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.     

Genetic and morphological analyses of tub gurnard Chelidonichthys lucerna populations in Turkish marine waters

Genetic and morphological structure of tub gurnard Chelidonichthys lucerna populations in Turkish marine waters were investigated with mtDNA sequencing of 16S rRNA and morphological characters. C. lucerna samples were collected from the Black Sea, Marmara, Aegean and northeastern Mediterranean coasts of Turkey. The lowest genetic diversity was found in the northeastern Mediterranean (Iskenderun Bay) population, while the highest was in the Marmara population with overall average value of genetic diversity within populations. A total of 14 haplotypes was found, and the highest haplotype diversity was in the Black Sea whereas the lowest was in the northeastern Mediterranean population (Iskenderun Bay). The Black Sea and Iskenderun Bay populations showed the least genetic divergence (0.001081), while the highest was between the Marmara Sea and northeastern Mediterranean (Antalya Bay) populations (0.002067). Pairwise comparisons of genetic distance revealed statistically significant differences (P < 0.05) between the Marmara and both the Aegean and northeastern Mediterranean (Antalya Bay) samples. Neighbour joining tree analyses clustered the northeastern Mediterranean populations (Antalya Bay and Iskenderun Bay) as genetically more interrelated populations, whereas the Aegean Sea population was clustered as most isolated one. Discriminant function analysis of morphological characters showed that only the Black Sea population is differentiated from the other populations.     

Identification of subspecies of European anchovy <Emphasis Type="Italic">Engraulis encrasicolus</Emphasis> (Engraulidae) in the wintering aggregations based on morphological parameters of otoliths

A new method of identification of the Sea of Azov and Black Sea subspecies of European anchovy Engraulis encrasicolus in mixed wintering aggregations is suggested based on discriminant analysis using a morphological parameter, the angle of the central groove of the otolith. This angle differs significantly in the subspecies, ranging between 92° and 148° and between 55° and 95° in the Azov and Black Sea subspecies, respectively. In the Azov anchovy, the difference between the eastern and western populations in the angle of the central groove is not registered. The classification equations obtained in the study can be used for subsequent assessment of the ratio between the abundances of each subspecies in mixed aggregations of European anchovy in the inshore zone of Crimea.     

The taxonomic status of the Black Sea and Marmara Sea populations of the Broadnosed Pipefish Syngnathus cf. argentatus Pallas (Teleostei: Syngnathidae) based on morphological and molecular characters

As the Black Sea and Marmara Sea population of the Broadnosed Pipefish Syngnathus cf. argentatus show some morphological differences from the Mediterranean Sea populations, some authors regard it as an endemic species Syngnathus argentatus Pallas, 1814, while others consider it as a synonym of S. typhle Linnaeus, 1758. The aim of this study is to compare the populations of the Black Sea and Sea of Marmara with the Aegean Sea population, using a combination of morphological and molecular characters, in order to clarify their taxonomic status. Sampling was carried out at three stations in the Black Sea, two in the Sea of Marmara and three in the Aegean Sea, and a total of 24 morphometric and 6 meristic characters were examined. Metric data were analysed by Principal Component Analysis (PCA) and phylogenetic relationships between the populations were analysed using both cytochrome oxidase subunit 1 (COI) gene and cytochrome b (cyt-b) gene sequences. Although constant differences were observed in snout depths between the Black Sea/Marmara Sea and the Aegean Sea populations, other morphological features and genetic analysis did not enable these populations to be differentiated. These findings indicate that S. argentatus is a synonym of S. typhle.     

Genetic and morphological divergence between populations of the flatfish Platichthys flesus (L.) (Pleuronectidae)

Populations of the flounder, Platichthys flesus , were screened for electrophoretically detectable protein variation at up to 37 loci. Atlantic and North Sea populations (subspecies flesus ) were genetically very similar to one another ( I >0.99) but different from Adriatic (subspecies italicus ) and Black Sea (subspecies luscus ) populations. The values for genetic identity between subspecies were around 0.9. Diagnostic loci enabled specimens of flesus, italicus and luscus to be differentiated from one another. Samples of flesus showed two to three times the heterozygosity levels of italicus or luscus , consistent with the greater population size of flesus . Morphological comparisons enabled populations to be categorized to subspecies. Platichthys flesus italicus and luscus are valid subspecies: the terms are not synonyms. The genetic distance data provide an estimate of divergence time of the three subspecies at about 2 My ago, an estimate that seems reasonable in the light of present knowledge concerning the recent geological histories of the Mediterranean and Black Sea basins.     

