Alejandro de Carlos

A Microsatellite Genetic Map of the Turbot (Scophthalmus maximus)

A consensus microsatellite-based linkage map of the turbot (Scophthalmus maximus) was constructed from two unrelated families. The mapping panel was derived from a gynogenetic family of 96 haploid embryos and a biparental diploid family of 85 full-sib progeny with known linkage phase. A total of 242 microsatellites were mapped in 26 linkage groups, six markers remaining unlinked. The consensus map length was 1343.2 cM, with an average distance between markers of 6.5 ± 0.5 cM. Similar length of female and male maps was evidenced. However, the mean recombination at common intervals throughout the genome revealed significant differences between sexes, ∼1.6 times higher in the female than in the male. The comparison of turbot microsatellite flanking sequences against the Tetraodon nigroviridis genome revealed 55 significant matches, with a mean length of 102 bp and high sequence similarity (81–100%). The comparative mapping revealed significant syntenic regions among fish species. This study represents the first linkage map in the turbot, one of the most important flatfish in European aquaculture. This map will be suitable for QTL identification of productive traits in this species and for further evolutionary studies in fish and vertebrate species.

Value of the Serum Alpha-L-Fucosidase Activity in the Diagnosis of Colorectal Cancer

Objectives: The purpose of this study was to assess the value of the serum levels of α-L-fucosidase activity in the diagnosis of patients with colorectal cancer. Methods: Using a fluorometric method we analyzed the α-L-fucosidase activity in preoperative sera from 137 colorectal cancer patients and in sera from 232 donors. Results: The enzymatic activity of α-L-fucosidase was significantly lower (p < 0.001) in patients (4.8 ± 3.09 U/ml) than in donors (10.5 ± 5.46 U/ml). Using the ROC curve, the ideal cut-off for the diagnostic value of α-L-fucosidase activity was determined to be 5.6 U/ml. The diagnostic efficiency for colorectal cancer of α-L-fucosidase activity was higher than that observed for carcinoembryonic antigen (cut-off 5.0 ng/ml), especially for tumors at an early stage. Conclusions: Our results suggest that preoperative serum α-L-fucosidase activity may be used as a cheap and easy complementary test, in addition to standard clinical procedures routinely used for the diagnosis of colorectal cancer.

Macrogeographic differentiation by mtDNA restriction site analysis in the S.W. European Mytilus galloprovincialis Lmk

The mussel Mytilus galloprovincialis Lmk. is distributed in Europe from the Mediterranean Sea to the English Channel and British Isles. Recently, an abrupt change at allozyme frequencies has been pointed out between Atlantic and Mediterranean Iberian populations. M. galloprovincialis samples on both sides of the allozyme discontinuity area were taken between 1992 and 1994. Four samples were from pure populations and one from the French hybrid zone of M. galloprovincialis and M. edulis L. A pure M. edulis sample was used as the control population. Mitochondrial DNA (mtDNA) restriction site analysis of EcoRI enzyme was applied. Significant differentiation at the haplotype frequencies between Atlantic and Mediterranean M. galloprovincialis populations was found, which represented a mean variation of about 27% for the FA haplotype frequencies. This mtDNA differentiation pattern was geographically concordant with that for allozyme loci previously reported. Moreover, significant differences at haplotype frequencies between M. galloprovincialis from the hybrid zone and from pure Atlantic populations were unexpectedly found, exhibiting a mean variation of about 27% for FA haplotype. FA frequencies for M. galloprovincialis from the hybrid zone (> 68%) were not intermediate between those for Atlantic pure populations (43%) and for M. edulis (< 25%). Evolutionary implications of these macrogeographic genetic differentiations are discussed.

Extensive studies on biomorphic SiC ceramics properties for medical applications

Biomorphic silicon carbide ceramics are light, tough and high-strengt h materials with interesting biomedical applications. The fabrication method of the biomor phic SiC is based in the infiltration of molten-Si in carbon preforms with open porosity. The fina l product is a biostructure formed by a tangle of SiC fibers. This innovative process allows the fabrication of complex shapes and the tailoring of SiC ceramics with optimised properties and cont rollable microstructures that will match the biomechanical requirements of the natural host tiss ue. An interdisciplinary approach of the biomorphic SiC fabricated from beech, sapelly and eucalyptus is presented. Their mechanical properties, microstructure and chemical composition were evaluated. The biocompatible behaviour of these materials has been tested in vitro .

Purification of human alpha-l-fucosidase precursor expressed in Escherichia coli as a glutathione S-transferase fusion protein

Alpha-L-fucosidase (FUC) is a glycosidase involved in the degradation of fucose-containing glycoconjugates. A cDNA representing the complete sequence of human FUC was inserted into the prokaryotic expression vector pGEX-2T. High levels of the glutathione S-transferase (GST) fusion protein were detected in Escherichia coli cells after induction with isopropyl thio-beta-D-galactopyranoside. The GST-FUC protein was mostly found as inclusion bodies and attempts to optimise its expression as a soluble form were unsuccessful. Nevertheless, the recombinant protein was purified by affinity chromatography on glutathione-sepharose and its fucosidase activity was characterised. After thrombin cleavage of the GST tag, the FUC precursor protein was purified by electro-elution.

