Genetic and morphological variation and differentiation of Raja clavata populations in the European seas: marker development and preliminary data

In this study the population structure and connectivity of the Mediterranean and Atlantic Raja clavata (L., 1758) were investigated by analyzing the genetic variation of six population samples (N = 144) at seven nuclear microsatellite loci. The genetic dataset was generated by selecting population samples available in the tissue databases of the GenoDREAM laboratory (University of Bologna) and of the Department of Life Sciences and Environment (University of Cagliari), all collected during past scientific surveys (MEDITS, GRUND) from different geographical locations in the Mediterranean basin and North-east Atlantic sea, as North Sea, Sardinian coasts, Tuscany coasts and Cyprus Island. This thesis deals with to estimate the genetic diversity and differentiation among 6 geographical samples, in particular, to assess the presence of any barrier (geographic, hydrogeological or biological) to gene flow evaluating both the genetic diversity (nucleotide diversity, observed and expected heterozygosity, Hardy- Weinberg equilibrium analysis) and population differentiation (Fst estimates, population structure analysis). In addition to molecular analysis, quantitative representation and statistical analysis of morphological individuals shape are performed using geometric morphometrics methods and statistical tests. Geometric coordinates call landmarks are fixed in 158 individuals belonging to two population samples of Raja clavata and in population samples of closely related species, Raja straeleni (cryptic sibling) and Raja asterias, to assess significant morphological differences at multiple taxonomic levels. The results obtained from the analysis of the microsatellite dataset suggested a geographic and genetic separation between populations from Central-Western and Eastern Mediterranean basins. Furthermore, the analysis also showed that there was no separation between geographic samples from North Atlantic Ocean and central-Western Mediterranean, grouping them to a panmictic population. The Landmark-based geometric morphometry method results showed significant differences of body shape able to discriminate taxa at tested levels (from species to populations).

Nuovi suoni di balenottera comune (Balaenoptera physalus) nel Mediterraneo occidentale

Fin whale (Balaenoptera physalus) is the only misticeto commonly observed in the Mediterranean. The males emit sounds, classic pulse products in sequences called songs, at 20Hz for sexual purposes: sounds are produced during the spring for migration to the Tirreno-Ligurian-Provençal basin, the summer feeding area, and during the autumn, when there is a migration to the south to meet the winter breeding season. This area in the Mediterranean sea is unknown. The east coast of the Iberian Peninsula is a migration area. The study was conducted by analyzing through Adobe Audition 3.0 and XBAT softwares files audio of 30 minutes recorded in 2006, in 2011 and 2012 at the level of the Columbretes Islands, in the western Mediterranean sea, using two hearing aids: the MARU, used in 2006 and the EAR, used in 2011 and 2012. From the analysis have emerged that, in addition to songs with pulses of 20 Hz, there are new sounds of fin whale never previously recognized: the VFPs (Variable Frequency Pulses), higher-frequency pulses emitted, between 50 and 120Hz and the ramps, a set of 7-8 pulses, pertaining to a particular song, of increasing frequency. Further studies are needed to understand the importance of these new sounds.

Inference of Raja miraletus population structure using mitochondrial and nuclear DNA: comparing the resolution power of molecular markers in exploring species and population boundaries

