Marine macroalgae surfaces constitute suitable substrata for bacterial colonization which are known to produce bioactive compounds. Thus, hereby we focused on heterotrophic aerobic bacteria species associated with coralline red alga Jania rubens (northern coast of Tunisia, southern Mediterranean Sea) and their inhibition against several microbial marine and terrestrial species. The whole collection (19 isolates, J1 to J19) was identified, based on their 16S ribosomal RNA gene sequences as Proteobacteria (14 strains), Bacteroidetes (4 strains) and Firmicutes (1 strain). Thirty-six percent of the isolates (J2, J9, J11, J13, J16, J17 and J18) were antibiotic-like producers with in vitro inhibition against Gram + and Gram − bacteria and the yeast Candida albicans. Highest level of inhibition was revealed for the isolates J2, J9 and J13 identified respectively as Bacillus, Aquimarina and Pseudomonas, with strong activity against Staphylococcus aureus, Micrococcus and C. albicans, with inhibition diameters of 25 to 35 mm shown by drop test assay on T soy agar plates. Furthermore, we tested inhibition of J. rubens crude organic extracts against human and marine bacteria as well as against all J. rubens isolates, in order to determine the degree of affinity of the epibionts to their proper host. The recovery of strains with antimicrobial activity suggests that J. rubens represent an ecological niche which harbors a specific microbial diversity worthy of further secondary metabolites investigation.