| Summary: | Sea cucumbers are highly marketable as food and medicinal product. This has resulted in an increasing overfishing and in a new interest in European species. In this work, the reproductive biology of <i>Holothuria forskali</i> and <i>Holothuria mammata</i> was performed by evaluating the gonadosomatic index and histological analyzes of the gonadal tubules. The biotechnological potential was assessed through the evaluation of the antioxidant, antimicrobial and antitumor potential. The antioxidant activity was evaluated through the quantification of the total phenolic content, DPPH radical scavenging activity and ORAC method. The antimicrobial activity was evaluated against <i>Staphylococcus aureus, Candida albicans, Saccharomyces cerevisiae, Bacillus subtilis, Salmonella enteritidis, Pseudomonas aeruginosa</i> and <i>Escherichia coli</i> through growth inhibition tests. The antitumor potential was performed on HepG-2 and MCF-7 human cells lines using the MTT and Calcein - AM methods. Finally, the fatty acid profile was evaluated through gas-chromatography analysis. The gonadosomatic index and histology revealed that the range from February to April corresponds to the peak of gonad maturation for both species. No significant antioxidant activity was detected. The methanolic fraction of <i>H. forskali</i> revealed the highest antimicrobial potential against <i>Candida albicans</i> with an IC50 of 233.2 µg ml-1 and also presented the highest cytotoxic and anti-proliferative activities through the MTT method in both cells lines, with an IC50 of 238.2 and 396.0 µg ml-1 for MCF-7 cells, respectively and 260.3 and 218.7 µg ml-1 for HepG-2 cells, respectively. Regarding the fatty acid profile, the total fat content was 1%, 3.36% and 4.83% for <i>H. forskali, H. mammata</i> and <i>S. regalis</i>, respectively and the highest values were obtained for C16:0 (9.96%) and ARA (20.36%) for H. forskali and C18:0 (12.43%), C18:1 n-7 (5.13%), EPA (12.49%) and DHA (7.35%) for S. regalis. These findings showed the potential for aquaculture and also the high biotechnological and seafood potential of the three species.
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