ABSTRACT: Human encroachment on nature grows constantly, increasing human-wildlife interactions. Flight initiation distance (FID, the distance at which animals flee from an approaching threat) is often used to measure antipredator behaviour and establish buffer zones to reduce human impact on wildlife. In this study, we approached 10 waterbird species on larger lakes and narrower rivers using a motorboat. We investigated whether water body type, season (winter/spring), approach starting distance, species body mass, and group size influenced bird FID. Average bird FID was 145 ± 92 m and differed between species. In general, FID of all species was larger on lakes than rivers and increased with increasing group size and approach starting distance. When analysed separately for the two most common species, common goldeneyes (Bucephala clangula) and mallards (Anas platyrhynchos), FID increased with increasing starting distance on rivers, but not lakes, likely because birds on lakes have enough time to evaluate the approaching object and take flight at great distance. Additionally, birds might perform different activities on lakes versus rivers, leading to varying energetic trade-offs between the two habitat types, which may affect the decision when to take flight. Finally, mallards aggregated in larger groups on lakes, which affected FID, likely due to enhanced visibility (i.e., earlier detection of the approaching boat) and detection probability (via increased group size) on lakes. Our results emphasize the importance of accounting for habitat characteristics, such as water body type, when studying waterbird FID, because they can affect the visibility of stimuli, group size and potentially animal behaviour, factors which should be taken into account when planning buffer zones for waterbirds in conservation areas.