Abstract:Anchialine caves contain haline bodies of water with underground connections to the ocean and limited exposure to open air. Despite being found on islands and peninsular coastlines around the world, the isolation of anchialine systems has facilitated the evolution of high levels of endemism among their inhabitants. The unique characteristics of anchialine caves and of their predominantly crustacean biodiversity nominate them as particularly interesting study subjects for evolutionary biology. However, there is presently a distinct scarcity of modern molecular methods being employed in the study of anchialine cave ecosystems. The use of current and emerging molecular techniques, e.g., next-generation sequencing (NGS), bestows an exceptional opportunity to answer a variety of long-standing questions pertaining to the realms of speciation, biogeography, population genetics, and evolution, as well as the emergence of extraordinary morphological and physiological adaptations to these unique environments. The integration of NGS methodologies with traditional taxonomic and ecological methods will help elucidate the unique characteristics and evolutionary history of anchialine cave fauna, and thus the significance of their conservation in face of current and future anthropogenic threats. Here we review previous contributions to our understanding of anchialine biodiversity and evolution, and discuss the potential of "speleogenomic" methods for future research in these threatened systems.