were used in binomial logistic and linear regression analyses, respectively, to determine factors associated with perceptions of biosecurity effectiveness. Variables with a p-value <0.05 in multivariable models were retained in the final models.