Introduction: In recent years, the province of Ontario has reported one of the highest Lyme disease incidence rates in Canada due to the abundance of habitat that favours the survival and reproduction of ticks and wildlife species that carry the LD pathogen (Borrelia burgdorferi). The aim of this study was to understand the acceptability and feasibility of various methods for LD prevention in the context of peri-urban settings in Ottawa, where LD is an emerging problem. Methods: This research used an exploratory sequential mixed methods design. We conducted 12 semi-structured key informant interviews with public health practitioners, land management officials, and community members to identify the most locally acceptable and feasible LD interventions. We used themes that emerged during the interviews to develop and refine a previously validated questionnaire and implement a population-representative cross-sectional survey to evaluate public acceptability and feasibility, which applied a 9-point acceptability scale to this context. The survey also collected information on frequency of tick bites, uptake of LD prevention behaviours, and explanatory variables including demographic information, property type and characteristics, and engagement with occupational or recreational outdoor activities. We distributed the survey to the Ottawa public via a web panel in 2024. Results: A total of 1015 participants responded to the survey. Findings from both key informant interviews and the population survey indicated higher acceptability of personal protection strategies and landscape management strategies. Twenty-nine (29) percent of survey respondents reported knowing someone with an LD diagnosis and 3% reported having received a diagnosis of LD at some point, while 22% and 16% of respondents reported having at least one tick encounter or tick bite in the last year, respectively. Only 10% reported consistently using some methods of protection against ticks while outdoors. Acceptability of interventions was more consistently associated with respondents sociodemographic characteristics than with their risk and exposure indices, which varied greatly in their contribution to acceptability across intervention types. Conclusions: In our study, participants generally found host-targeted and chemical strategies to be the least acceptable type of intervention to prevent LD, while personal prevention strategies, land management, and natural alternatives to chemicals were found to be the most acceptable in the Ottawa context. The factors associated with higher acceptability varied by intervention type. This information will be helpful in producing high quality, targeted messaging to promote the uptake of different LD interventions. Overall, these findings will help to better inform policy on feasible and acceptable LD prevention practices that respond to local population preferences and context.