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The eastern part of the Black Sea and its tributaries are suitable habitats for several sturgeon species, among which Acipenser gueldenstaedtii, A. stellatus, A. nudiventris, A. persicus, A. sturio, and H. huso are well documented. However, different threats have led these species to a dramatic decline, all of them are currently listed as Critically Endangered, and some Locally Extinct, in that area. We tested 94 wild sturgeon samples from the Black Sea and Rioni River by analyzing the mitochondrial Control Region and nuclear markers for hybrid identification. The data analyses (1) assessed mitochondrial diversity among samples, (2) identified their species, as well as (3) indicated instances of hybridization. The data collected, besides confirming a sharp decrease of catches of Beluga and Stellate sturgeon in recent years, also revealed four juvenile hybrids between Russian and Stellate sturgeon, providing the first evidence of natural interspecific hybridization in the Rioni. The present communication raises concerns about the status of sturgeon species in this area and underlines the urgent need for conservation programs to restore self-sustaining populations.

Aim: The species of the genus Chionodraco (Notothenioidei) are the most abundant icefish on the continental shelf of the Weddell Sea. While previous studies indicated that only Chionodraco hamatus and Chionodraco myersi inhabit the Weddell Sea, the third Chionodraco species, Chionodraco rastrospinosus, was recently sampled in the area. As C. rastrospinosus is supposed to be found only at the Antarctic Peninsula and Scotia Arc, this study aimed at confirming the species classification of C. rastrospinosus by molecular methods and identifying its putative source population. Given the documented evidence of introgression among the three species, we tested whether the newly found C. rastrospinosus shared any genetic variability with the other Chionodraco species. To explain the pattern of distribution of the Chionodraco species, we aimed at estimating the hydrodynamic connectivity between the Antarctic Peninsula and the Weddell Sea. Location: Antarctic Peninsula, southern Scotia Arc and the south-eastern Weddell Sea. Methods: We genotyped 19 microsatellites and sequenced the mitochondrial D-loop for 560 Chionodraco individuals. We simulated the dispersal of more than 3 million drifters (Lagrangian model). Results: The molecular analyses support the presence of C. rastrospinosus in the Weddell Sea and its homogeneity with C. rastrospinosus from the Antarctic Peninsula. Bayesian clustering identifies three putative hybrids among C. rastrospinosus and the other congenerics. Lagrangian simulations do not support connectivity driven by the oceanographic features of the Antarctic Peninsula and Weddell Sea via passive larval dispersal only. Main conclusions: This study documents, for the first time, the presence of C. rastrospinosus in the Weddell Sea unveiling more biodiversity than previously known in this region. The sympatry of the three Chionodraco species explains the occurrence of occasional, ongoing events of hybridization in the genus. Alternative possible hypotheses need to be tested in future studies about the mechanisms maintaining the interspecific connectivity in Chionodraco spp.

China has recently become the leader country for sturgeon aquaculture and caviar production, deeply changing the traditional geography of this market in few years. As a consequence, some species originating from the Far East Asia increased their economic relevance, joining the ones traditionally harvested for caviar. In this context, the possibility to reliably and promptly identify these species on the market has increasing importance for the enforcement of control actions against illegal trade or commercial frauds. The present study focuses on two commercially relevant species, massively reared in China not only as pure species but also as reciprocal hybrids: the Amur ( Acipenser schrenckii ) and Kaluga ( Huso dauricus ) sturgeons. We assess the identification power of two putatively diagnostic markers isolated from two predicted introns of the nuclear coding gene Ribosomal Protein L 8 . The markers were tested on tissue or caviar of 508 individuals of the two species and 31 hybrids. In order to compare results across loci, most individuals were also checked at two already published microsatellite markers, with a good, even if incomplete, identification efficiency for the two species. No marker showed fixed alternative alleles between Amur and Kaluga sturgeons, confirming the difficulty of distinguishing these two sympatric species in spite of the marked morphological differences and the consequent classification into different genera. So far, the multi-locus panel here used represents the more effective tool for the genetic identification of pure Amur and Kaluga sturgeons and resulted to be fully efficient to validate caviar and tissues obtained from hybrids between the two species.