Genetic variability of anchovy in the Azov-Black Sea basin

The intraspecific structure of the European anchovy (Engraulis encrasicolus) was studied on the basis of mitochondrial cytochrome b gene (cytb) fragment variability in 84 individuals from seven localities of the Black Sea and the Sea of Azov. The data on haplotype and nucleotide diversity and the values of neutrality tests suggested expansive growth of anchovy populations in the Azov-Black Sea basin. All samples from anchovy populations demonstrated a high level of haplotype diversity (Hd = 0.962). Two dominant haplotypes were identified, the frequencies of which were not directional, and they were present in all localities. Sequence analysis of the mitochondrial cytb gene fragment showed no differentiation between the Sea of Azov and the Black Sea subspecies.     

How an invasive alga species (<Emphasis Type="Italic">Caulerpa taxifolia</Emphasis>) induces changes in foraging strategies of the benthivorous fish <Emphasis Type="Italic">Mullus surmuletus</Emphasis> in coastal Mediterranean ecosystems

The foraging behaviour variability of three striped red mullet (Mullus surmuletus L. 1758) populations with respect to the vegetation cover was examined along the French Mediterranean coasts. We tested both the carrying capacity of different habitats and the hypothesis of a food segregation by the invasive alga Caulerpa taxifolia (Vahl) C. Agardh, on the functioning of benthic fish populations. The results indicated a significant site effect on M. surmuletus foraging behaviour and movements. The vegetation cover may play an important role in modifying the striped red mullet cost of foraging (sampling effort) and thus its strategy of prey capture. As long as the cover of marine phanerogams and/or macrophyte algae (Caulerpaceae) increases, the foraging budget and the distances covered in search of prey decrease significantly. Similarly, the striped red mullet increases the prospected sample periods to augment its foraging success, swimming above the bottom. These changes are related to the development of a dense superficial network of plagiotropic rhizomes and stolons, and to the reduction of space between fronds that limits the accessibility to resources and may increase intra-specific food competition. The role of C. taxifolia does not differ from that of other marine phanerogams but induces significant changes in the structure of Mullidae populations at the local level. The rapid expansion of Caulerpaceae in the Mediterranean Sea could constitute a real threat for the balance of the marine coastal biodiversity and the ecology of M. surmuletus, which is considered a flagship species for coastal Mediterranean demersal fisheries.     

Invasion Pathway of the Ctenophore Mnemiopsis leidyi in the Mediterranean Sea

Gelatinous zooplankton outbreaks have increased globally owing to a number of human-mediated factors, including food web alterations and species introductions. The invasive ctenophore Mnemiopsis leidyi entered the Black Sea in the early 1980s. The invasion was followed by the Azov, Caspian, Baltic and North Seas, and, most recently, the Mediterranean Sea. Previous studies identified two distinct invasion pathways of M. leidyi from its native range in the western Atlantic Ocean to Eurasia. However, the source of newly established populations in the Mediterranean Sea remains unclear. Here we build upon our previous study and investigate sequence variation in both mitochondrial (Cytochrome c Oxidase subunit I) and nuclear (Internal Transcribed Spacer) markers in M. leidyi, encompassing five native and 11 introduced populations, including four from the Mediterranean Sea. Extant genetic diversity in Mediterranean populations (n = 8, N _a = 10) preclude the occurrence of a severe genetic bottleneck or founder effects in the initial colonizing population. Our mitochondrial and nuclear marker surveys revealed two possible pathways of introduction into Mediterranean Sea. In total, 17 haplotypes and 18 alleles were recovered from all surveyed populations. Haplotype and allelic diversity of Mediterranean populations were comparable to populations from which they were likely drawn. The distribution of genetic diversity and pattern of genetic differentiation suggest initial colonization of the Mediterranean from the Black-Azov Seas (pairwise F _ST = 0.001–0.028). However, some haplotypes and alleles from the Mediterranean Sea were not detected from the well-sampled Black Sea, although they were found in Gulf of Mexico populations that were also genetically similar to those in the Mediterranean Sea (pairwise F _ST = 0.010–0.032), raising the possibility of multiple invasion sources. Multiple introductions from a combination of Black Sea and native region sources could be facilitated by intense local and transcontinental shipping activity, respectively.     

