Between 1992 and 2015, nearly 148 million hectares (Mha) within biodiversity hotspots - biologically rich but threatened terrestrial regions - worldwide underwent land-cover changes, equating to 6% of the total areal extent of hotspots. Forest losses in hotspots amounted to 54 Mha (-7% of the forest area present in 1992), driven primarily by agricultural expansion (38 Mha); shrubland or savanna also declined by 23 Mha (-8%). Over the same time, urban areas expanded by 10 Mha (+108%). Major losses in forest areas occurred in Sundaland (11 Mha, -13% relative to 1992), Indo-Burma (6 Mha, -6%), and Mesoamerica (5 Mha, -7%). Approximately 7.5 Mha of forest loss occurred within protected areas (-5% of the respective forest area in 1992), of which 3.9 Mha was cleared between 2000 and 2015, with similar to 1 Mha alone converted in the 5 years after 2010. More stringent and effective land-based policies are urgently needed to prevent additional landscape fragmentation and preserve existing species richness in the world's biodiversity hotspots.