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Embryo transfer procedures have been widely implemented in bovine all around the world. These techniques help to accelerate the increase in the genetic merit and to keep the level of inbreeding under control, which is especially important in breeds, such as Japanese Black cattle, in areas outside of Japan, where there are scarce individuals and few herds. Our study describes an adequate embryo productivity of Japanese Black heifers under Spanish management and environmental conditions, like that which has been previously reported, demonstrating the adequate adaptation capacity of these animals. No effect on embryo production or quality was found due to plasma metabolic parameters of the donors, suggesting an optimal nutritional and body conditioning of the donors’ herd. Pregnant recipients after embryo transfer showed significantly higher levels of cholesterol-related parameters, glucose, and urea, which may be related to higher availability of nutrients for the future pregnancy. Heat stress at embryo transfer negatively impacted conception rates as expected, while larger parity and ET number corresponded to numerically higher conception rates., Multiple ovulation and embryo transfer (MOET) systems have been intensively implemented in Japanese Black cattle in Japan and to create Japanese Black herds out of these areas. Environmental conditions influence MOET efficiency. Thus, we describe results of 137 in vivo, non-surgical embryo flushings performed between 2016–2020, in a full-blood Japanese Black herd kept in Spain and the possible effects of heat, year, bull, donor genetic value, and metabolic condition. Additionally, 687 embryo transfers were studied for conception rate (CR) and recipient related factors. A total of 71.3% of viable embryos (724/1015) were obtained (5.3 ± 4.34/flushing). Donor metabolites did not affect embryo production (p > 0.1), although metabolite differences were observed over the years, and by flushing order, probably related to the donor age. CR was not affected by embryo type (fresh vs. frozen), recipient breed, and whether suckling or not suckling (p > 0.1). CR decreased significantly with heat (44.3 vs. 49.2%; (p = 0.042)) and numerically increased with recipient parity and ET-number. Pregnant recipients showed significantly higher levels of cholesterol-related metabolites, glucose, and urea (p < 0.05). Therefore, adequate MOET efficiency can be achieved under these conditions, and heat stress should be strongly avoided during Japanese Black embryo transfers. Moreover, recipients’ metabolites are important to achieve pregnancy, being probably related to better nutrient availability during pregnancy

The Trans-Mexican Volcanic Belt (TMVB), located in central Mexico, is an area for which low to moderate seismic risk is considered. This is based on the limited instrumental data available, even though large historical earthquakes have damaged some urban centers in the past. However, site effects is an aspect that must be considered in estimating risk, because there are some instances of important amplifications that have been documented with serious effects. In this work, ambient noise and earthquake records from 90 seismic permanent and temporary stations are used to analyze site response in the TMVB. The results obtained show a heterogeneous range in the value of the fundamental frequency. When possible, a comparison was made of the results obtained from ambient noise and earthquake records. In almost all these comparisons, no significant differences were observed in terms of the fundamental frequency. However, there were some stations with a flat average HVSR ambient noise curve that contradicted earthquake data results, which showed peaks at some frequencies. Our results are a first step towards categorizing the different site responses in the TMVB but in order to provide finer details, it is necessary to improve the actual monitoring conditions.

Some diatom species of the genus Pseudo-nitzschia produce the toxin domoic acid. The depuration rate of domoic acid in Pecten maximus is very low, for this reason, king scallops generally contain high levels of domoic acid in their tissues. A transcriptomic approach was used to identify the genes differentially expressed in the P. maximus digestive gland after the injection of domoic acid. The differential expression analysis found 535 differentially expressed genes (226 up-regulated and 309 down-regulated). Protein–protein interaction networks obtained with the up-regulated genes were enriched in gene ontology terms, such as vesicle-mediated transport, response to stress, signal transduction, immune system process, RNA metabolic process, and autophagy, while networks obtained with the down-regulated genes were enriched in gene ontology terms, such as response to stress, immune system process, ribosome biogenesis, signal transduction, and mRNA processing. Genes that code for cytochrome P450 enzymes, glutathione S-transferase theta-1, glutamine synthase, pyrroline-5-carboxylate reductase 2, and sodium- and chloride-dependent glycine transporter 1 were among the up-regulated genes. Therefore, a stress response at the level of gene expression, that could be caused by the domoic acid injection, was evidenced by the alteration of several biological, cellular, and molecular processes.

[EN] The sensitive state of conservation of several endemic fish species such as Iberian toothcarp (Aphanius iberus) and Valencia toothcarp (Valencia hispanica) has led governments to consider the implementation of conservation measures to preserve their populations. However, limited knowledge about the reproductive biology of these species makes it necessary to investigate different aspects of their reproductive cycle. In this sense, the main objectives of this work were i) to advance knowledge of the breeding biology of both species, and ii) to develop protocols for the conservation of gametes for the future management and conservation. During the spring of 2019 a temporal series of samplings were carried out in different places in the Comunitat Valenciana. Sperm samples were collected and sperm motion parameters were assessed for the first time in both species. Kinetic patterns were similar showing high motility and velocity values during the first 30 s, and a rapid decrease from that point. At the same time, an in-depth morphometric analysis was carried out using computer-assisted sperm analysis software. Spermatozoa from A. iberus and V. hispanica showed similar sizes and shapes to other external fertilizers belonging to Cyprinodontiformes, with small spherical heads, uniflagellated and without acrosomes. In addition, a new cryopreservation protocol was designed for cryobanking the sperm of these threatened species. Cryopreserved samples showed lower motility than fresh samples but reaching acceptable percentages of motile cells after thawing of around 20 and 25% (A. iberus and V. hispanica, respectively). This study is the first of its kind to successfully achieve gamete cryopreservation of these two endemic and endangered species from the Iberian Peninsula, providing new and useful tools to complement the management and conservation programs that are being developed for both species., This project has received funding from the Generalitat Valenciana (GV; Valencia, Spain) under the "Subvenciones para la realizacion de proyectos de I+D+I desarrollados por grupos de investigacion emergentes (GV/2019/130)". VG has a postdoc grant from the Ministerio de Economia, Industria y Competitividad (MINECO; Madrid, Spain) under the project IJCI-2017-34200. We would like to thank all the technicians of the CCEDCV for their work and effort during the samplings, specially to J. Velazquez, J. Hernandez and A. Pradillo.