Contemporary genetic structure and postglacial demographic history of the black scorpionfish,Scorpaena porcus,in the Mediterranean and the Black Seas

Understanding the distribution of genetic diversity in the light of past demographic events linked with climatic shifts will help to forecast evolutionary trajectories of ecosystems within the current context of climate change. In this study, mitochondrial sequences and microsatellite loci were analysed using traditional population genetic approaches together with Bayesian dating and the more recent approximate Bayesian computation scenario testing. The genetic structure and demographic history of a commercial fish, the black scorpionfish, Scorpaena porcus, was investigated throughout the Mediterranean and Black Seas. The results suggest that the species recently underwent population expansions, in both seas, likely concomitant with the warming period following the Last Glacial Maximum, 20 000 years ago. A weak contemporaneous genetic differentiation was identified between the Black Sea and the Mediterranean Sea. However, the genetic diversity was similar for populations of the two seas, suggesting a high number of colonizers entered the Black Sea during the interglacial period and/or the presence of a refugial population in the Black Sea during the glacial period. Finally, within seas, an east/west genetic differentiation in the Adriatic seems to prevail, whereas the Black Sea does not show any structured spatial genetic pattern of its population. Overall, these results suggest that the Black Sea is not that isolated from the Mediterranean, and both seas revealed similar evolutionary patterns related to climate change and changes in sea level.     

Importance of morphological traits and coloration for diagnostics of species of the genus <Emphasis Type="Italic">Mullus</Emphasis> (Mullidae,Perciformes), assessment of the taxonomic status of populations,and revision of ranges

Comparative analysis was performed of morphological characters and coloration elements in goatfish of the genus Mullus from collections of the Zoological Museum (Moscow State University, Russian Academy of Sciences), on the basis of which individuals from Atlantic waters off the coasts of western Africa were identified as M. argentinae. Brief characteristics of all valid species of the genus are presented; the main diagnostic characters include: ratio between the length of the barbels and pectoral fins, number of pyloric caeca, and the presence of a dark spot (or stripe) in the upper part of dorsal fin. The Mediterranean populations of M. surmuletus are separated as a special subspecies M. surmuletus fuscatus, and M. argentinae from eastern Atlantic is described as a new subspecies M. argentinae africanus.     

Phylogeography of the planktonic chaetognath Sagitta setosa reveals isolation in European seas

Numerous planktonic species have disjunct distribution patterns in the world's oceans. However, it is unclear whether these are truly unconnected by gene flow, or whether they are composed of morphologically cryptic species. The marine planktonic chaetognath Sagitta setosa Müller has a discontinuous geographic distribution over the continental shelf in the northeastern Atlantic, Mediterranean Sea, and Black Sea. Morphological variation between these populations has been described, but overlaps and is therefore unsuitable to determine the degree of isolation between populations. To test whether disjunct populations are also genetically disjunct, we sequenced a 504-bp fragment of mitochondrial DNA comprising the cytochrome oxidase II region of 86 individuals. Sequences were highly variable; each represented a different haplotype. Within S. setosa, sequence divergence ranged from 0.2 to 8.1% and strong phylogeographic structure was found, with four main groups corresponding to the northeastern Atlantic, Mediterranean Sea (including Ligurian Sea, Tyrrhenian Sea and Gulf of Gabes), Adriatic Sea, and Black Sea. Two of these (Atlantic and Black Sea) were resolved as monophyletic clades, thus gene flow between disjunct populations of S. setosa has been extremely limited and lineage sorting has taken place. The deepest divergence was between Atlantic and Mediterranean/Black Sea populations followed by a split between Mediterranean and Black Sea populations. The Mediterranean/Black Sea clade comprised three groups, with the Adriatic Sea as the most likely sister clade of the Black Sea. These data are consistent with a colonization of the Black Sea from the Mediterranean. Furthermore, a possible cryptic species was found in the Black Sea with 23.1% sequence divergence from S. setosa. Two possibilities for the evolutionary origin of this species are proposed, namely, that it represents a relict species from the ancient Paratethys, or that it represents another chaetognath species that colonized the Black Sea more recently. Even though the exact timing of disjunction of S. setosa populations remains unclear, on the basis of the geological and paleoclimatic history of the European basins and our estimates of net nucleotide divergence, we suggest that disjunct populations arose through vicariance resulting from the cyclical changes in temperature and sea levels during the Pleistocene. We conclude that these populations have remained disjunct, not because of limited dispersal ability, but because of the inability to maintain viable populations in suboptimal, geographically intermediate areas.     

