DNA barcoding taxonomy and diversity of the Mediterranean sharks and rays

Elasmobranchs are an important by-catch of commercial fisheries targeting bony fishes. Fisheries targeting sharks are rare, but usually almost all specimen bycatched are marketed. They risk extinction if current fishing pressure continues (Ferretti et al., 2008). Accurate species identification is critical for the design of sustainable fisheries and appropriate management plans, especially since not all species are equally sensitive to fishing pressure (Walker & Hislop 1998). The identification of species constitutes the first basic step for biodiversity monitoring and conservation (Dayrat B et al., 2005). More recently, mtDNA sequencing has also been used for species identification and its use has become widespread under the DNA Barcode initiative (e.g. Hebert et al. 2003a, 2003b; Ward et al. 2005, 2008a; Moura et al 2008; Steinke et al. 2009). The aims of this work were: 1) identify sharks and skates species using DNA barcode; 2) compare species of different provenance; 3) use DNA barcode for misidentified species. Using DNA barcode 15 species of sharks (Alopias vulpinus, Centrophorus granulosus, Cetorhinus maximus, Dalatias licha, Etmopterus spinax, Galeorhinus galeus, Galeus melastomus, Heptranchias perlo, Hexanchus griseus, Mustelus mustelus, Mustelus punctulatus, Oxynotus centrina, Scyliorhinus canicula Squalus acanthias, Squalus blainville), 1 species of chimaera (Chimaera monstrosa) and 21 species of rays/skayes (Dasyatis centroura, Dasyatis pastinaca, Dasyatis sp., Dipturus nidarosiensis, Dipturus oxyrinchus, Leucoraja circularis, Leucoraja melitensis, Myliobatis aquila, Pteromylaeus bovinus, Pteroplatytrygon violacea, Raja asterias, Raja brachyura, Raja clavata, Raja miraletus, Raja montagui, Raja radula, Raja polystigma, Raja undulata, Rostroraja alba, Torpedo marmorata, Torpedo nobiliana, Torpedo torpedo) was identified.

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.

Climatological analysis of temperature and salinity fields in the Mediterranean sea

A climatological field is a mean gridded field that represents the monthly or seasonal trend of an ocean parameter. This instrument allows to understand the physical conditions and physical processes of the ocean water and their impact on the world climate. To construct a climatological field, it is necessary to perform a climatological analysis on an historical dataset. In this dissertation, we have constructed the temperature and salinity fields on the Mediterranean Sea using the SeaDataNet 2 dataset. The dataset contains about 140000 CTD, bottles, XBT and MBT profiles, covering the period from 1900 to 2013. The temperature and salinity climatological fields are produced by the DIVA software using a Variational Inverse Method and a Finite Element numerical technique to interpolate data on a regular grid. Our results are also compared with a previous version of climatological fields and the goodness of our climatologies is assessed, according to the goodness criteria suggested by Murphy (1993). Finally the temperature and salinity seasonal cycle for the Mediterranean Sea is described.

Temporal variation of genetic diversity and differentiation of albacore, Thunnus alalunga, in the Mediterranean Sea

This study is on albacore (Thunnus alalunga, Bonnaterre 1788), an epi- and mesopelagic oceanic tuna species cosmopolitan in the tropical and temperate waters of all oceans including the Mediterranean Sea, extending in a broad band between 40°N and 40°S. What it’s known about albacore population structure is based on different studies that used fisheries data, RFLP, mtDNA control region and nuDNA markers, blood lectins analysis, individual tags and microsatellite. At the moment, for T. alalunga six management units are recognized: the North Pacific, South Pacific, Indian, North Atlantic, South Atlantic and Mediterranean stocks. In this study I have done a temporal and spatial comparison of genetic variability between different Mediterranean populations of Thunnus alalunga matching an historical dataset ca. from 1920s composed of 43 individuals divided in 3 populations (NADR, SPAIN and CMED) with a modern dataset composed of 254 individuals and 7 populations (BAL, CYP, LIG, TYR, TUR, ADR, ALB). The investigation was possible using a panel of 94 nuclear SNPs, built specifically for the target species at the University of Basque Country UPV/EHU. First analysis done was the Hardy-Weinberg, then the number of clusters (K) was determined using STRUCTURE and to assess the genetic variability, allele frequencies, the average number of alleles per locus, expected (He) and observed (Ho) heterozygosis, and the index of polymorphism (P) was used the software Genetix. Historical and modern samples gives different results, showing a clear loss of genetic diversity over time leading to a single cluster in modern albacore instead of the two found in historical samples. What this study reveals is very important for conservation concerns, and additional research endeavours are needed.