The natural production of the native flat oyster Ostrea edulis and the production of seed in hatcheries are insufficient to meet demand. Several projects have recently been developed to optimise flat oyster production and to establish a reliable protocol for broodstock conditioning in hatcheries. In this sense, the positive effect of a gradient of temperature and a gradient of daylight on the gonadal development, spawning and larval production of O. edulis during winter and autumn conditioning was previously confirmed. No other study has yet been optimised the effect of other factors on flat oyster conditioning. This work examines the combined effect of two important factors on broodstock conditioning, namely the food ration and the water flow rate, for the first time. Furthermore, the bacteriological load of oyster broodstock and its influence on O. edulis conditioning had not been studied before. The results showed that the broodstock conditioned with a food ration of 3% produced the highest number of viable larvae. Oysters conditioned at a flow rate of 3 L h−1 oyster−1 had the highest mortality rate, coinciding with the highest levels of vibrio and heterotrophic bacteria in both meat and intervalvar fluid. The best larval production results were obtained when the 3% food ration was combined with a water flow rate of 2 L h−1 oyster−1.

[EN] In teleosts, vitellogenin (Vtg) is a phospholipoglycoprotein synthesized by the liver, released into the blood circulation and incorporated into the oocytes via endocytosis mediated by the Vtg receptor (VTGR) to form the yolk granules. The VTGR is crucial for oocyte growth in egg-laying animals but is also present in non-oviparous vertebrates, such as human. The VTGR belongs to the low-density lipoprotein receptor superfamily (LDLR) and is also named very-low-density lipoprotein receptor (VLDLR). In this study, we identified and phylogenetically positioned the VTGR of a basal teleost, the European eel, Anguilla anguilla. We developed quantitative real-time PCR (qRT-PCR) and investigated the tissue distribution of vtgr transcripts. We compared by qRT-PCR the ovarian expression levels of vtgr in juvenile yellow eels and pre-pubertal silver eels. We also analyzed the regulation of ovarian vtgr expression throughout vitellogenesis in experimentally matured eels. The Vtg plasma level was measured by homologous ELISA experimental maturation. Our in silico search and phylogenetical analysis revealed a single vtgr in the European eel, orthologous to other vertebrate vtgr. The qRT-PCR studies revealed that vtgr is mainly expressed in the ovary and also detected in various other tissues such as brain, pituitary, gill, fat, heart, and testis, suggesting some extra-ovarian functions of VTGR. We showed that vtgr is expressed in ovaries of juvenile yellow eels with no higher expression in pre-pubertal silver eels nor in experimentally matured eels. This suggests that vtgr transcription already occurs during early pre-vitellogenesis of immature eels and is not further activated in vitellogenic oocytes. European eel Vtg plasma level increased throughout experimental maturation in agreement with previous studies. Taken together, these results suggest that vtgr transcript levels may not be a limiting step for the uptake of Vtg by the oocyte in the European eel., M.M., A.G.L. and S.D. were granted with Short Term Scientific Missions by the COST Office (COST Action FA1205: AQUAGAMETE). S.D. was also awarded with a grant from the UPV's School of Doctorate (Accion para la Internacionalizacion de los Programas de Doctorado) in 2017. We thank E. Feunteun and colleagues, MNHN, Dinard, for the ovarian samples from eels of the Fremur, France.

Source at https://doi.org/10.1186/s12864-019-5969-6. © The Author(s). 2019 Background: The impossibility of closing the life cycle of the European eel (Anguilla anguilla) in captivity troubles the future of this critically endangered species. In addition, the European eel is a highly valued and demanded resource, thus the successful closing of its life cycle would have a substantial economic and ecological impact. With the aim of obtaining the highest gamete quality, the study of the effects of environmental factors, such as temperature, on reproductive performance may prove valuable. This is especially true for the exposure to cold water, which has been reported to improve sexual development in multiple other Actinopterygii species. Results: European eel males treated with cold seawater (10 °C, T10) for 2 weeks showed an increase in the proliferation and differentiation of spermatogonial cells until the differentiated spermatogonial type A cell stage, and elevated testosterone and 11-ketotestosterone plasma levels. Transcriptomes from the tissues of the brain-pituitary-gonad (BPG) axis of T10 samples revealed a differential gene expression profile compared to the other experimental groups, with clustering in a principal component analysis and in heat maps of all differentially expressed genes. Furthermore, a functional analysis of differentially expressed genes revealed enriched gene ontology terms involved in the regulation of circadian rhythm, histone modification, meiotic nuclear division, and others. Conclusions: Cold seawater treatment had a clear effect on the activity of the BPG-axis of European eel males. In particular, our cold seawater treatment induces the synchronization and increased proliferation and differentiation of specific spermatogonial cells. In the transcriptomic results, genes related to thermoception were observed. This thermoception may have caused the observed effects through epigenetic mechanisms, since all analysed tissues further revealed differentially expressed genes involved in histone modification. The presented results support our hypothesis that a low temperature seawater treatment induces an early sexual developmental stage in European eels. This hypothesis is logical given that the average temperature experienced by eels in the early stages of their oceanic reproductive migration is highly similar to that of this cold seawater treatment. Further studies are needed to test whether a cold seawater treatment can improve the response of European eels to artificial hormonal treatment, as the results suggest.

