Gene expression profiling reveals functional specialization along the intestinal tract of a carnivorous teleostean fish (Dicentrarchus labrax)

High-quality sequencing reads from the intestine of European sea bass were assembled, annotated by similarity against protein reference databases and combined with nucleotide sequences from public and private databases. After redundancy filtering, 24,906 non-redundant annotated sequences encoding 15,367 different gene descriptions were obtained. These annotated sequences were used to design a custom, high-density oligo-microarray (8 × 15 K) for the transcriptomic profiling of anterior (AI), middle (MI), and posterior (PI) intestinal segments. Similar molecular signatures were found for AI and MI segments, which were combined in a single group (AI-MI) whereas the PI outstood separately, with more than 1900 differentially expressed genes with a fold-change cutoff of 2. Functional analysis revealed that molecular and cellular functions related to feed digestion and nutrient absorption and transport were over-represented in AI-MI segments. By contrast, the initiation and establishment of immune defense mechanisms became especially relevant in PI, although the microarray expression profiling validated by qPCR indicated that these functional changes are gradual from anterior to posterior intestinal segments. This functional divergence occurred in association with spatial transcriptional changes in nutrient transporters and the mucosal chemosensing system via G protein-coupled receptors. These findings contribute to identify key indicators of gut functions and to compare different fish feeding strategies and immune defense mechanisms acquired along the evolution of teleosts.This work was funded by the EU seventh Framework Programme by the ARRAINA (Advanced Research Initiatives for Nutrition and Aquaculture; KBBE-2011-288925) project. It does not necessarily reflect the views of the EU and in no way anticipates the Commission's future policy in this area. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. Additional funding was obtained from the Spanish Ministerio de Economía y Competitividad through the MI2-FISH project (Unraveling Metabolic, Intestinal and Immunopathological Fish Status; AGL2013-48560) and Generalitat Valenciana (PROMETEO FASE II-2014/085).Peer Reviewe

Advances in comparative endocrinology : vol. VIII

Ponències presentades al 10th Congress of the Iberian Association of Comparative Endocrinology (AIEC), celebrat a la Universitat Jaume I, els dies 23 al 25 de setembre de 2015Les diverses comunicacions presentades al 10è Congrés de la Asociación Ibérica de Endocrinología Comparada (23-25 setembre 2016, Castelló) s'agrupen en aquest volum. Les intervencions han aportat els darrers avenços en àrees científiques com ara reproducció, metabolisme, estrés, resposta immune, creixement, mineralització i pigmentació...Las diversas comunicaciones presentadas en el 10º Congreso de la Asociación Ibérica de Endocrinología Comparada (23-25 septiembre 2016, Castellón) se agrupan en este volumen. Las intervenciones han aportado los últimos adelantos en áreas científicas como por ejemplo reproducción, metabolismo, estrés, respuesta inmune, crecimiento, mineralización y pigmentación...The present volume of Advances in Comparative Endocrinology collects the contributions of the participants at the 10th Congress of the Iberian Association of Comparative Endocrinology (AIEC). Eighteen years after the foundational meeting of our Association in Peñíscola, the return of this Congress to Castellón highlights the growing success of this initiative to foster the research and scientific development in the field of comparative endocrinology developed in the Iberian Peninsula. AIEC meetings have proven to be a way to keep in contact among research groups with common interests. Some of the participants in this last meeting were also present in the foundational one, others members came after and keep assisting every time. As one of the aims of AIEC has been to encourage students to participate, we are particulary proud of those young students and doctors from the first editions that have gained more permanent positions and continue participating in the AIEC meetings with new students

Modulation of the IgM gene expression and IgM immunoreactive cell distribution by the nutritional background in gilthead sea bream (Sparus aurata) challenged with Enteromyxum leei (Myxozoa)

