The landscape between the Laurentian Mountains in Québec and the Adirondack Mountains in New York State is one of three north-south wildlife movement linkages that connect natural areas in Southeastern Canada with Northeastern United States. This region boasts a wide variety of habitats that still maintain a high degree of ecological integrity and are rich in biodiversity. However, increasing development is putting the area under heightened risk of habitat loss and fragmentation. The objective of this study was to quantify the degree of human modification that has occurred within the region over the past 26 years. Habitat loss was measured by comparing changes in area and proportion of land use and land cover classes and landscape fragmentation was calculated using the Effective Mesh Size fragmentation metric as well as measuring changes in the length and density of the road network. Between 1992 and 2018 developed areas expanded by 1056.9 km2 (69.5 percent), whereas, wetlands decreased by -1365.1 km2 (-68.9 percent), and forests types decreased by -1363.2 km2. Habitat fragmentation increased at all scales and the road network increased by 2587.6 km. Although 75.5 percent of the remaining landscape is habitat, it is made up of 67,790 fragments. Of these fragments, only 8201 are greater than 1 km2 in size. MRC/Counties with the lowest levels of fragmentation were those that contained or were adjacent to protected areas. Thus, conservation planning at the scale of the MRC/County where local municipal governments can work together to create in situ conservation plans is recommended.