Introducción. El análisis de ADN antiguo (ADNa) a diferencia de otro tipo de investigaciones, requiere de la implementación de condiciones de infraestructura y metodológicas especializadas que garanticen la autenticidad de los resultados. Uno de los criterios de autenticidad para este tipo de muestras, corresponde a la cuantificación del material genético, en la cual es común el uso de la PCR cuantitativa en tiempo real dada su sensibilidad y especificidad. La implementación de estas metodologías y condiciones necesarias para el cumplimiento de los requisitos de autenticidad hace dispendioso y costoso este tipo de investigación.Objetivo. Generar una estrategia de cuantificación del ADN mitocondrial de muestras altamente degradadas mediante un sistema sencillo y de fácil implementación Materiales y métodos. El sistema está basado en el uso de primers que permiten la amplificación especifica de fragmentos cortos del ADN mitocondrial. La posterior purificación de este fragmento permite generar una curva estándar con concentraciones acordes al estado de degradación de la muestra.Resultados. Se detectó ADN antiguo proveniente de restos óseos y tejidos momificados datados de diferente cronología. Adicionalmente el sistema permitió detectar la presencia de agentes inhibidores del ADN.Conclusión. La estrategia aquí planteada no solo es sencilla de implementar sino que puede reducir los costos de la investigación, adicionalmente permite la detección de ADNa en muestras altamente degradadas y la discriminación de muestras que no poseen material genético de aquellas que poseen agente inhibidores.

La presente investigación tuvo como objetivo analizar, a partir de diversos aportes teóricos y dialógicos, las influencias e impactos del liderazgo informal en la productividad y competitividad de las organizaciones. Para lo cual, se utilizó un enfoque hermenéutico reflexivo de la investigación bibliográfica, que permitió la comprensión e interpretación de investigaciones consultadas en bibliotecas y bases de datos electrónicas de reconocimiento mundial. Además, se consultó la opinión de expertos académicos del campo administrativo en Colombia para recopilar las voces y percepciones sobre el tema. Los principales resultados evidencian que el liderazgo informal, como una dinámica propia de los grupos informales, al vincularla al factor humano de la organización, puede influir de manera positiva o negativa en la productividad y competitividad. Se concluye que la gestión administrativa de este fenómeno organizacional se convierte en una herramienta para lograr ventajas competitivas en las organizaciones a través del mejoramiento de las variables funcionales de la empresa y el establecimiento de relaciones sociales más eficientes.

New specific European eel (Anguilla anguilla) recombinant gonadotropins (aarGths) produced in the ovarian cells of Chinese hamsters (CHO) were used to induce maturation in captive male eels. In the first experiment, five different hormonal treatments were assayed: one group was given a constant dose of recombinant European eel follicle-stimulating hormone (aarFsh; 4 μg/fish) for 9 weeks, and the second group received a constant dose of recombinant European eel luteinizing hormone (aarLh; 2 μg/fish) also for 9 weeks. The other three groups were injected with different combinations of both aarGths (some doses constant, some variable). All five treatments stimulated androgen synthesis, but the increase was more pronounced in the fish treated with a combination of both aarGths. Unlike aarLh, aarFsh alone was able to induce spermiation, the best results were achieved in the fish that were treated with a constant dose of aarFSH and an increasing dose of aarLH, with spermiation being induced (20% motile cells) despite the fact that these fish were immature at the start of the experiment. In order to improve sperm quality, a second experiment was performed. Immature males received three constant doses of aarFsh (2.8, 1.4 or 0.7 μg/fish) and increasing doses of aarLh (every 3 weeks; 1, 2, 6 μg/fish). All the treatments induced spermiation, however the best sperm quality (with ≥50% motile cells) was observed in the males treated with the highest dose of aarFsh. In conclusion, these specific recombinant gonadotropins have demonstrated their capacity to induce spermatogenesis and spermiation in vivo in a teleost fish, the European eel, This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 642893 (ETN IMPRESS), the Spanish Ministry of Economy, Industry and Competitiveness (Project REPRO-TEMP; AGL2013-41646-R), Generalitat Valenciana (PROMETEOII/2014/051), VLC/CAMPUS Program (SP20140630) and the COST Office (COST Action FA1205: AQUAGAMETE). VG has a postdoc grant from the UPV (PAID-10-14).

Genomic scale duplication of genes generates raw genetic material, which may facilitate new adaptations for the organism. Previous studies on eels have reported specific gene duplications, however a species-specific large-scale gene duplication has never before been proposed. In this study, we have assembled ade novoEuropean eel transcriptome and the data show more than a thousand gene duplications that happened, according to a 4dTv analysis, after the teleost specific 3R whole genome duplication (WGD). The European eel has a complex and peculiar life cycle, which involves extensive migration, drastic habitat changes and metamorphoses, all of which could have been facilitated by the genes derived from this large-scale gene duplication.Of the paralogs created, those with a lower genetic distance are mostly found in tandem repeats, indicating that they are young segmental duplications. The older eel paralogs showed a different pattern, with more extensive synteny suggesting that a Whole Genome Duplication (WGD) event may have happened in the eel lineage. Furthermore, an enrichment analysis of eel specific paralogs further revealed GO-terms typically enriched after a WGD. Thus, this study, to the best of our knowledge, is the first to present evidence indicating an Anguillidae family specific large-scale gene duplication, which may include a 4R WGD.