Mitochondrial phylogeography of the European sprat (Sprattus sprattus L., Clupeidae) reveals isolated climatically vulnerable populations in the Mediterranean Sea and range expansion in the northeast Atlantic

We examined the genetic structure of the European sprat ( Sprattus sprattus ) by means of a 530-bp sequence of the mitochondrial control region from 210 fish originating from seven sampling localities of its distributional range. Phylogeographical analysis of 128 haplotypes showed a phylogenetic separation into two major clades with the Strait of Sicily acting as a barrier to gene flow between them. While no population differentiation was observed based on analysis of molecular variance and net nucleotide differences between samples of the Baltic Sea, the North Sea and the Bay of Biscay nor between the Black Sea and the Bosporus, a strong population differentiation between these samples and two samples from the Mediterranean Sea was found. Further, the biggest genetic distance was observed within the Mediterranean Sea between the populations of the Gulf of Lyon and the Adriatic Sea, indicating genetic isolation of these regions. Low genetic diversities and star-like haplotype networks of both Mediterranean Sea populations point towards recent demographic expansion scenarios after low population size, which is further supported by negative F _S values and unimodal mismatch distributions with a low mean. Along the northeast Atlantic coast, a northwards range expansion of a large and stable population can be assumed. The history of a diverse but differentiated Black Sea population remains unknown due to uncertainties in the palaeo-oceanography of this sea. Our genetic data did not confirm the presently used classification into subspecies but are only preliminary in the absence of nuclear genetic analyses.     

Effect of salinity on vegetative growth and sexual reproduction of algae from the genus <Emphasis Type="Italic">Ardissonea</Emphasis> de Notaris (Bacillariophyta)

Effect of salinity on vegetative growth and sexual reproduction was investigated in laboratory cultures of a benthic diatom Ardissonea crystallina (C. Agardh) Grunow from the Black Sea and vegetative growth of Ardissonea sp. from the coast of Martinique, the Atlantic Ocean. Typical water salinity in the places where the populations inhabit differs two times. It was shown that the clones of both populations have a broad tolerance and great ability to adapt to changes in salinity. Cells of different size (i.e., in different stages of the life cycle) responded differently to desalination. Salinities optimal for vegetative growth of these species were not the same. Ardissonea from the coast of Martinique was more sensitive to low levels of salinity as compared with A. crystallina from the Black Sea. It was surprising that salinity optimal for vegetative growth and sexual reproduction of the Black Sea species did not coincide with the real salinity level of the Black Sea. Position of physiological optimums indicates an oceanic (or Mediterranean) origin of the Black Sea population of A. crystallina.     

The Taxonomic Status and Intraspecific Differentiation of the Black Sea Horse Mackerel, <Emphasis Type="Italic">Trachurus mediterraneus ponticus</Emphasis> (Aleev, 1956) (Carangidae)

The taxonomic status of the horse mackerel, Trachurus mediterraneus ponticus (Aleev, 1956), which inhabits the Black Sea, has been clarified based on the analysis of variation in the nucleotide sequence of the mtDNA cytochrome b (Cyt-b) locus. A comparison of this species with the horse mackerel from the Mediterranean basin and the World Ocean has shown that the Black Sea horse mackerel is neither an independent species nor a subspecies and can only be qualified as the species Trachurus mediterraneus (Steindachner, 1868) distributed throughout the Mediterranean Sea basin. A hypothesis has been made about the possible cause of the origin and existence of two forms, “large” and “small”, of the horse mackerel. By using the technique of calculating ontogenetic trajectories, it has been found that these are two morpho-ecological forms, whose differentiation is most likely determined by the type of feeding.     

