Otolith shape analysis is an efficient fish stock identification tool. However, most applications used left and right otoliths or only one of them arbitrarily chosen without testing for biases resulting from potential directional bilateral asymmetry (DA) in otolith shape, i.e. a unimodal population-level deviation form bilateral symmetry between right and left otolith shapes. In this study, 560 bogues (Boops boops) were sampled from 11 geographical locations from the Canary Islands to the Aegean Sea and elliptical Fourier descriptors were used to describe their otoliths’ shape. First, a significant otolith DA was observed at the global scale with an average amplitude of 2.77%. However, at the scale of sampling locations, DA was not always significant and varied in amplitude and direction. Second, population structure was investigated using the shape of either right otoliths or left otoliths or both together. Analyses based on right otoliths or both otoliths together, suggested three stock units: a North-Western Mediterranean Sea stock, an Eastern Mediterranean Sea stock, and a Central-Eastern Atlantic Ocean and South-Western Mediterranean Sea stock. In contrast, no coherent geographical pattern was found based on left otoliths. Our results highlight the importance of accounting for potential otolith DA in otolith shape-based stock identification.

The European sardine (Sardina pilchardus, Walbaum 1792) is indisputably a commercially important species. Previous studies using uneven sampling or a limited number of makers have presented sometimes conflicting evidence of the genetic structure of S. pilchardus populations. Here, we show that whole genome data from 108 individuals from 16 sampling areas across 5000 km of the species’ distribution range (from the Eastern Mediterranean to the archipelago of Azores) support at least three genetic clusters. One includes individuals from Azores and Madeira, with evidence of substructure separating these two archipelagos in the Atlantic. Another cluster broadly corresponds to the center of the distribution, including the sampling sites around Iberia, separated by the Almeria–Oran front from the third cluster that includes all of the Mediterranean samples, except those from the Alboran Sea. Individuals from the Canary Islands appear to belong to the Mediterranean cluster. This suggests at least two important geographical barriers to gene flow, even though these do not seem complete, with many individuals from around Iberia and the Mediterranean showing some patterns compatible with admixture with other genetic clusters. Genomic regions corresponding to the top outliers of genetic differentiation are located in areas of low recombination indicative that genetic architecture also has a role in shaping population structure. These regions include genes related to otolith formation, a calcium carbonate structure in the inner ear previously used to distinguish S. pilchardus populations. Our results provide a baseline for further characterization of physical and genetic barriers that divide European sardine populations, and information for transnational stock management of this highly exploited species towards sustainable fisheries.

The otolith, found in both inner ears of bony fish, has mainly been used to estimate fish age. Another application that has been developing significantly in recent years, however, is the use of otolith shape as a tool for stock identification. Often, studies have directly used the shape asymmetry between the right and left otoliths. We tested the magnitude of directional asymmetry between the sagittal otoliths (left vs. right) of 2991 individuals according to their catch locations, and we selected species to evaluate whether directional asymmetry may itself be a tool to evaluate stock boundaries. Elliptical Fourier descriptors were used to describe the otolith shape. We used a flatfish, the common sole (Solea solea, n = 2431), from the eastern English Channel and the southern North Sea as well as a roundfish, the bogue (Boops boops, n = 560), from the Mediterranean Sea. Both species showed significant levels of directional asymmetry between the testing locations. The bogue otoliths showed significant asymmetry for only 5 out of 11 locations, with substantial separation between two large areas: the Algerian coast and the western part of the Italian coast. The sole otoliths showed significant asymmetry in the shape analysis (3.84–6.57%), suggesting a substantial separation between two large areas: the English and French parts of the English Channel and the southern North Sea. Consequently, directional bilateral asymmetry in otolith shape is a potential new method for stock identification.