Introducción: La incapacidad temporal (IT) constituye un proceso de origen multifactorial. Objetivo: Evaluar el proceso de IT en relación con la enfermedad, estado de salud, satisfacción laboral, perfil de locus de control y características sociodemográficas. Método: Estudio observacional de cohortes prospectivo sobre 404 pacientes en IT. Realizamos un análisis de supervivencia describiendo la evolución de los participantes en cuanto a su permanencia en IT. Resultados: La permanencia media en IT fue significativamente superior (p

espanolFundamentos: La incapacidad temporal (IT) constituye un importante problema sanitario, economico y social. El objetivo del estudio es describir los motivos de IT, satisfaccion laboral y caracteristicas sociosanitarias de los sujetos que permanecen en IT durante un periodo prolongado. Metodos: Estudio descriptivo transversal, sobre 404 pacientes en IT. Se recogieron datos sobre: diagnostico de IT, satisfaccion laboral, salud autopercibida, caracteristicas sociodemograficas, estado de salud y perfil de locus de control. Resultados: Entre las personas en IT predominan los varones (53,2%), con edad media de 48,1 anos, casados (64,1%), con bajo nivel de instruccion (36,1%) y pertenecientes a clases sociales inferiores (27,2%). Las enfermedades predominantes son las enfermedades del aparato locomotor (46,5%), seguidas de problemas psicologicos (18,6%) y enfermedades cardiovasculares (6,2%). Los pacientes muestran un nivel aceptable de satisfaccion laboral, siendo esta superior en hombres (76,5 ± 12,9DE), en pertenecientes a categorias sociales superiores (80,9 ± 14,9 DE en categorias I-II) y en trabajadores con nivel de instruccion elevado (80,5 ± 14,5 DE en personas con estudios universitarios). La mayoria califica su salud como buena (57,9%), y sin un perfil predominante de locus (37,6%). La mayoria tienen episodios previos de IT, generalmente inferiores a 6 meses (74,5%). Conclusiones: Varones de mediana edad, con enfermedades del aparato locomotor, bajo nivel de instruccion, buena salud autopercibida, satisfechos con su trabajo y pertenecientes a clases sociales inferiores conforman el perfil mas comun del paciente que permanece en IT durante un periodo prolongado, no observandose entre ellos un perfil de locus de control predominante. EnglishFundamentals: The temporary disability (TD) constitutes a major health, economic and social problem. The objective of the study is to describe the reasons for IT, job satisfaction, and features social and health care of subjects that remain in IT for a long time. Methods: Descriptive study cross, about 404 patients in IT. Data were collected on: diagnosis of IT, job satisfaction, self-perceived health, features sociodemographic, health status and profile of locus of control. Results: Among the people in IT dominate males (53.2%), with average age of 48.1, married (64.1%), with low level of education (36.1%) and lower social classes (27.2%). Predominant diseases are diseases of the locomotor system (46.5%), followed by psychological problems (18.6%) and cardiovascular diseases (6.2%). Patients show an acceptable level of job satisfaction, which is higher in men (76.5 ± 12, US), in belonging to higher social categories (80.9 ± 14.9 of in categories I-II) and workers with high level of instruction (80.5 ± 14.5 of in people with University studies). Most qualifies as good health (57.9%), and without a predominant profile of locus (37.6%). Most have previous episodes of IT, generally less than 6 months (74.5%). Conclusions: Men of middle age, with diseases of the musculoskeletal, low level of instruction, good health self-perceived, satisfied with his work and belonging to the lower social classes comprise the most common profile of the patient who stays in IT for an extended period, not observed including a profile of locus of control predominantly.

[EN] Vitrification was successfully applied to the sperm of two fish species, the freshwater Eurasian perch (Perca fluviatilis) and marine European eel (Anguilla anguilla). Sperm was collected, diluted in species specific non-activating media and cryoprotectants and vitrified by plunging directly into liquid nitrogen without pre-cooling in its vapor. Progressive motility of fresh and vitrified-thawed sperm was evaluated with computer-assisted sperm analysis (CASA). Additional sperm quality parameters such as sperm head morphometry parameters (in case of European eel) and fertilizing capacity (in case of Eurasian perch) were carried out to test the effectiveness of vitrification. The vitrification method for Eurasian perch sperm resulting the highest post-thaw motility (14 +/- 1.6%) was as follows: 1:5 dilution ratio, Tanaka extender, 30% cryoprotectant (15% methanol + 15% propylene-glycol), cooling device: Cryotop, 2 mu l droplets, and for European eel sperm: dilution ratio 1:1, with 40% cryoprotectant (20% MeOH and 20% PG), and 10% FBS, cooling device: Cryotop, with 2 mu l of sperm suspension. Viable embryos were produced by fertilization with vitrified Eurasian perch sperm (neurulation: 2.54 +/- 1.67%). According to the ASMA analysis, no significant decrease in head area and perimeter of vitrified European eel spermatozoa were found when compared to fresh spermatozoa., The work was funded by the NKFI (previously OTKA) project number K-109847 and by a Short-term Scientific Mission awarded to E. Kasa by the COST Office (Food and Agriculture COST Action FA1205: Assessing and improving the quality of aquatic animal gametes to enhance aquatic resources. The need to harmonize and standardize evolving methodologies, and improve transfer from academia to industry; AQUAGAMETE). The work was supported by the project Research Center of Excellence - 9878-3/2016/FEKUT of the Ministry of Human Resources of Hungary and the project EUREKA_HU_12-1-2012-0056 (PERCAHATCH).

