Both growth estimation and otoliths analysis represent a valuable source of information that can be used to reconstruct life-history traits of species, to investigate its trophic interactions, and delineate its stock structure. In this study, we focused on analysing the growth pattern and otolith’s shape of the spotted flounder Citharus linguatula (Linnaeus, 1758) with the aim of preliminary assessing the possible presence of biological and morphological variations between two population of specimens caught in a bottom trawling-banned area (Gulf of Patti, Sicily, law n.25, art.9 of the Sicilian Region, emitted in the 7th August 1990) and a highly fishing impacted area (GFCM Geographic Sub Area 11, Sardinia). A total of 517 right sagittal otoliths (290 collected in Sicily and 227 in Sardinia) was analysed. From the results of a multi-model growth analysis emerged that the Sicilian population seems to benefit the absence of bottom trawling, being capable to reach age up to 6 years, while in the whole Sardinian area no specimen above 3 years have been caught. In support of these initial findings, the shape analysis further highlighted some differences in the otoliths’ contour between the antirostrum and postantirostrum and in the postrostrum region, as confirmed by the ANOVA test and the Canonical Analysis of Principal Coordinates (CAP). Both the distinct age structure found between the areas and the variations in otolith’s shape underline how deeply the presence of human-related stress could impact the life history traits of demersal species. In conclusion, this study highlights the potential of growth estimation and otolith shape analysis as a cost-effective preliminary method for stock status evaluation. The resulting data are crucial for implementing sustainable fisheries management strategies, thereby addressing important ecological challenges.