The aim of the present work was to determine if a plant protein-based diet containing vegetable oils (VO) as the major lipid source could alter the distribution of IgM immunoreactive cells (IRCs) and the IgM expression pattern in the intestine and haematopoietic tissues of gilthead sea bream (GSB) (. Sparus aurata) challenged with the myxosporean Enteromyxum leei. In a first trial (T1), GSB fed for 9 months either a fish oil (FO) diet or a blend of VO at 66% of replacement (66VO diet) was challenged by exposure to parasite-contaminated water effluent. All fish were periodically and non-lethally sampled to know their infection status. After 102 days of exposure, samples of intestine and head kidney were obtained for IgM expression and immunohistochemical detection (IHC). Additional samples of spleen were taken for IHC. Fish were categorized as control (C, not exposed), and early (E), or late (L) infected. The 66VO diet had no effect on the number of IgM-IRCs in any of the tissues or on IgM expression in C fish, whereas the infection with E. leei had a strong effect on the intestine. A combined time-diet effect was also observed, since the highest expression and IRCs values were registered in the posterior intestine (Pi) of E-66VO fish. A positive correlation was found between IgM expression and the presence of IgM-IRCs in the Pi. The effect of the time of infection was studied more in detail in a second trial (T2) in which samples of Pi were taken at 0, 24, 51, 91 and 133 days after exposure to the parasite. A significant increase of the IgM expression was detected only in parasitized fish, and very late after exposure. These results show that the duration of the exposure to the parasite is the most determinant factor for the observed intestinal IgM increased phenotype which gets magnified by the feeding of a high VO-based diet. © 2012 Elsevier Ltd.This work was funded by EU through projects AQUAMAX (FOOD-CT-2006-16249; Sustainable Aquafeeds to Maximise the Health Benefits of Farmed Fish for Consumers) and ARRAINA (Advanced Research Initiatives for Nutrition & Aquaculture, FP7/2007/2013; grant agreement n° 288925), and by the Spanish Ministry of Science and Innovation (MICINN) through the project AGL2009-13282-C02-01. Additional funding was obtained from the “Generalitat Valenciana” (research grant PROMETEO 2010/006). I. E. received a Spanish PhD fellowship (FPI) from MICINN.Peer Reviewe

A novel miniaturized biosensor for monitoring atlantic salmon swimming activity and respiratory frequency

Good fish welfare is one of the prerequisites for sustainable aquaculture. Knowing how fish respond to the production conditions would allow us to better understand their biology and to further optimize production. The new miniaturized biosensor AEFishBIT was successfully used to monitor individual physical activity and respiratory frequency of two Mediterranean farmed fish species (gilthead sea bream and European sea bass). In this study, we aimed to test the use of AEFishBIT to monitor the performance of Atlantic salmon under experimental conditions. An adapted tagging procedure for salmon was developed and used to record salmon responses to handling and changing light conditions. AEFishBIT data showed a stabilization of swimming activity 8 h after handling and tagging with changes in activity or activity and respiratory quotient after changes in light intensity regimes. The results of this study supported the use of AEFishBIT to generate new behavior insights in Atlantic salmon culture.publishedVersio

Un estudio sobre la utilidad de la realización de prácticas de laboratorio en el proceso de aprendizaje y su influencia en los resultados de la evaluación del alumno

En el presente trabajo, realizado en la Escuela Técnica Superior de Informática Aplicada (ETSIAp) de la Universidad Politécnica de Valencia, se ha analizado el efecto que la realización de las prácticas de la asignatura de Estadística tiene sobre la nota final del examen. Para dicho estudio se han empleado dos técnicas estadísticas complementarias

Transcriptional Assessment by Microarray Analysis and Large-Scale Meta-analysis of the Metabolic Capacity of Cardiac and Skeletal Muscle Tissues to Cope With Reduced Nutrient Availability in Gilthead Sea Bream (Sparus aurata L.)