[EN] The role of seminal plasma sodium and activation media sodium on sperm motility was examined by selectively removing the element from these two media, in European eel sperm. Sperm size (sperm head area) was also measured using an ASMA (Automated Sperm Morphometry Analyses) system, in the different conditions. Intracellular sodium [Na+](i) was quantitatively analyzed by first time in the spermatozoa from a marine fish species. Measurement of [Na+](i) was done before and after motility activation, by Flow Cytometry, using CoroNa Green AM as a dye. Sperm motility activation induced an increase in [Na+](i) from 96.72 mM in quiescent stage to 152.21 mM post-activation in seawater. A significant decrease in sperm head area was observed post activation in seawater. There was a notable reduction in sperm motility when sodium was removed from the seminal plasma, but not when it was removed from the activation media. Sodium removal was also linked to a significant reduction in sperm head area in comparison to the controls. Our results indicate that the presence of the ion Na+ in the seminal plasma (or in the extender medium) is necessary for the preservation of sperm motility in European eel, probably because it plays a role in maintaining an appropriate sperm cell volume in the quiescent stage of the spermatozoa. (C) 2016 Elsevier Inc All rights reserved., Funded from the SPERMOT project (Spanish Ministry of Science and Innovation, MICINN; AGL2010-16009). M.C. Vilchez has a predoctoral grant from UPV PAID Program (2011-S2-02-6521), Marina Morini has a predoctoral grant from Generalitat Valenciana (Programa Grisolia, GRISOLIA/2012/006), Victor Gallego has a postdoctoral contract from UPV (PAID-10-14), and David S. Penaranda was supported by MICINN (PTA2011-4948-I) and UPV (PTA2011-4948-I). Grants to attend meetings were received from COST Office (Food and Agriculture COST Action FA1205: AQUAGAMETE).

The settlements produced by the load transmitted to the structure on expansive soils, and those calculated by the classical theories of soil mechanics, are different because the swelling pressure acts inversely to the applied stress. In this paper we describe a procedure to determine a volumetric variation coefficient by hydration (αh ) which considers the expansive soil behaviour. In order to do this, it is necessary to know the soil’s initial water content, the swelling pressure, and the applied stress. Soil suction and swelling pressure were measured with filter paper technique and a mechanical oedometer, respectively. Unsaturated undisturbed samples of expansive soil were used. The water content was varied, starting from 0%, with increments of 5.5% to reach 38%. Furthermore, we present a set of curves that show the magnitude of the coefficient (αh ) associated to a water content, and the relationship between applied stress and swelling pressure. The results show that the variation of the coefficient under different initial water contents ranges between 0% to 22%. This is because water is strongly attracted by clay minerals, but this attraction decreases as water layers are father from the surface of clay minerals, thus decreasing its swelling potential.

The ancestry of the Colombian population comprises a large number of well differentiated Native communities belonging to diverse linguistic groups. In the late fifteenth century, a process of admixture was initiated with the arrival of the Europeans, and several years later, Africans also became part of the Colombian population. Therefore, the genepool of the current Colombian population results from the admixture of Native Americans, Europeans and Africans. This admixture occurred differently in each region of the country, producing a clearly stratified population. Considering the importance of population substructure in both clinical and forensic genetics, we sought to investigate and compare patterns of genetic ancestry in Colombia by studying samples from Native and non-Native populations living in its 5 continental regions: the Andes, Caribe, Amazonia, Orinoquía, and Pacific regions. For this purpose, 46 AIM-Indels were genotyped in 761 non-related individuals from current populations. Previously published genotype data from 214 Colombian Natives from five communities were used for population comparisons. Significant differences were observed between Native and non-Native populations, among non-Native populations from different regions and among Native populations from different ethnic groups. The Pacific was the region with the highest African ancestry, Amazonia harboured the highest Native ancestry and the Andean and Orinoquían regions showed the highest proportion of European ancestry. The Andean region was further sub-divided into 6 sub-regions: North East, Central West, Central East, West, South West and South East. Among these regions, the South West region showed a significantly lower European admixture than the other regions. Hardy-Weinberg equilibrium and variance values of ancestry among individuals within populations showed a potential stratification of the Pacific population. COL0035429

[EN] Contents Sperm cryopreservation is a useful tool in captive fish reproduction management, that is to synchronize gamete production, especially in the case of species as the European eel, where the time of female spawning readiness is unpredictable. Several protocols to cryopreserve sperm of this species have been described, but until recently fertilization trials were not feasible. This study evaluated the effect of cold storage of diluted sperm prior to fertilizations and tested whether a previously defined protocol for European eel sperm cryopreservation can be successfully applied in fertilization trials to produce viable offspring. In our experiment, the sperm motility was evaluated after the extraction and the best samples were selected and pooled. Until stripping of eggs and fertilization, diluted sperm samples were maintained at either 4 or 20 degrees C, or cryopreserved, following existing protocols. Fertilization of two egg batches was attempted. Diluted sperm caused a similar percentage of fertilized eggs and a similar number of embryos and larvae, independently of storage temperature (4 or 20 degrees C). The cryopreserved sperm resulted in a lower percentage of fertilized eggs, but embryos developed and a few larvae (cryolarvae') were obtained 55h after fertilization in one of the two egg batches. This result evidences that the tested cryopreservation protocol is applicable for eel reproduction management, although improvements will be required to enhance fertilization success., This study was part of the project: 'Reproduction of European Eel: Towards a Self-sustained Aquaculture' (PRO-EEL) European Community's 7th Framework Programme under the Theme 2 'Food, Agriculture and Fisheries, and Biotechnology', Grant Agreement no245257. Juan F. Asturiano and Luz Perez had a grant to staying in Denmark from Programa de Apoyo a la Investigacion y Desarrollo (PAID-00-11) of the Universitat Politecnica de Valencia. We thank Christian Graver and Lars B. Tybjerg for assistance during

