The invertebrates of the Macocha Abyss, Moravian Karst, Czech Republic, were collected in 2007–2008 and 222 species were identified in total. The relative abundance of individual taxa of land snails, harvestmen, pseudoscorpions, spiders, millipedes, centipedes, terrestrial isopods, beetles, and ants was evaluated. The cold-adapted mountain and subterranean species inhabit the bottom and lower part of the abyss, whereas the sun-exposed rocky margins were inhabited by thermophilous species. Macocha harbors several threatened species that are absent or very rare in the surrounding habitats. In the forest landscape, the Macocha Abyss represents a natural habitat with a distinct microclimatic gradient, and is an excellent refuge area for psychrophilous as well as thermophilous species, which significantly contributes to maintenance of landscape biodiversity.