A questionnaire for farm managers was designed, to obtain information regarding biosecurity on Ontario commercial broiler chicken and turkey operations, and then pre-tested. The questions that could be validated were verifiable by seeing the facility, by using farm records or by interviewing technical personnel other than the survey respondent. The survey was validated using a convenience sample of 24 farms from two companies. For 15 questions with dichotomous responses, the sensitivity ranged from 16.7 to 100%; the specificity ranged from 0 to 100%. For example, fences and gates seen during the farm visit were not accurately reported on the survey (poor sensitivity). Chance-corrected agreement was low (kappa < 0.4) for 34 questions, fair to good (0.4 < kappa < 0.8) for 25 questions, and excellent (kappa > 0.8) for seven questions. The percent agreement for questions where only one of the possible options was observed on validation ranged from 60.9 to 100%. Five questions with continuous numeric variables were analysed. A difference was observed (P < 0.1) between the survey and validation data for three questions regarding the number of birds, the bird sources and the downtime between flocks. In spite of pre-testing, the lack of clear wording and the absence of definitions for technical terms appeared to reduce validity. Response bias seems to be an issue with biosecurity surveys. The value of validating questionnaires before their use in epidemiologic research is confirmed.