Background: UNAIDS estimated the adult HIV prevalence rate in Trinidad and Tobago was 1.5% in 2010 – the fourth highest in the Caribbean. Global and Caribbean evidence demonstrates that areas of high tourism activity can be sites of high-risk sexual interaction between tourists and locals, and this may contribute to the growing HIV transmission. The aim of this study is to explore and understand the factors that influence the adoption of HIV practices within hotels in Trinidad and Tobago. By examining hotels’ (lack of) response to national policies, recommendations are made to inform future policies of a major public health concern within the private sector. Methodology: The study uses policy analysis approaches to understand why hotels adopt HIV prevention and control practices, and an ecological lens to categorise these factors. Qualitative data were collected using semi-structured interviews with representatives from 54 hotels, government agencies and hotel and tourism associations, together with a review of relevant policy documents. Results: Commonly reported practices in the tourism sector include non-discrimination towards HIV-positive staff and guests; use of gloves to clean rooms; preventing staff-guest interactions; and HIV training and sensitisation. Commonly reported barriers included HIV not being considered a priority; limited hotel resources; and staff attitudes and resistance, while important facilitators included existing hotel systems and procedures; and management being directly affected by HIV. Conclusion: Recommendations include increased HIV awareness for both hotel management and staff using hotels’ already existing systems; increased leadership among hotel and tourism associations in the adoption of HIV policies and programmes through sharing of information and peer collaboration; and further operational research on HIV in the tourism industry to help inform the decisions of hotel owners/managers and policy makers.