SNP genotyping revealed a hybrid zone between Mytilus chilensis and M. platensis in southern South America (the Strait of Magellan, Isla Grande de Tierra del Fuego and the Falkland Islands).

Hybrid zones among mussel species have been extensively studied in the northern hemisphere. In South America, it has only recently become possible to study the natural hybrid zones, due to the clarification of the taxonomy of native mussels of the Mytilus genus. Analysing 54 SNP markers, we show the genetic species composition and admixture in the hybrid zone between M. chilensis and M. platensis in the southern end of South America. Bayesian, non-Bayesian clustering and re-assignment algorithms showed that the natural hybrid zone between M. chilensis and M. platensis in the Strait of Magellan, Isla Grande de Tierra del Fuego and the Falkland Islands shows clinal architecture. The hybrid zone can be divided into three different areas: the first one is on the Atlantic coast where only pure M. platensis and hybrid were found. In the second one, inside the Strait of Magellan, pure individuals of both species and mussels with variable degrees of hybridisation coexist. In the last area at the Strait in front of Punta Arenas City, fjords on the Isla Grande de Tierra del Fuego, and at the Beagle Channel, only M. chilensis and a low number of hybrids were found. According to the proportion of hybrids, bays with protected conditions away from strong currents would give better conditions for hybridisation. We do not find evidence of any other mussel species such as M. edulis, M. galloprovincialis, M. planulatus or M. trossulus in the zone.

Development and Validation of a Multi-Locus PCR-HRM Method for Species Identification in Mytilus Genus with Food Authenticity Purposes.

DNA-based methods using informative markers such as single nucleotide polymorphism (SNPs) are suitable for reliable species identification (SI) needed to enforce compliance with seafood labelling regulations (EU No.1379/2013). We developed a panel of 10 highly informative SNPs to be genotyped by PCR-High resolution melting (HRM) for SI in the Mytilus genus through in silico and in vitro stages. Its fitness for purpose and concordance were assessed by an internal validation process and by the transference to a second laboratory. The method was applicable to identify M. chilensis, M. edulis, M. galloprovincialis and M. trossulus mussels, fresh, frozen and canned with brine, oil and scallop sauce, but not in preserves containing acetic acid (wine vinegar) and tomato sauce. False-positive and negative rates were zero. Sensitivity, expressed as limit of detection (LOD), ranged between 5 and 8 ng/µL. The method was robust against small variations in DNA quality, annealing time and temperature, primer concentration, reaction volume and HRM kit. Reference materials and 220 samples were tested in an inter-laboratory assay obtaining an "almost perfect agreement" (κ = 0.925, p < 0.001). In conclusion, the method was suitable for the intended use and to be applied in the seafood industry.

Carminic Acid Linked to Silica Nanoparticles as Pigment/Antioxidant Bifunctional Excipient for Pharmaceutical Emulsions.

The incorporation of pigments and natural polyphenols into inorganic matrices, resulting in a hybrid material that improves the resistance and chemical stability of the pigments and the antioxidant capacity of the materials, has been of great interest to the pharmaceutical, chemical and food industries. The aim of this work was to prepare and characterize a bifunctional pigment-antioxidant nanomaterial-based carminic acid-decorated solid core-mesoporous shell silica nanoparticles, evaluating its properties as a pigment, its antioxidant capacity and its properties as a chemical stabilizer of emulsions. The chemical stability of oil-in-water (O/W) Pickering emulsions was evaluated determining the stability of vitamin E solubilized in the oil phase. Carminic acid was attached through the action of coupling ethylcarbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS) agents, and the resulting spherical and homogeneous nanoparticles showed a diameter close to 175 nm. A notorious change of emulsion color was observed by the addition of the nanomaterial. Emulsions showed an attractive pink color, and when the pH was adjusted to pH 3 and pH 9, a change in color was observed, analogous to carminic acid in solution. The nanomaterial incorporation also improved chemical stability, decreasing vitamin E consumption to 9.26% of the initial value, demonstrating an important antioxidant effect of the developed nanomaterial.

Comparison between single and multi-locus approaches for specimen identification in Mytilus mussels.

Mytilus mussels have been the object of much research given their sentinel role in coastal ecosystems and significant value as an aquaculture resource appreciated for both, its flavour and nutritional content. Some of the most-studied Mytilus species are M. edulis, M. galloprovincialis, M. chilensis and M. trossulus. As species identification based on morphological characteristics of Mytilus specimens is difficult, molecular markers are often used. Single-locus markers can give conflicting results when used independently; not all markers differentiate among all species, and the markers target genomic regions with different evolutionary histories. We evaluated the concordance between the PCR-RFLP markers most commonly-used for species identification in mussels within the Mytilus genus (Me15-16, ITS, mac-1, 16S rRNA and COI) when used alone (mono-locus approach) or together (multi-locus approach). In this study, multi-locus strategy outperformed the mono-locus methods, clearly identifying all four species and also showed similar specimen identification performance than a 49 SNPs panel. We hope that these findings will contribute to a better understanding of DNA marker-based analysis of Mytilus taxa. These results support the use of a multi-locus approach when studying this important marine resource, including research on food quality and safety, sustainable production and conservation.

High resolution melting analysis for identification of commercially-important Mytilus species.

