Spread of Microcosmus squamiger (Ascidiacea: Pyuridae) in the Mediterranean Sea and adjacent waters

The Mediterranean Sea is subject to an ever-increasing arrival of non-indigenous marine organisms. Microcosmus squamiger is a solitary ascidian that inhabits shallow rocky littoral habitats. It probably originated in Australia and it has shown great invasive potential in other parts of the world. In the Mediterranean, M. squamiger has only been reported at a few sites in Spain and Italy. However, the closely related species Microcosmus exasperatus has been reported in several areas of the western Mediterranean. As these species can be easily confused, we re-examined most of the material from previous studies and our personal collections. In addition, sampling was done at several sites along the western Mediterranean and Atlantic coasts. The results showed that the majority of the M. exasperatus reports correspond to M. squamiger, and that M. squamiger is common on the Atlantic shores. This suggests that M. squamiger has entered the Mediterranean through the Gibraltar Strait, while the restricted distribution in the eastern Mediterranean of M. exasperatus suggests that this species is probably a Lessepsian migrant. In the Mediterranean Sea, M. squamiger has the ability to occupy extensive areas of hard substrata and to outcompete native species. Further studies are necessary to assess what impacts this invasive species have on native communities.     

On the occurrence of the fireworm Eurythoe?complanata complex (Annelida,Amphinomidae) in the Mediterranean Sea with an updated revision of the alien Mediterranean amphinomids

The presence of two species within the Eurythoe complanata complex in the Mediterranean Sea is reported, as well as their geographical distributions. One species, Eurythoe laevisetis, occurs in the eastern and central Mediterranean, likely constituting the first historical introduction to the Mediterranean Sea and the other, Eurythoe complanata, in both eastern and Levantine basins. Brief notes on their taxonomy are also provided and their potential pathways for introduction to the Mediterranean are discussed. A simplified key to the Mediterranean amphinomid genera and species of Eurythoe and Linopherus is presented plus an updated revision of the alien amphinomid species reported previously from the Mediterranean Sea. A total of five exotic species have been included; information on their location, habitat, date of introduction and other relevant features is also provided.     

Morphokaryological variability and divergence of stargazers (Uranoscopus, perciformes) from the Mediterranean Sea basin: I. Divergence and taxonomic state of the Black Sea Stargazer

Based on museum collections, the variability of morphological characters of stargazers (Uranoscopus, Uranoscopidae) from different seas of the Mediterranean Sea basin have been studied. On the basis of the obtained results and data on the karyological divergence of the previously studied populations, stargazers of the Black Sea are regarded as an independent subspecies, Uranoscopus scaber anostomus, differing from the Mediterranean Sea populations in relatively long ventral fins, whose length usually exceeds half of the head length, the presence in most individuals of not less than 12 pyloric caeca, chromosome polymorphism with 2n = 28?C32, NF = 50, and the presence in the karyotype of a pair of marker small metacentric chromosomes. Morphological characters suitable for analysis of morphological divergence of populations of the Mediterranean region have been revealed.     

Octopus vulgaris (Cuvier, 1797) in the Mediterranean Sea: Genetic Diversity and Population Structure

The common octopus, Octopus vulgaris Cuvier 1797, is a largely exploited cephalopod species in the Mediterranean Sea and the Atlantic Ocean, as well as along the coasts of Africa, Brazil and Japan, where its taxonomic identity is still debated. The assessment of its genetic structure is a pressing need to correctly manage the resource and to avoid overfishing and collapsing of local stocks. Here we analysed genetic variation and population structure of O. vulgaris using thirteen microsatellite loci in seven sampling localities from the Mediterranean Sea and one from the Atlantic Ocean. We also used a DNA barcoding approach by COI gene fragment to understand the phylogenetic relationships among the specimens here investigated and the ones whose sequences are available in literature. Our results reveal high levels of allelic richness and moderate heterozygosity in all samples investigated, and a pronounced differentiation of the Atlantic and Sicilian specimens. This latter aspect seems to support the isolation of the biota within the Strait of Messina. A certain degree of differentiation was detected among the other geographic samples within the Mediterranean Sea, which is more compatible with an island model than isolation by distance. The occurrence of null alleles affected more genetic diversity indices than population structure estimations. This study provides new insights about the genetic diversity and structure of O. vulgaris in the area of interest, which can be used as guidelines for a fisheries management perspective.     

