Deforestation in the biosphere reserves, which are key Protected Areas has negative impacts on biodiversity, climate, carbon fluxes and livelihoods. Comprehensive study of deforestation in biosphere reserves is required to assess the impact of the management effectiveness. This article assesses the changes in forest cover in various zones and protected areas of Nilgiri Biosphere Reserve, the first declared biosphere reserve in India which forms part of Western Ghats-a global biodiversity hotspot. In this study, we have mapped the forests from earliest available topographical maps and multi-temporal satellite data spanning from 1920's to 2012 period. Mapping of spatial extent of forest cover, vegetation types and land cover was carried out using visual interpretation technique. A grid cell of 1 km × 1 km was generated for time series change analysis to understand the patterns in spatial distribution of forest cover (1920-1973-1989-1999-2006-2012). The total forest area of biosphere reserve was found to be 5,806.5 km(2) (93.8 % of total geographical area) in 1920. Overall loss of forest cover was estimated as 1,423.6 km(2) (24.5 % of the total forest) with reference to 1920. Among the six Protected Areas, annual deforestation rate of >0.5 was found in Wayanad wildlife sanctuary during 1920-1973. The deforestation in Nilgiri Biosphere Reserve is mainly attributed to conversion of forests to plantations and agriculture along with submergence due to construction of dams during 1920 to 1989. Grid wise analysis indicates that 851 grids have undergone large-scale negative changes of >75 ha of forest loss during 1920-1973 while, only 15 grids have shown >75 ha loss during 1973-1989. Annual net rate of deforestation for the period of 1920 to 1973 was calculated as 0.5 followed by 0.1 for 1973 to 1989. Our analysis shows that there was large-scale deforestation before the declaration of area as biosphere reserve in 1986; however, the deforestation has drastically reduced after the declaration due to high degree of protection, thus indicating the secure future of reserve in the long term under the current forest management practices. The present work will stand as the most up-to-date assessment on the forest cover of the Nilgiri Biosphere Reserve with immediate applications in monitoring and management of forest biodiversity.