Ticks are significant vectors of pathogens in human and veterinary medicine and have been identified as (re)emerging health threats. The primary objective of this study was to collect new data on the fauna of hard ticks within the region of Istria with a focus on spatial distribution using a geographical information system (GIS). All tick specimens were collected over three years (2020–2023), and this research included all 41 self-government units of Istria and Brijuni Islands National Park. Ticks were collected using the flagging/dragging method and manually from hosts (humans, domestic, or wild animals). In addition, morphological identification using tick keys was performed. The obtained data were used to create maps and feed models and to predict risk assessments. Collected data reveal the predominant presence of Ixodes ricinus, accounting for (n = 446) or 48.1% of the tick population. Rhipicephalus sanguineus (Ixodida: Ixodidae) follows with (n = 253) or 27.23%, and Hyalomma marginatum represents (n = 136) or 14.64% of the tick species collected using the host method in the region. Tick–host relationships are complex and influenced by a range of ecological and environmental factors. The results of this research will contribute to a better understanding, identification, and prediction of the changes in their geographic ranges and help in the prevention and control of zoonosis transmitted to humans by ticks. The obtained results mapped using GIS support the first study on the spatial distribution of ticks in the region of Istria in Croatia.