Species identification of archived fish bones collected from the Mediterranean Sea 100 years ago using molecular techniques

With the discovery that DNA can be successfully recovered from museum collections, a new source of genetic information has been provided to extend our comprehension of the evolutionary history of species. However, historical specimens are often mislabeled or report incorrect information of origin, thus accurate identification of specimens is essential. Due to the highly damaged nature of ancient DNA many pitfalls exist and particular precautions need to be considered in order to perform genetic analysis. In this study we analyze 208 historical remains of pelagic fishes collected in the beginning of the 20th century. Through the adaptation of existing protocols, usually applied to human remains, we manage to successfully retrieve valuable genetic material from almost all of the examined samples using a guanidine and silica column-based approach. The combined use of two mitochondrial markers cytochrome-oxidase-1(mtDNA COI) and Control Region (mtDNA CR), and the nuclear marker first internal transcriber space (ITS1) allowed us to identify the majority of the examined specimens using traditional PCR and Sanger sequencing techniques. The creation of primers capable of amplifying heavily degraded DNA have great potential for future uses, both in ancient and in modern investigation. The methodologies developed in this study can in fact be applied for other ancient fish specimens as well as cooked or canned samples.

A study of air-sea interaction processes on water mass formation and upwelling in the Mediterranean sea

Air-sea interactions are a key process in the forcing of the ocean circulation and the climate. Water Mass Formation is a phenomenon related to extreme air-sea exchanges and heavy heat losses by the water column, being capable to transfer water properties from the surface to great depth and constituting a fundamental component of the thermohaline circulation of the ocean. Wind-driven Coastal Upwelling, on the other hand, is capable to induce intense heat gain in the water column, making this phenomenon important for climate change; further, it can have a noticeable influence on many biological pelagic ecosystems mechanisms. To study some of the fundamental characteristics of Water Mass Formation and Coastal Upwelling phenomena in the Mediterranean Sea, physical reanalysis obtained from the Mediterranean Forecating System model have been used for the period ranging from 1987 to 2012. The first chapter of this dissertation gives the basic description of the Mediterranean Sea circulation, the MFS model implementation, and the air-sea interaction physics. In the second chapter, the problem of Water Mass Formation in the Mediterranean Sea is approached, also performing ad-hoc numerical simulations to study heat balance components. The third chapter considers the study of Mediterranean Coastal Upwelling in some particular areas (Sicily, Gulf of Lion, Aegean Sea) of the Mediterranean Basin, together with the introduction of a new Upwelling Index to characterize and predict upwelling features using only surface estimates of air-sea fluxes. Our conclusions are that latent heat flux is the driving air-sea heat balance component in the Water Mass Formation phenomenon, while sensible heat exchanges are fundamental in Coastal Upwelling process. It is shown that our upwelling index is capable to reproduce the vertical velocity patterns in Coastal Upwelling areas. Nondimensional Marshall numbers evaluations for the open-ocean convection process in the Gulf of Lion show that it is a fully turbulent, three-dimensional phenomenon.

Citizen science as a tool for the environmental quality assessment of the Mediterranean coastal habitats

The growing need to assess the environmental status of the Mediterranean coastal marine habitats and the large availability of data collected by Reef Check Italia onlus (RCI) volunteers suggest the possibility to develop innovative and reliable indices that may support decision makers in applying conservation strategies. The aims of this study were to check the reliability of data collected by RCI volunteers, analyse the spatial and temporal distribution of RCI available data, resume the knowledge on the biology and ecology of the monitored species, and develop innovative indices to asses the ecological quality of Mediterranean subtidal rocky shores and coralligenous habitats. Subtidal rocky shores and coralligenous were chosen because these are the habitats more attractive for divers; therefore mlst data are referring to them, moreover subtidal rocky bottom are strongly affected by coastal urbanisation, land use, fishing and tourist activities, that increase pollution, turbidity and sedimentation. Non-indigenous species (NIS) have been recognized as a major threat to the integrity of Mediterranean native communities because of their proliferation, spread and impact on resident communities. Monitoring of NIS’ spreading dynamics at the basin spatial scale is difficult but urgent. According to a field test, the training provided by RCI appears adequate to obtain reliable data by volunteers. Based on data collected by RCI volunteers, three main categories of indices were developed: indices based on species diversity, indices on the occurrence non-indigenous species, and indices on species sensitive toward physical, chemical and biological disturbances. As case studies, indices were applied to stretches of coastline defined according to management criteria (province territories and marine protected areas). The assessments of ecological quality in the Tavolara Marine Protected Area using the species sensitivities index were consisten with those previously obtained with traditional methods.

