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In addition to their nutritional value and their wellness effects, Molluscs in general and specially Mussels are a potential source of several bioactive compounds that have gained much more importance over the last decade due to their protein richness and their various uses as nutraceutical, pharmaceutical, therapeutic and functional food agents. This study aims to assess the antimicrobial, antioxidant and cytotoxicity activities of peptide fraction extracted from hemolymph of Algerian blue mussel, Mytilus galloprovincialis. Antimicrobial activity of hemolymph peptide fraction (HPF) against a panel of Gram-positive and Gram negative pathogenic bacteria, lactic acid bacteria (LAB) and fungi strains was determined with well diffusion method. Antioxidant activity of HPF was determined using DPPH and FRAP tests, as well as its ability to scavenge hydroxyl radicals (OH). HPF toxicity towards several tumor cells was evaluated with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) test. The growth of most pathogenic bacteria and Candida albicans tested were inhibited by HPF of M. galloprovincialis (inhibition zone diameters ranging from 10 to more than 24 mm); whereas that of LAB was not affected. Antioxidant activity of 3 mg HPF/mL was about 78.14±2.59, 53.86±1.25 and 84.86±5.68 %, respectively, when determined with DPPH, FRAP, and OH tests. HPF of M. galloprovincialis did not exert any toxicity against any normal cell line, human skin keratinocyte, cancer cell lines, human lung carcinoma, human liver hepatoblastoma, human fibrosarcoma, HPV-16 E7-expressing mouse lung epithelial cell (- MHC class I). According to the results of mouse macrophage cell line (preosteoclast) (RAW264.7), HPF had a slight immunosuppressant activity; while results on human stomach adenocarcinoma (AGS) have shown a marginal cancer cytotoxicity of HPF.