<p>The rapid recovery and expansion of the Eurasian beaver (<i>Castor fiber</i>) across Europe have led to increasing human–wildlife conflicts and the implementation of targeted management measures, including regulated culling. However, systematic evaluations of beaver culling at the national scale remain limited. This study provides the first comprehensive analysis of Eurasian beaver culling in the Czech Republic since the introduction of the Beaver Management Programme in 2013. Data on issued derogations and the number of individuals culled were collected from all administrative regions and analysed to assess their spatial and temporal patterns. Between 2013 and 2023, a total of 497 individuals were culled under 72 derogations and general binding measures. Culling intensity increased after 2016 and was strongly concentrated in the western and southeastern parts of the Czech Republic, particularly in areas with high beaver occurrence and increased potential for conflicts with human activities. Most culling occurred in Zone B, which functions as a buffer between strictly protected areas and high-conflict zones. Despite ongoing population expansion, culling remains spatially selective and is primarily applied in response to local conflicts. This study provides the first systematic overview of beaver culling in the Czech Republic and highlights the increasing need for active management of human–beaver conflicts.</p>