Sea level trends in the Mediterranean from tide gauges and satellite altimetry

Sea level variation is one of the parameters directly related to climate change. Monitoring sea level rise is an important scientific issue since many populated areas of the world and megacities are located in low-lying regions. At present, sea level is measured by means of two techniques: the tide gauges and the satellite radar altimetry. Tide gauges measure sea-level relatively to a ground benchmark, hence, their measurements are directly affected by vertical ground motions. Satellite radar altimetry measures sea-level relative to a geocentric reference and are not affected by vertical land motions. In this study, the linear relative sea level trends of 35 tide gauge stations distributed across the Mediterranean Sea have been computed over the period 1993-2014. In order to extract the real sea-level variation, the vertical land motion has been estimated using the observations of available GPS stations and removed from the tide gauges records. These GPS-corrected trends have then been compared with satellite altimetry measurements over the same time interval (AVISO data set). A further comparison has been performed, over the period 1993-2013, using the CCI satellite altimetry data set which has been generated using an updated modeling. The absolute sea level trends obtained from satellite altimetry and GPS-corrected tide gauge data are mostly consistent, meaning that GPS data have provided reliable corrections for most of the sites. The trend values range between +2.5 and +4 mm/yr almost everywhere in the Mediterranean area, the largest trends were found in the Northern Adriatic Sea and in the Aegean. These results are in agreement with estimates of the global mean sea level rise over the last two decades. Where GPS data were not available, information on the vertical land motion deduced from the differences between absolute and relative trends are in agreement with the results of other studies.

On the Mediterranean conveyor belt system

The Mediterranean Sea is a semi-enclosed sea, connected to the Atlantic Ocean through the Gibraltar Strait and subdivided in two different regions by the Sicily Strait. The geographical extension of the basin, the surface heat flux, and the water inflow from the Gibraltar Strait are some of the basic factors determining its horizontal and vertical circulation. In the Mediterranean strong salinity and temperature zonal gradients contribute to maintain the zonal-vertical circulation, while meridional-vertical cells are equally forced by winds and deep water mass formation in three regions, the Gulf of Lyon, the southern Adriatic and the Cretan Sea areas. The objective of this thesis is to study how these cells combine together to form the Mediterranean conveyor belt system. This has never been attempted before so the conclusions are necessarily preliminary. In the first part we discuss the vertical zonal and meridional circulation by reconstructing the Wust Mediterranean vertical salinity and temperature structures in an attempt to evaluate the water mass structure consistent with modern data. Our results confirm that Wust depicted vertical circulation from scarce data is reproduced by the past 27 years observations. The structure of both meridional and zonal circulations was discussed using velocity vertical streamfunctions with two different methods. The first one, eulerian, allowed us to observe vertical structures that were already reported in the literature. Recent studies in the Atlantic Ocean have shown that gyres and eddies have an important influence in the isopycnal vertical circulation. This is called the residual circulation which was computed in this study for the first time. A possible interpretation of horizontal connection between the meridional and zonal cells was discussed using horizontal streamfunction. In the last part of the thesis we have been developing an idealized numerical model to study the vertical circulation in the Mediterranean.

Analysis of the Mediterranean tropical-like cyclone Numa from a numerical model simulation