Oocytes development,maturity classification,maturity size and spawning season of the red mullet (Mullus barbatus barbatus Linnaeus, 1758)

The present study was designed to identify maturity of female red mullet, Mullus barbatus barbatus, using microscopic criteria, offering an accurate determination of the ovarian cycle and estimation of maturity size. Histological analysis of 499 females from 13 monthly samplings was applied (Thermaikos Gulf, N. Aegean Sea, Greece). Oocyte development was divided into five stages, with a mean oocyte size range from 56 to 363 μm in diameter. Ovarian maturity was defined by the maturation stage of the most advanced oocytes and divided into four phases. By using the monthly changes in the percentage frequency of ovarian phases, the ovarian cycle of M. barbatus was divided into three periods: a long period of early oogenesis (November to February), a short period of vitellogenesis (February to April) and a spawning period from April through June with peak activity in May. Both gonadosomatic (up to 7.5%) and hepatosomatic indices (up to 2%) can be used together to predict the spawning period. Red mullet should be classified as a multiple spawning species. Females reached 50% maturity (Lm₅₀) at 11.4 cm and 95% maturity (Lm₉₅) at 15.5 cm TL. The Lm₅₀ values for female red mullet populations across the Mediterranean Sea tended to show differences ranging from 11 to 14.4 cm FL or 11.4–14 cm TL. The current European Union fisheries management plan stipulates a minimum landing size (MLS) of 11 cm TL for Mullidae spp. fisheries in the Mediterranean. This MLS is at the lower part of the Lm₅₀ range, thus allowing an extensive removal of immature juveniles. A revision of this MLS value is recommended to help ensure that red mullet stocks remain within safe biological limits.     

Historic speciation and recent colonization of Eurasian monkey gobies (Neogobius fluviatilis and N. pallasi) revealed by DNA sequences,microsatellites, and morphology

Aim Hidden diversity within an invasive ‘species’ can mask both invasion pathways and confound management goals. We assessed taxonomic status and population structure of the monkey goby Neogobius fluviatilis across Eurasia, comparing genetic variation across its native and invasive ranges. Location Native populations were analysed within the Black and Caspian Sea basins, including major river drainages (Dnieper, Dniester, Danube, Don and Volga rivers), along with introduced locations within the upper Danube and Vistula river systems. Methods DNA sequences and 10 nuclear microsatellite loci were analysed to test genetic diversity and divergence patterns of native and introduced populations; phylogenetic analysis of mtDNA cytochrome b and nuclear RAG‐1 sequences assessed taxonomic status of Black and Caspian Sea lineages. Multivariate analysis of morphology was used to corroborate phylogenetic patterns. Population genetic structure within each basin was evaluated with mtDNA and microsatellite data using F_ST analogues and Bayesian assignment tests. Results Phylogenetic analysis of mitochondrial and nuclear sequences discerned a pronounced genetic break between monkey gobies in the Black and Caspian Seas, indicating a long‐term species‐level separation dating to c. 3 million years. This pronounced separation further was confirmed from morphological and population genetic divergence. Bayesian inference showed congruent patterns of population structure within the Black Sea basin. Introduced populations in the Danube and Vistula River basins traced to north‐west Black Sea origins, a genetic expansion pattern matching that of other introduced Ponto‐Caspian gobiids. Main conclusions Both genetic and morphological data strongly supported two species of monkey gobies that were formerly identified as subspecies: N. fluviatilis in the Black Sea basin, Don and Volga Rivers, and the Kumo‐Manych Depression, and Neogobius pallasi in the Caspian Sea and Volga River delta. Genetic origins of introduced N. fluviatilis populations indicated a common invasion pathway shared with other introduced Ponto‐Caspian fishes and invertebrates.     

Multilocus approach reveals an incipient differentiation process in the Stone-curlew, <Emphasis Type="Italic">Burhinus oedicnemus</Emphasis> around the Mediterranean basin

The Stone-curlew Burhinus oedicnemus, a steppe bird species, is mainly distributed in the Mediterranean and Macaronesian regions, which are considered hotspots of biodiversity with priority for animal and plant species richness conservation. In this study, we investigated the genetic diversity of the Stone-curlew in the Mediterranean basin and in the Canary Islands by applying a multilocus approach. We analysed mitochondrial and nuclear markers in order to evaluate the genetic structure and the congruence between morphological subspecies and geographic samples. We found a significant level of genetic differentiation between Mediterranean and Canary Island populations with all markers. Both in the Mediterranean basin and in the Canary Islands, we found a significant level of genetic diversity with nuclear markers only. We identified seven population groups, including insular populations. The four subspecies described for the Western Palaearctic were confirmed with some changes in distribution range. In spite of habitat fragmentation and negative population trend, the Stone-curlew showed a significant level of genetic diversity and gene flow among continental populations. However, islands constitute important reservoirs of genetic diversity and a potential for the evolution of the species.