Alexander Triantafyllidis

Mitochondrial DNA variation in Northeast Atlantic and Mediterranean populations of Norway lobster, Nephrops norvegicus

Analysis of the genetic structure of the Norway lobster (Nephrops norvegicus), a marine crustacean with high commercial value, was undertaken to gain information regarding the differentiation of Atlantic from Mediterranean populations of marine invertebrates. Restriction fragment length polymorphism analysis of two mitochondrial DNA segments, 3.6 kilobases in total, was performed. Twelve populations from the North Sea, Irish Sea, Portuguese coast and Aegean Sea were analysed. Low levels of differentiation were found among them (FST = 0.018, P < 0.001) and there were no signs of an Atlantic–Mediterranean divide or of an isolation‐by‐distance scheme of differentiation. Possible reasons for these low levels of differentiation can be found in the recent expansion of N. norvegicus populations. This is supported by the mismatch distribution of pairwise haplotype differences, as well as by the high mean haplotype diversity (h = 0.93) combined with medium nucleotide diversity (π = 0.0057) (in comparison to values for marine crustaceans or teleosts) found in this study. This combination of high levels of haplotype diversity with moderate to low levels of nucleotide diversity has also been frequently attributed to a recent time of divergence for various marine species. No evidence was found for a Mediterranean refugium during glaciation periods, separate from the Atlantic, as has been reported for some marine species. The Irish Sea population was the most differentiated as a result of reduced levels of diversity. Results are also discussed in the light of future management of N. norvegicus stocks.

High Level of Mislabeling in Spanish and Greek Hake Markets Suggests the Fraudulent Introduction of African Species

DNA analysis of hake products commercialized in southern European (Spanish and Greek) market chains have demonstrated more than 30% mislabeling, on the basis of species substitution. Tails and fillets were more mislabeled than other products, such as slices and whole pieces. African species were substitute species for products labeled as American and European species, and we suggest it is a case of deliberate economically profitable mislabeling because real market prices of European and American hake products are higher than those of African in Spanish market chains. The presented results suggest fraud detection that disadvantages African producers. Government-mandated genetic surveys of commercial hakes and the use of subsequent statements of fair trade on labels of seafood products could help to reduce fraud levels in a global market of increasingly conscious consumers sensitive to ethical issues.

Is polyploidy a persevering accident or an adaptive evolutionary pattern? The case of the brine shrimp Artemia

Asexual organisms are confronted with substantial drawbacks, both immediate and delayed, threatening their evolutionary persistence. Yet, genetic associations with asexuality may refresh the gene pool promoting adaptation of clonal lineages; polyploidy is one of them. Parthenogenesis itself and/or polyploidy are responsible for the maintenance and spread of clones in Artemia, a sexual-asexual genus of halophilic anostracans. We applied flow cytometry, microsatellite genotyping, and mtDNA sequencing to 23 asexual populations. Artemia parthenogens have evolved multiple times either through hybridization or spontaneously. Nine out of 23 populations contained clones of mixed ploidy (2n, 3n, 4n). Most clones were diploid (20/31) while two and nine clones were triploid and tetraploid, respectively. Apomictic triploids and tetraploids formed two distinct groups of low genetic diversity compared with the more divergent automictic diploids. Polyploidy is also polyphyletic in Artemia, with triploids and tetraploids having independent origins from different sexual ancestors. We discern a pattern of geographical parthenogenesis with all clonal groups being more widespread than their closest sexuals. In favour of a specialist model, asexual diploids are restricted to single locations and are strikingly segregated from generalist triploids and tetraploids occupying a variety of sites. This is a rare pattern of mixed life-history strategies within an asexual complex.

The outcome of sperm competition is affected by behavioural and anatomical reproductive traits in a simultaneously hermaphroditic land snail.

Sperm competition is important in species with reproductive strategies that involve multiple mating and prolonged sperm storage such as the simultaneously hermaphroditic land snail Cornu aspersum. Double mating trials in this species have revealed that mating order and courtship behaviour affect paternity success. We investigated the effect of behavioural and anatomical reproductive traits on paternity success from triple mating trials. Triple mating resulted in triple fertilization in 58% of the cases whereas zero paternity was observed in 16% of sperm donors. Third sperm donors achieved higher paternity followed by first and second sperm donors. Snails with a longer epiphallus, the spermatophore forming organ, sired more offspring regardless of their mating order. Genetic compatibility between sperm donor and recipient did not influence paternity success. The results of the present study identified mating order and epiphallus length, as traits affecting the outcome of sperm competition in this species.

Genetic structure and phylogeography of European catfish (Silurus glanis) populations

The genetic structure of Silurus glanis (Europe’s largest freshwater fish species) across most of its natural distribution was investigated using 10 microsatellite loci. The revealed levels of genetic diversity were much higher than previous allozyme and restriction fragment length polymorphism mitochondrial DNA analyses had shown; relative levels of variability among populations were however, in good agreement with the previous studies. Populations from large basins (Volga and Danube rivers) were the most polymorphic, while samples from the smaller Greek rivers, which are more prone to genetic bottleneck, exhibited the lowest levels of genetic diversity. Microsatellite multilocus genotyping permitted the assignment of individual fish to their population of origin with a score as high as 98.3%. Despite the great genetic differentiation of S. glanis populations, no consistent pattern of geographical structuring was revealed, in contrast to previous studies of European freshwater fish species. A model of isolation by distance seems more probable and a hypothesis of recent dispersion from only one glacial refugium is proposed. The discovery of the highest levels of microsatellite and mitochondrial diversity in the Volga sample and the presence of river connections, during the Pleistocene, between this area and all major areas of the present catfish distribution, place this refugium around the Ponto‐Caspian region. Combining these data with those from previous studies, a number of markers are now available to monitor wild and hatchery populations even at the individual level.

