Rita Cannas

Persistence of Pristine Deep-Sea Coral Gardens in the Mediterranean Sea (SW Sardinia)

Leiopathes glaberrima is a tall arborescent black coral species structuring important facies of the deep-sea rocky bottoms of the Mediterranean Sea that are severely stifled by fishing activities. At present, however, no morphological in vivo description, ecological characterization, age dating and evaluation of the possible conservation actions have ever been made for any population of this species in the basin. A dense coral population was reported during two Remotely Operated Vehicle (ROV) surveys conducted on a rocky bank off the SW coasts of Sardinia (Western Mediterranean Sea). L. glaberrima forms up to 2 m-tall colonies with a maximal observed basal diameter of nearly 7 cm. The radiocarbon dating carried out on a colony from this site with a 4 cm basal diameter revealed an approximately age of 2000 years. Considering the size-frequency distribution of the colonies in the area it is possible to hypothesize the existence of other millennial specimens occupying a supposedly very stable ecosystem. The persistence of this ecosystem is likely guaranteed by the heterogeneous rocky substrate hosting the black coral population that represents a physical barrier against the mechanical impacts acted on the surrounding muddy areas, heavily exploited as trawling fishing grounds. This favorable condition, together with the existence of a nursery area for catsharks within the coral ramifications and the occurrence of a meadow of the now rare soft bottom alcyonacean Isidella elongata in small surviving muddy enclaves, indicates that this ecosystem have to be considered a pristine Mediterranean deep-sea coral sanctuary that would deserve special protection.

Population structure, demographic history, and selective processes: Contrasting evidences from mitochondrial and nuclear markers in the European spiny lobster Palinurus elephas (Fabricius, 1787)

The European spiny lobster Palinurus elephas (Fabricius, 1787) is an ecologically and economically important species inhabiting a wide geographic range that extends from the North-east Atlantic and Azores to the Eastern Mediterranean. We investigated the population structure and evolutionary history of this species by both mitochondrial and microsatellite markers. Ten population samples covering a large part of the species distribution range (three samples from the Atlantic Ocean and seven from the Mediterranean Sea) were analyzed for a portion of the mitochondrial control region and seven polymorphic microsatellite loci. Both markers rejected the hypothesis of panmixia identifying two differentiated gene pools. The control region clearly distinguished the Atlantic and Mediterranean populations in two genetically separated groups. Microsatellites, also revealed two groups roughly associated to the Atlantic-Mediterranean separation, however, the Azores sample did not conform to this geographic scheme. Discrepancy between mitochondrial and nuclear markers emerged also when reconstructing the history of the species. Neutrality tests of the mitochondrial sequences indicated a departure from mutation-drift equilibrium that, combined to the mismatch analysis, pointed toward a sudden population expansion in both Atlantic and Mediterranean gene pools. Unexpectedly, microsatellites did not identify any signal of population expansion neither in the Atlantic pool nor in the Mediterranean one.

Major and 5S ribosomal sequences of the largemouth bass Micropterus salmoides (Perciformes, Centrarchidae) are localized in GC-rich regions of the genome

Major and 5S ribosomal genes have been localized in the chromosomes of Micropterus salmoides. By C-banding, Ag-staining, CMA3-staining and 45S and 5S fluorescence in-situ hybridization (FISH), we demonstrate that the 45S and 5S ribosomal genes are clustered in two different chromosome pairs and both are located in heterochromatic GC-rich regions. PCR amplification and sequencing of the 5S intergenic non-transcribed sequences have allowed us to identify variability essentially due to a trinucleotide tandem repeat (GCT).

Improving the Conservation of Mediterranean Chondrichthyans: The ELASMOMED DNA Barcode Reference Library

Cartilaginous fish are particularly vulnerable to anthropogenic stressors and environmental change because of their K-selected reproductive strategy. Accurate data from scientific surveys and landings are essential to assess conservation status and to develop robust protection and management plans. Currently available data are often incomplete or incorrect as a result of inaccurate species identifications, due to a high level of morphological stasis, especially among closely related taxa. Moreover, several diagnostic characters clearly visible in adult specimens are less evident in juveniles. Here we present results generated by the ELASMOMED Consortium, a regional network aiming to sample and DNA-barcode the Mediterranean Chondrichthyans with the ultimate goal to provide a comprehensive DNA barcode reference library. This library will support and improve the molecular taxonomy of this group and the effectiveness of management and conservation measures. We successfully barcoded 882 individuals belonging to 42 species (17 sharks, 24 batoids and one chimaera), including four endemic and several threatened ones. Morphological misidentifications were found across most orders, further confirming the need for a comprehensive DNA barcoding library as a valuable tool for the reliable identification of specimens in support of taxonomist who are reviewing current identification keys. Despite low intraspecific variation among their barcode sequences and reduced samples size, five species showed preliminary evidence of phylogeographic structure. Overall, the ELASMOMED initiative further emphasizes the key role accurate DNA barcoding libraries play in establishing reliable diagnostic species specific features in otherwise taxonomically problematic groups for biodiversity management and conservation actions.