Looking for long-term changes in hydroid assemblages (Cnidaria,Hydrozoa) in Alboran Sea (South-Western Mediterranean): a proposal of a monitoring point for the global warming

In the last 20–30 years, the temperature of the Mediterranean Sea has increased and global warming is allowing the establishment of tropical-affinity species into more temperate zones. Sessile communities are particularly useful as a baseline for ecological monitoring; however, a lack of historical data series exists for sessile marine organisms without commercial interest. Hydroids are ubiquitous components of the benthic sessile fauna on rocky shores and have been used as bio-indicators of environmental conditions. In this study on the benthic hydroid assemblages of the Chafarinas Islands (Alboran Sea, South-Western Mediterranean), we characterized the hydroid assemblages, identified the bathymetric gradients, and compared them with a previous study carried out in 1991. Hydroid assemblages showed a significant difference both between year and among depths. Furthermore, eight species not present in 1991 were found, including two possible new species and the tropical and subtropical species Sertularia marginata. Due to its strategic position at the entrance of the Mediterranean and the existence of previous data on hydroid assemblages, the Chafarinas Islands are proposed as a possible monitoring point for entrance of Atlantic tropical species into the Mediterranean Sea.     

Extant silicoflagellates from the Northeast Aegean (eastern Mediterranean Sea): Morphologies and double skeletons

Silicoflagellate abundance, vertical distribution and morphology were studied during spring (March 2014) at three sampling stations located in the Northeast Aegean Sea adjacent to the Dardanelles Strait and characterized by a variable influx of cold, low-salinity Black Sea water. The silicoflagellate assemblage was dominated by Dictyocha stapedia and Stephanocha speculum with minor contribution of D. aculeata and Octactis pulchra. While specimens of D. stapedia were represented by the typical morphologies described in other areas of the Mediterranean Sea, populations of S. speculum displayed peculiar characters: they were large, predominantly 7-sided, with a small apical ring as well as apical ring spines, concave basal ring sides and non-rotated apical structure. Some of these features have been described for S. speculum at high latitudes, but the combined characters make these specimens slightly different from the high latitude populations. Similar morphologies have been observed in the western Black Sea, thus we can infer that the peculiar specimens detected in the Northeast Aegean are associated with the influx of Black Sea water masses.     

Phylogenetic relationships of Cochlodinium polykrikoides Margalef (Gymnodiniales,Dinophyceae) from the Mediterranean Sea and the implications of its global biogeography

Although the diversity of dinoflagellates has been intensively studied in several locations in the Mediterranean Sea since the 1950s, it is only during the last two decades that the morphotype of the toxic unarmoured dinoflagellate Cochlodinium polykrikoides Margalef has been detected, coinciding with its apparent worldwide expansion in marine coastal waters. In this study, vegetative cells of C. polykrikoides morphotype from the Catalan coast (NW Mediterranean Sea) were detected and isolated, and the DNA from collected cells was sequenced. While in the Mediterranean Sea, detections are scarce and C. polykrikoides is consistently present at low concentrations, we reported exceptional blooms of this species, in which the maximum abundance reached 2 × 10⁴ cells L⁻¹. Partial LSU rDNA region sequences showed that most C. polykrikoides populations from the Catalan coast formed a new differentiated ribotype, but others were included within the ‘Philippines’ ribotype, demonstrating their coexistence in the Mediterranean Sea. Thus, the current biogeographic nomenclature of the ribotypes is likely to be invalid with respect to the available information from populations comprising the ‘Philippines’ ribotype. The phylogeny suggests the existence of cryptic species that should be evaluated for species-level status. Accordingly, the ribotype determination must be carefully evaluated for all detections and bloom events, since accurate characterization of the morphology, ecophysiology and distribution of the ribotypes are not well resolved.     

The presence of exotic Hypnea flexicaulis (Rhodophyta) in the Mediterranean Sea as indicated by morphology, rbcL and cox1 analyses

Here we report the first finding of Hypnea flexicaulis Yamagishi and Masuda in the Mediterranean Sea (Lagoon of Venice, Italy), identified through molecular analyses using the plastid ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) and the mitochondrial protein-coding cytochrome c oxidase subunit I (cox1) genes. The phylogenetic reconstruction, based on rbcL + cox1 multiple alignment, showed that all specimens of H. flexicaulis from Venice, Korea, Philippines and Taiwan were included in a monophyletic group supported by a bootstrap value of 100%.It is highly probable that H. flexicaulis has been introduced from Indo-Pacific populations, in particular the Korean one, probably via ship traffic or shellfish transfers.The use of DNA barcoding combined with morphological observations was, in this case, a rapid way to identify this allochthonous species.     

