Haemolymph-associated microbiota of marine bivalves was explored for antibacterial activity against important aquaculture pathogens. A collection of 843 strains were cultured from the haemolymph of four bivalve species (Crassostrea gigas, Mytilus edulis, Pecten maximus and Tapes rhomboides) collected by deep-sea diving in the Glenan Archipelago (France). Cell-free culture supernatants were investigated for antibacterial activity using the well-diffusion assay. About 3% of haemolymph-associated cultivable bacteria displayed antibacterial activity toward Gram-negative pathogens. Among the active bacteria, Pseudoalteromonas strains exhibited the highest antibacterial activity. The cell-free culture supernatant of one of them, named hCg-51, was able to inhibit the growth of bacterial pathogens even after drastic dilution (1 : 1024). Hemocyte survival was not significantly altered in the presence of the haemolymph-associated strains assayed. Moreover, a dose-dependent beneficial effect on hemocyte survival rates was observed with the hCg-51 strain. These results suggest that haemolymph microbiota may participate in bivalve protection and therefore confer a health benefit on the host. As a result, the results highlight bivalve haemolymph microbiota as a promising novel source for aquaculture probiotics. This work also gives a first insight into the contribution of the haemolymph-associated microbiota as part of the bivalve ‘hologenome’.