Homing and orientation of Palinurus elephas (Fabricius) in three no-take areas of the central-western Mediterranean: implications for marine reserve design

The homing and orientation skills of Palinurus elephas were investigated in three no-take areas of the central-western Mediterranean in order to inform future reserve design. In general, P. elephas did not show a particular ability to orient homeward. A considerable portion of tagged lobsters were recaptured in the same direction as the capture point but, the points of capture and recapture were at such a distance from each other as to exclude any possible return to the original den. Homing ability seemed to be constrained to where lobsters were only displaced a short distance. For distances longer than 0.5km, the lobster movements seem to become nomadic, without a particular direction. The movement pattern suggests that it will be important that any restocking of marine reserves must be performed with lobsters collected in adjacent zones at distance more than 0.5km. Lobsters captured in a fishing zone <0.5km outside the reserve will return to the point of capture whilethose collected from further away will not go back to their original place. Our results indicate that, for species like P. elephas, small reserves can protect most small subpopulations with a consequent benefit for the surrounding commercial areas.

Habitat constraints and self-thinning shape Mediterranean red coral deep population structure: implications for conservation practice.

The Mediterranean red coral, Corallium rubrum, is one of the most precious corals worldwide. Below 50 m depth, C. rubrum populations are generally characterised by large and sparse colonies, whereas shallow populations (above 50 m depth) show high densities of small colonies. We show here instead that populations dwelling between 80 and 170 m depth exhibited a continuous range of population density (from 2 to 75 colonies per 0.25 m2), with less than 1% of variance explained by water depth. An inverse relationship between maximum population density and mean colony height was found, suggesting that self-thinning processes may shape population structure. Moreover, demographically young populations composed of small and dense colonies dominated along rocky vertical walls, whereas mature populations characterised by large and sparsely distributed colonies were found only in horizontal beds not covered by sediment. We hypothesise that, in the long term, shallow protected populations should resemble to present deep populations, with sparsely distributed large colonies. Since the density of red coral colonies can decay as a result of self-thinning mechanisms, we advise that future protection strategies should be based also on a measure of red coral spatial coverage instead of population density.

Egg clutch, sperm reservoirs and fecundity of Neorossia caroli (Cephalopoda: Sepiolidae) from the southern Sardinian sea (western Mediterranean)

First observations on reproduction of Neorossia caroli from the southern Sardinian sea are reported, based on the analyses of an egg clutch and specimens collected by bottom trawl on sandy and muddy grounds. The egg clutch, consisting of 13 white eggs of 9 mm length with embryos at an advanced developmental stage, was found at 1200 m depth, attached to a piece of hard substrate. Adult specimens were collected in a depth range between 400 and 1600 metres. DNA sequences indicated that embryos, spermatangia and adults belonged to the same species, the carol bobtail squid, N. caroli. Fecundity analysis made on 40 females showed a variable number of eggs (20-611) in the ovaries, with the combined presence of advanced and developing oocytes, and few smooth eggs ready to be spawned in the oviducts of mature females. Several sperm reservoirs (spermatangia) were embedded in the mantle of some of the females, in the anterior ventral area overlying the oviduct.

Replication banding in two Mediterranean moray eels: chromosomal characterization and comparison

Early and late replication bandings have been obtained by in vitro BrdU incorporation in the Mediterranean Muraenidae species Muraena helena and Gymnothorax unicolor, and used to characterize their karyotypes. A comparative analysis of the banding patterns allowed to point out high karyotype similarity as well as chromosome rearrangements that occurred in karyotype evolution between these species.

Genetic characterization and artificial reproduction attempt of endemic Sardinian trout Salmo trutta L., 1758 (Osteichthyes, Salmonidae): Experiences in captivity

Abstract Native species of Sardinian brown trout (Salmo trutta) have declined drastically because of the introduction of non-native salmonids, overharvesting, and habitat degradation. One approach to conservation of declining species is to establish new populations through repopulation with genetically certified fish. In this study, samples of brown trout from seven rivers in Sardinia were analysed to look for the presence of pure individuals of the endemic Sardinian trout. Analyses of the mitochondrial DNA control region and the LDH-C1* gene revealed the presence of non-endemic and hybrid individuals in four of the seven rivers analysed. In addition, we reared a stock of native Sardinian trout at the experimental fish farm of Sadali and had them reproduce artificially. This experiment provided more knowledge about the incubation phase, hatching, reabsorption of the vitelline sac and the initial phases of growth, which can be used to develop more optimal management of this resource.

Abnormal rostrum in Polycheles typhlops Heller, 1862 (Decapoda: Polychelidae) from the Central Western Mediterranean

Abstract Anomalies of the number of rostral spines have been recorded in blind lobsters, Polycheles typhlops, caught in Sardinian waters (Central Western Mediterranean). The abnormal specimens, 66 out a total of 789 individuals analysed, showed all the morphometric characteristics of P. typhlops except for the unusual presence of two rostral spines instead of the canonical one. The analysis of partial nucleotide sequences of the mitochondrial cytochrome oxidase subunit I gene (COI) showed that the individuals with two spines were not genetically different from the normal specimens of P. typhlops, while both were quite dissimilar from the other Mediterranean species, P. sculptus. Our results suggest the need of an update of the morphological classification guides for this family combined with a wider genetic study.