Brown algae of the genus Ericaria are habitat formers on Mediterranean rocky shores supporting marine biodiversity and ecosystem functioning. Their population decline has prompted attempts for restoration of threatened populations. Although epilithic microbial biofilms (EMBs) are determinant for macroalgal settlement, their role in regulating the recovery of populations through the recruitment of new thalli is yet to be explored. In this study, we assessed variations in microbial biofilms composition on the settlement of Ericaria amentacea at sites exposed to different human pressures. Artificial fouling surfaces were deployed in two areas at each of three study sites in the Ligurian Sea (Capraia Island, Secche della Meloria and the mainland coast of Livorno), to allow bacterial biofilm colonization. In the laboratory, zygotes of E. amentacea were released on these surfaces to evaluate the survival of germlings. The EMB's composition was assessed through DNA metabarcoding analysis, which revealed a difference between the EMB of Capraia Island and that of Livorno. Fouling surfaces from Capraia Island had higher rates of zygote settlement than the other two sites. This suggests that different environmental conditions can influence the EMB composition on substrata, possibly influencing algal settlement rate. Assessing the suitability of rocky substrata for E. amentacea settlement is crucial for successful restoration.