This study assessed the implementation of a virtual assistant chatbot, EpiBot, in an online epidemiology course using the Reach, Effectiveness, Adoption, Implementation, and Maintenance framework to evaluate acceptability, feasibility, and effectiveness. EpiBot was piloted by 29 public health undergraduate students at a Hispanic Serving Institution on the West Coast of the United States. EpiBot's use and accuracy were monitored via the Amazon Web Services platform and an electronic questionnaire, based on the Technology Acceptance Model, was used to evaluate participant perceptions of usefulness, ease of use, attitudes, intentions, self-efficacy, and preferences. Evaluation of EpiBot's sustainability in the course was guided by the Program Sustainability Assessment Tool. EpiBot accurately answered 93.7% of 1607 student submitted queries (89.5% epidemiology content; 10.5% course administration). Participants perceived EpiBot as useful and easy to use, reported a positive impact on their grades (65.5%), used EpiBot as their primary source of information (55.2%), and experienced minimal barriers to use. Key implementation and maintenance challenges included time for development and non-developer training; platform limitations; and operational costs. Strengths supporting sustainability included capacity for adaptation, supportive climate, and interdisciplinary collaboration.