Microsatellite analysis of the genetic population structure of native and translocated Aristotle's catfish (Silurus aristotelis)

Abstract The genetic structure of two native Silurus aristotelis (one of Europes oldest surviving freshwater fish species) populations was investigated using eight microsatellite loci. Average values of population heterozygosity were very high, even approaching values reported for marine fishes, possibly due to stable population sizes and the prolonged undisturbed conditions prevailing in the lakes that represent the native distribution of S. aristotelis. The success of an attempt to introduce the species into a new environment was also evaluated. No loss of genetic variability in the introduced population was detected. Additionally, assignment tests and trees based on genetic distances among individuals indicated the low differentiation of the translocated population from its donor population, and, differentiated the two autochthonous populations successfully. Comparisons with previous allozyme and mitochondrial RFLP studies were also made and showed that results on relative levels of genetic variability among populations were in good agreement among all methods. However, microsatellite analysis exhibited higher power of statistical tests for differentiation among population samples compared to allozymes.

Microsatellite variability of wild and farmed populations of Sparus aurata

The genetic diversity of Greek wild and farmed populations of Sparus aurata was investigated using seven microsatellite markers. Selective breeding programmes and founder effects have altered the composition of farmed populations leading to significant population differentiation between wild and cultured populations and lower allelic richness in farmed populations.

Identification of Cultured Brachionus Rotifers Based on RFLP and SSCP Screening

The marine finfish industry worldwide depends greatly on the mass culture of Brachionus rotifers. Recently, molecular data have revealed a more complicated view about the species status of Brachionus rotifers than previous mainly morphological assessments. Under this view, Brachionus rotifers are comprised of many morphologically similar, albeit genetically differentiated, cryptic members of larger groups. A redefinition of the cultured rotifer species/biotypes is therefore needed if aquaculture is to reach higher levels of standardization and predictability. In this work, restriction fragment length polymorphism (RFLP) and single-strand conformational polymorphism (SSCP) methods are applied to the COI and 16S rRNA mitochondrial genes. A detailed COI restriction map was constructed, using sequence data from all known representatives of Brachionus phylogroups. Therefore, it is the first time that such an extended restriction database has been produced. Several restriction endonucleases are proposed for the discrimination of the different Brachionus species/biotypes. Furthermore, eight different SSCP gel alleles are described for the 16S region. Using these data, five Brachionus species/biotypes were identified in 78 samples collected from laboratories and hatcheries around the world.

Morphological and taxonomic demarcation of Brachionus asplanchnoidis Charin within the Brachionus plicatilis cryptic species complex (Rotifera, Monogononta)

Three well-defined groups, consisting of 15 species, have recently been ascribed to organisms historically identified as the Brachionus plicatilis species complex. One of these groups, the large clade, is composed of two named species (Brachionus plicatilis s.s. and Brachionus manjavacas) and two species identifiers (B. ‘Nevada’ and B. ‘Austria’). B. ‘Austria’ has been confirmed to be B. asplanchnoidis. As no type specimen exists for this species, and the original taxonomic description is lacking in detail, we give a detailed account of this species using material from Obere Halbjochlacke in Austria where B. ‘Austria’ was first identified genetically. Our analysis of B. asplanchnoidis populations was of global scope, an approach that revealed a great degree of morphological variability. However, combining aspects of both the dorsal and ventral surfaces clearly discriminated B. asplanchnoidis from the rest of the large-type members. This approach may prove useful in taxonomic studies of other cryptic species with relatively few morphological features. We also observed a geographic pattern of genetic divergence within B. asplanchnoidis. Average uncorrected COI divergences for a 554-bp fragment of the COI gene ranged from 3.9% within species to 17.5% between species of the large clade and indicate deep divisions within the cryptic species complex.

A Multiplex PCR Method for Rapid Identification of Brachionus Rotifers

Cryptic species are increasingly being recognized in many organisms. In Brachionus rotifers, many morphologically similar yet genetically distinct species/biotypes have been described. A number of Brachionus cryptic species have been recognized among hatchery strains. In this study, we present a simple, one-step genetic method to detect the presence of those Brachionus sp. rotifers that have been found in hatcheries. With the proposed technique, each of the B. plicatilis sensu stricto, B. ibericus, Brachionus sp. Nevada, Brachionus sp. Austria, Brachionus sp. Manjavacas, and Brachionus sp. Cayman species and/or biotypes can be identified with polymerase chain reaction (PCR) analysis. Based on 233 cytochrome c oxidase subunit I sequences, we reviewed all the available cryptic Brachionus sp. genetic polymorphisms, and we designed six nested primers. With these primers, a specific amplicon of distinct size is produced for every one of the involved species/biotypes. Two highly sensitive protocols were developed for using the primers. Many of the primers can be combined in the same PCR. The proposed method has been found to be an effective and practical tool to investigate the presence of the above six cryptic species/biotypes in both individual and communal (bulk) rotifer deoxyribonucleic acid extractions from hatcheries. With this technique, hatchery managers could easily determine their rotifer composition at the level of cryptic species and monitor their cultures more efficiently.