Marine bioconstructions are biodiversity-rich, three-dimensional biogenic structures, regulating key ecological functions of benthic ecosystems worldwide. Tropical coral reefs are outstanding for their beauty, diversity and complexity, but analogous types of bioconstructions are also present in temperate seas. The main bioconstructions in the Mediterranean Sea are represented by coralligenous formations, vermetid reefs, deep-sea cold-water corals, Lithophyllum byssoides trottoirs, coral banks formed by the shallow-water corals Cladocora caespitosa or Astroides calycularis, and sabellariid or serpulid worm reefs. Bioconstructions change the morphological and chemicophysical features of primary substrates and create new habitats for a large variety of organisms, playing pivotal roles in ecosystem functioning. In spite of their importance, Mediterranean bioconstructions have not received the same attention that tropical coral reefs have, and the knowledge of their biology, ecology and distribution is still fragmentary. All existing data about the spatial distribution of Italian bioconstructions have been collected, together with information about their growth patterns, dynamics and connectivity. The degradation of these habitats as a consequence of anthropogenic pressures (pollution, organic enrichment, fishery, coastal development, direct physical disturbance), climate change and the spread of invasive species was also investigated. The study of bioconstructions requires a holistic approach leading to a better understanding of their ecology and the application of more insightful management and conservation measures at basin scale, within ecologically coherent units based on connectivity: the cells of ecosystem functioning.

Aim Marine bioconstructions such as coralligenous formations are hotspot of biodiversity and play a relevant ecological role in the preservation of biodiversity by providing carbon regulation, protection and nursery areas for several marine species. For this reason, the European Union Habitat Directive included them among priority habitats to be preserved. Although their ecological role is well established, connectivity patterns are still poorly investigated, representing a limit in conservation planning. The present study pioneers a novel approach for the analysis of connectivity in marine bioconstructor species, which often lack suitable genetic markers, by taking advantage of next‐generation sequencing techniques. We assess the geographical patterns of genomic variation of the sunset cup coral Leptopsammia pruvoti Lacaze‐Duthiers, 1897, an ahermatypic, non‐zooxanthellate and solitary scleractinian coral species common in coralligenous habitats and distributed across the Mediterranean Sea. Location The Italian coastline (Western and Central Mediterranean). Methods We applied the restriction site‐associated 2b‐RAD approach to genotype over 1,000 high‐quality and filtered single nucleotide polymorphisms in 10 population samples. Results The results revealed the existence of a strongly supported genetic structure, with highly significant pairwise FST values between all the population samples, including those collected about 5 km apart from each other. Moreover, genomic data indicate that the strongest barriers to gene flow are between the western (Ligurian–Tyrrhenian Sea) and the eastern side (Adriatic Sea) of the Italian peninsula. Main conclusions The strong differentiation found in L. pruvoti is similar to that found in other species of marine bioconstructors investigated in this area, but it strongly contrasts with the small differences found in many fish and invertebrates at the same geographical scale. All in one, our results highlight the importance of assessing connectivity in species belonging to coralligenous habitats as, due to their limited dispersal ability, they might require specific spatial conservation measures.

Marine bioconstructions are biodiversity-rich, three-dimensional biogenic structures, regulating key ecological functions of benthic ecosystems worldwide. Tropical coral reefs are outstanding for their beauty, diversity and complexity, but analogous types of bioconstructions are also present in temperate seas. The main bioconstructions in the Mediterranean Sea are represented by coralligenous formations, vermetid reefs, deep-sea cold-water corals, Lithophyllum byssoides trottoirs, coral banks formed by the shallow-water corals Cladocora caespitosa or Astroides calycularis, and sabellariid or serpulid worm reefs. Bioconstructions change the morphological and chemicophysical features of primary substrates and create new habitats for a large variety of organisms, playing pivotal roles in ecosystem functioning. In spite of their importance, Mediterranean bioconstructions have not received the same attention that tropical coral reefs have, and the knowledge of their biology, ecology and distribution is still fragmentary. All existing data about the spatial distribution of Italian bioconstructions have been collected, together with information about their growth patterns, dynamics and connectivity. The degradation of these habitats as a consequence of anthropogenic pressures (pollution, organic enrichment, fishery, coastal development, direct physical disturbance), climate change and the spread of invasive species was also investigated. The study of bioconstructions requires a holistic approach leading to a better understanding of their ecology and the application of more insightful management and conservation measures at basin scale, within ecologically coherent units based on connectivity: the cells of ecosystem functioning.