Distribution, ecology and morphology of Lytocarpia myriophyllum (Cnidaria: Hydrozoa), a Mediterranean Sea habitat former to protect

In the Mediterranean Sea, as well as in other parts of the word, intense bottom trawling threatens deep and mesophotic assemblages, compromising mainly the survivorship of erect organisms and of the habitat complexity they shape. Protection of species able to affect their habitats, by increasing spatial complexity and enhancing interspecific interactions, is crucial for biodiversity conservation. It is urgent to highlight the occurrence of those species which act as ecosystem engineers and/or habitat former to enhance awareness on their ecological role and to develop focused conservation strategies. Lytocarpia myriophyllum is the largest Leptomedusan hydroid of the Mediterranean Sea, with colonies up to 1 m high, and the most abundant Aglaopheniid in the eastern part of the North Atlantic Ocean. This species creates wide forests on soft bottoms stabilizing sediments, providing refuge and food for several other associated organisms and could be defined both a habitat former and an ecosystem engineer. Thanks to trimix diving here we report on new insights on the morphological, biological and ecological features of L. myriophyllum meadows from the Mediterranean Sea furnishing a baseline for protection plans focused on these facies. This work demonstrates that direct studies of mesophotic habitats allow to collect far more detailed information than grabs, ROVs, or towed camera arrays and highlights the urgent need to redefine the vertical extension of several marine protected areas.     

Posidonia oceanica in the Marmara Sea

The seagrass Posidonia oceanica is a stenohaline species endemic to the Mediterranean Sea, where it normally lives at a salinity of between 36.5 and 39.5 ppt. Surveys carried out at the North-eastern distribution limits revealed large beds in the Dardanelles Strait and isolated beds in the Marmara Sea, where the salinity ranges between 21.5 and 28 ppt. Microsatellite analysis of these low-salinity tolerant P. oceanica beds, show different signs of genetic isolation: excess of heterozygosity and a presence of fixed alleles. These particularities are rarely found in the whole distributional range of the species. Moreover, all the populations considered in the analysis have a very low genetic diversity in comparison with most of the meadows sampled throughout the Mediterranean Sea. Taking into consideration the genetic data, rhizome expansion rate and the actual extent of the isolated beds in the Marmara Sea and knowing the reproductive rate and dissemination characteristics of P. oceanica, we hypothesize that the isolated population of the Marmara Sea has been established since the Middle Holocene, before the catastrophic intrusion of brackish water into the Marmara Sea and the strong and persistent flow coming from the Black Sea.     

Morphological and molecular tools in identifying the Mediterranean limpets Patella caerulea, Patella aspera and Patella rustica

Allozyme electrophoresis, a partial nucleotide sequence of a mitochondrial gene for cytochrome oxidase I (COI) and discriminant analysis of shell morphometric characters were used to study the relationships among the Sicilian marine gastropods of the Patella genus. Allozyme and mtDNA markers unequivocally distinguished the species and were very useful markers in correctly classifying the different species when morphological characters overlapped each other. Several allozyme loci and many nucleotide positions were diagnostic of species. In contrast, the discriminant analysis of simple morphometric shell characters failed to adequately discriminate the species, suggesting that environmental factors influence colouration and morphological patterns in the Patella species. Our results underline the importance of a genetic approach, as compared to a morphological approach, in discriminating the Mediterranean Patella species.     

Ulva (Chlorophyta,Ulvales) Biodiversity in the North Adriatic Sea (Mediterranean,Italy): Cryptic Species and New Introductions

Ulva Linnaeus (Ulvophyceae, Ulvales) is a genus of green algae widespread in different aquatic environments. Members of this genus show a very simple morphology and a certain degree of phenotypic plasticity, heavily influenced by environmental conditions, making difficult the delineation of species by morphological features alone. Most studies dealing with Ulva biodiversity in Mediterranean waters have been based only on morphological characters and a modern taxonomic revision of this genus in the Mediterranean is not available. We report here the results of an investigation on the diversity of Ulva in the North Adriatic Sea based on molecular analyses. Collections from three areas, two of which subject to intense shipping traffic, were examined, as well as historical collections of Ulva stored in the Herbarium Patavinum of the University of Padova, Italy. Molecular analyses based on partial sequences of the rbcL and tufA genes revealed the presence of six different species, often with overlapping morphologies: U. californica Wille, U. flexuosa Wulfen, U. rigida C. Agardh, U. compressa Linnaeus, U. pertusa Kjellman, and one probable new taxon. U. californica is a new record for the Mediterranean and U. pertusa is a new record for the Adriatic. Partial sequences obtained from historical collections show that most of the old specimens are referable to U. rigida. No specimens referable to the two alien species were found among the old herbarium specimens. The results indicate that the number of introduced seaweed species and their impact on Mediterranean communities have been underestimated, due to the difficulties in species identification of morphologically simple taxa as Ulva.     

