The aim of the current study was to evaluate the occurrence of actinomycetes in the Cost of Bejaia City using different isolation methods, as well as to evaluate their bioactivity. Methods and Results: Different selective media and methods were used, leading to the isolation of 103 actinomycete strains. The number of strains was influenced by isolation procedures and their interactions based on a three-way ANOVA and a post-hoc Tukey test, which revealed that using M2 medium, dilution of samples followed by moderate heat treatment, and sampling sediment at 10-20 m yielded the highest numbers of actinomycetes. The isolates were screened for their antimicrobial activity against human pathogenic microorganisms using agar and well diffusion methods. Of all the isolates, ten displayed activity against at least one Gram-positive bacterium, of which P21 showed the highest activity against Staphylococcus aureus, Methicillin-resistant S. aureus, and Bacillus subtilis, with a diameter of 32, 28 and 25 mm, respectively. Subsequently, active isolates were assigned to Streptomyces spp. and Nocardiopsis spp. based on 16S rRNA gene sequencing, including a putative new Streptomyces species (S3). The phenotypic characteristics of the P21 strain were determined, and interesting enzymatic capacities were shown. Conclusion: The recovery of actinomycetes along the coast of Bejaia City was influenced by the isolation procedure. Ten strains displayed interesting antibacterial activity against Gram-positive bacteria, of which the P21 strain was selected as the most active strain. Significance and Impact of the Study: This work provides a new insight into the occurrence of actinobacteria in the Coast of Bejaia using selective isolation. It suggests also that polluted environments such as Bejaia Bay could provide access to interesting actinomycetes as sources of antibiotic leads.