Chamelea gallina is a valuable commercial species in the Mediterranean Sea. The strong fishing pressure on C. gallina in the northern and central Adriatic Sea has paralleled a clear-cut decrease in clam population density and the occurrence of several irregular mortality events. Despite the commercial interest in this species, nothing is known about its genetic sub-structuring at the geographic and/or temporal scale, nor its levels of genetic variability. Analyzing microsatellite genotypes for samples collected in the Adriatic Sea, we detected large geographic genetic homogeneity with gene flow guided by broad scale circulation in the north-south direction. Our results also indicate weak genetic differentiation among size classes at the local and temporal scale. These small genetic differences might be determined by variability of local circulation and reproductive success, partial overlapping generations and high larval mortality rates as suggested by our estimates of the effective number of breeders. In fact, global effective population size estimates over multiple generations are medium-high, but a low number of breeders are responsible for the small clams size class recruitment. Notwithstanding, it was not possible to detect signatures of bottleneck. Future efforts in fishery management should aim to maintain genetic diversity – essential to the long-term sustainability of the resource – and limit effective population size fluctuations while considering the need to improve water quality to avoid mass mortality events.

Ninety-five microsatellite loci were isolated from the output of two Next Generation Sequencing projects: a genomic Illumina RADSeq and a transcriptomic Roche 454 cDNA sequencing of the Adriatic sturgeon (Acipenser naccarii). From a total of 7697 loci identified, 72 were characterized and 57 were polymorphic in the tetraploid A. naccarii, providing new markers suitable for parental allocations to be performed for future conservation actions. Loci isolated from the transcriptome showed a higher level of polymorphism, which could be related to the different sequencing approaches used. Notably, a higher rate of duplication was observed in the transcribed loci, pointing to a preferential retention of sequences with a functional role after whole genome duplications occurred in sturgeons. Cross-species amplification of these markers was also assessed in the diploid beluga sturgeon (Huso huso), with the aim of setting up a panel of markers to be applied in future conservation programmes regarding this species.

Sturgeons are well known for the delicacy of their eggs, the caviar, one of the most valuable products on the food market. The high price of caviar led in the past to a severe overharvest of wild sturgeon species and to an increase in trade of counterfeit products sold with impunity in spite of the strict trade limitations. A priority in the effort to reduce illegal trading is the development of genetic tools in order to identify the species of traded products using a standardized, cheap and rapid approach. We developed the first genetic nuclear marker for the identification of the Beluga sturgeon ( Huso huso ), the most sought-after caviar producer. We explored the interspecific variability at the second intron of the nuclear S6 Ribosomal Protein (RP2S6), selected among 1867 introns, predicted by aligning the transcriptome of 3 sturgeon species with 3 complete fish genomes. The Beluga-specific SNP was identified by cloning and sequencing RP2S6 in 65 individuals of 11 species, validated on 341 additional individuals and tested on 18 caviar samples. Diagnostic primers designed on the SNP successfully amplified the expected band in all Beluga specimens while no PCR product was obtained from other pure species. The marker can also contribute to the identification of interspecific hybrids in which the Beluga is one of the parent species, such as in the case of the Bester, which produces one of the most mislabeled caviars in trade. The complete identification power on this highly relevant species and the proved efficacy on caviar samples represent an essential progress towards a standardized panel of nuclear markers for the control of illegal poaching, smuggling and mislabeling of sturgeons and their products.

Connectivity between populations influences both their dynamics and the genetic structuring of species. In this study, we explored connectivity patterns of a marine species with long‐distance dispersal, the edible common sea urchin Paracentrotus lividus, focusing mainly on the Adriatic–Ionian basins (Central Mediterranean). We applied a multidisciplinary approach integrating population genomics, based on 1,122 single nucleotide polymorphisms (SNPs) obtained from 2b‐RAD in 275 samples, with Lagrangian simulations performed with a biophysical model of larval dispersal. We detected genetic homogeneity among eight population samples collected in the focal Adriatic–Ionian area, whereas weak but significant differentiation was found with respect to two samples from the Western Mediterranean (France and Tunisia). This result was not affected by the few putative outlier loci identified in our dataset. Lagrangian simulations found a significant potential for larval exchange among the eight Adriatic–Ionian locations, supporting the hypothesis of connectivity of P. lividus populations in this area. A peculiar pattern emerged from the comparison of our results with those obtained from published P. lividus cytochrome b (cytb) sequences, the latter revealing genetic differentiation in the same geographic area despite a smaller sample size and a lower power to detect differences. The comparison with studies conducted using nuclear markers on other species with similar pelagic larval durations in the same Adriatic–Ionian locations indicates species‐specific differences in genetic connectivity patterns and warns against generalizing single‐species results to the entire community of rocky shore habitats.