9 páginas, 1 tabla, 7 figuras, By first time, 2DE protein profile of European eel seminal plasma has been determined. 14 different proteins corresponding to 9 major families were identified in seminal plasma, through hormonal treatment. Some of them play a part in sperm maturation, including carbonic anhydrase which is responsible for modulating the pH of seminal plasma, and warm temperature acclimation protein, which may play an important role in the final maturation of this species, due to the warm temperature of their spawning ground (in the Sargasso Sea). Sperm samples were classified into three motility categories depending on the percentage of motile cells, I: 0–25%, II: 25–50% and III: > 50%. Different protein profiles were observed depending on the sperm motility categories, specifically, with the apolipoproteins and complement C3. Higher numbers of proteins from the apolipoprotein family were registered at lower motilities; whereas the complement C3-like family was higher in the samples with the highest percentage of motile cells. These results suggest that the proteins linked to the transportation of lipids (apolipoprotein) and to the immune system (complement C3) may carry out their functions at different stages of spermatogenesis. Using SDS-PAGE analysis, 13 bands were identified, most of which migrated between 20 to 60 kDa. In the last weeks of treatment significant increases were observed in the percentage of motile spermatozoa, curvilinear velocity and beat cross frequency. This improvement in sperm quality coincided with a higher amount of proteins located at 19 KDa, therefore, this protein could be involved in sperm motility of the European eel., Funded from the REPRO-TEMP project (Spanish Ministry of Science and Innovation, MICINN; AGL2013-41646-R). and COST Office (COST Action FA1205: AQUAGAMETE). M.C.V. and V.G. have pre- and post-doctoral grants from UPV PAID Programme (2011-S2-02-6521 and 10-14 respectively).

[EN] Positive effects of probiotics on fish reproduction have been reported in several species. In the present study, 40 male European eels were weekly treated with recombinant hCG for 9 weeks and with three different concentrations (10(3), 10(5), and 10(6) CFU/mL) of probiotic Lactobacillus rhamnosus IMC 501 (Sinbyotec, Italy). The probiotics were daily added to the water from the sixth week of the hCG treatment. Males from the treated and control groups were sacrificed after 1, 2, and 3 weeks of probiotic treatment (seventh ninth weeks of hCG treatment); at this point, sperm and testis samples were also collected. Sperm volume was estimated, and motility was analyzed by computer-assisted sperm analysis software. Alternations in transcription of specific genes involved in reproductive process such as activin, androgen receptors alpha and beta (ar alpha and ar beta), progesterone receptor 1 (pr1), bone morphogenetic protein 15 (bmp15), and FSH receptor (fshr) were analyzed in the testis. After 2 weeks of probiotic treatment, sperm production and sperm motility parameters (percentage of motile cells and percentage of straight-swimming spermatozoa) were increased in the European eel treated with 105 CFU/mL compared to controls or to the other probiotic doses. These changes were associated with increases in messenger RNA expression of activin, ar alpha, ar beta, pr1, and fshr. Conversely, after 3 weeks, activin and pr1 expression decreased. No significant changes were observed on bmp15 expression throughout the duration of the treatment with 10(5) CFU/mL dose. The lowest and highest probiotic dose (10(3) and 10(6) CFU/mL, respectively) inhibited the transcription of all genes along all the experiment, except for ar alpha and ar beta after 1 week of probiotic treatment when compared to controls. The changes observed by transcriptomic analysis and the sperm parameters suggest that a treatment with L rhamnosus at 10(5) CFU/mL for 2 weeks could improve spermatogenesis process in Anguilla anguilla. (C) 2015 Elsevier Inc. All rights reserved., This study was funded by the European Community’s 7th FP (grant agreement no. 245257, PRO-EEL) and COST Office (Food and Agriculture COST Action FA1205: AQUAGAMETE) Victor Gallego and M. Carmen Vilchez have predoctoral grants from MINISTERIO DE ECONOMIA Y COMPETITIVIDAD (BES-2009-020310) and UNIVERSIDAD POLITECNICA DE VALENCIA PAID Program (2011-S2-02-6521), respectively. Fondo Ateneo 2012 to Oliana Carnevali.

A hybrid model for soils, which combines percolation theory and finite element method is presented. The internal soil structure is modelled via the finite element method, and percolation networks are used for analyzing its mechanical behaviour. Through a microscopic characterization of elastic properties of soil grains, the model is generated. The effective percolation threshold obtained is lower than that of the network geometric percolation. The effective mechanical properties predicted are successfully compared to published experimental results.

