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The scientific evaluation of landscape fragmentation and connectivity is important for habitat conservation. It is strongly influenced by the spatial resolution of source maps, particularly in urban environments. However, there is limited comprehensive investigation of the spatial grain effect on urban habitat and few in-depth analysis across different urban gradients. In this paper, we scrutinize the spatial grain effects of urban green space (UGS) cover maps (derived from remote sensing imagery and survey data) with respect to evaluating habitat fragmentation and connectivity, comparing among different urban gradient scenarios (downtown, urban periphery, and suburban area) in Hangzhou, a megacity in China. The fragmentation was detected from three indices, including Entropy, Contagion, and Hypsometry. Then morphological spatial pattern analysis (MSPA) was applied for the landscape element identification. The possibility of connectivity (PC) and patch importance (dPC) were proposed for measuring the landscape connectivity based on Cores and Bridges from MSPA results. The results indicate that the farther the location is from downtown, the less sensitive the landscape element proportion to the spatial resolution. Among the three fragmentation indices, the overall hypsometry index has the lowest sensitivity to the spatial resolution, which implies this index’s broader application value. Considering connectivity, high spatial resolution maps are appropriate for analyzing highly heterogeneous urban areas, while medium spatial resolution maps are more applicable to urban periphery and suburban area with larger UGS patches and less fragmentation. This study suggests that the spatial resolution of UGS maps substantially influence habitat fragmentation and connectivity, which is critical for decision making in urban planning and management.

Spatial Grain Effects of Urban Green Space Cover Maps on Assessing Habitat Fragmentation and Connectivity