NORTHERNMOST RECORDS OF PAGRUS AURIGA (ACTINOPTERYGII: PERCIFORMES: SPARIDAE) AND POMADASYS INCISUS (ACTINOPTERYGII: PERCIFORMES: HAEMULIDAE) IN THE EASTERN ATLANTIC

Pagrus auriga Valenciennes, 1843 (Sparidae), and the bastard grunt, Pomadasys incisus (Bowdich, 1825) (Haemulidae), were caught in Galician waters (NW Spain) by spear fishermen (Fig. 1). The aim of this note was to describe the unusual records of these two southern species found in Galician coastal waters. Pagrus auriga is a demersal marine fish inhabiting various types of sea bottom, especially rocky bottoms, from the shore to depths of about 170 m (the closer to the shore the specimens are younger). It is distributed along the eastern Atlantic Ocean, from Portugal to Angola, and around the Madeira, Canary Islands, and Cape Verde Islands. It is also found throughout the Mediterranean Sea, but is very rare along the northern shores of this sea (Bauchot and Hureau 1986, 1990). Pomadasys incisus is a coastal demersal species inhabiting marine and brackish waters, usually near sandy or muddy substrates, at depths ranging from 10 to 100 m but most often around 50 m (Kapiris et al. 2008). It is distributed along the eastern Atlantic Ocean, from Portugal to Angola, and in South Africa, around the Azores, Canary Islands, and Cape Verde Islands, as well as in the Mediterranean Sea (Bodilis et al. 2013). ACTA ICHTHYOLOGICA ET PISCATORIA (2014) 44 (4): 323–327 DOI: 10.3750/AIP2014.44.4.07

Biomorphic Silicon Carbide Ceramics Coated with Bioactive Glass for Medical Applications

There is a need to develop new tough bioactive materials capable to withstand high loads when implanted in the body and with improved fixation, which led to the production of bioactive coatings on metallic substrates. A new approach, which consists of biomorphic silicon carbide (SiC) coated with bioactive glass, was recently presented. This new material joins the high mechanical strength, lightness and porosity of biomorphic SiC, and the bioactive properties of PLD glass films. In this work, a multiple evaluation in terms of biocompatibility of this new material was carried out starting from the biomorphic SiC morphology and porosity, following with the bioactivity of the coatings in simulated body fluid, and ending with a deep biocompatibility study with MG-63 cells. Different ranges of porosity and pore size were offered by the biomorphic SiC depending on the starting wood. The PLD glassy coatings had a high bioactivity in vitro and both the biomorphic SiC coated and uncoated presented high levels of biocompatibility.

Halosaur fishes (Notacanthiformes: Halosauridae) from Atlantic Spanish waters according to integrative taxonomy

From 2009 to 2011 thirty-five specimens belonging to six halosaurid species of the family Halosauridae were captured in two different locations in the northern waters of Spain. The specimens were identified as belonging to the genera Halosauropsis Collett, 1896, Halosaurus Johnson, 1864 and Aldrovandia Goode & Bean, 1896, including the following species: Halosauropsis macrochir (Günther, 1878), Halosaurus ovenii Johnson, 1864, Halosaurus johnsonianus Vaillant, 1888, Aldrovandia affinis (Günther, 1877), Aldrovandia phalacra (Vaillant, 1888) and Aldrovandia oleosa Sulak, 1977. The morphometric measurements and meristic characters of these specimens are given. As a result, a new northern limit of distribution of A. oleosa from the northeastern Atlantic is reported. Using a taxonomical integrative approach, the mitochondrial DNA COI gene sequences from all individuals where determined and their comparison with morphological characters showed no incongruities. Among these specimens, the highest genetic distance within species was 0.8% while the lowest value between species was 3.3%. This ample barcoding gap has allowed the delimitation and assignment of all species reported in a way that matches the traditional taxonomical methods previously employed.

DNA barcoding of deep‐water notacanthiform fishes (Teleostei, Elopomorpha)

Notacanthiformes Goodrich, 1909, is an order of deep‐sea teleost fishes with a leptocephalus larval stage whose biology and systematics are not widely known. The aim of this work was to apply the DNA barcode standard, based on COI sequence variation, to the delimitation of the species of this order, which is composed by the families Halosauridae (halosaurus) and Notacanthidae (spiny eels). The sequence data used for the analyses were obtained from two sources: 71 samples collected during oceanographic surveys in the North Atlantic Ocean (including the rare species Lipogenys gillii Goode & Bean, 1895) and 95 sequences collected from the BOLD data set. The neighbor‐joining analysis of the barcodes was successful in identifying 96% of the specimens, representing 9 of 16 and 9 of 10 of the recognised species of halosaurus and spiny eels, respectively, including all the current genera. The comparison between the Atlantic and the BOLD data sets also flagged the possibility of occurrences of misidentification and cryptic species. A modern molecular tool like the DNA barcoding supports the previously morphological‐based systematics of the order Notacanthiformes and will provide better access to the taxonomic knowledge of these deep‐water fishes.

Taxonomic study of Bathygadidae fishes (Gadiformes) from Atlantic Spanish waters combining morphological and molecular approaches.

From 2009 to 2011 eleven specimens belonging to four bathygadid species of the family Bathygadidae were captured in two different locations in the northern waters of Spain. The morphometric measurements and meristic characters of these specimens are given. The specimens were identified as belonging to the genera Gadomus Regan, 1903, and Bathygadus Günther, 1878, including the following species: Gadomus dispar (Vaillant, 1888), Gadomus longifilis (Goode & Bean, 1885), Gadomus arcuatus (Goode & Bean, 1886) and Bathygadus melanobranchus Vaillant, 1888. As a result, a new northern limit of distribution of G. arcuatus from the northeastern Atlantic is reported. The first molecular identification and genetic interrelationships of Bathygadidae species, based on the mitochondrial COI nucleotide sequences -DNA barcodes- is reported. Sequences corresponding to specimens from the same species were identical and the overall mean genetic diversity (uncorrected p-distance) was 0.096 ± 0.008. Based on a morphological and meristic examination of the specimens, as well as on the available literature, an updated key of the members of the family Bathygadidae from the north-eastern Atlantic Ocean is provided.