This study focused on the role of oceanographic discontinuities and the presence of transitional areas in shaping the population structure and the phylogeography of the Raja miraletus species complex, coupled with the test of the effective occurrence of past speciation events. The comparisons between the Atlantic African and the North-Eastern Atlantic-Mediterranean geographic populations were unravelled using both Cytochrome Oxidase I and eight microsatellite loci. This approach guaranteed a robust dataset for the identification of a speciation event between the Atlantic African clade, corresponding to the ex Raja ocellifera nominal species, and the NE Atlantic-Mediterranean R. miraletus clade. As a matter of fact, the origin of the Atlantic Africa and the NE Atlantic-Mediterranean deep split dated about 11.74MYA and was likely due to the synergic influence currents and two upwelling areas crossing the Western African Waters. Within the Mediterranean Sea, particular attention was also paid to the transitional area represented by Adventura and Maltese Bank, that might have contributed in sustaining the connectivity of the Western and the Eastern Mediterranean geographical populations. Furthermore, the geology of the easternmost part of Sicily and the geo-morphological depression of the Calabrian Arc could have driven the differentiation of the Eastern Mediterranean Sea. Although bathymetric and oceanographic discontinuity could represent barriers to dispersal and migration between Eastern and Western Mediterranean samples, a clear and complete genetic separation among them was not detected. Results produced by this work identified a speciation event defining Raja ocellifera and R. miraletus as two different species, and describing the R. miraletus species complex as the most ancient cryptic speciation event in the family Rajidae, representing another example of how strictly connected the environment, the behavioural habits and the evolutionary and ecologic drivers are.

Ruolo della gorgonia Paramuricea clavata (Risso 1826) nei popolamenti epibentonici coralligeni dell'Isola d'Elba

The red gorgonian Paramuricea clavata is considered a key species in Mediterranean coralligenous habitats and, when present at high density, its colonies may develop “gorgonian forests”, which are three-dimensional structures that increase habitat complexity. Mediterranean coralligenous habitats support high biodiversity but the structure and heterogeneity of communities can be strongly modified by several kinds of human-derived impacts. The global environmental change and human activity could threaten the coralligenous habitats and during the last few decades, Mediterranean suspension feeders have been involved in mass mortality events, in which P. clavata was one of the most affected. Without the complex three-dimensional structures formed by P. clavata, many benthic species may have not adequate sheltering and feeding opportunities. The aim of the present study was to investigate the ecological role of Paramuricea clavata in coralligenous habitats by comparing epibenthic assemblages established inside and outside gorgonian forests. This study was carried out in four sites randomly chosen, along the south-east coast of Elba island (north-western Mediterranean). The structure of the epibenthic assemblages were significantly different between area with and without gorgonians. Main differences concern very important taxa in the coralligenous bioconstruction processes like the encrusting red algae, Peyssonnelia spp. and Halimeda tuna, which were more abundant in presence of gorgonians. On the contrary, algal turf, mucilaginous algae and the invasive seaweed Caulerpa cylindracea were more abundant in areas without P. clavata. The presence of gorgonians may have a positive effect also on species diversity. The results of this study highlight the ecological role of P. clavata in the coralligenous habitats.

Genetic variability and differentiation in the Mediterranean endemic starry ray Raja asterias (Delaroche, 1809)

The present study deal with the population structure and connectivity of the Mediterranean endemic starry ray Raja asterias (Delaroche, 1809) in the Western and Eastern Mediterranean basin. A panel of eight microsatellite loci which cross-amplify in Rajidae (El Nagar, 2010) was used to assess population connectivity and structure. Those aims were investigated by analyzing the genetic variation of 9 population sample for a total of 185 individuals collected during past scientific surveys (MEDITS, GRUND), commercial trawling and also directly at fish markets. The purpose of this thesis is to estimate the genetic divergence occurring between the Mediterranean populations and, in particular, to assess the presence of any barrier (geographic, hydrogeological and biological) to gene flow for this species. Different statistical approaches were performed to reach this aim evaluating both the genetic diversity (nucleotide diversity, allelic richness, observed and expected heterozygosity and Hardy-Weinberg equilibrium test) and the population differentiation patterns (pairwise Fst estimated and population structure analysis). The results obtained from the analysis of the microsatellite dataset suggest a geographic and genetic separation between the starry ray populations of the Mediterranean basin into three or four distinct groups: Western and Eastern Mediterranean basins and Sicilian coast always clustering as an independent group and Algeria which could be or not considered another separate group. The data were discussed from both an evolutionary and a conservation point of view and in relation to previous results obtained by the analysis of mitochondrial marker. A comparison with other Mediterranean demersal skate species was performed in order to better contextualise our results. Finally, our results could offer useful information to protect vulnerable species as R. asterias and developing effective conservation plans in the Mediterranean.