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The relationship between cities and biodiversity is extremely complex in Latin America. The region is simultaneously the world’s most urbanized, has some of the world’s largest social and economic inequities, and hosts some of the world’s most biodiversity-rich ecosystems, including several biodiversity hotspots. As cities in Latin America are expected to continue to expand, partly on areas harboring valuable biodiversity hotspots, there is an urgent need to understand how biodiversity and ecosystem services interplay in and around cities. This assessment aims to describe urbanization trends in Latin America and the related impacts on urban biodiversity and ecosystem services, complementing the general framework with shorter case studies of four cities around the region. It also explores the potential for city planning to provide support for biodiversity and ecosystem services. The study found that cities in Latin America exhibit extreme social and economic differences, which generate a complex mosaic of urban settlement structures and ecosystem management systems. Low-income neighborhoods are typically either interspersed with the local ecosystems in peri-urban areas or completely lacking green spaces. High-income neighborhoods have a higher concentration of green areas, but are usually dominated by nonnative species. It also found that conservation of ecosystems and biodiversity, and ecosystem services provisioning, are low priorities in urban planning; they are not acknowledged as key elements for the quality of life of the city inhabitants and human well-being. The knowledge base is also limited, as research on the …