[EN] This research investigated the regulation of aromatase and androgen receptor gene expression in the brain–pituitary– gonad (BPG) axis of male and female European eels (Anguilla anguilla) during induced sexual maturation. Complete A. anguilla aromatase (aa-cyp19a1) and partial androgen receptor a and b (aa-ara and aa-arb) sequences were isolated, and qPCR assays were validated and used for quantification of transcript levels for these three genes. Expression levels of the genes varied with sex, tissue and stage of maturation. aa-arb was expressed at higher levels than aa-ara in the pituitary and gonad in both sexes, suggesting aa-arb is the physiologically most important androgen receptor in these tissues. In the female brain, a decrease in aa-ara and an increase in aa-cyp19a1 were observed at the vitellogenic stage. In contrast, a progressive increase in all three genes was observed in the pituitary and ovaries throughout gonadal development, with aa-arb and aa-cyp19a1 reaching significantly higher levels at the vitellogenic stage. In the male pituitary, a decrease in aa-arb and an increase in aa-cyp19a1 were observed at the beginning of spermatogenesis, and thereafter remained low and high, respectively. In the testis, the transcript levels of androgen receptors and aa-cyp19a1 were higher during the early stages of spermatogenesis and decreased thereafter. These sex-dependent differences in the regulation of the expression of aa-ara, aa-arb and cyp19a1 are discussed in relation to the role of androgens and their potential aromatization in the European eel during gonadal maturation, This work was funded by the European Community's 7th Framework Programme under the Theme 2 'Food, Agriculture and Fisheries, and Biotechnology', grant agreement no 245257 (PRO-EEL). D. S. P. received a postdoc grant from UPV (CEI-01-10), a mobility grant from UPV (PAID-00-11) and has also been supported by a contract cofinanced by MICINN and UPV (PTA2011-4948-I). V. G. and I. M. received predoctoral grants from the Spanish Ministry of Science and Innovation (MICINN) and Generalitat Valenciana, respectively. F.-A. W. received funding from the Norwegian School of Veterinary Science. The fish farm Valenciana de Acuicultura, S. A. supplied the male eels used in the experiments. The English revision was carried out by Professor Lucy Robertson (Lucy Robertson Writing Services, Norway).

[EN] The objective of this study was evaluating the effects on the growth rate and flesh quality of separating larvae according to their larval width in cultured gilthead seabream (Sparus aurata, L.). Progeny from two broodstocks (A and B) were divided into two groups according to larval width: heads (being the larger specimens) and tails. After 18months both groups were analysed to evaluate growth, carcass traits and proximal composition. The head specimens reached a larger size and showed a greater level of well-being and degree of nourishment. Total body and fillet composition in general were found to be as expected for this species. Some differences were found in total body composition between head and tail specimens in both batches, and in the fillet composition in batch B (higher fat and lower moisture content in the head specimens). Sensory analyses were carried out using untrained panelists, who were unable to distinguish between the head and tail samples in batch A, whereas differences were noticeable in batch B. Samples from the head group were judged to be tastier and juicier, a consequence of their higher fat content. Hence, fish separation according to larval width is an effective tool to separate progeny into slow and fast growing groups, whereas the total body and fillet analyses and a sensory test ensure that the selection does not generate negative effects on the product quality., Funded by the Coordinated Project SELECTBREAM (Spanish Ministry of Science and Innovation; AGL2007-64060-C03-00, including European Regional Development Funds); I. Mazzeo was supported by a FPI scholarship financed by Generalitat Valenciana; V. Gallego was supported by a FPI scholarship financed by the Spanish Ministry of Science and Innovation; C. Garcia-Fernandez was supported by a FPU scholarship financed by the Spanish Ministry of Science and Innovation.

[EN] Although it is widely accepted that osmolality and ion fluxes are the main factors triggering sperm motility in fish, a complex universal mechanism for sperm motility activation does not exist in fish, and studies of marine fish species are even more scarce. Therefore, the main goal of this study was to estimate the intracellular variations in the main ions involved in sperm activation for the first time in European eel, in order to provide additional new data about this little-known process. It was observed that levels of intracellular Ca2+ and K+ sperm ions increased significantly 30 s after the hyperosmotic shock compared to baseline levels, and remained at this level until 120 s post-activation. In contrast, the intracellular pH remained constant during the first 30 s, and decreased gradually at 60 and 120 s post-activation. Our data agree with the current main theory for explaining motility activation in marine fish, in which internal fluctuations of Ca2+ and K+ seem to participate in sperm activation. In addition, fluorescent images showed that both Ca2+ and K+ were concentrated in the apical area of the sperm head, which corresponds to the location of the eel sperm mitochondria, suggesting this organelle plays an important role in sperm motility activation. (C) 2013 Elsevier B.V. All rights reserved., Funded from the European Community's 7th Framework Programme under the Theme 2 "Food, Agriculture and Fisheries, and Biotechnology", grant agreement no 245257 (Pro-Eel) and the Spanish Ministry of Science and Innovation (MICINN; AGL2010-16009). Victor Gallego has a predoctoral grant (MICINN; BES-2009-020310) and has been granted a fellowship (EEBB-I-12-05858) of the Spanish Personnel Research Training Programme to carry out this study in the Universidad de Leon (Leon, Spain). Ilaria Mazzeo had a predoctoral grant from GVA. David S. Penaranda has a contract co-financed by MICINN and UPV (PTA2011-4948-I). F. Martinez-Pastor was supported by the Ramon y Cajal program (RYC-2008-02560, MICINN).