Summary The Adriatic sturgeon (Acipenser naccarii Bonaparte, 1836), endemic to the North Adriatic region, was recently reclassified by IUCN as ‘Critically endangered and possibly extinct in the wild’ since no natural spawning has been recorded in the last 20 years. Its survival relies on restocking activities originating from a single captive broodstock collected by a private aquaculture plant (V.I.P.) in the 1970s and is currently reduced to 13 individuals. Few alternative brood-stocks of F1 animals have been retained for use in the near future. Thus far, brood-stocks, as well as all stocks released in the wild, have been randomly chosen without regard to their genetic composition. The consequence of breeding programmes with no genetic input was evaluated in the Ticino River Park (TRP) in Italy. A parental allocation procedure based on microsatellite markers useful for tetraploids was used following a Band-Sharing approach. Levels of relatedness within the TRP F1 captive breeders (Stock_1) and among animals released by TRP in the past (Stock_2) were explored and compared with the stock of wild origin. The pronounced decrease in genetic diversity observed in the analysed sample suggests the need for complete reorganization and coordination of conservation efforts for this species, paying particular attention to the long-term preservation of the genetic diversity. Also identified is the only potentially suitable stock of F1 animals that should be used as source of future breeders.

Summary The parental allocation (PA) in aquaculture allows the association between progeny and broodstock stocks even if the different families of offspring are reared in the same tank. Fingerprinting is needed for PA, and microsatellites are the codominant and polymorphic markers mainly used for this purpose. The simplest method of PA is the exclusion approach, based on the Mendelian mode of inheritance. The rejection of a parent-offspring hypothesis is based on incompatibility between DNA-profiling of offspring and parents. When the exclusion method is not possible a likelihood approach can be used, therefore the progeny is assigned to parents which are not excluded using the likelihood scores derived from their genotypes. Parentage is ambiguous when more parents present the highest non zero likelihood. The software for PA is disposable for diploids, but not for polyploids like sturgeons, in particular Acipenser naccarii which is a functional tetraploid species. To solve the problems concerning to the PA in tetraploids a Windows-based software was developed, using the exclusion method and likelihood approach in the case of multiple allocations. A simulation computer-procedure was also developed to validate the software algorithms. The total results obtained were: correct allocations 93.7%; ambiguous allocations 2.1% and erroneous allocations 4.2%.

Overexploitation of wild populations due to the high economic value of caviar has driven sturgeons to near extinction. The high prices commanded by caviar on world markets have made it a magnet for illegal and fraudulent caviar trade, often involving low-value farmed caviar being sold as top-quality caviar. We present a new molecular approach for the identification of pure sturgeon species and hybrids that are among the most commercialized species in Europe and North America. Our test is based on the discovery of species-specific single nucleotide polymorphisms (SNPs) in the ribosomal protein S7, supplemented with the Vimentin gene and the mitochondrial D-loop. Test validations performed in 702 specimens of target and nontarget sturgeon species demonstrated a 100% identification success for Acipenser naccarii, A. fulvescens, A. stellatus, A. sinensis and A. transmontanus. In addition to species identification, our approach allows the identification of Bester and AL hybrids, two of the most economically important hybrids in the world, with 80% and 100% success, respectively. Moreover, the approach has the potential to identify many other existing sturgeon hybrids. The development of a standardized sturgeon identification tool will directly benefit trade law enforcement, providing the tools to monitor and regulate the legal trade of caviar and protect sturgeon stocks from illicit producers and traders, hence contributing to safeguarding this group of heavily threatened species.

In species with long life cycles and discontinuous availability of individuals to reproduction, implementing a long-term captive breeding program can be difficult or impossible. In such cases, managing diversity among familiar groups instead of individuals could become a suitable approach to avoid inbreeding and increase the possibility to accomplish a breeding scheme. This is the case of several sturgeon species including the Adriatic sturgeon, whose recovery depends on the management of a few captive stocks directly descended from the same group of wild parents. In the present study, relatedness among 445 potential breeders was inferred with a novel software for pedigree reconstruction in tetraploids ("BreedingSturgeons"). This information was used to plan a breeding scheme considering familiar groups as breeding units and identifying mating priorities. A two-step strategy is proposed: a short-term breeding program, relying on the 13 remaining F0 individuals of certain wild origin; and a long-term plan based on F1 families. Simulations to evaluate the loss of alleles in the F2 generation under different pairing strategies and assess the number of individuals to breed, costs and logistical aquaculture constraints were performed. The strategy proposed is transferable to the several other tetraploid sturgeon species on the brink of extinction.

