Urbanization impacts biodiversity, yet few studies examine general impacts of urbanization on insects. Furthermore, few studies examine availability and limitation of potential cavity nesting sites for ants, an important regulating factor in ant communities that may vary in different urban habitats. We compared three urban habitat types (gardens, vacant lots, and forests) in Toledo, OH, to examine availability and ant preferences for different cavity nesting resources (small and large hollow twigs and cavities). We added 72 artificial large hollow twigs (83 by 6 mm), small hollow twigs (140 by 2 mm), and spherical hollow cavities (6.52–31.1 cm3 in volume, 1-mm opening) to six sites from May to August 2007 to determine whether nest-site limitation impacts ant communities. We collected natural nests to compare natural abundance and occupancy of cavity nests in different urban habitats. We opened artificial and natural nests to calculate the percentage occupied by cavity-nesting ants. Across all habitats, small twigs represented 81.1% of natural nests, spherical nests represented 10.1%, and large twigs 8.2%. Ants occupied 8.1% of natural large twigs, 14.6% of cavities, and 4.1% of small twigs. For artificial nests, 21.5% of large twigs, 1% of small twigs, and 1% of spheres were occupied. The high percentage of occupied artificial large twigs could imply this is a preferred and limiting resource in urban habitats. The results show that certain types of nesting resources may be an important factor mediating ant communities in urban green spaces.