Marine organisms are vast sources of bioactive components, with  holothorians, or sea cucumbers being one of these sources. Therefore, sea cucumber specimens were examined as a promising source of natural substances that can be used as antibacterial, antifungal, anti-inflammatory, antioxidant, antiviral, and antitumor agents. Holothuria papillifera wascollected fromZaafarana, Ras Ghareb, and Altur stations and employed as a biological weapon against some pathogens. H. papillifera extracts showed high activity as antibacterial extracts against Staph. aureus and Enterococcus faecalis (Gram+ve), Escherichia coli, Salmonella typhi, and Klebsiella pneumoniae (Gram-ve). It also exhibited positive antifungal efficacy against Candida albicans and revealed moderate activity against liver cancer cells and a weak activity against breast cancer cells. Alternatively, H. papillifera crude extract has weak antiviral activity against CoxB4 and very weak antiviral efficacy against HAV (hepatitis A virus). Moreover, our results proved that H. papillifera extract had anti-inflammatory effects and exhibited antioxidant power. In conclusion, H. papillifera has broad antimicrobial activity against many pathogens, as well as antioxidant, antitumor, anti-inflammatory, among others. It is recommended to explain and characterize the composition of the bioactive compounds of H. papillifera to be included in the pharmaceutical industry