This study described the development of an interactive euthanasia training program and its potential to improve dairy workers' perceived euthanasia decision-making skills and awareness of timely euthanasia by using a survey instrument before and after the program. Training material encompassed euthanasia information over 2 production stages (calves and cows or heifers) and material was delivered on-farm in a case-scenario format (14 cases). During a 3-mo period, 30 different dairy farms were visited and 81 participants were enrolled in this study. Each participant was required to complete a survey pretraining, to complete the case studies from the production stage in which their job responsibility was more closely aligned with (estimated completion time of 1 h), and to complete a survey post-training. Surveys contained 8 statements regarding participants' perceived knowledge of euthanasia practices. The questions were answered on a 5-point scale: (1) strongly disagree, (2) disagree, (3) neither agree nor disagree, (4) agree, or (5) strongly agree. Multivariable mixed-effects logistic regression models were created for each question to investigate the effect of age, sex, dairy experience, farm size, role at the farm, race, previous experience with euthanasia, veterinarian degree, and production stage in the score change, defined by the presence or absence of an increase in the 5-point scale score. Upon completion of the training, respondents were more confident in identifying compromised animals (score change = 0.35), determining when an animal should be euthanized (score change = 0.64), and understanding the importance of timely euthanasia (score change = 0.26). Age and euthanasia experience were significantly associated with the respondents' perceived knowledge; suggesting that younger, less-experienced caretakers on-farm should be prioritized to receive training. The proposed interactive case-based euthanasia training program has proven to be valuable to dairy participants and veterinarians as it provides a means to improve dairy welfare.