The effects of nutrient availability on the transcriptome of cardiac and skeletal muscle tissues were assessed in juvenile gilthead sea bream fed with a standard diet at two feeding levels: (1) full ration size and (2) 70 % satiation followed by a finishing phase at the maintenance ration. Microarray analysis evidenced a characteristic transcriptomic profile for each muscle tissue following changes in oxidative capacity (heart > red skeletal muscle > white skeletal muscle). The transcriptome of heart and secondly that of red skeletal muscle were highly responsive to nutritional changes, whereas that of glycolytic white skeletal muscle showed less ability to respond. The highly expressed and nutritionally regulated genes of heart were mainly related to signal transduction and transcriptional regulation. In contrast, those of white muscle were enriched in gene ontology (GO) terms related to proteolysis and protein ubiquitination. Microarray meta-analysis using the bioinformatic tool Fish and Chips (http://fishandchips.genouest.org/index.php) showed the close association of a representative cluster of white skeletal muscle with some of cardiac and red skeletal muscle, and many GO terms related to mitochondrial function appeared to be common links between them. A second round of cluster comparisons revealed that mitochondria-related GOs also linked differentially expressed genes of heart with those of liver from cortisol-treated gilthead sea bream. These results show that mitochondria are among the first responders to environmental and nutritional stress stimuli in gilthead sea bream, and functional phenotyping of this cellular organelle is highly promising to obtain reliable markers of growth performance and well-being in this fish species. © 2014 Springer Science+Business Media New York.This work was funded by the EU seventh Framework Programme by the AQUAEXCEL (Aquaculture Infrastructures for Excellence in European Fish Research, FP7/2007-2012; grant agreement no. 262336) project. Additional funding was obtained from the Generalitat Valenciana (research grant PROMETEO 2010/006) and the Spanish Government through AQUAGENOMICS project (Consolider-Ingenio-2010 Programme).Peer Reviewe

Effect of ivermectin on the liver of gilthead seabream Sparus aurata: a proteomic approach

23 p., 2 tablas, 4 figuras y bibliografíaGilthead sea bream Sparus aurata is the most commercialized Mediterranean aquacultured fish species. Ivermectin has recently(experimentally) started to be used to control ectoparasitic infestations in Mediterranean cultured marine fish. The potential hepatotoxicity of ivermectin was investigated in gilthead sea bream juveniles (35g) following oral administration at the recommended dose of 0.2mg kg-1 fish for 10d. Difference Gel Electrophoresis Technology (DIGE) was used to study the effect of this treatment in gilthead sea bream liver protein profile under routine culture conditions. The 2D-DIGE protein maps obtained were analyzed using the DeCyder 6.5 software. The results obtained showed significant changes in the expression of 36 proteins respect to the control group. Among these proteins, six increased in abundance, and 30 decreased. Spot showing differential expression respect to the control were analyzed by mass spectrometry and database search, which resulted in three positive identifications corresponding to hepatic proteins involved in lipid metabolism (apoA-I), oxidative stress responses and energy generation (beta-globin, ATP synthase subunit beta). These proteins have not been previously associated to invermectin effect.This work was funded by projects GV06B-351 “Generalitat Valenciana” and CTM 2006-14279-CO2-01/MAR MEC-FEDER. This research was performed within the framework of a concerted action between Spain and Greece (HG-2004-0016). I. Varó was a recipient of a “Ramón y Cajal” contract at the University of Valencia from the “Ministerio de Educación y Ciencia (Spain)”.Peer reviewe

Untargeted metabolomics approach for unraveling robust biomarkers of nutritional status in fasted gilthead sea bream (Sparus aurata)

