Metazoan gill parasites of the Atlantic bluefin tuna Thunnus thynnus (Linnaeus) (Osteichthyes: Scombridae) from the Mediterranean and their possible use as biological tags.

The gills of 63 specimens of the Atlantic bluefin tuna Thunnus thynnus (Linnaeus) (Osteichthyes: Scombridae) from three localities of the Mediterranean (Sardinian, Tyrrhenian and Levantine Seas) were examined for metazoan parasites. The parasite fauna of T. thynnus from the Sea of Sardinia included 11 species: five didymozoid trematodes, three capsalid and one hexostomid monogeneans, and one caligid and one pseudocycnid copepods. Four didymozoids were found in fish from the Levantine Sea and only one didymozoid was recorded in fish from the Tyrrhenian Sea. Dividing the hosts into four size-groups (small, medium-sized, large and extra large), the pairwise comparison of prevalence and mean abundance of the new and literary data) showed differences according to host size. The differences in the composition of the parasitic faunas and in the prevalence of parasites, observed between the small tunas from the Tyrrhenian Sea and the medium-sized tunas from the Adriatic Sea, Levantine Sea and the North-East (NE) Atlantic Ocean, indicated that these groups form discrete units. The parasite fauna of the large tunas from the Sea of Sardinia is the richest among the bluefin tuna populations of the Mediterranean and the NE Atlantic, due to the presence of species not found elsewhere in bluefin tunas, such as Caligus coryphaenae Steenstrup et Lütken, 1861, Capsala magronum (Ishii, 1936) and C. paucispinosa (Mamaev, 1968). This fact and the prevalence of some parasites of this group (lower than those of medium-sized fish from the NE Atlantic and higher than the small and medium-sized tunas from the Mediterranean) suggest that the large-sized tuna group in the western Mediterranean is formed by Mediterranean resident tunas (poorly infected), and by tunas migrating from the Atlantic Ocean (heavily infected).

Comparative cytogenetics in four species of Palinuridae: B chromosomes, ribosomal genes and telomeric sequences.

The evolutionary pathway of Palinuridae (Crustacea, Decapoda) is still controversial, uncertain and unexplored, expecially from a karyological point of view. Here we describe the South African spiny lobster Jasus lalandii karyotype: n and 2n values, heterochromatin distribution, nucleolar organizer region (NOR) location and telomeric repeat structure and location. To compare the genomic and chromosomal organization in Palinuridae we located NORs in Panulirus regius, Palinurus gilchristi and Palinurus mauritanicus: all species showed multiple NORs. In J. lalandii NORs were located on three chromosome pairs, with interindividual polymorphism. In P. regius and in the two Palinurus species NORs were located on two chromosome pairs. In the two last species 45S ribosomal gene loci were also found on B chromosomes. In addition, the nature and location of telomeric repeats were investigated by FISH in J. lalandii, P. gilchristi, P. mauritanicus Palinurus elephas, and P. regius (Palinuridae, Achelata), and in Scyllarus arctus (Scyllaridae, Achelata): all these Achelata species showed the (TTAGG)n pentameric repeats. Furthermore, in J. lalandii these repeats occurred in all the telomeres and in some interstitial chromosomal sites, associated with NORs.

fshr: a fish sex-determining locus shows variable incomplete penetrance across flathead grey mullet populations.

Whole-genome sequencing data were produced from a single flathead grey mullet female and assembled into a draft genome sequence, whereas publicly available sequence data were used to obtain a male draft sequence. Two pools, each consisting of 60 unrelated individuals, respectively, of male and female fish were analyzed using Pool-Sequencing. Mapping and analysis of Pool-Seq data against the draft genome(s) revealed >30 loci potentially associated with sex, the most promising locus of which, encoding the follicle-stimulating hormone receptor (fshr) and harboring two missense variants, was genotyped on 245 fish from four Mediterranean populations. Genotype data showed that fshr represents a previously unknown sex-determining locus, although the incomplete association pattern between fshr genotype and sex-phenotype, the variability of such pattern across different populations, and the presence of other candidate loci reveal that a greater complexity underlies sex determination in the flathead grey mullet.

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.

The hemocyanin of the shamefaced crab Calappa granulata: structural-functional characterization.

Arthropod hemocyanins (Hcs) transport and store oxygen and are composed of six subunits, or multiples thereof depending on the species. Calappa granulata Hc is found as a mixture of dodecamers (95%) and hexamers (5%). Removal of calcium ions and alkaline pH induce an incomplete partially reversible dissociation of dodecameric Hc. Two-dimensional electrophoretic pattern of dissociated Hc indicated a large heterogeneity in Hc subunit: most differences are likely to be explained by post-translational modifications. Dodecameric Hc showed a large Bohr effect (Deltalog P50/DeltapH = -0.95) and a normal cooperativity (h50 values = 2.7 +/- 0.2) in the presence of 10 mM CaCl2. The hexameric molecule displayed lower Bohr effect and cooperativity than the dodecamer. Lactate effect on the oxygen affinity (Deltalog P50 = 0.55) and the increase of lactate concentrations in animals kept in emersion were related to the increased oxygen requirements that occur during hypoxia in vivo. Calcium affects oxygen affinity only at high concentrations: this Hc appeared to lack the calcium high-affinity binding sites found in other species. The effect of temperature on both oxygen affinity and cooperativity was measured in the absence and presence of 10 mM lactate, allowing calculation of the exothermic contribution of lactate binding (DeltaH = -25 kJ mol(-1)).

Distribution and assessment of marine debris in the deep Tyrrhenian Sea (NW Mediterranean Sea, Italy).

Marine debris is a recognized global ecological concern. Little is known about the extent of the problem in the Mediterranean Sea regarding litter distribution and its influence on deep rocky habitats. A quantitative assessment of debris present in the deep seafloor (30-300 m depth) was carried out in 26 areas off the coast of three Italian regions in the Tyrrhenian Sea, using a Remotely Operated Vehicle (ROV). The dominant type of debris (89%) was represented by fishing gears, mainly lines, while plastic objects were recorded only occasionally. Abundant quantities of gears were found on rocky banks in Sicily and Campania (0.09-0.12 debris m(-2)), proving intense fishing activity. Fifty-four percent of the recorded debris directly impacted benthic organisms, primarily gorgonians, followed by black corals and sponges. This work provides a first insight on the impact of marine debris in Mediterranean deep ecosystems and a valuable baseline for future comparisons.

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.

Isolation and characterization of 14 polymorphic microsatellite markers for the blue and red shrimp, Aristeus antennatus (Crustacea, Decapoda).

Eighteen microsatellite loci were isolated and characterized from the blue and red shrimp, Aristeus antennatus Risso 1816, a commercially exploited marine crustacean widely distributed throughout the Mediterranean Sea and in the eastern Atlantic. Polymorphism was assessed in a population (n = 20) from the southwestern Sardinian seas; 14 loci resulted polymorphic and showed from three to 13 alleles. The observed heterozygosity ranged from 0.2 to 0.85. These microsatellites will be potentially useful for the study of A. antennatus population genetic structure.