[EN] Little is known about the role of fat and fatty acids in European eel spermatogenesis. The aim of this research was to study the changes in fat content and to carry out a quantitative analysis of the fatty acid composition of the muscle, liver and gonad of European male eels during hormonally induced sexual maturation. Three different thermal regimes were used (two variable: T10 and T15; and one constant: T20) to replicate the changes in temperature that these fish experience during their transoceanic reproductive migration. Spermatogenesis was reached earlier in treatment T20, suggesting that spermatogenesis in the European eel is closely regulated by water temperature. Although eels lose body mass due to the period of fasting that accompanies gonadal growth, no significant changes were found in the fat and fatty acid content of the muscle during the experimental period. With regard to the liver, the levels of palmitic, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids increased significantly at the start of the spermiation process in all the thermal treatments. In the testis, levels of EPA, arachidonic acid (ARA) and DHA remained constant during the maturation process whereas the levels of the rest of the fatty acids decreased significantly. The stability of the ARA and EPA levels in the testis may have a physiological significance, whereas the stability of the DHA levels may have a structural significance. The results suggest that the progression of spermiation is influenced by water temperature and demonstrate the importance of EPA, ARA and DHA in European eel reproduction. This study demonstrates that complementary research focusing on the lipid composition of commercial diets could improve the sperm quality of this species., Funded by the European Community's 7th Framework Programme under the Theme 2 "Food, Agriculture and Fisheries, and Biotechnology", grant agreement no. 245257 (PRO-EEL). Victor Gallego, Ilaria Mazzeo and M. Carmen Vilchez had predoctoral grants from the Spanish Ministry of Science and Innovation (MICINN), Generalitat Valenciana, and UPV PAID Programme (2011-S2-02-6521), respectively. David S. Penaranda was supported by a contract co-financed by MICINN and UPV (PTA2011-4948-I). Rosa Baeza was supported by contract financed by PRO-EEL. The authors want to thank Vicente Javier Moya Salvador for his technical assistance with gas chromatography analyses.

[EN] The first goal of this study was the development of a short-term storage method for pufferfish (Takifugu niphobles) sperm. In this respect, the best results were obtained by diluting the sperm in a seminal-like solution and keeping it in a Petri dish in chilled storage (4 degrees C). This method was successful in preserving sperm quality parameters without resulting in differences in fresh sperm for a relatively long-term period (7 days), for use in aquaculture matters. The addition of bovine serum albumin (BSA) to the medium did not improve the results. On the other hand, both the osmolality and the ion composition of the media are essential factors which can modulate the sperm motility parameters. The osmolality of the activating medium was the most important factor in triggering pufferfish sperm motility, and osmolalities of 750-825 mOsm/kg were necessary to initiate this process, demonstrating that it appears to be a dose-independent mechanism. Regarding the ion composition of the activation media, this study has shown that despite the spermatozoa being able to initiate movement without any ion in the activation medium, the absence of ions can negatively affect the kinetic parameters of the spermatozoa. Finally, in natural conditions (seawater), the activation of sperm motility generates intracellular increases in Ca2+ and K+, suggesting that these ions play an essential role in the activation mechanism of pufferfish sperm. (C) 2013 Elsevier B.V. All rights reserved., This study was funded by the Spanish Ministry of Science and Innovation (MICINN; AGL2010-16009). Victor Gallego has a predoctoral grant (MICINN; BES-2009-020310) and has been granted a fellowship (EEBB-I-12-05858) from the Spanish Personnel Research Training Programme to carry out this study in the Misaki Marine Biological Station (Miura, Japan).

[EN] We examined the effect of temperature in European silver eels during their maturation induced by injections of carp pituitary extract on endocrine parameters: pituitary fshß and lhß expression, plasma 17ß-estradiol (E2) and vitellogenin, estrogen receptor 1 (esr1), and vitellogenin 2 (vtg2) expression in liver. A variable thermal regime (T10) that increased from 10° to 14° and 17 °C was compared with a constant 20 °C regime (T20) during 12. weeks. T10 caused a faster development until week 8, higher fshß, lhß, esr1 expression, and higher E2 levels. The results strongly suggest that T10 is inducing a higher endogenous FSH level which increases the E2 circulating level during vitellogenesis. A variable thermal regime induced an fshß expression and E2 profile in vitellogenic hormonally matured eel females that were more similar to the profile observed in other naturally maturing fish. © 2011 Elsevier Inc., This study was funded from the European Community’s 7th Framework Programme under the Theme 2 ‘‘Food, Agriculture and Fisheries, and Biotechnology’’, Grant Agreement No. 245257 (Pro-Eel) and Generalitat Valenciana through the GV/2007/202 project and ACOMP/2011/229. D.S. Peñaranda has a postdoc grant from the Universitat Politècnica de València (CE-01-10). The authors would like to thank M. Nieveen for technical assistance.

International audience; European eel males can be artificially matured (1.5IU hCG/g fish), but the regulatory mechanisms of their reproductive development are practically unknown. Spermatogenic stages (S1-S6), biometric characters [eye index (EI), gonadosomatic index (GSI), hepatosomatic index (HSI)] and sperm quality parameters (motility, viability and head spermatozoa morphometry) were analysed. Moreover, the present study evaluated the expression of GnRHs (mammal and chicken II Gonadotropin Release Hormone I) and gonadotrophins (FSHbeta and LHbeta) during hormonal treatment, as well as 11-ketotestosterone (11-KT) and 17,20beta-dihydroxy-4-pregnen-3-one (17,20beta-P) plasma levels. One week was enough to observe the S2 of gonad development, but it was necessary to reach the 7th week of treatment to obtain animals that presented the most advanced stage of development (S6). Differential regulation of the two GnRH expressions was found, supporting the main role of mGnRH in the control of gonadotrophin release. One hCG injection was enough to dramatically decrease the FSHbeta expression, being close to zero during the rest of the treatment. LHbeta expression and 17,20beta-P registered a significant increase in the same stage of development, S3/4, confirming the role of this gonadotrophin in the last steps of maturation and 17,20beta-P in the spermatozoa maturation. The 11-KT increased with GSI, and the highest 11-KT values coincided with the advanced steps of spermatogenesis prior to spermiation. Being consistent with the known role of the steroid in these processes. Furthermore, this study supports a role for 11-KT in stimulating eye growth, presenting high values when EI increased. Sperm production was obtained from the 4th week of treatment, but it was in the 8th week when a significant increase was observed in sperm quality [viability, high motility (>75%)].