Historical records of Ateles chamek (black-faced black spider monkey) suggest that the species range extends further south of the known species distribution, within an ecotonal region between the Amazonia, Cerrado and Pantanal biomes in Brazil. Ecotones are zones of habitat transition with high species richness that remain undersampled as conservationists often prioritize biodiversity hotspots. Thus, distribution ranges may be inaccurately measured when species occur in ecotonal zones. We report the first precise records of A. chamek in 24 new localities surveyed in the ecotonal zone of the Upper Paraguay River Basin, and we present subgroup encounter rates in the 11 largest patches (>70 ha) along 207 km of the line transects surveyed. The new records represent an expansion of the distribution of A. chamek approximately 200 km to the south, increasing the known extent of its occurrence by 10.8%. Local tributaries may not be barriers for spider monkeys, which are able to swim and cross slow-moving rivers. However, the dry forests of the Cerrado and the flooded areas of the Pantanal, formed by grassland and scarce trees, may be habitat barriers for A. chamek. The populations living in this ecotonal zone are relatively abundant (1.1-6.67 subgroup sightings/10 km) compared to the heavily hunted continuous forests of northern Amazonia. Furthermore, these values are similar to those for other Ateles spp. inhabiting forests with low or no hunting pressure. We highlight the need for specific conservation action to protect the spider monkeys living in these landscapes, which are threatened by agriculture expansion.