Mytilus are edible mussels, including commercially-significant species such as M. chilensis, M. galloprovincialis and M. edulis. The scientific name of the species must be indicated on commercial products to satisfy labelling and traceability requirements. Species identification using morphological criteria is difficult due the plasticity of these characteristics and the absence of shells in processed products, and conventional PCR-based methods are laborious and time-intensive. As alternative, we propose high resolution melting (HRM) analysis as a simple tool to detect and identify SNP (single nucleotide polymorphisms) and length polymorphisms in Mytilus spp. We designed HRM-specific primers for the Mytilus genus to identify M. chilensis, M. galloprovincialis, M. edulis and their hybrids through clearly-distinguishable melting curves. HRM analysis showed high sensitivity (0.9639), specificity (1.0000) and precision (1.0000) compared to a conventional PCR-RFLP test. HRM is a fast and low cost method, being a reliable tool for species identification within the Mytilus genus.

Adaptive genetic variation distinguishes Chilean blue mussels (Mytilus chilensis) from different marine environments.

Chilean mussel populations have been thought to be panmictic with limited genetic structure. Genotyping-by-sequencing approaches have enabled investigation of genomewide variation that may better distinguish populations that have evolved in different environments. We investigated neutral and adaptive genetic variation in Mytilus from six locations in southern Chile with 1240 SNPs obtained with RAD-seq. Differentiation among locations with 891 neutral SNPs was low (FST = 0.005). Higher differentiation was obtained with a panel of 58 putative outlier SNPs (FST = 0.114) indicating the potential for local adaptation. This panel identified clusters of genetically related individuals and demonstrated that much of the differentiation (~92%) could be attributed to the three major regions and environments: extreme conditions in Patagonia, inner bay influenced by aquaculture (Reloncaví), and outer bay (Chiloé Island). Patagonia samples were most distinct, but additional analysis carried out excluding this collection also revealed adaptive divergence between inner and outer bay samples. The four locations within Reloncaví area were most similar with all panels of markers, likely due to similar environments, high gene flow by aquaculture practices, and low geographical distance. Our results and the SNP markers developed will be a powerful tool supporting management and programs of this harvested species.

Estudio de las modificaciones en proteínas de reineta (Bramas australis): sometidas a congelación y almacenamiento a 18°C y 30°C / Stydy of the reineta protein modifications (Brama australis): put under freezing and storage to 18°C and 30°C

El presente trabajo tuvo como objetivo estudiar las propiedades funcionales y térmicas en carne de reineta (Brama australis) congelada, mediante los análisis de capacidad de retención de agua (CRA), capacidad formadora de gel (CFG), textura, capacidad emulsionanate (CE) y calorometría diferencial de barrido (DSC). Para este estudio se utilizaron filetes de reinetas obtenidos y extraídos bajo las mismas condiciones, los cuales fueron trozados, envasados, congelados y almacenados a temperaturas de 18°C y 30°C durante siete meses. Los resultados para todos los tratamientos térmicos empleados no mostraron diferencias estadísticamente significativas entre individuos. Para los pescados congelados a 18°C y 30°C, los valores de proteínas totales fructuaron entre 23,5 ± 0,0 y 25,4 ± 1,0 por ciento respectivamente. Para el caso de CRA los valores se encontraron en un rango de 0,45 ± 0,1 y 1,59 ± 0,0 g agua/ g proteínas. En cuanto a la CFG sólo hubo formación de gel para la reineta fresca, existiendo producción de agregados de protéicos para las muestras almacenadas. Por otra parte los valores de CE fluctuaron entre 960 a 1400 g de aceite/g proteína, con una tendencia al aumento a medida que el tiempo de almacenamiento fue mayor. Para el caso del DSC los valores de temperatura y desnaturalización (T) y entalpía de desnaturalización (?H) de miosina fluctuaron entre 39,2 ± 0,5 y 44,8 ± 0,8°C y entre 1,12 ± 0,3 y 0,52 ± 0,2 J/g. Para la actina los valores fluctuaron entre 71,0 0,6 y 75,3 ± 0,5°C y entre 0,5 ± 0,1 y 0,7 ± 0,1 J/g, la cooperatividad disminuyó a medida que pasó el tiempo, lo cual está demostrando un cierto grado de desplazamiento de las proteínas. Los valores encontrados para propiedades térmicas presentan una directa relación con respecto a los valores de propiedades funcionales estudiadas, presentando ambos una disminución en el tiempo

Efecto de tratamientos térmicos sobre el contenido de lisina disponible en carne de jaiba mora (Homalaspis plana) / Effect of thermal treatment on the available lysine content of mora crab (Homalaspis plana) meat

The effect of different high temperatures treatments on the available lysine content of mora crab meat, was studied. Fresh pincer meat from mora crab (from the V region) was extracted and kept in ice until the thermal process of the canned product. A 3(2) statistical design was applied, considering the following variables temperature (80 degrees C, 100 degrees C and 121 degrees C) and time (15, 30 and 45 minutes). Nine conditions temperature-time were obtained. Nutritional properties from available lysine were studied. A decrease from 8.33 (in raw meat) to 6.01 g/g protein in the most drastic thermal conditions, was observed. It ca be concluded that the content of available lysine in mora crab meat is more affected by time than by the temperature of the thermal treatments. Therefore the nutritive quality can be maintained applying high temperature and short time treatments.