Population Genetic History of Aristeus antennatus (Crustacea: Decapoda) in the Western and Central Mediterranean Sea

Aristeus antennatus is an ecologically and economically important deep-water species in the Mediterranean Sea. In this study we investigated the genetic variability of A. antennatus sampled from 10 sampling stations in the Western and Central Mediterranean. By comparing our new samples with available data from the Western area, we aim to identify potential genetic stocks of A. antennatus and to reconstruct its historical demography in the Mediterranean. We analyzed two regions of mitochondrial DNA in 319 individuals, namely COI and 16S. We found two main results: i) the genetic diversity values consistent with previous data within the Mediterranean and the absence of barriers to gene flow within the Mediterranean Sea; ii) a constant long-term effective population size in almost all demes but a strong signature of population expansion in the pooled sample about 50,000 years B.P./ago. We propose two explanation for our results. The first is based on the ecology of A. antennatus. We suggest the existence of a complex meta-population structured into two layers: a deeper-dwelling stock, not affected by fishing, which preserves the pattern of historical demography; and genetically homogeneous demes inhabiting the fishing grounds. The larval dispersal, adult migration and continuous movements of individuals from “virgin” deeper grounds not affected by fishing to upper fishing areas support an effective ‘rescue effect’ contributing to the recovery of the exploited stocks and explain their genetic homogeneity throughout the Mediterranean Sea. The second is based on the reproduction model of this shrimp: the high variance in offspring production calls for a careful interpretation of the data observed under classical population genetics and Kingman’s coalescent. In both cases, management policies for A. antennatus will therefore require careful evaluation of the meta-population dynamics of all stocks in the Mediterranean. In the future, it will be particularly relevant to sample the deepest ones directly.     

Recognition of a new species of Kallymenia (Gigartinales,Rhodophyta) from Croatia (Mediterranean Sea) based on morphology and DNA barcode

The Mediterranean red algal flora is diverse but current knowledge of its diversity is at best fragmentary. Here, a new species of Kallymenia from Croatia is described based on morphological and molecular data. Members of the genus Kallymenia share similar morphology making their generic identification relatively easy, whereas species level identification is notoriously difficult. In this paper an integrative systematics study using three gene markers, cox1 (COI), rbcL and nuclear LSU, allowed us to (i) confirm the identity of four existing members of this genus inhabiting the Mediterranean Sea: K. feldmannii, K. lacerata, K. patens and K. requienii; (ii) detect the presence of K. reniformis only in the Atlantic, and (iii) reveal the presence of a new member of Kallymenia from the Mediterranean Sea, K. ercegovicii Vergés & Le Gall, sp. nov., which is described based on distinctive morphological and molecular characters. Kallymenia ercegovicii is distinguished, using three molecular markers, from all Kallymenia species for which these sequences are available. Morphologically, the new species can be distinguished from other polycarpogonial Kallymenia by a set of characters including a deeply lobed thallus, large inner cortical cells, stellate but non-glanglionic medullary cells and non-ostiolate cystocarps surrounded by a filamentous net composed of elongated cells forming fascicles. An initial phylogeny of the genus Kallymenia was inferred from cox1 (COI), rbcL and nuclear LSU sequences. Kallymenia ercegovicii was resolved with full support within the same lineage as K. reniformis (the generitype), K. feldmannii and K. patens, whereas K. lacerata and K. requienii were only distantly related.     

Nocardiosis in Mediterranean bivalves: First detection of Nocardia crassostreae in a new host Mytilus galloprovincialis and in Ostrea edulis from the Gulf of Naples (Italy)

In this work M. galloprovincialis and O. edulis specimens were surveyed for a pathological study in the Gulf of Naples (Mediterranean sea, Campania Region, southern Italy). Clusters of Nocardia sp.-like cells were observed in histological slides. PCR amplification, sequencing and in situ hybridization were carried out in order to corroborate Nocardia species identification for both hosts. Blast results showed a 99% of maximum identity with Nocardia crassostreae sequences in Genbank. This is the first report of N. crassostreae in the new host M. galloprovincialis and, in a new area, the Mediterranean Sea.