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Halophilic microorganisms inhabiting hypersaline environments such as salt lakes, Dead Sea, or salt evaporation ponds, have acquired specific cell adaptation to grow within stressful conditions. In this study, we isolated heterotrophic and autotrophic microorganisms from several saltern ponds located at the Natural Reserve “Saline di Trapani e Paceco”, Sicily, Italy. The aim of the study was to investigate the biotechnological potential of new microbial strains from saltern ponds, by capturing their biological and chemical diversity. After the isolation and identification of the sampled strains, their growth capacity was determined under low and high salinity conditions. The metabolomic profiles of heterotrophs and pigments production of photosynthetic organisms were analyzed. In parallel, antiproliferative tests on human cell lines were conducted with total extracts coming from the microorganism cultures, together with …