A metabolomic study has been performed to identify sensitive and robust biomarkers of malnutrition in farmed fish, using gilthead sea bream (Sparus aurata) as a model. The metabolomic fingerprinting of serum from fasted fish was assessed by means of ultra-high performance liquid chromatography coupled to quadrupole time-offlight mass spectrometry. More than 15,000 different m/z ions were detected and Partial Least Squares–Discriminant analysis allowed a clear differentiation between the two experimental groups (fed and 10-day fasted fish) with more than 90% of total variance explained by the two first components. The most significant metabolites (up to 45) were elucidated on the basis of their tandem mass spectra with a broad representation of amino acids, oligopeptides, urea cycle metabolites, L-carnitinerelated metabolites, glutathione-related metabolites, fatty acids, lysophosphatidic acids, phosphatidylcholines as well as biotin- and noradrenaline-related metabolites. This untargeted approach highlighted important adaptive responses in energy and oxidative metabolism, contributing to identify robust and nutritionally-regulated biomarkers of health and metabolic condition that will serve to assess the welfare status of farmed fish.This work was supported by the Spanish MINECO (MI2-Fish, AGL2013-48560) and Plà de promoció de la investigació de la Universitat Jaume I (P1•1B2015-59) and the Generalitat Valenciana as research group of excellence (PROMETEO II/2014/023, PROMETEO II/2014/085, Collaborative Research on Environment and Food-Safety, ISIC/2012/016)

Dietary supplementation of heat-treated Gracilaria and Ulva seaweeds enhanced acute hypoxia tolerance in gilthead sea bream (Sparus aurata)

Intensive aquaculture practices involve rearing fish at high densities. In these conditions, fish may be exposed to suboptimal dissolved O2 levels with an increased formation of reactive O2 species (ROS) in tissues. Seaweeds (SW) contain biologically active substances with efficient antioxidant capacities. This study evaluated the effects of dietary supplementation of heat-treated SW (5% Gracilaria vermiculophylla or 5% Ulva lactuca) on stress bioindicators in sea bream subjected to a hypoxic challenge. 168 fish (104.5 g average weight) were distributed in 24 tanks, in which eight tanks were fed one of three experimental diets for 34 days: (i) a control diet without SW supplementation, (ii) a control diet supplemented with Ulva, or (iii) a control diet with Gracilaria. Thereafter, fish from 12 tanks (n=4 tanks/dietary treatment) were subjected to 24 h hypoxia (1.3 mg O2 l-1) and subsequent recovery normoxia (8.6 mg O2 l-1). Hypoxic fish showed an increase in hematocrit values regardless of dietary treatment. Dietary modulation of the O2-carrying capacity was conspicuous during recovery, as fish fed SW supplemented diets displayed significantly higher haemoglobin concentration than fish fed the control diet. After the challenge, survival rates in both groups of fish fed SW were higher, which was consistent with a decrease in hepatic lipid peroxidation in these groups. Furthermore, the hepatic antioxidant enzyme activities were modulated differently by changes in environmental O2 condition, particularly in sea bream fed the Gracilaria diet. After being subjected to hypoxia, the gene expression of antioxidant enzymes and molecular chaperones in liver and heart were down regulated in sea bream fed SW diets. This study suggests that the antioxidant properties of heat-treated SW may have a protective role against oxidative stress. The nature of these compounds and possible mechanisms implied are currently being investigated.Fil: Magnoni, Leonardo Julián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Universidad de Porto; PortugalFil: Martos Sitcha, Juan Antonio. Consejo Superior de Investigaciones Científicas; EspañaFil: Queiroz, Augusto. Universidad de Porto; PortugalFil: Calduch Giner, Josep Alvar. Consejo Superior de Investigaciones Científicas; EspañaFil: Magalhaes Gonçalves, Jose Fernando. Universidad de Porto; PortugalFil: Rocha, Cristina M.R.. Universidad de Porto; PortugalFil: Abreu, Helena T.. ALGAplus; PortugalFil: Schrama, Johan W.. Wageningen University; Países BajosFil: Ozorio, Rodrigo O.A.. Universidad de Porto; PortugalFil: Perez Sanchez, Jaume. Consejo Superior de Investigaciones Científicas; Españ