I Medicane sono rari cicloni che si sviluppano sul Mar Mediterraneo e presentano caratteristiche dei cicloni tropicali, come la forma a spirale delle bande di nubi, un occhio privo di venti e nubi, venti intensi nella banda che circonda l’occhio e la presenza di un nucleo caldo. Nel presente lavoro, è stata compiuta un’analisi del Medicane Numa, verificatosi nel novembre del 2017, utilizzando gli output della simulazione del modello RAMS-ISAC. L’obiettivo di questa tesi è l’identificazione e la descrizione delle caratteristiche tropicali di Numa, focalizzandosi sulla descrizione dei diversi stadi di sviluppo del ciclone. Il sistema di bassa pressione è stato identificato utilizzando la pressione sul livello del mare, mentre l’anomalia termica e il vento orizzontale hanno permesso una descrizione della struttura del Medicane e del suo nucleo caldo. Un’illustrazione della struttura a spirale delle bande di nubi e dell’occhio è stata ottenuta con il grafico dei rapporti di mescolanza delle idrometeore di nubi e pioggia. Questi parametri hanno consentito di ricavare il diametro dell’occhio, pari a 75 km, mentre il diametro del Medicane è risultato 230 km. Numa ha registrato una velocità massima del vento in superficie di 20 m/s nella banda adiacente all’occhio del ciclone. Il diagramma di Hart dello spazio delle fasi del ciclone ha confermato la natura simil-tropicale di Numa e ne ha descritto l’evoluzione, identificando la transizione da sistema a caratteri tropicali a sistema ibrido. La traiettoria nello spazio delle fasi ha consentito l’identificazione delle sottofasi dell’evoluzione di Numa, confermate dai grafici dell’evoluzione temporale dei parametri menzionati in precedenza. L’analisi ha mostrato il ruolo cruciale della presenza di una struttura organizzata nel determinare l’intensità e la durata delle caratteristiche tropicali. Tutti i parametri hanno evidenziato la simmetria della struttura durante la persistente fase matura di Numa.

Assessment and Protection of maritime-vernacular architectural Heritage on the Mediterranean Coasts. Documentation and Preservation Strategies for the Dry Docks on the Island of Formentera.

The importance of vernacular architecture as an integral part of our cultural heritage is often undervalued, and the management of rural constructions of heritage value often requires the application of more flexible and adjusted preservation principles compared to monumental assets. For vernacular architecture, the preservation and consolidation concern not only their physical substance but also their intangible values and purpose in society. More than other heritage values, the vernacular raises the question of prospective functions it can fulfill in contemporary societies without undermining its legacy. This work analyzes the topic by studying the case study of the dry docks on the Balearic Island of Formentera, including abundant documentation of traditional construction techniques and materials realized through a field study, followed by the assessment of threats to and opportunities for vernacular architecture on Formentera, closing with suggestions for the maintenance and potential functions of the dry docks. For this, a comparative case study was introduced: the capanni da pesca at the Adriatic coast of Italy. The suggestions focus on the importance of maintenance for rural heritage, expressed through the creation of a guide for owners of protected dry docks highlighting good and bad practice examples and recurrent works of care. Furthermore, the thesis seeks to raise awareness for the significance of landscape and social factors in the discourse about vernacular heritage sites. Ultimately, different outlines of tourism development are proposed. The idea is to explore rural and sustainable tourism as a tool for territory development and enhancement of the cultural heritage value by helping to prevent the destination's decline through careful evaluation of its limits of acceptable change and identifying beneficial, sustainable scenarios for the future of the heritage asset and the respective community.

The routes of the Mediterranean Sea circulation

The Mediterranean Sea is a semi-enclosed basin connected to the Atlantic Ocean through the narrow and shallow Strait of Gibraltar and further subdivided in two different sub-basins, the Eastern Mediterranean and the Western Mediterranean, connected through the Stait of Sicily. On annual basis, a net heat budget of −7 W/m2, combined with exceeding evaporation over precipation and runoff together with wind stress, is responsible for the antiestuarine character of the zonal thermoaline circulation. The outflow at Gibraltar Strait is mainly composed of Levantine Intermediate Water (LIW) and deep water masses formed in the Western Mediterranean Sea. The aim of this thesis is to validate and quantitatively assess the main routes of water masses composing the ouflow at Gibraltar Strait, using for the first time in the Mediterranean Sea a lagrangian interpretation of the eulerian velocity field produced from an eddy-resolving reanalysis dataset, spanning from 2000 to 2012. A lagrangian model named Ariane is used to map out three-dimensional trajectories in order to describe the pathways of water mass transport from the Strait of Sicily, the Gulf of Lyon and the Northern Tyrrhenian Sea to the Gibraltar Strait. Numerical experiments were carried out by seeding millions of particles in the Strait of Gibraltar and following them backwards in time to track the origins of water masses and transport exchanged between the different sections of the Mediterranean. Finally, the main routes of the intermediate and deep water masses are reconstructed from virtual particles trajectories, which highlight the role of the Western Mediterranean Deep Water (WMDW) as the main contributor to the Gibraltar Strait outflow. For the first time, the quantitative description of the flow of water masses coming from the Eastern Mediterranean towards the Gibraltar Strait is provided and a new route that directly links the Northern Tyrrhenian Sea to Gibralatr Strait has been detected.