Changes in land use in Brazil, particularly agricultural expansion, have significant impacts on ecosystems and their associated services. This study aimed to evaluate the temporal and spatial dynamics of ecosystem services (ES) in the Pedras River basin (PRB), the main water supply for Guarapuava, southern Brazil, between 2002, 2018 and 2023. The analysis was based on the Ecosystem Services Identification Matrix, integrating land use and land cover (LULC) maps and field observations of environmental and socioeconomic characteristics in the area. This approach allowed the classification and assessment of the potential supply of provisioning, regulating, and cultural ES in the basin. The findings between 2002 and 2018 revealed a general decline in most regulating and cultural services, with notable reductions in genetic resources, local climate regulation, air quality regulation services, and medicinal resources. Conversely, provisioning services related to food provision, energy provision, and soil quality regulation showed increases. Between 2018 and 2023, food provision, water quality regulation, soil quality, and pollination declined, along with decreases in cultural services. In contrast, the increase occurred in water supply, raw materials, genetic resources, ornamental resources (18.16%), and air quality and climate regulation services. Agricultural expansion resulted in a trade-off, increasing food provision while simultaneously degrading other ES. The study emphasizes that sustainable land management is necessary to preserve ecosystem functions and services, thereby preventing the degradation of their potential.