The Mediterranean bath sponge Spongia officinalis is an iconic species with high socio-economic value and imperiled present and future status due to unregulated harvesting, mortality incidents and lack of established knowledge regarding its ecology. This study aims to assess genetic diversity and population structure of the species at different geographic sectors and levels of geographic distance along its distribution. For this purpose, 11 locations in the eastern Mediterranean (Aegean Sea), western Mediterranean (Provence coast), and the Strait of Gibraltar were sampled; specimens were analysed using partial mitochondrial cytochrome oxidase subunit I (COI) sequences, along with a set of 8 microsatellite loci. According to our results (i) no genetic differentiation exists among the acknowledged Mediterranean morphotypes and presumably S. officinalis can be viewed as a single, morphologically variable species; (ii) a notable divergence was recorded in the Gibraltar region, indicating the possible existence of a cryptic species; (iii) restriction to gene flow was evidenced between the Aegean Sea and Provence giving two well-defined regional clusters, thus suggesting the existence of a phylogeographic break between the two systems; (iv) low levels of genetic structure, not correlated to geographic distance, were observed inside geographic sectors, implying mechanisms (natural or anthropogenic) that enhance dispersal and gene flow, promoting population connectivity; (v) the genetic diversity of S. officinalis is maintained high in most studied locations despite pressure from harvesting and the influence of devastating epidemics. These findings provide a basis towards the effective conservation and management of the species.