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The degradation of aquatic habitats has increasingly become one of the most important factors influencing the distribution of freshwater species worldwide. We analysed the occurrence of three crayfish species, Astacus astacus, Austropotamobius torrentium and Orconectes limosus, in relation to habitat degradation status (based on Directive 2000/60/ES), stream morphology, geographical characteristics and land cover. In total, we analysed 6,768 sites within the Czech Republic (Central Europe), of which 6,187 sites lacked crayfish; among the remainder, A. astacus was present in 507 sites, O. limosus occurred in 44 sites and A. torrentium was present in only 30 sites. The analysis revealed that A. astacus preferred streams of better water quality that were not surrounded by agricultural land or settlements. This species also preferentially occurred in smaller streams with stony bottom substrata that were located at higher altitudes. Austropotamobius torrentium occurrence was associated with the natural character of the water body and the presence of protected areas at higher altitudes. Conversely, the non-native crayfish species O. limosus was typically recorded at lower altitudes in downstream reaches surrounded by agricultural land and with deteriorated water quality. The degree of environmental (human) pressure besides differences in habitat characteristics at sites with ICS and NICS may be of general importance for conservation strategies aimed at central European native crayfish species, primarily because the habitat-driven co-occurrence pattern (and its possible changes in the future) may strongly influence interspecific relationships, such as direct competition and the spread of infectious diseases between crayfish species.