Sturgeons are a group of Condrostean fish with very high evolutionary, economical and conservation interest. The eggs of these living fossils represent one of the most high prized foods of animal origin. The intense fishing pressure on wild stocks to harvest caviar has caused in the last decades a dramatic decline of their distribution and abundance leading the International Union for Conservation of Nature to list them as the more endangered group of species. As a direct consequence, world-wide efforts have been made to develop sturgeon aquaculture programmes for caviar production. In this context, the characterization of the genes involved in sex determination could provide relevant information for the selective farming of the more profitable females. The 454 sequencing of two cDNA libraries from the gonads and brain of one male and one female full-sib A. naccarii, yielded 182,066 and 167,776 reads respectively, which, after strict quality control, were iterative assembled into more than 55,000 high quality ESTs. The average per-base coverage reached by assembling the two libraries was 4X. The multi-step annotation process resulted in 16% successfully annotated sequences with GO terms. We screened the transcriptome for 32 sex-related genes and highlighted 7 genes that are potentially specifically expressed, 5 in male and 2 in females, at the first life stage at which sex is histologically identifiable. In addition we identified 21,791 putative EST-linked SNPs and 5,295 SSRs. This study represents the first large massive release of sturgeon transcriptome information that we organized into the public database AnaccariiBase, which is freely available at http://compgen.bio.unipd.it/anaccariibase/ . This transcriptomic data represents an important source of information for further studies on sturgeon species. The hundreds of putative EST-linked molecular makers discovered in this study will be invaluable for sturgeon reintroduction and breeding programs.

We report the discovery of a new putatively active Tc 1-like transposable element ( Tana 1) in the genome of sturgeons, an ancient group of fish considered as living fossils. The complete sequence of Tana 1 was first characterized in the 454-sequenced transcriptome of the Adriatic sturgeon ( Acipenser naccarii ) and then isolated from the genome of the same species and from 12 additional sturgeons including three genera of the Acipenseridae ( Acipenser , Huso , Scaphirhynchus ). The element has a total length of 1588 bp and presents inverted repeats of 210 bp, one of which partially overlapping the 3′ region of the transposase gene. The spacing of the DDE motif within the catalytic domain in Tana 1 is unique (DD38E) and indicates that Tana 1 can be considered as the first representative of a new Tc 1 subfamily. The integrity of the native form (with no premature termination codons within the transposase), the presence of all expected functional domains and its occurrence in the sturgeon transcriptome suggest a current or recent activity of Tana 1. The presence of Tana 1 in the genome of the 13 sturgeon species in our study points to an ancient origin of the element that existed before the split of the group 170 million years ago. The dissemination of Tana 1 across sturgeon genomes could be interpreted by postulating vertical transmission from an ancestral Tana 1 with a particularly slow evolutionary rate Horizontal transmission might have also played a role in the dissemination of Tana 1 as evidenced by the presence of a complete copy in the genome of Atlantic salmon. Vertical and horizontal transmission are not mutually exclusive and may have concurred in shaping the evolution of Tana 1.

The Adriatic sturgeon is a highly endangered tetraploid species whose conservation depends up ex-situ management of the remnant genetic variability. Understanding whether the species follows a tetrasomic or a disomic inheritance pattern is of primary importance to set up a parental allocation procedure and to establish a long-term breeding plan. Moreover, comprehending the inheritance modality can strongly contribute to understanding the origin of tetraploidy in this species. For this purpose, microsatellite inheritance patterns were analyzed in 7 complete families and at 7 loci for a total of 12 family/locus combinations. For each available family, a preliminary selection of loci was performed, in order to avoid ambiguities due to allele dosage, null alleles or interference between parental contributions. Results allowed to unambiguously reject a strict disomic inheritance pattern and to suggest tetrasomy as the more likely model. Accordingly, parental chromosomes can be expected to pair in the gametes in all possible combinations, though a certain degree of preferential pairing could not be excluded for the limited statistical power reached. This study represents the first investigation of the inheritance pattern in the Adriatic sturgeon and provides relevant information for the correct management of its residual genetic diversity.