AbstractAim Land use intensity has been recognized as one of the major determinants of native species declines. The re‐expansion of species previously constrained by habitat degradation has been rarely investigated. Here, we use site occupancy models incorporating imperfect detection to identify the land use drivers of the re‐expansion of the Eurasian otter (Lutra lutra).Location Czech Republic.Methods We applied multi‐season occupancy models to otter presence–non‐detection data collected in three national surveys (1992, 2000, 2006) at 552 sites (11.2 × 12 km grid cells). Model parameters included site occupancy, colonization and extinction probabilities, and detection probability at a sub‐site level. We modelled changes in occupancy over time as a function of agricultural, urban and industrial land use and change in the extent of agricultural land use.Results Under the best fitting model, occupancy was estimated to be 34.6% in 1992, 51.3% in 2000 and 83.7% in 2006. Detection probability was neither perfect nor constant. Occupancy probability in 1992 was negatively related to land use gradients. Colonization was more likely to occur where a reduction in agricultural land was larger. Variation in extinction and colonization rates along land use gradients resulted in increased occupancy in industrial and especially urban landscapes. Conversely, occupancy remained almost unchanged along agricultural gradients.Main conclusions Dynamics of otter expansion were strongly associated with the two main patterns of the rapid environmental transition that has taken place in the Czech Republic since the early 1990s. Results show that a reduction in intensive agricultural land use led to an increase in otter distribution, providing evidence of the impact of agricultural land use on stream ecosystems. Moreover, otters recolonized urban and industrial landscapes, probably as a result of extensive reduction in water pollution from point sources. Our results suggest that active conservation of otter populations should focus on restoration of freshwater habitat at large scales, especially in agricultural landscapes.