The European roe deer (Capreolus capreolus) is the most common deer species in Europe. The species can be a reservoir of some tick-borne diseases but it is primarily recognized for its contribution as an amplifier host. In Central Europe, two roe deer ecotypes are living in adjacent areas: field and forest. We investigated differences in tick load and species composition on these two ecotypes. We collected ticks from 160 (80 the forest ecotype and 80 the field ecotype) roe deer culled in Wielkopolska Region (West-Central Poland). The most common was Ixodes ricinus (n = 1610; 99%) followed by Ixodes hexagonus (n = 22; 1%). The dominant life stage of the ticks was female. Prevalence was higher for forest roe deer. Mean number of ticks found on the forest ecotype was almost fivefold higher than on the field ecotype (3.75 ± 0.83 vs. 0.77 ± 0.20 ticks). The mean probability of tick occurrence was threefold higher in the forest (0.915 ± 0.050) than the field ecotype (0.279 ± 0.125). The most infested body parts of roe deer from both ecotypes were the neck and the head.